Trace Element 2

The Journal of Trace Elements in Experimental Medicine 17:181–272 (2004)
DOI: 10.1002/jtra.10028
International Society
For Trace Element Research
In Humans (ISTERH)
Seventh International Conference
‘‘TRACE ELEMENT NUTRITION AND HUMAN DISEASE’’
Bangkok, Thailand
November 7 12, 2004
Host of ISTERH 2004

Songsak Srianujata
ISTERH 2004 Planning Committee Chair

Curtiss D. Hunt, USA
Plenary Speakers and Symposia Committee

Harold H. Sandstead, Chair
George J. Brewer
Curtiss D. Hunt
Abstracts, Platform and Poster Sessions Committee

Monica Nordberg, Chair
Ole Andersen, George Brewer, John Duffus,
Jeanne Freeland-Graves, Rosalind Gibson, Edward Harris,
Mary L’Abbe, Forrest Nielsen, Ananda Prasad, Anne Roussel,
Hiramu Sakurai, J. J. Strain, Leslie Woodhouse
Student Travel Award Committee

George J. Brewer, Chair
ISTERH 2004 Local Organizer
The Institute of Nutrition
Mahidol University (INMU)
Conference Secretariat
Agora Communication, Québec City, Canada
Communicated by A. Klug
September 7 12, 2002
This work is reprinted from: Te EMBO Journal 1985;4:1609 1614, with permission
2004 Wiley-Liss, Inc.

President
Harold H. Sandstead, USA
Vice President
Mohammed Abdulla, Sweden
Treasurer
Curtiss D. Hunt, USA
Secretary
Muriel Bost, France
Past President
George J. Brewer, USA
ISTERH Council
Mary L’Abbe, Canada
Jan Aaseth, Norway
Ayhan Cavdar, Turkey
Jeanne Freeland-Graves, USA
Rosalind Gibson, New Zealand
Forrest Nielsen, USA
Monica Nordberg, Sweden
Manuel Ruz, Chile
Hiramu Sakuri, Japan
Noel Solomons, Guatemala
Anne Roussel, France
Swapan Kumar Roy, Bangladesh
Yoji Takagi, Japan
Neil Ward, United Kingdom
ISTERH Abstracts 183
Abstracts
Category 1: Bioavailability of Trace Elements and Dietary Patterns
1.01
BLOOD SELENIUM IN SMOKERS AND NON-SMOKERS IN SOUTHERN
NORWAY
Dag Ellingsen [1], Yngvar Thomassen [1], Jon Efskind [2], Jan Aaseth* [3]
National Institute of Occupational Health, Dep, N-0033 Oslo, NORWAY [1]; The
Health Dept., Borregaard industries, N-1700 Sarpsborg, NORWAY [2]; Dept. of
Internal Medicine, Kongsvinger Hospital, N-2226 Kongsvinger, NORWAY [3]
The object of this study was to investigate blood selenium values as influenced by
smoking habits. The population under investigation (n = 298) were recruited from work
places in the southern part of the country. The included attendants were apparently
healthy individuals. Roughly half of the participants (47 %) turned out to be smokers.
The smoking group could be classified as heavy, medium or light smokers. Blood sele-
nium was determined by electrothermal atomic absorption, that gave results in agree-
ment with the certified values of reference materials. The blood selenium values in non-
smokers and smokers were 1.52 and 1.38 umol/l, respectively, in the eastern part of the
region, and 1.33 and 1.23 umol/l, respectively, in the western part of the southern region
of Norway. Thus, it is clear that habitual smoking depressed the blood selenium values
statistically significantly, by 10 % or more. The activity of the selenoenzyme glutathione
peroxidase was depressed as well. The effect of smoking turned out to be dose-depen-
dant, as heavy smoking exerted a more pronounced effect than light smoking. Fur-
thermore, it appeared that the average blood selenium levels in the southern areas of
Norway had decreased from values around 1.6 umol/L in 1980 to an average below 1.4
umol/l at our examination twenty years later. Whereas about 60 % of the grains con-
sumed in Norway in 1980 were imported from high selenium areas in America, the
consumption of domestic grain has increased considerably during the last 20 years. The
low selenium content of Scandinavian grain may explain the decreasing selenium levels
observed in the population.
1.02
MACRO- AND TRACE ELEMENTS IN PLASMA OF INJURED MILITARIES
Igor Bakulin* [1,2], Vladislav Novozhenov [1], Margarita Skalnaya [2]
Municipal clinical hospital 29, Moscow, RUSSIA [1]; Russian Society of Trace Elements
in Medicine (RUSTEM), Moscow, RUSSIA [2]
The multielement plasma profiles of militaries, injured during the anti-terrorist action in
North Caucasus by ICP-OES (K, Mg, Na, Ca, P, Fe) and ICP-MS (Zn, Cu, Cr, Se, Mn,
I, Co, Sn, Pb, As, Cd, Ni, Ti, V) methods were investigated. All patients had severe mines
injuries. There were divided in 2 groups, depending on term of injuries. The 1-st group
included persons, injured 1-6 days before investigation. The patients, injured in term 14-
20 days before investigation, were included in 2-nd group. In the 1-st group there were
found statistically (p<0.05) increased K, Mg, Fe, Cr, Co, Sn, I and decreased Na, Se,
plasma levels. In later period after injury (group 2) the elevated plasma level of Ca, P,
Mg, Zn, Cu, I, Cr, Se and low one of Na and Fe were found. Obtained data suggests the
significant difference in elemental plasma patterns of injured patients, depending on term
after injuries and enteral or parenteral administration of macro- and trace elements for
184 ISTERH Abstracts
the improvement of treatment results. The use of different formulas for enteral nutrition
shows the significant positive changes in elemental status of injured patients and guides to
the further clinical investigations.
1.03
INTERACTION BETWEEN IRON, IODINE AND ZINC STATUS IN A REGIONAL
SAMPLE OF MOROCCAN WOMEN
Rekia Belahsen* and Imane Bahbouhi
Chouaib Doukkali University, Training and Research Unit on food sciences, Laboratory
of Physiology Applied to Nutrition and Feeding, School of Sciences, El Jadida 24000,
MOROCCO
Trace element nutrition is an important public health problem for many populations of
Morocco. Deficiencies of iron, iodine, and vitamin A continue to be prevalent in the
Moroccan population. Also there are many programmes enrolled to fight against iron,
vitamin A and iodine deficiencies. However these nutrient deficiencies continue to be the
most common deficiency diseases in Morocco. No data were available on zinc status or
prevalence of zinc deficiency in the Moroccan population. Zinc deficiency association
with iron deficiency are areas of investigation and zinc supplementation improve other
micronutrients deficiency diseases. In this survey the aim was to investigate the interac-
tion of zinc with iron and iodine deficiencies. A sample of childbearing women was
selected from rural and urban area. 24hours dietary recall and food frequency were used
to assess dietary intakes and dietary habits. Blood and urine samples were collected for
biochemical determination and, socio demographic data were also collected using a
questionnaire. The blood determinants and dietary data of iron, iodine and zinc evalu-
ated show that a deficiency in the micronutrients investigated was prevalent in the
population sample. Conclusion: Zinc intakes were generally inadequate in this regional
sample of Moroccan population. The zinc intake was associated to other micronutrient
deficiency. The data were discussed in relation to age, socio demographic and other
micronutrients status. The results will provide useful information in establishing or
updating public health policies related to trace element nutrition. The study brings to the
knowledge the challenges facing public health officials on trace element nutrition in
North Africa. (R. Belahsen serves on the National Micronutrient Committee in Mor-
occo.)
1.04
GEOPHAGIA, ANEMIA, AND BIOAVAILABLE-IRON
Momoko Chiba* [1], Taeko Shimoda [2], Satoshi Sasaki [3], Osamu Kunii [4], Masahiro
Hashizume Dept. Epidemiology and Environmental Health, Juntendo Univ. School of
Medicine, Tokyo 113-8421, JAPAN [1]; Faculty of Home Economics, Kyushu Women’s
University, Kitakyushu 807-8586, JAPAN. [2]; National Institute of Health and Nutri-
tion, Tokyo 162-8636, JAPAN [3]; Economic Cooperation Bureau, Ministry of Foreign
Affairs, Tokyo 100-8919, JAPAN [4]; Dept. International Community Health, Graduate
School of Medicine, The Tokyo University, Tokyo 113-0033, JAPAN [5]
A total 964 school-age children were randomly selected from the resident lists in two (K
and Z) districts in Kazakhstan. K is near the Aral Sea and Z is about 600 km east from
K-district. 815 provided blood specimens. 62% in K-district and 39% in Z-district were
defined as anemia. Prevalence of geophagia was high, 15.7% in K-distract and 16% in Z-
district. Hemoglobin values were significantly difference between geophagia and non-
geophagia groups (average hemoglobin values 11.46 g/dl (n=128), and 11.97 g/dl
(n=681). Iron concentrations were significantly lower in geophagia group. In addition
magnesium and phosphorus concentrations in blood were significantly lower in geo-
phagia group. 799 responded to the food frequency questionnaire, and the results showed
that fish consumption was much higher in K-district, and fruits and vegetable con-
sumption was higher in Z-district. Bioavailable-iron was calculated using the results of
24-hour recall of 114 children and Kazakh Food Composition Table taking account with
enhancers and inhibitors of iron absorption. Bioavailable-iron was significantly lower in
anemia children than non-anemia children. (M. Chiba serves on this project as a chief
member.)
1.05
TRACE ELEMENTS CONCENTRATION IN HUMAN NAILS USING ATOMIC
ABSORPTION SPECTROMETRY
F. A. El-Amri* [1] and R. I. Damja [2]
United Arab Emirate Universits,Faculty of Scienc Chem.Dept P.O.Box 17551, Al-Ain,
UNITED ARAB EMIRATES [1]; University of Al-Fathi Faculty of Science Chem.Dept.
P.O.Box 13361 Tripoli, LIBYA [2]
Evaluation of trace elements in human tissues such as hair and nails has proven useful in
the studies pertaining to chronic body exposure. These may have also been considered as
indexes to evaluate environmental exposure. Nail samples were collected from healthy
people age 5-60 year. The samples were digested with nitric and perchloric acid, the
elements were then determined by atomic absorption spectrometry. These elements are
calcium, iron, magnesium, and zinc. Results show that calcium has the highest concen-
tration than other elements. Elemental concentrations are compared to literature values
and chronological variation is illustrated. A pattern of the concentration of elements in
nails samples were analyzed regarding to age and sex.
1.06
BIOAVAILABILITY OF ZINC FROM STAPLE CEREALS AND PULSES
S. Hemalatha*, Platel Kalpana, and K. Srinivasan
Department of Biochemistry & Nutrition, Central Food Technological Research
Institute, Mysore-570 020, INDIA
Zinc deficiency-mediated health problems are of growing concern in the developing
world. Although animal foods are good sources of zinc, it has to be mainly derived
from plant foods in countries such as India. In view of the limited information on the
bioavailability of zinc from staple food grains, the same was examined from cereals
and pulses commonly consumed in India. An in vitro dialyzability method involving
simulated gastrointestinal digestion was employed to study the bioavailability. The
foods tested were: Rice (Oryza sativa); Ragi (Eleucina coracana); Jowar (Sorghum
vulgare); Wheat (Triticum aestivum); Maize (Zea mays) Bengal gram (Cicer arietinum)
whole & decorticated; Blackgram (Phaseolus aureus) decorticated; Cowpea (Vigna
catjang); Green gram (Phaseolus mungo) whole & decorticated; Red gram (Cajanus
cajan) decorticated; and Rajmah (Phaseolus vulgaris). The total zinc content of cereals
ranged from 1.08 mg/ 100 g in rice to 2.24 mg/ 100 g in jowar. The zinc content of
pulses was between 2.03 mg/ 100 g (whole Bengal gram) and 2.68 mg/ 100 g (dec-
orticated Bengal gram). Bioavailability of zinc from pulses was generally higher than
that from cereals. Decorticated Bengal gram had the highest amount of bioavailable
zinc (56% of inherent zinc). Zinc bioavailability of these food grains seemed to have
an inverse relationship with their phytate content. Pressure-cooking the food grains as
carried out in domestic food processing generally reduced the bioavailability of zinc
substantially (4.6 to 62 %), unlike the bioavailability of iron, which was generally
increased.
1.07
ZINC ABSORPTION FROM FORTIFIED RICE FLOUR IN SRI LANKAN
CHILDREN
Manjula Deshapriya Hettiarachchi* [1], David Carl Hilmers [2], Kariyawasam Don
Chandrani Elizabeth Liyanage [1], Steven Allan Abrams [2]
Nuclear Medicine Unit, Faculty of Medicine, University of Ruhuna, P. O. Box 70,
Karapitiya, Galle, Sri Lanka [1]; U.S. Department of Agriculture/ARS Children’s
Nutrition Research Center, Department of Pediatrics, Baylor College of Medicine and
Texas Children’s Hospital, Houston, TX, 77030 [2]
Rice flour has been proposed as a vehicle for fortification in Sri Lanka to decrease the
prevalence of iron and zinc deficiency. While widely consumed, rice flour has not been
previously evaluated as a fortified food, and the absorption of added zinc is unknown.
Determination of the bioavailability of zinc is a critical step prior to commencing a
fortification program. After informed consent was obtained, 24 Sri Lankan schoolchil-
dren ages 7 to 10 years were randomized to consume a fortified rice flour meal labeled
with 1.5 mg Zinc Oxide (67ZnO) and either ferrous sulfate (FeSO4) or ferrous sulfate
Disodium ethylenediaminetetraacetic acid (FeSO4-Na2EDTA). An intravenous dose of
0.5 mg Zinc Oxide (70Zn) was simultaneously given to allow calculation of Zinc
absorption by mass spectrometry after urine collection 72 hours later. The subjects re-
ceived a local dish prepared with 25 g of fortified flour. Twenty-three subjects completed
the trial. Fractional absorption of zinc was 13.5 % ± 6.0 in the FeSO4Na2EDTA group
and 8.8 % ± 2.0 in the FeSO4 group. The difference between the two groups was
significant (p=0.037). Total absorption from added zinc in the meal was 202.5 lg and
132 lg in the two groups respectively. This constitutes about 17% and 11% of the 1200 lg
absorbed zinc needs of an 8-year old child. While zinc absorption was low, this study
verified the benefit of Na2EDTA in improving zinc absorption. Additional strategies such
as dephytinization or an increase in the zinc fortification (60 ppm in this trial) may need
to be considered.
1.08
EFFECT OF CARNOSINE ON THE ABSORPTION OF IRON AND VARIOUS
TRACE ELEMENTS IN RATS
Kaori Igarashi* [1], Rieko Hirunuma [1], Hiromitsu Haba [1], Shuichi Kimura[1,2],
Shuichi Enomoto [1]
Cyclotron Center, RIKEN, Wako Saitama, 351-0198, JAPAN [1]; Graduate School of
Human Life Sciences, Showa Women’s University, Setagaya Tokyo, 154-8533, JAPAN
[2]
It is well established that the bioavailability of nonheme iron from food is enhanced by
meat, poultry and fish. However, the factor that improves iron absorption in these
foods is yet to be clarified. Carnosine (b-alanyl-L-histidine) is found in millimolar

concentrations in the skeletal muscles and brains of animals. One of the major func-
tions of carnosine is as a natural antioxidant. Furthermore, carnosine has a property of
chelating metals. Thus, to investigate the effect of carnosine on the absorption of iron
and other trace elements, we determined the uptake rate of various trace elements in
rats administered carnosine using a multitracer technology. Rats were administered the
multitracer solution prepared in the RIKEN Ring Cyclotron with or without carnosine
(dose, 10 mg/rat) into the stomach through a tube. The rats were sacrificed 1, 3, 6 and
24 h after the multitracer administration, and then tissues and blood were collected.
These samples were placed in a Ge detector, and the radioactivities of the multitracers
were measured. The uptake rates of iron into the blood and intestine in the carinosine-
administered rats were significantly lower than that in the control rats after treatment
for only 3 h. Additionally, no significant difference in the uptake rates of iron into
other tissues was observed between the control and carnosine-administered rats. The
uptake rates of zinc into the blood and tissues in the carnosine-administered rats were
significantly lower than those in the control rats. Similarly, the carnosine-administered
rats exhibited a significant decrease in the uptake rates of cobalt and selenium into the
blood and tissues compared with the control rats, suggesting that carnosine adminis-
tration decreases the uptake rates of zinc, cobalt, and selenium into the body. These
results suggest that carnosine may influence the absorption of zinc, cobalt, and sele-
nium compared with that of iron. However, it is considered that the results of this
study were affected by a substantial difference in administration volume between car-
nosine and the trace elements used, and by a higher affinity of carnosine for zinc,
cobalt, copper and manganese than for iron.
1.09
STUDY OF INTERELEMENTAL EFFECTS OF MINERAL BIOAVAILABILITY
USING IN VITRO CONTINUOUS-FLOW DIALYSIS WITH INDUCTIVELY
COUPLED PLASMA OPTICAL EMISSION SPECTROMETRIC METHOD
Kunchit Judprasong,* Mathuros Ornthai, Sopon Purawatt, Atitaya Siripinyanond, and
Juwadee Shiowatana
Department of Chemistry, Faculty of Science, Mahidol University, Rama VI Rd.,
Bangkok, 10400, THAILAND
A continuous-flow dialysis method with an on-line inductively coupled plasma optical
emission spectrometric (ICP-OES) simultaneous multielement measurement for the
study of in vitro mineral bioavailability was developed. The method was based on a
simulated gastrointestinal digestion in a batch system followed by a continuous-flow
intestinal digestion. The simulated intestinal digestion was performed in a dialysis bag
placed inside a channel of flowing stream of dialyzing solution (NaHCO3). The
mineral (Ca, Mg, P, Fe, Zn) concentrations in dialysate were monitored by ICP-OES
at wavelengths (in nm) 396.847 and 317.933 for Ca, 279.553 and 280.270 for Mg,
177.495 for P, 259.940 and 238.204 for Fe, 202.548 and 213.856 for Zn. Internal
standards (Sc, Y) were used for removal of interference effects. The overlaid dialysis
profiles of elements obtained were used to understand the interelemental effects of
essentials elements in various food. The newly developed dialysis system and the
capability of ICP-OES for simultaneous multielement detections and adequate sensi-
tivity for important essential elements make the technique a useful tool for nutritional
studies.
1.10
MULTIELEMENT HAIR PROFILE OF PROFESSIONAL FOOTBALL PLAYERS
Anatoly Skalny [1], Artem Katulin* [1,2]
Russian Society of Trace Elements in Medicine, P.O. Box 56, Moscow, 125047, Russia
[1]; Moscow Scientific-Practical Center of Sport Medicine, Moscow, Russia [2]
Concentration of 24 chemical elements (K, Na, Ca, Mg, P, Co, Cr, Cu, Fe, Mn, Zn, Se,
As, Li, Sn, V, Si, Ti, Ni, Al, Cd, Pb, Hg, Sr) in hair of 149 professional football players
was investigated by ICP-AES and ICP-MS methods. In comparison with control group
of non-sporting men, in sportsmen increased level of most elements was found. The Ca,
Mg, Na, hair concentrations in sportsmen were respectively 1.8, 1.6, 1.7 times higher as
compared to control. Also, there are significantly elevated Na/K, Ca/P ratios in football
players. These deviations can be due to decreased body mass index in sportsmen
(Demidov, 2001). Among the trace elements there were significantly elevated hair Mn,
Co, Cu, Fe (2.1, 1.6, 1.5, 1.3 times, respectively), but Zn, Se concentration were similar to
control. This fact suggests the opinion about the relative deficiency of Zn and Se in
football players. Relation between hair elemental portrait and playing specialization of
the sportsmen was also studied. Goalkeepers were found to have the highest hair content
of Mg, Ca, K, Na, Mn, Si, Cr, Ti and the lowest Co; back players highest Cu, Co, Zn,
Se, Al, As, lowest K, Pb, Sn; forwards highest Fe, Li, Sn, lowest Zn and Si; halfbacks
were characterized by minimal concentration of most elements: Ca, Mg, Na, P, Mn, Cr,
Fe, Ni, Li, Ti, Al, As, Cd. These data demonstrated the necessity of individual approach
to nutritive support of football players for the improvement of health conditions and
sportive results.
1.11
BIOAVAILABILITY OF VARIOUS FORMS OF IRON FORTIFICANTS IN FRESH
WHEAT NOODLES USING IN VITRO DIGESTION TECHNIQUE
Ratchanee Kongkachuichai*, Sooksan Pirapatdit, Visith Chavasit, Rin Charoensiri
Institute of Nutrition Mahidol University, Salaya Campus, Phutthamonthon, Nakhon-
Pathom, 73170, THAILAND
Fresh wheat noodle was used as a model food for iron fortification to compare the
bioavailability of various iron fortificants, the ferrous sulfate (FS), ferrous fumarate (FF),
encapsulated H-reduced elemental iron powder (EEI) and ferric sodium EDTA (Na-
FeEDTA), using in vitro digestion method. The fortification dosage was 5 mg per 100g
serving of cooked noodles equivalent to 1/3 of the Thai Recommended Daily Intake
(RDI) for iron. The data showed that the iron content was closed to 5 mg per serving of
FS, FF, and EEI fortified cooked noodle (5.07 ± 0.36, 4.86 ± 0.10, and 3.77 ± 0.72 mg
respectively), except the NaFeEDTA fortified noodles which was 2.80 ± 0.27 mg. The
test indicated that the iron bioavailability of NaFeEDTA (32%) in fortified noodle was
four to six times higher than FS (5%), FF (8%) and EEI (7%) fortified noodles. The FS
and FF significantly changed the color of noodle dough sheet when compared with the
control. During 96 h storage, all iron fortified dough sheets had changed in color, more
when stored at 30oC than at 4oC. No change in color was observed when cooked noodles
were combined with pork or tomyam flavored soups. Although the addition of FS and
NaFeEDTA may induce metallic odor in raw noodles, the overall acceptability scores of
all fortified raw noodles were ‘‘like slightly’’ and for cooked noodles were ‘‘neither like
nor dislike’’. The cost of iron fortification in fresh wheat noodle was minimal, <0.025
US$ per kg.
1.12
EFFECT OF ZINC AND IRON DEFICIENCY ON RECTAL TEMPERATURE AND
IRON PARAMETERS IN RATS
Aki Konomi*, Katsuhiko Yokoi
Department of Human Nutrition, Seitoku University Graduate School, 550 Jwase,
Matsudo, Chiba 271-8555, JAPAN
To investigate the effect of zinc (Zn) and iron (Fe) deficiency on rectal temperature and
Fe parameters, forty 4-week-old male Sprague-Dawley rats were assigned into 4 dietary
treatment groups of 10 for the 4-week 2 x 2 factorially arranged study: Zn-deficient group
(4.5 mg Zn and 35 mg Fe/kg; Zn-), Fe-deficient group (30 mg Zn/kg, no supplemental Fe;
Fe-), Zn/Fe-deficient group (4.5 mg Zn/kg, no supplemental Fe; Zn-Fe-), and control
group (AIN-93G; Cont). Water intake determined at day 19 was similar among all
treatment groups. At day 24-27, rectal temperature was measured. Rectal temperature of
Zn- was significantly lower than Cont (p < 0.01). Zn deprivation decreased rectal tem-
perature only in Fe-adequate rats. Although rectal temperature of Zn-Fe- was similar to
Cont, TSH of Zn-Fe- was significantly lower than Cont. At day 27-28, bioimpedance was
measured with impedance analyzer. Total body water/body weight ratio of Fe- was
highest among all groups. Intracellular water/extracellular water ratio was significantly
increased in Zn- (p < 0.05). Compared to Cont, hematocrit was significantly decreased in
both Fe- and Zn-Fe-, and was significantly increased in Zn- (p < 0.01). Transferrin
saturation of Fe- and Zn-Fe- was significantly lower than Cont (p < 0.01). Transferrin
saturation of Zn- was highest among all groups. These results suggest that thermogenic
effect of thyroid hormones has little significance in combined deficiency of Zn and Fe.
Increased hematocrit by Zn deprivation in rats may be due to redistribution of body
water and iron.
1.13
BIOAVAILABLE IRON IN SELECTED SPECIES OF GRASSHOPPERS
Virginia Melo*, Maritza Garcı̀a, Rosa Casillas, José Salas, Norma Nogueda
Universidad Autónoma Metropolitana, Xochimilco, Calz. Del Hueso 1100, México, D.
F. 04960, MEXICO
Iron deficiency anemia is still a major nutritional problem in vulnerable population
groups worldwide, mainly during infancy because it can led to negative changes in psy-
chomotor and mental development, wich may be irreversible. The capacity of food to
meet iron requirements depends on both iron content and bioavailability. Iron bio-
availability is the proportion of metal in a food available for intestinal absorption in a
form that is physiologically active. Bioavailability often is categorized as either heme
animal (organic) iron or nonheme (inorganic) iron, these two forms are absorbed by
different pathways, bioavailability of heme iron is relatively high and is not much affected
by diet composition, and nonheme iron is determined by solubility of it in the upper
gastrointestinal tract. Solubility varies widely and depends on pH and chemical form of
the element. The aim of this research is to analyze heme iron content in three species of
grasshoppers: Melanoplus femorrubrum D, Brachystola magna G and Sphenarium pur-
purascens Ch. regularly consumed by different social groups. Iron assessment of the
samples by digest ashes in HCl, then dissolve in deionized distilled water and performed
by atomic absorption spectrophotometry Perkin Elmer 303 with a graphite furnace HGA
400. Results so obtained mg/100 g dry basis were: Melanoplus femurrubrum D 7.3, Bra-
chystola magna G 6.1, Sphenarium purpurascens Ch 2.3. Amounts of heme iron in
grasshoppers represent a good source of the trace element however cooking method or
insect processing might be affected by food combination.
1.14
INFLUENCES ON BIOCHEMICAL AND SEXUAL FUNCTIONS OF ZINC
SUPPLEMENTATION IN MEN
Tomohide Koizumi [1], Kanako Hattori [1], Kazuya Murakami* [1], Aki Konomi [2],
Katsuhiko Yokoi [2]
Tomita Pharmaceutical Co., Ltd., Akinokami, Seto-cho, Naruto, Tokushima 771-0360,
JAPAN [1]; Human Nutrition, Seitoku University Graduate School, Iwase, Matsudo
271-8555, Chiba, JAPAN [2]
Twenty men, ages 24-49, were screened for serum zinc (Zn) levels and divided into 2
groups: hypozincemic group (LZn) and normozincemic group (NZn). The subjects
completed the 2-week baseline and 8-week randomized placebo-controlled, double-blind
crossover trial to test the hypothesis that Zn supplements administering 115 mg of Zn
gluconate (15 mg as Zn) per day are beneficial for male sexual function. Blood and urine
samples were collected at the end of each period. Semen samples were collected 3 times in
each period; a 1st sample 7 days before the end of each period; a 2nd and 3rd sample over
2 consecutive days after a 3-day abstinence. In LZn, serum Zn concentrations increased
significantly (p<0.05) b Zn, while there were no significant changes in other parameters.
In contrast, there were no significant changes in biochemical indices in NZn through all
periods. The volumes of semen and the numbers of spermatozoon of both groups were
unchanged throughout this experiment. However, the number of days per week for sexual
behaviors, such as ejaculation by coitus and self-stimulation in Zn-supplementation
period was significantly more than the baseline (p<0.05). In addition, most of the sub-
jects reported good physical conditions during the Zn-supplementation period. These
results suggest that Zn supplementation is an effective method to enhance male
sexual functions. (K. Murakami works as a senior researcher of Tomita Pharmaceutical
Co., Ltd.)
1.15
ZINC INTAKE AMONG A SELECTED GROUP OF FEMALE
UNDERGRADUATES IN SRI LANKA
Thushangi Pathiraja* [1], Pulani Lanerolle [2], Sunethra Atukorala [2]
Department of Chemistry, Faculty of Science, University of Colombo, SRI LANKA [1];
Department of Biochemistry, Faculty of Medicine, University of Colombo, SRI LANKA
[2]
There have been no studies on zinc intakes among women of reproductive age in Sri
Lanka. A cross sectional study was conducted among thirty female undergraduates
(mean age: 24.8 ± 0.62 years) to assess their daily zinc intake and dietary patterns.
Representative meal samples were obtained from all three main meals on one day, from
each subject. The zinc concentration was determined by atomic absorption spectroscopy
(AAS) following wet digestion and the daily zinc intake was calculated. Nutrient intake
and dietary patterns were evaluated using a 24 hour dietary recall questionnaire and a
food frequency questionnaire. Height and weight was measured and body mass index
(BMI) was calculated. The mean daily zinc intake was 8.20 ± 2.93 mg/day. The intake,
expressed as a percentage of the recommended daily allowance (RDA) was low (54.6 ±
19.6 %). Only one subject had a daily zinc intake that met the RDA. Daily zinc intakes
were significantly positively correlated with protein (r = 0.653, p < 0.01) and energy (r
= 0.397, p < 0.01). The mean ratio of phytate:zinc was 4.20 ± 1.40. The percentage of
subjects consuming red meats and fruits once a day was < 40% and < 30% respectively.
No association was observed between zinc and BMI. Inadequacy of zinc intake and the
observed association between zinc, protein and energy consumption in this group of
subjects warrants further study on zinc status in women of reproductive age in Sri Lanka.
In these subjects phytate levels were unlikely to affect bioavailability of zinc. However,
undesirable dietary patterns which lead to an inadequate frequency of consumption of
zinc containing foods and enhancers, is a cause for concern.
1.16
DIETARY AND BIOCHEMICAL IRON STATUS OF STUNTED AND
NON-STUNTED SCHOOL CHILDREN IN NE THAILAND
Tippawan Pongcharoen* [1], A. Boonpraderm [1], S. Gorwachirapan [1], R. Cook [2], R.
Gibson [2], P. Egkantrong [1] P. Winichagoon [1]
Institute of Nutrition, Mahidol University, Salaya, THAILAND [1]; Department of
Human Nutrition, University of Otago, NEW ZEALAND [2]
In 1995, the anemia prevalence was 24% among primary school children in NE
Thailand. Nutritional iron deficiency (ID) is assumed to be a major aetiological factor,
induced by rice-based diets and low intakes of animal source foods. We compared the
dietary and biochemical iron status of stunted (S) (n=56; 9.7 ± 1.5 y) and non-stunted
(NS) (n=172; 9.3 ± 1.7 y) children from 10 schools in Ubon Ratchathani. Dietary
intakes were based on single 24-hr recalls and Thai food composition values. Bio-
chemical iron indices investigated were: hemoglobin (Hb), serum ferritin, transferrin
receptor, Hb type. Mean (SE) age-adjusted intakes for the S and NS children were:
energy (kcal) 1200±53.3 vs1303±30.3; protein (g) 36.8 ± 1.9 vs 41.8±1.1; animal
protein (g) 17.5 ± 1.4 vs 21.7 ± 0.8; Fe (mg) 5.4 ± 0.4 vs 5.7 ± 0.2; Fe from animal
protein (mg) 2.6 ± 0.3 vs 2.9 ± 0.2; vitamin C (mg) 10.3 vs 8.6. Intakes of total
protein (g/d, per 1000 kcal) and animal protein (g/d) were lower (p<0.05) in S com-
pared to NS children. Of the S vs NS children, 31 vs 29% were anemic, of whom12.5 vs
20% had Hb type EE, 37 vs 34% had AE, and 50 vs 46% had AA. After controlling for
Hb type and age, mean Hb tended to be lower (112.5 vs116.4 g/L; p=0.09) and mean
serum transferrin receptor higher (8.13 vs 7.17 mg/L;p=0.10) in S vs NS. However,
prevalence of ID anemia (based on low Hb + serum ferritin <12 lg/L in absence of
infection) was very low in both groups (0 and 3%), despite iron intakes markedly below
the Thai RDA. This discrepancy warrants further investigation. (Supported by Mi-
cronutrient Initiative and University of Otago.)
1.17
A NOVEL CONTINUOUS-FLOW DIALYSIS SYSTEM WITH ON-LINE
ELECTROTHERMAL ATOMIC ABSORPTION SPECTROMETRIC AND
pH MEASUREMENTS FOR IN VITRO IRON BIOAVAILABILITY STUDY
Jeerawan Promchan*, Wutthika Kitthikhun, and Juwadee Shiowatana
Bangkok 10400, THAILAND
A continuous-flow dialysis (CFD) method with on-line electrothermal atomic absorption
spectrometric (ETAAS) and pH measurements for the study of in vitro mineral bio-
availability was developed. The method is based on a simulated gastrointestinal digestion

in a batch system followed by a continuous-flow intestinal digestion. The simulated
intestinal digestion was performed in a dialysis bag placed inside a channel of flowing
stream of dialyzing solution. The mineral concentration and pH of dialysate were
monitored by ETAAS and a pH meter for 2.5 h. The percentage of dialyzed minerals
(dialyzability) was used to express the bioavailability. The dialysis profile and pH change
can be used to interpret the differences in dialyzability of various samples. The developed
system was also applied to estimate iron dialyzability of various kinds of milk. Iron
dialyzability for cow, corn, cereal, soy, and fortified soy milk was found to be 1.4, 0.0,
20.6, 22.5, and 34.5 % respectively. The developed CFD-ETAAS-pH system was a
simple, rapid and inexpensive tool for mineral bioavailability studies, especially for
minerals in ultratrace levels.
1.18
IRON SUPPLEMENTATION DECREASES THE SIZE OF THE RAPIDLY
EXCHANGEABLE ZINC POOL IN WOMEN
Manuel Ruz* [1], K Michael Hambidge [2], Nancy F Krebs [2], Lei Sian [2], Jamie L
Westcott [2], Juana Codoceo [1], Annabella Rebolledo [1], Jorge Inostroza [1], Fernando
Pizarro [3], Manuel Olivares [3]
Department of Nutrition, Faculty of Medicine, University of Chile, Santiago, Chile [1];
Section of Nutrition Pediatrics, UCHSC, Denver, Co 80220, USA [2]; INTA, University
of Chile, Santiago, CHILE [3]
We evaluated the changes of the size of the rapidly exchangeable zinc pool (EZP) and
fractional zinc absorption (FAZ) during five months of iron supplementation in non-
anemic women consuming ferrous sulfate. Fifty-four apparently healthy women (average
age: 26.9 ± 5.0 y) received: 80 mg elemental Fe/d (Group A, n=14); 40 mg elemental Fe/
d (Group B, n=14); 80 mg elemental Fe/week (Group C, n=14); and a placebo (Control,
n=12). FAZ was determined from a single standard meal labeled with 0.8 mg 67Zn; and
from an aqueous solution labeled with 2 mg 68Zn. FAZ and EZP were determined by an
extrinsic labeling with enriched zinc stable isotope preparations and a urine enrichment
technique. Initial zinc absorption from an aqueous solution was 72.8±14.1% and from a
standard meal was 25.3 ± 6.3%. Two-factor repeated measures ANOVA showed no
significant Time X Group effects. Initial EZP was 159.3 ± 32.5 mg. After five months,
EZP was reduced by 23% (Group A), 26% (Group B), and 11% (Group C), the control
group remained unchanged. Time X Group effect p= 0.002. Results suggest points of
iron-zinc interaction beyond the intestinal wall. (Supported by Fondecyt, Research
project 1000896)
1.19
CHARACTERIZATION OF MOLECULAR WEIGHT AND SIZE DISTRIBUTION
OF ESSENTIAL ELEMENTS IN MILK USING FIELD-FLOW FRACTIONATION-
INDUCTIVELY COUPLED PLASMA SPECTROMETRY
Usarat Kamtabtim, Sudarat Saeseaw*, Atitaya Siripinyanond, and Juwadee Shiowatana
Department of Chemistry, Mahidol University, Rama VI Rd., Bangkok 10400, THAI-
LAND
Elemental size distribution of proteins and colloidal particles in milk was investigated
using a combined technique of field-flow fractionation (FFF) and inductively coupled
plasma (ICP) spectrometry. Two FFF sub-techniques, flow FFF and sedimentation
FFF, were used in this study. With the former, molecular weight distribution of pro-
teins (5 kDa to 1,000 kDa) was observed, whereas with the latter, size distribution
information of colloidal particles (100 nm to 1,000 nm) was obtained. Aluminum and
Cu binding to milk proteins were observed using flow FFF coupled with ICP-mass
spectrometry (ICP-MS). Calcium and Fe binding to milk aggregate and colloidal
particles were detected using sedimentation FFF coupled with ICP-atomic emission
spectrometry (ICP-AES). The information obtained from this study was used to predict
dialyzability of Al, Ca, Cu, and Fe in milk. Moreover, effect of added organic acids,
i.e., citric and ascorbic acid, on the alteration of elemental size distribution profile was
examined. The shift of elemental size distribution profile was observed, implying that
the dialyzability of aforementioned elements changed with addition of organic acids.
This information demonstrates the capability of FFF-ICP spectrometry for nutritional
study.
1.20
EVALUATION OF TWO HOME FORTIFICATION APPROACHES
AMONG PRESCHOOL CHILDREN - EFFECT ON THE
HEMATOLOGICAL MARKERS
Pratibha Dhingra [1], Sunil Sazawal* [1,2], Usha Dhingra [2], Archana Sarkar [1],
Deepak Marwah [1], Saikat Deb [1], Venugopal P Menon [1], Venketesh Iyengar [3],
Olivier Fontaine [4], Robert E Black [2]
Center for Micronutrient Research, Annamalai University, India [1]; Department of
International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore,
USA [2]; Division of Nutrition IAEA Vienna [3]; AUSTRIA; CAHD WHO Geneva,
SWITZERLAND [4]
There have been demonstrated benefits of zinc supplementation from controlled trials.
Daily supplementation is not a feasible approach, We evaluated two home based for-
tification approaches use of a sprinkle (added to regular food of child) or fortified
weaning food (given as single feed per day). Study was conducted among 6-24mo
children (n=226) in peri urban New Delhi. Cluster sampling with 12 clusters of 25
households each were split into 3 matched groups. These were randomly allocated to
receive Sprinkle (Sp, n= 70), Fortified Weaning Food (Wf= 80) and Nutrition Edu-
cation (Ed=76) as the Control Group. Intervention was continued for 6 months. A
blood sample was taken at baseline and end study. Baseline characteristics of the
subjects among three groups were similar. The compliance in the food group was
marginally better than sprinkle. Data on hematological markers indicated that there
was a significant increase in mean hemoglobin Sp. 0.50 (95% CI 0.22-0.77; p=0.001),
Wf 1.28 (95%CI 0.93-1.64; p<0.001). Proportion of anemic children also significantly
decreased in both fortified groups. There was increase in mean Hct Sp 0.95 (95%CI
0.26-1.66);p=0.01, Wf 3.09 (95%CI 2.11-4.07); p<0.001. RDW significantly decreased
in both groups Sp -0.78 (-1.46, -2.27) p=0.03, Wf -1.54 (-0.99, -2.09);p<0.001. Edu-
cation group was unchanged for all these markers. These data indicated efficacy of both
approaches both in terms of improvement in hemoglobin and iron stores, effect size was
higher for fortified weaning food as compared to sprinkle. Using a single feed of
fortified weaning food seems to be a promising strategy to effectively deliver zinc and
iron.
1.21
DIETARY FACTORS AFFECTING TRACE ELEMENT BIOAVAILIBILITY FROM
HUMAN MILK, COW’S MILK AND INFANT FORMULAS IN MOROCCO
F. Zaida [1], N. Lekouch [1], D. Bouglé [2], F. Bureau [2], P. Arhon [2], and A. Sedki* [1]
Laboratory of Ectoxicology, Faculty of Science Semlalia, Marrakech, MOROCCO [1];
Laboratory of Digestive and Nutritional Physiology, CHU de Caen, FRANCE [2]
Human milk is the main source of nutriments to infants during the early stages of their
life. However many deficiencies of minerals and trace elements appear when breast
feeding is discontinued very early by an introduction of cow’s milk or infant formula
which are not adapted to needs of babies nursing. This study presents the results of a
survey focusing on a sample of 228 infants 0-6 months old from Marrakech City
(Morocco). Among this sample, 44 % of the cases exclusively nurse mother’s milk, 37 %
had a partial breast feeding with cow’s milk and the remainder had a breast feeding
with infant formula. Through this study we have raised the impact of socio-economic
family level on both infant nutritional state and milk source feeding. In vitro model was
applied in order to predict the Fe, Zn and Ca bioavailability from breast milk, cow’s
milk and first age infant formula found in Marrakech City. The technique consists on
an intraluminal digestion phase adapted to the gastrointestinal microenvironment of
young infant followed by a dialysis phase with a membrane of molecular mass cut-off
1000 Da. After dialysis, Fe, Zn and Ca contents in the samples and dialysates were
analyzed by Atomic Absorption Spectrometry. The availability of each element is ex-
pressed as a percentage of the total amount of element disappeared from the samples.
Results show that Fe availability is lower in cow’s (18% ± 11%) than in human milk
(23% ± 7%) but it is higher in infant formula (37% ± 5%, P< 0.01). Availability of
Zn is very lower in cow’s milk (7% ± 5%, P<0.001), lower in infant formulas (33% ±
12%) but higher in human milk (48% ± 12%, P<0.001) whereas Ca availability is
higher in infant formula and cow’s milk (respectively 76% ± 8% and 71 % ±4%) but
lower in human milk (64% ±10%, P<0.05). This differences in bioavailability observed
between various type of milk can be explains by the distribution of other components
of milk such as calcium and casein, and also by the influence of the antagonistic
interactions between metals. This study shows the potential risk of iron and zinc de-
ficiency when the cow’s milk is exclusively used during the first months of life while
infant formula seems to be an acceptable vehicle for preventing trace element defi-
ciencies during infant and early childhood. Human milk is considered, however, as a
standard and ideal food for infant.
1.22
MOBILE FORMS OF ZINC, COPPER AND MANGANESE IN ARABLE SOILS
FROM VARIOUS RUSSIAN REGIONS AND CHILDREN’S HAIR CONTENT
Margarita Skalnaya* [1], Viktor Tutelian [2]
Russian Society of Trace Elements in Medicine (RUSTEM), Moscow, Russia [1]; Insti-
tute of Nutrition, Russian Academy of Medical Sciences, Moscow, Russia [2]
For the investigation of supposed relation between the Zn, Cu and Mn concentration in
soils and humans the data of long-term observation of trace elements in soil mobile form
obtained from agrochemical studies with data on 1445 1-6 year old children hair analyses
were compared. The soil trace element content from 19 regions of Russia was measured
by AAS and hair one was measured by ICP-AES. All regions were divided in 3 groups
according to relatively low, optimal or high Zn, Cu, Mn levels in soil. It is found that
children, living in areas with low (group 1) Zn soil levels (<0.7 ppm) have the lowest
average hair Zn concentration (107.8 ± 3.8 ppm, p<0.05), as compared to optimal
(group 2) and high Zn in soil (group 3) (128.3 ± 7.3 and 121.3 ± 4.6 ppm, respectively).
The similar case for Mn was obtained: children from group 1 (Mn<30 ppm) had the
lowest hair Mn concentration (1.08 ± 0.21 ppm). Children from the groups 2 (Mn=30-
70 ppm in soil) and 3 (Mn>70 ppm in soil) had relatively higher Mn hair levels (1.19 ±
0.09 and 1.28 ± 0.14 ppm, respectively). But there were observed no significant depen-
dence between Cu in soil and hair. So, on the basis of epidemiological study it can be
concluded that children hair levels of Zn, and, possibly, Mn reflect a pattern of mobile
form distribution of these elements in soil. This approach can be used for preliminary
estimation of Zn-deficient regions and for investigation of Zn status of population.
1.23
MACRO- AND TRACE ELEMENTS CONTENT OF TEMPORAL AND
OCCIPITAL HAIR SAMPLES
Vladislav Tkachev [1], Vladimir Skalny* [1,2], Evgeny Serebryansky [1]
Russian Society of Trace Elements in Medicine (RUSTEM), Moscow, RUSSIA [1];
Institute of Bioelementology at Orenburg State University, Orenburg, RUSSIA [2]
Multielement analysis of temporal and occipital hair, simultaneously sampled in 66 18-
36 y.o. women according to IAEA recommendations, was provided by ICP-AES (Ca,
Mg, K, Na, P, Fe, Zn, Si) and ICP-MS (Se, Cu, Cr, Mn) (Skalny, Serebryansky, 2002;
2003). All observed women had no serious chronic somatic or psychic diseases, lived in
Moscow and Moscow Region and working in housekeeping or offices. But all of them
suffered from initial symptoms of androgenetic alopecia. Significant (P < 0.05) dif-
ference between this group of women and relatively healthy controls (n = 3748), living
in Moscow Region (Skalny, Bykov, 2003) in cases of Ca, Cu, Zn, Si (elevated in
comparison to controls), and K, Mn, Cr, Se (decreased) was found. The comparison of
elemental content of hair samples, obtained in each woman simultaneously from
temporal and occipital parts of scalp, showed no significant difference. According to
these data, we concluded that there is no significant influence of the location of scalp
hair, used for Ca, Mg, K, Na, P, Fe, Zn, Cu, Mn, Se, Si and Cr analysis. This is why
in the cases of small amounts of hair for analysis, for example in alopecia patients or
in short cutting subjects, the hair from temporal and occipital parts of scalp can be
mixed.
1.24
ESTIMATION OF CALCIUM IN VITRO BIOAVAILABILITY OF VEGETABLES
USING A NOVEL CONTINUOUS-FLOW DIALYSIS SYSTEM
Sutthinun Taebunpakul*, Upsorn Sottimai, and Juwadee Shiowatana
Bangkok 10400, THAILAND
A novel continuous-flow dialysis method with flame AAS measurements was developed
for the estimation of minerals bioavailability in food by determination of their dialyz-
ability. The method involves a simulated gastric digestion with pepsin, followed by
dialysis occurring during a continuous flow of dialyzing solution (NaHCO3). The con-
tinuous flow dialysis was performed in a dialysis chamber for 2.5 h while dialysate was
collected continuously. The optimum concentration of dialyzing solution to obtain a pH
change of the original gastric pH of 2.0 to 5.0 of the intestinal pH within 30 min was
obtained from titratable acidity of the sample. The percentage of dialyzed minerals (di-
alyzability) was used to express the bioavailability. Using the proposed system, calcium
dialyzability in various vegetables was found to vary between 5 40%. The effect of
organic acids on calcium dialyzability for vegetables, fresh and in cooked Thai dishes (for
example; Tom Yam) was studied. It was found that organic acids containing in Thai
dishes can promote calcium dialyzability for vegetables of high dialyzability but not for
those of low dialyzability.
1.25
CHROMIUM CONCENTRATIONS IN HUMAN PLASMA DURING HOME
PARENTERAL NUTRITION AND INFUSUION PRODUCTS IN JAPAN
Yoji Takagi* [1], Takeo Kikuchi [2], Daisuke Ikeda [2], Seiji Nishida [2], Makoto Sato [2]
Dept. of Maternal and Child Nursing, School of Allied Health Sciences, Faculty of Med.,
Osaka Univ., JAPAN [1], Pharm. Research Center, Nipro Co., JAPAN [2]
To determine the necessity of chromium supplementation during long-term Home
Parenteral Nutrition (HPN), we compared plasma chromium concentrations of HPN
patients with those of healthy volunteers, and also measured the chromium contents in
several Total Parenteral Nutrition (TPN) products. The plasma was obtained from 20
male healthy volunteers, and 17 HPN patients. The concentrations of chromium in
plasma and TPN products were determined by graphite furnace atomic absorption
spectrophotometry. Average mean value of plasma chromium in male healthy subjects
was 0.184 lg/L (95% confidence limits: 0.061 0.654 lg/L). Meanwhile, its mean value
of HPN patients(average HPN periods: 105.8 months) was 0.197 lg/L(range:
0.091 0.671 lg/L), which was almost normal range. Chromium was not released from
infusion line with membrane filter. However, the amount of chromium derived from
infusion products was 1.34 11.03 lg/day, which might supply fairly amount of chro-
mium and might contribute to maintain physiological chromium levels in long-term
HPN patients (AMA’s recommendation: 10 15 lg/day). These results suggested the
one of the reasons why plasma chromium concentration does not decrease during long-
term HPN might be due to contamination of chromium in commercially available TPN
products in Japan.
1.26
BLOOD CONCENTRATIONS OF ESSENTIAL TRACE ELEMENTS IN PATIENTS
RECEIVING HOME PARENTERAL NUTRITION IN JAPAN
Kiyomi Tanaka* [1], Kinya Sando [1], Masafumi Wasa [1], Masahiro Fukuzawa [1], Yoji
Takagi [2]
Dept. Pediatric Surgery, Graduate School of Medicine, Faculty of Medicine, Osaka
University, Suita, Osaka, PO Box 565-0871, JAPAN [1], Dept. Maternal and Pediatric
Nursing, School of Allied Health Sciences, Faculty of Medicine, Osaka University, Suita,
Osaka, PO Box 565-0871, JAPAN [2]
A proper balance of essential trace elements is important for the patients receiving
home parenteral nutrition (HPN). However, their intravenously recommended daily
dosages vary widely by various reports. In Japan, commercially available trace elements
solutions for total parenteral nutrition (TPN) contain 2mg of iron(Fe), 3.9mg of
zinc(Zn), 0.3mg of copper(Cu), 50lg of manganese(Mn) and 120lg of iodine(I) in each
ampule which are considered to include daily recommended dosages for adult patients.
If needed, the solutions for each trace element prepared in our hospital are added.
Selenium(Se) solution is also prepared in our hospital and daily 100lg of Se are rou-
tinely given for long-term HPN patients, because there are not yet commercially
available Se solutions. There are few reports about prolonged investigation for trace
elements concentrations during HPN. So, concentrations of Zn, Cu, Fe in serum and
Mn and Se in whole blood were investigated in 21 HPN patients. Zn concentrations in
serum showed within normal range in all patients, although 8 patients needed addi-
tional Zn supply. Se concentrations in whole blood showed lower than normal in all 7
patients without Se supplementation and in 13 out of 19 patients with supplementation
of 100lg of Se. They increased in 3 patients after supply of additional 100lg (totally
200lg) of Se. Mn concentrations in whole blood showed within normal range in 18 and
higher in 1 patient. Cu concentrations in serum showed normal in 16 and higher in 3
patients. Fe concentrations in serum showed normal in 11 and lower in 7 patients. In
conclusions, daily 3.9mg of Zn and 100lg of Se may still require further additions to
TPN solutions to maintain within normal blood level. Commercially available solutions
for each Se, Zn and Cu seem to be indispensable in Japan. (K. Tanaka serves on home
medical division as a pharmacist.)
1.27
IRON ABSORPTION BY HUMAN SUBJECTS FROM DIFFERENT IRON
FORTIFICATION COMPOUNDS ADDED TO THAI FISH SAUCE
Siriporn Tuntipopipat* [1], Thomas Walczyk [2], Christophe Zeder [2], Emorn Wasan-
twisut [1], and Richard F Hurrell [2]
Institute of Nutrition, Mahidol University, Putthamonthon, Nakhon Pathom, THAI-
LAND [1]; Laboratory of Human Nutrition, Swiss Federal, Institute of Technology
(ETH) Zurich, Rüschlikon, SWITZERLAND [2]
Fish sauce has been suggested as a food vehicle for iron fortification in Southeast
Asia. Iron absorption from citric acid with added fish sauce and fortified with ferrous
sulfate, ferric ammonium citrate or ferrous lactate was measured in healthy Thai
women. In addition, the effect of citric acid on iron absorption from fish sauce
fortified with ferrous sulfate was evaluated. Three separate absorption studies were
conducted in adult women (10 women per study) each consumed a basic test meal of
rice and vegetable soup seasoned with isotopically labeled fish sauce (57Fe or 58Fe) on
two consecutive days using a randomized cross over design. Iron absorption was
determined via erythrocyte incorporation of isotopic labels (57Fe or 58Fe) 14 days after
intake of labeled meal. Iron absorption was significantly lower from ferrous lactate
and from ferric ammonium citrate fortified fish sauce than from ferrous sulfate for-
tified fish sauce. Fractional iron absorption (geometric mean; plus1SD, minus1SD)
was 8.7(3.6; 21.4) percent for ferrous lactate compared to 13.0(5.4; 31.4) percent from
ferrous sulfate, p=0.003 (study 1) and 6.0(2.5; 14.3) percent from ferric ammonium
citrate relative to 11.7(4.4; 30.7) percent from ferrous sulfate, p<0.001, in study 2.
Citric acid added at a molar ratio of about 2.5 to iron had no effect on iron ab-
sorption from ferrous sulfate (study 3). Iron absorption in presence of citric acid was
14.1(6.4; 30.8) percent compared to 12.0(5.8; 24.7) percent in its absence (p=0.26).
Iron absorption was 50-100 percent higher from ferrous sulfate fortified fish sauce
than from fish sauce fortified with ferric ammonium citrate or ferrous lactate. In the
presence of citric acid as a chelator, ferrous sulfate would appear to be a useful
fortificant for fish sauce.
1.28
ZINC, IRON, CALCIUM, AND MAGNESIUM METABOLISM WITH HIGH
COPPER INTAKE
Judith R. Turnlund, Joseph M. Domek, and William R. Keyes
Western Human Nutrition Research Center, USDA/ARS, University of California, One
Shields Avenue, Davis, CA 95616, USA
A study was conducted in men to determine the effect of long-term high copper (Cu)
intake on the metabolism and status of zinc (Zn), iron (Fe), calcium (Ca), and mag-
nesium (Mg). Nine men were confined to a metabolic research unit (MRU) for 18 d
and fed a diet with an average of 1.6 mg/d Cu. They continued the study under free-
living conditions for 129 d, supplementing their usual diets with 7 mg/d Cu. The men
returned to the MRU for another 18 d when Cu intake was 7.8 mg/d. Dietary intake
of the other minerals did not change and was 11.7 mg/d Zn, 15.6 mg/d Fe, 740 mg/d
Ca, and 255 mg/d Mg. There were several changes in Cu metabolism due to high Cu
intake. Minor increases were found in some Cu status indices and some indices of
immune function and antioxidant status declined. Cu absorption and retention were
affected, with retention of over 0.6 mg/d with high intake. Indices of mineral status
were determined at the end of each treatment. Absorption of Zn and Fe were deter-
mined using stable isotopes and mineral balances were determined during each MRU
period. Zn, Fe, Ca, and Mg plasma levels did not change and balances were not
significantly different between treatments. Hemoglobin and hematocrit levels increased
significantly (14.5 vs 15.0 g/dL and 42.6 vs 45.1 %) with high copper intake and
average serum ferritin was lower (90.0 vs 74.8 ug/mL), but not significantly so. Neither
Zn nor Fe absorption changed significantly (Zn 35.7 vs 38.7 %, Fe 9.6 vs 9.0 %). The
results suggest that long-term high Cu intake does not have marked effects on the
metabolism of most other minerals studied.
1.29
DO MULTIPLE TRACE ELEMENT DEFICIENCIES COEXIST AMONG SCHOOL
CHILDREN IN NE THAILAND?
Pattanee Winichagoon* [1], Tippawan Pongcharoen [1], S Gorwachirapan [1], A Boon-
praderm [1], Rosanne Cook [2], Karl Bailey [2], Rosalind Gibson [2]
Institute of Nutrition, Mahidol University, Salaya, THAILAND [1]; Department of
Human Nutrition, University of Otago, NEW ZEALAND [2]
Concurrent trace element deficiencies in developing countries are probably widespread.
Their etiology is multifactorial: inadequate intakes, parasitic infections, genetic haemo-
globinopathies, malaria, and infectious diseases may all play a role. Such deficiencies may
contribute to impairments in growth, immune competence, mental and physical devel-
opment in childhood. As part of a larger study, we have investigated the prevalence of
anemia and deficiencies of iron, zinc, iodine, and vitamin A in a selected sample of rural
children in NE Thailand. Non-fasting venipuncture blood samples were collected from
567 children aged 6-12 y and attending 10 schools in Ubon Ratchathani province for the
assessment of serum ferritin, transferrin receptor, zinc, retinol, C-reactive protein (CRP),
and urinary iodine from casual urine samples; haemoglobin (Hb) type and complete
blood cell count were also measured. Children with CRP > 10 mg/L (n=12) were
excluded. Of the children, 57% had low serum zinc levels (based on IZiNCG cutoffs), 40%
had urinary iodine < 100 ug/L, and 24% had serum retinol <1.05 umol/L, of which only
4% were <0.7 umol/L. By contrast, only 2% had iron deficiency (ID) based on a low
serum ferritin (ie <12 ug/L) and low Hb, despite an overall anaemia prevalence of 31%.
Haemoglobin concentrations were significantly associated with hemoglobin type, serum
retinol, and age. In summary, among 474 children, 68% were at risk of two or more
concurrent micronutrient deficiencies, principally iodine and zinc; ID was rare in this
population. (Funded by Micronutrient Initiative Fund and University of Otago.)
1.30
FACTORS AFFECTING ZINC KINETICS IN PREMENOPAUSAL WOMEN
Katsuhiko Yokoi* [1], Norman G. Egger [2], V. M. Sadagopa Ramanujam [2], Nancy W.
Alcock [2], Hari H. Dayal [2], James G. Penland [3], Harold H. Sandstead [2]
Dept. Human Nutrition, Seitoku University, Matsudo, Chiba 271-8555, JAPAN [1];
Dept. Preventive Medicine & Community Health, University of Texas Medical Branch,
Galveston, TX 77555-1109, USA [2]; USDA ARS Grand Forks Human Nutrition Re-
search Center, Grand Forks, ND 58202-9034,USA [3]
Factors affecting zinc (Zn) kinetics were examined in 50 premenopausal women including
15 oral contraceptive (OC) users and 35 OC nonusers. Kinetic parameters of Zn were
determined by tracing disappearance over 24 hours of 2 mg intravenous dose of Zn-67
using ICP-MS. Calculation was based on the three-compartment mammillary model.
OCs significantly (p < 0.05) elevated the central Zn pool (Q1, the plasma) per fat free
mass. The rate constant from Q1 to the lesser peripheral Zn pool (Q2) was significantly
higher in the OC users than the nonusers. The relationship between Zn and iron (Fe)
status was analyzed in the OC nonusers. In hypozincemic subjects with plasma Zn below
700 lg/L, Q2 and serum Fe were significantly lower than in normozincemic subjects.
Food frequency correlated with serum ferritin (SF) and Zn kinetics. Beef positively
predicted and bran cereal negatively predicted both SF and Q2. SF significantly and
positively correlated with the Zn pool sizes. In the broken line model, the breakpoint for
Q2 was at SF 17.8 lg/L. In summary, 1) OCs affected Zn kinetics; 2) Frequency of beef
and bran cereal consumption affected both Fe and Zn status; 3) Low Zn nutriture was
associated with SF less than 20 lg/L. (Support: DAMD 17-95-C-5112 & USPHS M01-
RR-00073.)
1.31
MATHEMATICAL APPROACH FOR ESTIMATING IRON REQUIREMENTS
FROM POPULATION DATA
Katsuhiko Yokoi*
Department of Human Nutrition, Seitoku University Graduate School, Matsudo, Chiba
271-8555, JAPAN
The established methods for estimating iron (Fe) requirements are the balance method
and factorial method. As an alternative, a mathematical approach for estimating Fe
requirements from the population data of Fe status (distribution of dietary Fe intake and
the prevalence of low Fe status) is proposed here. The prevalence (D) of the inadequate
nutritional status of the particular nutrient is the proportion of the individuals with the
inadequate nutritional status: D = (Sum of individuals whose dietary intake <
requirement) / (total number of individuals). Assuming that the dietary intake and the
requirement obey a certain distribution function such as bivariate normal distribution
function, the estimated average requirement (EAR) is obtained as the solution of the
equation derived from the above relationship. This situation can be applicable to Fe
requirements for certain populations excluding premenopausal women. For premeno-
pausal women, the convolution integral between basal Fe loss and menstrual Fe loss is
computed to obtain total Fe loss. The Fe status data reported in the 6th edition of
Recommended Dietary Allowances for Japanese (1999) were analyzed as an illustration.
EAR was affected by forms of distribution function, relative standard deviations of
requirements, and correlation coefficients between Fe intakes and requirements. We
conclude that mathematical approach can be very useful to estimate Fe requirements and
to elucidate dietary recommendations of Fe. This approach may contribute to deter-
mining requirements of other nutrients as well as Fe.
1.32
IRON ABSORPTION IN A MINERAL SUPPLEMENTED OAT-BASED DRINK
Huan-Mei Zhang* [1], Gunilla Önning [1], Rickard Öste [2], Elisabeth Gramatkovski [3],
Lena Hulthen [3]
Biomedical Nutrition, Lund University, Chemical Center, P.O. Box 124, SE22100 Lund,
SWEDEN [1]; CEBA Foods AB, Scheelevagen 18, SE22363 Lund, SWEDEN [2]; Dept.
Clinical Nutrition, Sahlgrenska Academy at Göteborg University, SE41345 Göteborg,
SWEDEN [3]
Oats-based drinks are introduced to human nutrition as alternatives to dairy prod-
ucts. However, the bioavailability of minerals in such drinks could be low due to the
processing loss and high content of phytates in oats. To increase the nutritive value
of such drinks, special treatments including iron supplementation, phytase hydrolysis
and citric-acid addition were performed. An oat drink was supplemented with iron
(ferric ammonium citrate or ferric pyrophosphate) and the iron absorption was
measured by extrinsic labeling with 55Fe and 59Fe and determined for whole-body
retention and the erythrocyte uptake of isotopes. The effect of adding citric acid and
phytate hydrolysis was also evaluated. There was no statistically significant difference
between the Fe absorption from ferric ammonium citrate supplemented and that
from ferric pyrophosphate supplemented drink. The addition of citric acid and
phytate hydrolysis improved Fe absorption in the supplemented oat drink. The
present study indicated that ferric ammonium citrate and ferric pyrophosphate are
well absorbed in a phytase-treated and citric-acid-added oat-based drink. Further
studies will be made to see if the drink can prevent Fe deficiency in vulnerable
groups.
1.33
EDIBLE INSECTS: SELENIUM SOURCE FOR MEXICAN RURAL
COMMUNITIES
Virginia Melo*, Maritza Garcı́a, Claudia Maass, Rosa Casillas, Norma Nogueda
Universidad Autónoma Metropolitana, Xochimilco, MEXICO
The role of selenium as a trace element is as a catalytic component in enzymes of
proteins. It is part of the glutation peroxidase enzyme that protects against oxidative
damage to erythrocytes and is part as well of other molecules involved in multiple
biochemical and physiological functions. The selenium amino acids are the main
dietetic source of the element in food. The selenomethionine proceeds of vegetables

and the selenocysteine of animals; both absorbs by the same mechanism of methionine
amino acid. The inorganic selenium absorption may be efficient. The principal sele-
nium sources are meat, shellfish Brasil nuts; foods with limited accessibility for the
rural communities in Mexico, that have a limited diet basis on beans, corn, insects,
chili and vegetables. The aim of the research is to analyze the selenium content in four
species of edible insects; grasshoppers (Sphenarium purpuranscens Ch, Sphenarium
histrio S) and jumiles forest bugs (Euchistus sulfutus S and Euchistus strenus D)
captured in the states of Guerrero, Morelos and Oaxaca. The organisms are consumed
in rural communities by cultural tradition. The studied insects were collected in the
consumption zones of the states mentioned above. Gathered insects were dehydrated
and ashed at 600 C to eliminate organic matter and to quantify trace element by
atomic absorption spectrophotometry. Data obtained for selenium (mg/100 g dry
basis) were similar for three states samples: Sphenarium purpuranscens Ch, 117 mg;
Sphenarium histrio S 115 mg; jumiles forest bugs: Euchistus sulfutus S, 109 mg; and
Euchistus strenus D, 101 mg. Selenium regulates many physiological functions and
according to RDA recommendations, adults requires 55 to 75 lg and children about
20 to 30 lg selenium/day. Average weight for sample unit was 0.17 g for grasshoppers
and 0.9 g for jumiles forest bugs. Mentioned insects represent a natural organic
selenium source. The intake of one or two insects a day meets the requirements for a
good human health.
Category 2: Trace Elements and Cancer
2.01
TRACE ELEMENTAL PROFILE: AN INDEX FOR DISEASE PROGRESSION
Anindita Chakraborty* [1], Sudarshan Mathummal [1], Bidhan Mohanta [2], Mandira
Barua [2], Amal Bhattacharya [3], Ajit Sinha [1]
Inter University Consortium for DAE Facilities, Calcutta Centre, Kolkata 700098,
INDIA [1]; Gauhati University, Guwahati 781014, INDIA [2], Calcutta University,
Kolkata 700019, INDIA [3]
Trace elements are known to play pivotal role in body metabolism. Alterations in
elemental status have been reported in different patho-physiological conditions
including several forms of malignancies. This has motivated interdisciplinary research
for quantitative determination of elemental constituents in biological tissues of subjects
suffering from any disease. In this perspective our research programs have been de-
signed to understand the role of trace elemental concentrations as a health index and
diagnostic tool for body homeostasis. These studies, carried out as collaborative re-
search involving a number of university departments, include determination of trace
elements in chemically induced carcinogenesis in animal models, chemopreventive po-
tential of medicinal plants and human subjects suffering from visceral leishmaniasis.
Our studies employ primarily PIXE complemented by EDXRF and AAS. Results
indicate chemopreventive potential of an essential micronutrient, vanadium and a
traditionally used medicinal plant, Vinca rosea for modulating the altered elemental
(Fe, Ca, Cu, Zn, Se, Mn) concentrations due to induced carcinogenesis. Data also
reflect definite correlation between elemental composition and progression of Leish-
maniasis. The observed alterations in Cu, Fe etc confirm the clinically reported path-
ogenicity associated with this disease. Our paper highlights correlation between ele-
mental profile and metabolic disorder.
2.02
ORAL CANCER-ITS RELATIONSHIP WITH SELENIUM AND ZINC: AN
EPIDEMIOLOGICAL STUDY
Nabakanta Jana
Dept. of Zoology, Charuchandra College, 22 Lake Road, Calcutta-700 029, INDIA
The incidence of oral cancer in different areas of West Bengal, India was recorded for
the past 10 years (1994 2003). Data based on epidemiological survey revealed that
there has been increased incidence of oral cancer from 107 to 340 per 1,00,000 of the
population. The results also indicate that the age group between 45 to 55 years among
male population was more susceptible to oral cancer wherein there is a positive
correlation between life style and personal habits with respect to oral hygiene and
cancer incidence. Trace elemental analysis, using Atomic Absorption Spectroscopy,
with special reference to Se and Zn in the hair, nail and serum was performed from
individuals residing in those areas. Results showed a positive correlation between the
Se, Zn levels and the incidence of oral cancer. Data were analyzed by routine sta-
tistical tests and multivariate stepwise regression analysis. Practice of proper oral
hygiene together with supplementation of essential trace elements in regular diet may
be a panacea to combat the challenges posed b cancer in future. Mass education,
awareness program and role of media are needed for the better solution of this
disease.
2.03
LOW FREQUENCY OF MALIGNANT TUMORS AMONG WORKERS FROM
BORON PROCESSING FACILITY 2: MALIGNANCY AMONG MINERS,
EMPLOYERS, AND SUBJECTS WITH BORON-UNRELATED OCCUPATION
Bekir Sıtkıur Sayli* [2]
Boron Research Center, Balıkesir University, Balikesir, TURKEY [1]; Dept. Orthopae-
dics and Traumatology, Fatih University Hospital, Ankara, TURKEY [2]
Although there is no experimental evidence on the carcinogenicity of boric acid, no
human data is available on the subject for the time being. In a field survey from 1999
to 2002 we observed two cases of malignant tumor among 2418 workers of all four
borate centers with large deposits and processing facilities, plus The Borax and Acid
Plants, in a country with the world’s largest borate deposits ever discovered. Another
two instances were found among 553 employers work-places of which were the same
or very close to these centers. Still two cases of malignancy involving 1014 subjects
with a boron-unrelated occupation from the same environment were revealed. Some
sufferers had a familial story no difference between subgroups was discovered
[p>0.05]. Even though this was not a formal prevalence study, these observations
with an indeed very few cases of a malignant disease in addition to the lack of
concentration in neither group lend support that the element shows ‘‘evidence of no-
carcinogenicity’’ for human beings.
Category 3: Trace Elements and Cardiovascular and Pulmonary Diseases
3.01
ZINC DEFICIENCY INDUCED CHANGES IN THE EXPRESSION OF VASCULAR
CELL PROTEINS USING PROTEOMICS
John H. Beattie* [1], Soo-Lim Lee [2], and In-Sook Kwun [2]
Division of Cellular Integrity, Rowett Research Institute, Bucksburn, Aberdeen AB21
9SB, UNITED KINGDOM [1]; Department of Food Science and Nutrition, Andong
National University, Andong, Kyungpook 760-749, SOUTH KOREA [2]
The role of zinc in protecting vascular tissue has not been widely considered, largely
because zinc status cannot be reliably measured and few epidemiological studies relating
status with disease have therefore been made. We have utilized proteomic techniques to
try and detect which vascular metabolic or signalling pathways are affected by physio-
logically relevant zinc deficiency. Three week old rats were fed semi-synthetic diets
containing adequate (35 mg/kg) or marginally deficient (3 mg/kg) levels of zinc for 43
days, and animals in a third group were pair-fed with deficient rats. Each group contained
9 animals. Two-dimensional gel electrophoresis of individual aorta samples using a pH
gradient of 3-10 and an acrylamide gradient of 8-16% revealed 1697 proteins, of which 18
showed levels significantly (p<0.01) lower in zinc deficiency as compared to adequacy.
Only one protein was significantly higher in zinc deficiency. Tryptic digestion and mass
spectrometry followed by peptide mass fingerprinting using the Mascot search engine
(http://www.matrixscience.com/) and the NCBInr database, revealed that many of the
zinc-regulated proteins were related to carbohydrate metabolism. Further work is in
progress to establish whether the identified proteins are affected in human endothelial
cells.
3.02
REGRESSION OF COPPER DEFICIENCY-INDUCED CARDIOMYOPATHY
Laila Elsherif [1], Youchun Jiang [1], Jack T. Saari [2], Y. James Kang* [1]
Dept. Medicine, Pharmacol. Toxicol., Univ. Louisville, Louisville, KY 40202, USA [1];
U.S. Department of Agriculture, Grand Forks Human Nutrition Research Center,
Grand Forks, ND 58202, USA [2]
Dietary copper deficiency (CuD) leads to cardiac hypertrophy in various animal models.
The present study was undertaken to determine whether CuD-induced cardiomyopathy is
reversible upon copper repletion (CuR). Dams of FVB mice were fed copper deficient diet
(0.3 mg/ kg) starting from day 3 post delivery and the weanling pups were fed the same
diet until CuR (6.0 mg/kg Cu in the diet) at 4 or 5 wk of age. A significant regression of
CuD-induced cardiac hypertrophy was observed in the CuR mice. Histopathological
examination revealed that CuR eliminated CuD-caused lipid deposition and reduced the
volume of the hypertrophied myocytes and electron microscopy demonstrated that CuR
reversed CuD-induced mitochondrial swelling and structural disarray of organelles.
Hemodynamic analysis showed that CuR completely reversed CuD-depressed systolic
and diastolic function. Furthermore, the CuD-blunted myocardial responses to b-
adrenergic agonist, isoproterenol, were also restored in the CuR mice. Microarray and
real-time RT-PCR analysis revealed that CuR corrected CuD-altered gene expressions in
the heart. This study thus demonstrates for the first time that CuR results in regression of
CuD cardiomyopathy as demonstrated by the reversal of depressed cardiac hemody-

namic and contractile function, the restored responsiveness to b-adrenergic stimulation
and the correction of the altered gene expressions. (Supported in by NIH grants HL63760
and HL59225 and USDA grant 2002-35200-11573.)
3.03
SELENIUM SUPPLEMENTATION AND BLOOD HEMORHEOLOGICAL
CHANGE
Hiroshi Koyama [1], Kaori Miyazaki [1], Misa Nara [2], Masami Murakami [2]
Dept. Public Health [1]; Dept. Clinical Laboratory Medicine [2], Gunma University
Graduate School of Medicine, 3-39-22 Showa, Maebashi, 371-8511 Japan
To investigate the effect of selenium supplementation on the blood fluidity, eleven healthy
adults were administered 200 microgram of selenium per day during one week. Before
and after the supplementation, blood fluidity of the subjects was measured with MC-
FAN (microchannel array-flow analyzer). The blood fluidity was indicated by the passage
time of the 100 microliter of heparinized whole blood through the narrow (id=7
micrometer) microchannel array. Before the supplementation the mean passage time was
44.0 (SD=5.7) seconds. And the time significantly (p<0.05) shortened to 37.5 (SD=2.8)
seconds after the supplementation. Serum selenium concentration and the activity of
serum glutathione peroxidase, a selenoenzyme, tended to be increased during the sup-
plementation. The factor analysis of the passage time and biochemical indices of the
subjects showed that the blood fluidity change was related to the metabolic modification
of lipoproteins during the supplementation. The results suggest that the selenium sup-
plementation improves the blood fluidity through the modification of lipoprotein meta-
bolism.
3.04
TRACE ELEMENT LEVELS AND REDOX MARKERS IN HUMAN
ATHEROSCLEROSIS: RELATIONSHIP WITH APO E POLYMORPHISM
Paula A. Lopes [1], Patrı́cia Napoleão [1], Teresa Pinheiro [2], Maria C. Santos [3], Maria
O. Rodrigues [4], Luciana Costa [4]; Fátima Ceia[1], Luis Vicente [1], Maria L. Pavão [5],
Jean Neve [6], Ana M. Viegas-Crespo* [1]
C.B.A, Fac. Ciências, Univ. Lisboa, 1749-016 Lisboa, PORTUGAL [1]; I.T.N, POR-
TUGAL [2]; C.Q.B., Fac. Ciências, Univ. Lisboa, PORTUGAL [3]; INSA, Lisboa,
PORTUGAL [4]; C.I.R.N, Univ. Açores, P. Delgada, PORTUGAL [5]; Inst. Pharm., U.
L. B., C. Plaine 205-5, Bruxelles, BELGIUM [6]
Oxidation of LDL in the vessel wall plays an important role in the development of
atherosclerotic lesions mediated by several mechanisms. Measurement of antioxidant/
oxidant-related parameters and trace element levels in serum in parallel with the allelic
profile of apo E may be useful in assessing the risk of atherogenesis in humans. Blood
activities of antioxidant enzymes, SOD and GPx, total thiols, caeruloplasmin,
a-tocopherol as well as products of oxidative damage, MDA and protein carbonyls were
evaluated in patients with diagnostic atherosclerosis in parallel with matched healthy
subjects. The elemental levels were assessed in plasma and blood cell fractions. The allelic
frequencies of apo E were similar to those referred in the literature. Significantly de-
creased K, Fe and Zn levels were found in plasma of the atherosclerotic group. A ten-
dency for a disruption in antioxidant enzyme status was observed in patients, although
serum caeruloplasmin and a-tocopherol contents were unchanged. Plasma protein car-
bonyls levels were decreased in patients. This study could contribute for a better
understanding of the relationship between genetic and redox balance markers, which is of
utmost importance for the prevention of atherosclerosis.
Category 4: Trace Elements and Diabetes Mellitus, Metabolic Syndrome X, and Obesity
4.01
STRESS INDUCED CHANGES IN TISSUE CHROMIUM, COPPER, AND ZINC
ARE NOT CORRELATED WITH CHANGES IN INSULIN REALATED
VARIABLES FOLLOWING THERMIC AGGRESSION
R. A. Anderson* [1], D. Aguay [3], N. A Bryden [1], C. Sandre [3], Y. Chancerelle [2],
M. M. Polansky [1], and A M. Roussel [3]
Beltsville Nutrition Center, Beltsville, MD, USA [1]; Centre de Recherche des Arme’s [2] ;
J Fourier Univ, La Tronche, FRANCE [3]
The functions of Cr, Cu and Zn are linked to those of insulin and factors that alter insulin
often lead to changes in these trace metals. The objective of the present study was to
determine the effects of thermic aggression on Cr, Cu, Zn and insulin related variables. A
third-degree burn injury involving 20% of total body surface was applied to 45 male
Wistar rats, divided into 9 groups. Study conformed to the guidelines established by the
NIH and the experiments were reviewed and approved by the French Army Ethical
Committee. There was a significant rise in corticosterone reaching a maximum the first
day of injury that was accompanied by significant increases in circulating insulin and
glucagon that were maximal after two days. Changes in IGF-1 were not significant. For
Cr, the most sensitive indicator of burn injury was a rapid decline in quadriceps Cr with
no significant changes for Cu and Zn. For Zn, the largest effects were observed in the liver
with a biphasic increase that was maximal within the first day of injury that decreased in
the next three days but then increased and was still increasing after 10 days. Changes in
tissue, plasma and urinary Cu concentrations were not significant. Trace metal concen-
trations in the brain were not altered by thermic aggression. These data demonstrate
independent changes in the tissue concentrations of Cr and Zn that cannot be directly
associated with the changes in insulin related variables.
4.02
EFFECT OF ZINC AND POLYPHENOLS TREATED DIABETIC RATS
Manuela Ciocoiu [1], Daniela Ciubotariu [1], Laurentiu Badescu [2], Magda Badescu* [1]
Dept. of Pathophysiology [1], Dept. of Cell Biology [2], University of Medicine and
Pharmacy ‘‘G. T. Popa’’ Iasi, Universitatii St. 16, Iasi 700116, ROMANIA
In this study, we investigated the in vivo effect of zinc (Zn) and polyphenols on plasma
antioxidant capacity of streptozotocin (STZ) treated diabetic rats, a model of diabetes
characterized by a major oxidative injury. Animals were divided into four groups (eight in
each group). One includes normal rats control group (C). Other animals were injected
intraperitoneal with STZ (55 mg/kg body weight). Diabetic rats were randomized
according to glycemia to: untreated diabetic (D); diabetic treated with polyphenols-rich
extract of grape seeds (D+P) and diabetic treated with Zn (D+Zn). The P and Zn were
administered by gavage for 12 weeks. Plasma antioxidant capacity was determined by the
total antioxidant status method of Randox. Untreated diabetic rats had a significantly
lower plasma antioxidant capacity than the nondiabetic control group. Treatment with
either P or Zn restored the plasma antioxidant capacity of diabetic animals to a level not
significantly different from that of nondiabetics. Glycemia, total cholesterol, aspartate-
aminotransferase (ASAT) and alanine-aminotransferase (ALAT) were significantly
(p<0.001) higher in diabetic (untreated or treated) than in nondiabetic rats. Our results
clearly indicate that both polyphenols and Zn restored plasma antioxidant capacity im-
paired by diabetes.
4.03
ANTI-DIABETIC ACTIVITY OF ZINC IN EXPERIMENTAL DIABETES
Magda Badescu* [1], Manuela Ciocoiu [1], Catalina Bohotin [1], Codruta Badescu [2]
Dept. of Pathophysiology [1], Dept. of Internal Medicine [2], University of Medicine and
Pharmacy ‘‘G. T. Popa’’ Iasi, Universitatii St. 16, Iasi 700115, ROMANIA
Zinc (Zn) was found to inhibit lipid peroxidation (LP), scavenge hydroxyl and superoxide
radicals in vitro. We investigated the effect of a high Zn diet in the presence and absence
of alloxan (Allx) induced diabetes mellitus on antioxidant status. The animals were
divided into four groups, each comprising ten young Wistar male rats. Zinc, administered
by gavage for six weeks, reduces glucose level in alloxan diabetic rats significantly
(p<0.005) at a dose level of 70 (mg/kg body weight) daily administration. The serum
LP level that was increased by alloxan was reduced significantly (p<0.001) by zinc
supplementation. Decreased liver glycogen, after alloxan administration showed a sig-
nificant (p<0.001) increase after zinc treatment. The zinc administration remakes the
catalase (CAT) and glutathione-peroxidase (GPx) activity in the hepatic tissue, decreases
the levels of malondyaldehyde (MDA) and produces o high synthesis of thyol groups
(glutathione-GSH). These results indicate that alteration in the glucose utilizing system
and oxidation status in rats increased by alloxan were partially reversed by zinc ad-
ministration. So, this fact proves that zinc acts like a free radical scavenger. Diabetic
evolution, especially in young people could be much better by adding certain soluble salts
to the basic treatment.
4.04
BIOMIMETIC CHROMIUM (III) COMPLEX AND FRUCTAN
SUPPLEMENTATION AFFECT INSULIN AND MEMBRANE GLUCOSE
TRANSPORT IN RATS
Bogdan Debski* [1], Zbigniew Krejpcio [2], Tomasz Kuryl [1], Rafal Wojciak [2], Maciej
Lipko [1]
Dept. Physiol. Sci., Vet. Med. Fac., Agricultural University, Nowoursynowska 159,
02-787 Warsaw, POLAND [1]; Dept. Human Nutrition and Hygiene, Agricultural
University, 31 Wojska Polskiego, 60-624 Poznan, POLAND [2]
In the 3-factorial experiment we studied the effect of Cr (III) and fructan supplementation
on plasma insulin concentration and glucose cross membrane glucose transport in red
blood cells in growing male Wistar rats. Animals were divided into 8 groups (of 7 animals
each) and fed ad libitum semi-synthetic AIN-93 diets composed of different type of
fructans (factor A: inulin or oligofructose), amount of fructan (factor B: 5% or 10% of
diet mass) and Cr (III) as [Cr3O(O2CCH2CH3)6(H2O)3]+ cation (factor C: 0.5 or 5 mg
Cr/kg diet) for 10 weeks. Rats were sacrificed to collect blood. Serum glucose (GL) was
estimated using enzymatic method, plasma insulin (INS) by radioimmuno-assay, glucose
transport (GL-T) was measured using 3H-deoxyglucose uptake in isolated erythrocytes.
Experimental variables did not affect serum GL concentration. Plasma INS level was
significantly reduced by high Cr diet by 15.6%. GL-T in RBC was markedly increased by
oligofructose by 5.7%, high fructan diet by 11.6% and high Cr diet by 9.6%. Also the
ABC-interaction significantly increased GL-T in RBC. Both non-digestable oligofructans
and biomimetic Cr (III) cation influence insulin activity and glucose utilization in rats,
thus possess therapeutic potential in regulation of carbohydrate metabolism. (The
experiment was an integral part of the Research Grant Nr 0528/P06/2002/23 financed by
KBN.)
4.05
SERUM SELENIUM AND GLUTATHIONE PEROXIDASE STATUS IN
METABOLIC SYNDROME AND OBESITY; EFFECTS OF DIETARY SELENIUM
AND STATIN THERAPY
Majid Ghayour-Mobarhan*, Andrew Taylor, David Lamb, Susan New, Callum Liv-
ingstone, Tim Wang, and Gordon Ferns
University of Surrey, Guildford, GU2 7XH, UNITED KINGDOM
Baseline serum selenium (Se) and glutathione peroxidase (GPx) status, and the effects of
statin therapy were investigated in subjects with metabolic syndrome (MS) (n=142) and
obesity (n=82). Patients were recruited from the lipid clinic (n=238). Control subjects
were recruited from university staff (n=192). Dietary Se was estimated using a validated
food frequency questionnaire. Dyslipidaemic patients had higher serum Se (p<0.05) and
dietary Se intake (p<0.05), and lower GPx (p<0.0001) than controls. Obese patients had
lower serum Se (p<0.05), and GPx (p<0.0001) compared to both controls and the non-
obese subgroup of these patients. Serum Se and GPx did not differ significantly between
the subgroup of patients with and without MS. However, serum Se decreased with ac-
cumulating features of the MS (p<0.05). Treatment with 10 mg/d of statin for 4 months
in dyslipidaemic patients (n=20) previously not treated with a lipid-lowering agent, did
not significantly alter serum Se and GPx. The increased serum Se in dyslipidaemic pa-
tients may be related to higher dietary intake of Se in patients than controls. The lower
serum Se and GPx in obesity and decreased serum Se with accumulating features of the
MS may be related to the presence of an atherosclerosis prone state and consequential
consumption of antioxidants by free radical interaction.
4.06
BIOMIMETIC CHROMIUM (III) COMPLEX AND FRUCTAN
SUPPLEMENTATION IMPROVE BLOOD VARIABLES IN STZ-INDUCED
DIABETIC RATS
Zbigniew Krejpcio* [1], Bogdan Debski [2], Rafal Wojciak [1], Tomasz Kuryl [2], Mal-
gorzata Tubacka [1]
Dept. Human Nutrition and Hygiene, Agricultural University, 31 Wojska Polskiego, 60-
624 Poznan, POLAND [1]; Dept. Physiol. Sci., Vet. Med. Fac., Agricultural University,
Nowoursynowska 159, 02-787 Warsaw, POLAND [2]
Diabetes type-1 was induced in male Wistar rats by intraperitoneal single injection of
STZ (60 mg/kg BW). Rats were subsequently divided into 8 groups (of 8 animals each)
and were fed ad libitum semi-synthetic AIN-93 diets composed of different type of
fructans (factor A: inulin or oligofructose), amount of fructan (factor B: 5% or 10% of
diet mass) and Cr (III) as [Cr3O(O2CCH2CH3)6(H2O)3]+ (factor C: 0.5 or 5 mg Cr/kg
diet) for 5 weeks. Rats were sacrificed to collect blood. Serum glucose (GLU) was esti-
mated using enzymatic method, plasma insulin (INS) by radioimmunoassay, serum TC
(total cholesterol), HDL, LDL, and TG levels were determined by appropriate methods.
It was found that oligofructose vs. inulin significantly increased plasma INS by 25% and
HDL by 15%. Higher amount of fructan in the diet (10% vs. 5%) markedly lowered
serum GLU, TC and LDL by 26%, 11% and 18%, respectively. Dietary Cr supplemen-
tation (up to 5 ppm Cr) significantly reduced serum GLU by 26%, and increased HDL by
14%. Apart from that interaction of experimental factors: ABC (for serum GLU), AC
(for plasma INS), AB and AC and ABC (for HDL) as well as AB (for TG) was noted. We
conclude that both Cr (III) and fructans are efficient dietary components able to improve
glucose and lipid metabolism in diabetes type-1. (The experiment was an integral part of
the Research Grant Nr 0528/P06/2002/23 financed by KBN.)
4.07
DIETARY BORON REDUCES SERUM INSULIN CONCENTRATIONS
REGARDLESS OF VITAMIN D STATUS IN RATS
Yeong Rhee* [1], Curtiss Hunt [2], Joseph Idso [2]
Dept. Health, Nutrition and Exercise Sciences, North Dakota State University, Fargo,
ND 58105, USA [1]; Grand Forks Human Nutrition Research Center, USDA/Agricul-
tural Research Service, P.O. Box 9034, Grand Forks, ND 58202, USA [2]
Our previous research indicated that dietary boron (B) decreased plasma insulin levels
and increased insulin sensitivity independent of other marginal nutritional stressors. To
investigate the effects of dietary boron on serum glucose (SG) and insulin (SI) levels
during severe vitamin D (VD) deficiency, 60 male weanling Sprague Dawley rats were
fed with diets depleted of VD and/or boron (0 IU VD/kg diet and/or 0 mg B/kg diet) or
diets adequate in VD and/or boron (1000 IU VD/kg diet and/or 2.5 mg B/kg diet) for 12
wks. The rats were challenged with a glucose tolerance test (GTT, 0.133% glucose of
body weight) at 6 wks. During the GTT, SI concentrations decreased and SG concen-
trations increased in VD depleted rats compared to those with adequate VD (SI: 0.285
vs. 0.561 ng/ml, p=0.0175, SG: 174 vs. 165 mg/dl, p=0.0224). During the GTT, dietary
boron did not affect either SG or SI levels. At 12 wks, dietary boron significantly
decreased SI levels (0.0886 vs. 0.1226 ng/ml, p=0.0041), but not SG nor body weight. As
expected, severe VD deficiency lowered SI concentrations below the normal range
(0.0628 vs. 0.148 ng/ml, p<0.0001) at 12 wks. The results suggest that dietary boron
affects insulin concentrations independent of VD status. Because dietary boron reduces
circulating insulin concentrations while maintaining serum glucose concentrations within
the normal range, dietary boron might help prevent pancreatic exhaustion caused by
chronic hyperinsulinemia.
4.08
STRUCTURE-ACTIVITY RELATIONSHIP OF VANADYL-PICOLINATE
COMPLEXES WITH ANITIDIABETIC ACTIVITY
Hiromu Sakurai*, Yusuke Adachi, Kojiroh Tayama, Naoko Yasumatsu and Hiroyuki
Yasui
Dep. Anal. Bioinorg. Chem., Kyoto Pharm. University, Kyoto 607-8414, JAPAN
Diabetes mellitus (DM) is one of the most important lifestyle- and food-dependent dis-
eases throughout the world in the 21th century. Development of new types of agents in
place of painful daily injections of insulin preparations and some synthetic therapeutics
with severe side effects to treat types 1 and 2 DM, respectively, have been anticipated.
Since 1990 we have developed many types of vanadyl (+4 oxidation state of vanadium)
complexes with different coordination modes around vanadyl ion such as VO(O4),
VO(S4), VO(N4), VO(N2O2), VO(N2S2) and VO(S2O2). In 1997, we found bis(6-
methylpicolinato)oxovanadium(IV) complex with effectiveness on dail administrations
and with long-term acting in treating both types of DM model animals. On the basis of
the results, we examined the structure-activity relationship of several vanadyl-picolinate
complexes using bis(picolinato)oxovanadium(IV) as a leading compound to find more
active complexes, and found that introduction of halogen atom or alkyl group on the
picolinate enhanced antidiabetic activity of the complexes in terms of both in vitro insulin
mimetic and in vivo blood glucose normalizing effects in experimental animals. The
obtained results will be discussed together with action mechanism examined with the
isolated rat adipocytes.
4.09
EFFECTS OF SELENIUM ON STRUCTURAL ALTERATIONS IN
DIABETES-INDUCED BRAIN CORTEX TISSUE
Yüksel Saran* [1], Belgin Can [1], Belma Turan [2]
Ankara University School of Medicine, Departments of Histology-Embryology [1], and
Biophysics, Ankara, TURKEY [2]
Diabetes affects the function of various organs as long term complications. It has been
suggested that oxidative stress can play an important role in tissue damage associated
with diabetes. Selenium compounds are very interesting compounds having some
beneficial effects on diabetic dysfunctions. The aim of this study was to investigate the
effects of selenium on structural alterations in diabetes-induced brain cortex tissue.
Diabetes was induced by a single intraperitoneal injection (ip) of STZ (50 mg/kg body
weight). All animals were fed with standard rat nutrients and water without restriction.
Diabetic animals randomly divided into two groups. Starting one week after the STZ
injection, one group received daily ip saline the other group received daily ip sodium
selenite (5 lmol/kg/day) injections for 4 weeks. Control animals were injected with
comparable volume of citrate buffered saline and divided in two groups. One group
received daily ip saline while other group received sodium selenite. Animals were sac-
rificed and sections taken from the cortical region of the frontal lobe were prepared for
microscopic investigations. In both control groups, neuronal and neuroglial structures
displayed normal appearance. In diabetic group, adjacent to normal neurons several
features of injured neurons were observed. Some of them contained less, barely visible
cytoplasm and the others showed dark shrunken neuronal perikarya and pyknotic
nucleus with lack of discernible nucleolus. The cortex showed spongiform appearance
containing microcystic spaces but showed no inflammation. Oligodendroglial cells
surrounded injured neurons. Most of the blood vessels and capilleries in piamater and
cortical region showed congestion. In selenite treated diabetic group, the normal and
injured neuronal structure were observed together. Oligodendroglial cells and microglial
cells were increased. The congested appearance in blood vessels of piamater and cap-
illaries displayed somewhat regression. The nervous tissue displayed some discernible
signs of injury probably due to microcirculation disturbance caused by diabetes. These
changes were accompanied by either neuronal damage or evident selective neuronal
death. (This work has been supported by Ankara University Research Fund No: 2003-
08-0998.)
4.10
PLASMA LEVELS OF ZINC WITH LEPTIN, GLUCOSE AND WEIGHT AS
PREDICTORS OF CIRCULATING GHRELIN LEVELS IN BABOONS
Saroja Voruganti* [1], Jeanne Freeland-Graves [1], Elizabeth Tejero [2], Michael Proffitt
[1,2], Guowen Cai [2], Shelley Cole [2], and Anthony Comuzzie [2,3]
Division of Nutritional Sciences, A2700, The University of Texas at Austin, Austin, TX
78712, USA [1]; Department of Genetics, Southwest Foundation for Biomedical Re-
search, San Antonio, TX, USA [2]; Southwest National Primate Research Center, San
Antonio, TX, USA [3]
Association of altered zinc status with type 2 diabetes and dysregulation of appetite has
been well documented in humans. The aim of this study was to use baboon as a model for
investigating the relationship of plasma zinc with parameters related to glucose metab-
olism and appetite. Subjects included 108 pedigreed baboons housed at Southwest
Foundation for Biomedical Research, San Antonio, Texas. Body weight and plasma
levels of zinc, ghrelin, peptide YY 3 36, cholecystokinin, glucose, insulin, leptin and
lipids were measured according to the standard procedures. Males had higher body
weight (32 vs. 21 kg, p < .05) and lower plasma zinc levels than females (.93 vs. 1.02 mg/l,
p < .05). Zinc was negatively correlated with both leptin (r = ).13, p < .05) and ghrelin
(r = ).28, p < .05). After adjustment for age and sex, zinc was negatively associated only
with ghrelin (r = ).41, p < .001). Stepwise regression analysis revealed a model in which
zinc, leptin, glucose and body weight were depicted as predictors for ghrelin (R = .6, p <
.0001). No association was found between zinc and peptide YY3-36, cholecystokinin,
glucose, insulin and lipids. These results suggest that zinc may be involved in appetite
regulation through an association with ghrelin. (Supported by grants HL28972, and
RR013986 from NIH.)
4.11
CONCENTRATIONS OF CALCIUM, COPPER, IRON, MAGNESIUM AND ZINC
IN YOUNG FEMALE HAIR WITH DIFFERENCE BODY MASS INDEX
Chin-Thin Wang* [1], Pou-An Liu [2], Li-Yun Liu [3], Wei-Tun Chang [4]
Department of Holistic Education Center, St. John’s and St. Mary’s Institute of Tech-
nology, Tam-sui, Taipei, TAIWAN ROC [1]; Mackay College of Nursing Pei-tau, Taipei,
TAIWAN ROC [2]; Department of Food Science, Nutrition, and Nutraceutical Bio-
technology Shin Chien University Taipei, TAIWAN ROC [3]; Department of Forensic
Science Central Police University, Tao-yuan, TAIWAN [4]
An investigation of the metal concentrations of calcium, copper, iron, magnesium and
zinc from three groups of young female hair (n=180), between the ages of 15 to 19, with
different Body Mass Index (BMI): BMI<18, BMI between 18 and 23, and BMI>24 was
performed by using atomic absorption spectroscopy (AAS). The hair samples are washed
with organic solvent (acetone: hexane: water; V/V; 4:2:1) to remove the external contents
and then are digesting by the microwave digester. Finally, the hair samples are analyzed
by using a Flame Atomic Absorption Spectrophotometer. We find that the BMI<18
group (n=45) have the highest concentration of calcium, copper, iron, magnesium and
zinc while the BMI between 18 and 23 group (n=75) have the second highest con-
centration. The BMI>24 group (n=60) have the lowest concentration. Further more,
when we compared the concentrations of calcium, copper, iron, magnesium and zinc
between the groups with BMI<18 and BMI>24, we find that there are significant dif-
ferences in the calcium and copper concentrations between these two groups (calcium,
2207.5±899.1 lg/g vs. 1665.5±722.2lg/g, respectively, with p<0.05; copper,

17.69±10.37 lg/g vs. 12.06±6.22 lg/g, respectively, with p<0.05. The differences are
extremely obvious in the iron and magnesium concentrations (iron, 52.17±26.09 lg/g vs.
28.47±14.16 lg/g, respectively, with p<0.0l; magnesium, 285.9±99.30 lg/g and
202.9±111.8 lg/g, respectively, with p<0.01). (Dr. Wang Chin-Thin severs on the office
of the technology cooperation director for the St. John’s and St. Mary’s Institute of
Technology.)
4.12
EFFECT OF SHORT-TERM WEIGHT LOSS ON ZINC STATUS IN
OVERWEIGHT ADULTS CONSUMING HYPOCALORIC DIETS WITH HIGH
DAIRY INTAKE, LOW DAIRY INTAKE OR CALCIUM SUPPLEMENTATION
Leslie R Woodhouse* [1], Erik R Gertz [1], Timothy L Radak [1], Dorothy Teegarden [2],
Michael B Zemel [3], Marta D Van Loan [1]
USDA ARS WHNRC, UCD, Davis, CA 95616, USA [1]; Dept. of Foods and Nutrition,
Purdue U, West Lafayette, IN 47907, USA [2]; Dept. of Nutrition, U of Tennessee,
Knoxville, TN 37996, USA [3]
Zinc (Zn) metabolism is altered in obesity, and improvements in Zn status may occur
with weight loss. A controlled intervention trial in obese adults was conducted for 12
weeks with an energy restriction of 500 kcal/d. Treatment groups were high dairy (HD
n=23), low dairy (LD n=25) and low dairy+calcium (CA n=17). HD consumed 3
dairy servings (DS) per day, LD consumed £1 DS per day + placebo capsule, and CA
consumed £1 DS per day + 900mg calcium. Fasting serum and a 24-hour urine sample
were collected at baseline (B) and 12 weeks. DEXA was conducted to determine fat mass
(FM) and lean body mass (LBM). Serum and urine were analyzed for Zn using ICP-AES.
Of the subjects who lost weight, % body weight loss was 5.9, 5.0, & 4.2 % for HD, LD &
CA, respectively. Serum Zn increased significantly (7.0%) from B to 12 weeks in the HD
group, with slight increases of 4.7 & 2.4% in LD & CA, respectively. Urine zinc decreased
significantly after 12 weeks in the CA group. The primary component of weight loss, FM,
was significantly higher in the HD group (96%) compared with 81% and 74% for LD and
CA. With caloric restriction resulting in weight loss, it appears that the HD diet, more
than the LD or the CA diets, may increase serum Zn as well as spare the loss of LBM
during weight loss, suggesting improved Zn status. (Support provided by the National
Dairy Council, USA.)
4.13
EFFECTS OF CHRONIC SELENIUM TREATMENT ON PLASMA TRACE
ELEMENT LEVELS OF STZ-INDUCED DIABETIC RATS
Nezahat Zaloglu* [1], Yalcin Günal [1], Ahmet Aydin [2], and Belma Turan [3]
Ankara University School of Medicine, Departments of Physiology [1], Biophysics [3],
Ankara, TURKEY; Gülhane Military Academy, Ankara, TURKEY [2]
Animal models in which diabetes is induced by streptozotocin (STZ) injection are being
used to study diabetic complications. There is ample evidence that selenium compounds
can restore some metabolic parameters in diabetes. In diabetic animals, trace elements,
such as selenium were found to have positive effects on the parameters of glucose
metabolism besides its effects on antioxidant defense mechanism. The aim of this study
was to investigate the effects of chronic selenium treatment on plasma trace elements
levels in STZ-induced diabetic rats. Diabetes was induced by a single intraperitoneal

injection (ip) of STZ (50 mg/kg body weight). All animals were fed with standard rat
nutrient and water without restriction. Diabetic animals randomly divided into two
groups. Starting one week after the STZ injection, one group received daily ip saline the
other group received daily ip sodium selenite (5 lmol/kg/day) injections for 4 weeks.
Control animals were injected with comparable volume of citrate buffered saline and
divided in two groups. One group received daily ip saline while other group received
sodium selenite (5 lmol/kg/day). Animals were sacrificed and blood samples were
collected, then plasma samples were stored till measurement of race element levels. STZ
could not induce any change in plasma selenium and magnesium levels of the rats
significantly but it did cause a significant increase in plasma manganese level
(14.00±1.55 ng/ml vs 7.58±1.39 ng/ml in diabetes vs control). Selenium treatment
could not affect this high manganese level (15.77±3.10 ng/ml), on the contrary, it could
increase this value in treated control group significantly (12.14±1.12 ng/ml). These data
suggest that both the plasma selenium level is not changed in STZ induced diabetic
rats and therapeutic level of selenium of supplementation of the diabetic and normal
rats can not alter this value. On the other hand, another trace element, manganese, can
significantly increase in the diabetic rat due to most probably altered antioxidant de-
fense system in the cell, and this amount of selenium treatment cannot normalize this
high manganese level. (This work has been supported by Ankara University Research
Fund No: 2003-08-0998.)
Category 5: Trace Elements and Disorders of Aging
5.01
COULD SELENIUM BE A PREDICTIVE FACTOR OF LONGEVITY?
Isabelle Hininger* [1], Josiane Arnaud [1], Tasmine Akbaraly [2], Claudine Berr [2], Alain
Favier, and Anne-Marie Roussel [1]
NVMC UFR Pharmacie, 38700 Grenoble, FRANCE [1]; INSERM E 036, Hôpital La
Colombière, 34093 Montpellier, FRANCE [2]
With ageing, early stages of pathologies are expected to increase. There is a need to
investigate the dietary intakes and antioxidant status in elderly as predictive factors of
health. Among trace elements, selenium (Se) seems of particular interest since Se status is
lower in oldest subject than in younger. However the evaluation of the decline of Se status
with ageing has never been evaluated. Our aim was to investigate the changes in Se status
in elderly healthy persons in a follow-up study in relation to their mortality. 1389 free-
living subjects aged 60 to 70 were recruited to participate in a 9-years longitudinal study
(The EVA study). For the first time, a decrease in Se plasma level was assessed in a
prospective study in individual healthy elderly subject. Furthermore, the mean Se plasma
level was significantly higher in the survival subjects at the end of the follow-up study
than in the subjects who died before the end (1.10±0.2 vs 1.01±0.2; p<10-4). All rela-
tionships were independent of risk factors tested and were consistent with the view that
plasma Se levels were higher in survival people compared to the others considering the 50
percentile or the 75 percentile. Consequently, our results might suggest that a low plasma
Se level could represent an early sign of risk of mortality. The direct implications of an
adequate Se status as protecting factor against degenerative processes known to be
especially high in elderly people should be further explored.
5.02
TRACE ELEMENTS IN CEREBROSPINAL FLUID IN SUBJECTS WITH
ALZHEIMER’ DISEASE AND NORMAL COGNITION
Nina Johansson* [1], Maria Eriksdotter-Jönhagen [2], Hans Basun 3], Ilja Rudshkin[4],
Monica Nordberg [1]
Inst. Environmental Medicine, Karolinska Institutet, SE-171 77 Stockholm[1]; Dept
Neurotec, Section of Clinical Geriatrics, Karolinska Institute and Karolinska Univer-
sity Hospital, Huddinge, Sweden [2] ; AstraZeneca, Clinical Science, R&D Södertälje,
SE-151 85 Södertälje and Dept. of Public Health / Geriatrics, Uppsala University
Hospital, SE-751 25 Uppsala, Sweden [3]; Analytica AB, Aurorum 10, S-977 75 Luleå,
SWEDEN [4].
Concentration of metals in cerebrospinal fluid (CSF) can serve as an index of brain
burden and affect cognitive function. Here we report the results from a pilot case control
study. The concentrations of Al27, P31, Ti47, V51, Cr52, Mn55, Co59, Ni60 and Sr88 in
CSF were analysed by ICP-OES and ICP-MS in 39 elderly (>65 years) subjects
undergoing dementia investigation at the Memory Clinic, Karolinska University Hos-
pital, Huddinge. 26 out of the 39 subjects (14 women / 12 men) was diagnosed with
Alzheimers disease (AD) and served as the case group. The remaining 13 subjects (5
women / 8 men) had no objective memory impairment and served as controls. The
subjects with AD displayed higher [Cr52] and [Mn55] (Students t-test: p=2.57E-5 and
p=0.0046 respectively) than the controls. We also found a higher [Al27] in women than
men suffering from AD (p=0.0008). Thus differences in certain metals in CSF between
Alzheimer patients and controls and between gender were found. Further analyses are
needed to elucidate the mechanisms behind these differences. Ethical committee KI: 364/
03 approved the study.
5.03
DISTURBANCES OF CERTAIN TRACE ELEMENT LEVELS IN PATIENTS WITH
BATTEN’S AND PARKINSON DISEASES
Tuomas Westermarck [*], Erland Johansson, Faik Atroshi
Rinnekoti Cesearch Center, Espoo, FI-02980; Applied Biochemistry Unit,
Huddinge Hospital, S-14186; Dep. Vet. Clinical Sciences, Pharmacology, University of
Helsinki, FINLAND
Disorders in which free radicals and antioxidants are implicated include Batten’s
disease (NCL) and Parkinson disease (PD). Recent evidence has shown the impor-
tance of nutrition in delaying the aging process and in protecting against many
degenerative and chronic diseases. Our growth in knowledge of reactive oxygen spe-
cies, oxidative damage, and the role that nutritional antioxidants which may play in
protection from this damage suggests that factors in our diet may be effective in
preventing or retarding the disease process. We have examined trace element levels in
platelets of patients with infantile NCL (n=3), juvenile type (n=3); and erythrocytes
from patients with PD (n=12). Metal ions from platelets were analysed by micro-
PIXE; metal ions from erythrocytes by ICP-MS. The concentration of Ca was sig-
nificantly (p < 0.01) increased, and Zn significantly (p<0.05) decreased in the pla-
telets in JNCL, but not in INCL, compared with the level of controls. Increased
concentration of Ag (p<.0.005), Cd (p<0.05), and Ni (p<0.05) was observed in the
erythrocytes from patients with PD.
Category 6: Trace Elements and Fetal Development and Pediatric Disorders
6.01
IODINE STATUS OF PREGNANT WOMEN PARTICIPATING IN THE
SEYCHELLES CHILD DEVELOPMENT NUTRITION STUDY
Maxine P. Bonham* [1], Emeir M Duffy [1], J. J. Strain [1], Julie Wallace [1], Paula J.
Robson [1], Conrad Shamlaye [2], Gary J. Myers [3]
Northern Ireland Centre for Food and Health (NICHE), University of Ulster, Coleraine
BT52 1SA, NORTHERN IRELAND [1]; Ministry of Health, SEYCHELLES [2];
Environmental Medicine, University of Rochester School of Medicine and Dentistry,
USA [3]
The Seychelles Child Development Nutrition Study (SCDNS) is a large prospective study
investigating the relationship between nutritional status and the effects of MeHg toxicity
on child development. Amongst the nutrients of interest is iodine which is an essential
component of the thyroid hormones triiodothyronine (T3) and thyroxine (T4) and
maternal thyroid function during early pregnancy is an important determinant of early
fetal brain development. Iodine status was examined in pregnant mothers (n=272) at
enrolment and 28 weeks gestation via assessment of thyroid stimulating hormone (TSH),
free T3 and free T4. Results indicate that levels of TSH, free T3 and free T4 are within
established reference ranges and follow the usual trends expected during pregnancy.
Mean concentrations of TSH increased by 20% from 1.0 mIU/L (SD 0.6) at enrolment to
1.2 mIU/L (SD 0.6) at 28 weeks gestation. Mean free T3 concentrations decreased from
5.0 pmol/L (SD 1.1) to 3.8 pmol/L (SD 0.5) and mean free T4 concentrations decreased
from 11.7 pmol/l (SD 1.3) to 8.6 pmol/L (SD 1.2). These are the first data on iodine status
emanating from the Seychelles and indicates iodine deficiency is not prevalent in this
cohort of pregnant women. (Funded by the National Institute of Environmental Health
Sciences [NIEHS].)
6.02
EFFICIENT STRATEGY FOR HAPLOTYPE ANALYSIS OF WILSON DISEASE
USING NEW POLYMORPHIC SHORT TANDEM REPEATS IN FRENCH
FAMILIES
Muriel Bost* [1,2], Giuseppe Sanna [3], Laetitia Boutrand [3], Philippe Latour [2], An-
toon Vandenberghe [2,3]
Trace Element - Institute for UNESCO [1]; Debrousse Hosp., Neurogenetics [2]; ISPB
Lyon, Human Molecular Genetics, FRANCE [3]
Wilson disease (WD) is an inherited autosomal recessive affection caused by mutations in
the ATP7B gene, encoding a copper transporting P-type ATPase. Due to the gene size (a
genomic region of about 80 kb), the 21 exons and the high number of disease causing
mutations (more than 300), a diagnosis by haplotype analysis is the most efficient in
identifying presymptomatic patients and heterozygous subjects. Presently, the laborato-
ries use amplification by PCR of polymorphic poly (AC) repeats. The frequent non-
informativeness of these markers led us to develop a new haplotype analysis with highly
polymorphic tri, tetra or pentanucleotide short tandem repeats (STRs) located within and
on each side of the WD gene. We collected overlapping sequences of clones, containing
atp7b gene, directly from the NCBI site. a contig of the WD gene was manually con-
structed using data bank searches with terminal sequences of the clones using the blast
program. Tri, tetra or pentanucleotide strs were assigned by tandem repeats finder pro-
gram and we tested them for polymorphic behavior and allelic distribution in 50 control
subjects and in 10 WD families. Alleles of interest were sequenced. We identified five new
polymorphic strs covering 1mb to each side of the atp7b gene. Combined use of these
markers allows a robust haplotype analysis with almost complete informativeness. In our
routine diagnosis of WD families, they have replaced or completed the use of poly (ac)
repeats.
6.03
ZINC ENHANCES THE EFFECTIVENESS OF ß-CAROTENE
SUPPLEMENTATION DURING PREGNANCY IN IMPROVING VITAMIN A
STATUS IN BOTH MOTHERS AND INFANTS
Marjoleine A. Dijkhuizen* [1], Frank T. Wieringa [1], Clive E. West [2], Muhilal [3]
Univ. Med. Center Nijmegen, Dept. Int. Med. PO Box 9101, 6500 HB, NETHER-
LANDS [1]; Dept Human Nutr, Univ Wageningen, NETHERLANDS [2]; Nutr Res Dev
Center Bogor, INDONESIA [3]
Deficiencies of vitamin A, iron and zinc are prevalent in developing countries. A ran-
domized double-blind supplementation trial in pregnant women (n=170), with ß-caro-
tene (4.5 mg), zinc (30 mg), both or placebo, in addition to iron (30 mg) and folic acid (0.4
mg), was done to investigate whether supplementation during pregnancy with ß-carotene
and zinc can improve micronutrient status of mothers and infants. Six mo post-partum,
plasma retinol concentrations were higher in women receiving zinc during pregnancy.
Infants from mothers receiving ß-carotene+zinc had higher plasma retinol concentra-
tions, with vitamin A deficiency reduced by >30%. Breast milk ß-carotene concentra-
tions were higher in all women receiving ß-carotene, but breast milk retinol
concentrations were higher only in women who received ß-carotene+zinc. Zinc con-
centrations did not differ among groups. Zinc supplementation during pregnancy im-
proved vitamin A status of mothers and infants post-partum, pointing to a specific role of
zinc in vitamin A metabolism. In developing countries, vitamin A deficiency is prevalent
even when diets are rich in carotenoids. This may be explained by impaired bioconversion
of provitamin A carotenoids to retinol due to zinc deficiency. Addition of both ß-carotene
and zinc to iron supplements during pregnancy could be effective to improve vitamin A
status of mothers and infants.
6.04
IRON STATUS OF PREGNANT WOMEN AND INFANTS PARTICIPATING IN
THE SEYCHELLES CHILD DEVELOPMENT NUTRITION STUDY
Emeir M. Duffy* [1], Maxine P. Bonham [1], J. J. Strain [1], Julie Wallace [1], Paula J.
Robson [1], Conrad Shamlaye [2] and Gary J. Myers [3]
Northern Ireland Centre for Food and Health (NICHE), University of Ulster, Coleraine
BT52 1SA, NORTHERN IRELAND [1]; Ministry of Health, SEYCHELLES [2];
Environmental Medicine, University of Rochester School of Medicine and Dentistry,
USA [3]
The Seychelles Child Development Nutritional Study (SCDNS) is a large prospective
study investigating the relationship between diet and the effects of MeHg, one such
nutrient is iron. Iron deficiency in infants may result in a permanent cognitive deficit
where mechanisms observed include abnormalities in neurotransmitter metabolism, de-
creased myelin formation and alterations in brain metabolism. Assessment of soluble
transferrin receptor (sTfR) status, haemoglobin (Hb) and ferritin has been carried out in
both maternal (n = 275) and cord (n = 234) blood samples. At enrolment, mean (SD)
hemoglobin (g/L), ferritin (nmol/L) and sTfR (nmol/L) were 10.5 (2.2), 52.2 (49.9) and
22.7 (6.8) respectively. At delivery sTfR (nmol/L) and ferrtin (ng/ml) were 36.3 (16.7),
39.2 (41.8) respectively. Cord blood mean (SD) hemoglobin (g/L), ferritin (nmol/L) and
sTfR (nmol/L) were 12.8 (4.1), 54.1 (19.8) and 119.3 (86.5) respectively. Plasma soluble
transferrin receptor (sTfR) and ferritin levels were generally within published reference
ranges; however, haemoglobins would suggest a marginal deficiency in both mothers and
infants. These are the first data representing iron status in pregnant Seychellois women
and highlight the need to monitor iron status in this population. (Funded by the National
Institute of Environmental Sciences.)
6.05
ZINC AND IODINE STATUS OF SCHOOLS CHILDREN FROM RURAL NORTH
EAST THAILAND
Sueppong Gowachirapant* [1], T Pongcharoen [1], A Boonpraderm [1], R Cook [2], R
Gibson [2], P Winichagoon [1]
Institute of Nutrition, Mahidol University, Salaya, Thailand [1]; Department of Human
Nutrition, University of Otago, NEW ZEALAND [2]
Zinc and iodine are essential micronutrients for maintaining good health. Deficiency of
these elements can lead to retardation of growth and cognitive function in children.
Therefore, this study aims to evaluate zinc and iodine status of school children in NE
Thailand by determining serum zinc and urinary iodine. Primary school children (286
girls, 281 boys) aged 6.00-12.99 years were recruited from 10 rural schools in Ubon
Ratchathani province. In this study, 57% of the children had low serum zinc concen-
trations. Boys had slightly lower serum zinc than girls in all age groups, but no statis-
tically significant difference was noted (p=0.84). In addition, there was no significant
difference in serum zinc concentration among age group of children. The median urinary
iodine excretion was 6.2 lg/dl for boys and 4.7 lg/dl for girls. There was significant
difference in the median urinary iodine between boys and girls (p=0.00). Using regres-
sion analysis, neither of these indicators were associated with height for age Z score.
There was an association between mid parental height and height for age Z score of
school children (p=0.00). This investigation indicates that the problem of zinc and iodine
deficiency still exists among school children in NE Thailand. Possibly, however, severity
was not great to compromise growth. Therefore, there is a need for effective strategies in
order to improve nutritional status of these children.
6.06
EPIDEMIOLOGICAL STUDY OF CHILDREN’S HAIR ELEMENT PATTERNS
FROM VARIOUS RUSSIAN CITIES
Yulia Lobanova [1], Vasily Demidov [1], Svetlana Notova* [2]
Center for Biotic Medicine, 125047, PB.56, Moscow, RUSSIA [1]; Institute of Bioele-
mentology at Orenburg State University, Orenburg, RUSSIA [2]
The combined multielement ICP-OES + ICP-MS analysis of 2569 samples of the Rus-
sian children’s hair, up to 7 years old has shown overall deficiency of such essential
elements, as Mg, Zn, Cu, and Co. At the same time, there is revealed a risk of adverse
effects on kids’ health due to the excess accumulation of the toxic elements (Hg, Cd, and
Pb), mainly for kid living in industrial cities. Young Moscow citizens feature relatively
low levels of hair Cd, Fe, Mn, Pb, Ca, Mg, Na and increased one of Cu comparing with
the mean data for Russia. As for the Moscow region’s children, there are found relatively
high hair levels of Fe, Cr, K, Mg, P, Pb and Ti. The study of children’s hair in Saint-
Petersburg has shown highest levels of As, Hg, Se and Sn over the all cities investigated
while the lack of essentials: Ca, Mg and Zn. It can be related to the environmental
factors, which keep imprint on the trace element status of children. For Novosibirsk’
children deviations from all-Russian data include relatively low hair levels of Mg and Ni
and high ones of Cd, Hg, K, Na, Pb, and Si. That can lead to trace element related
disorders of immune and endocrine systems of children. Accordingly to the data present,
the environmental, social and economical factors govern the children’s essential and toxic
element status in Russia. Investigated data are intended to be a base for a public and
political activity in the kid’s health prevention in Russia.
6.07
SEX DEPENDENT PECULIARITIES OF SOME IMPORTANT CHEMICAL
ELEMENT RATIOS IN HAIR OF SCHOOLCHILDREN
Andrei Grabeklis [1], Vasily Demidov [1], Svetlana Notova* [2]
Center for Biotic Medicine, P.O. Box 56, Moscow 125047, RUSSIA [1]; Institute of
Bioelementology at Orenburg State University, Orenburg, RUSSIA [2]
Hair samples of 6028 relatively healthy teenagers (2734 boys, 3294 girls) of 7-14 years old,
residing in different regions of Russia, were analyzed by ICP-AES method (see: Grab-
eklis, Skalny, 2003). Ca/P, Ca/Mg, K/Na, Zn/Cu, Mn/Cu, Al/Ca, Al/Mg, Pb/Ca, Pb/Mg,
Be/Mg, Cd/Cu, Cd/Zn, Pb/Zn, Ni/Zn, As/Se, Hg/Se ratios were estimated. Comparison
of the essential elements ratios in hair, such as Ca/P, Ca/Mg, K/Na, Zn/Cu, Mn/Cu,
showed that only K/Na ratio can be considered as sex-independent. Ca/P, Zn/Cu and
Mn/Cu ratios were found to be significantly higher in girls while the Ca/Mg one was
higher in boys (P < 0.05). The difference in Ca/P ratio was detected to 1.5 fold rise with
age from 7 to 14 y.o.; on the other hand, difference in Ca/Mg ratio was the most distinct
in early age while in children of 14 years old it became identical at the level of 14.5. Zn/Cu
and Mn/Cu ratios were most different in the middle of the investigated age range while at
its ends the values were more similar. The investigated ratios of toxic elements to the
essential ones were generally higher in boys’ hair. For Al/Ca, Al/Mg, Pb/Ca, Pb/Mg, Be/
Mg, Cd/Cu, Cd/Zn, Pb/Zn ratios the difference was found to be significant (P < 0.05),
for the As/Se Hg/Se ones it was the tendency. The only exception was the Ni/Zn ratio
which was higher in girls due to higher hair Ni level typical for this sex. In most cases
boys were characterized by less alteration of elements ratios in hair in comparison to girls
both within separate age groups and along the age as a whole.
6.08
HIGH RESOLUTION ICP-MS A USEFUL TOOL TO STUDY MANGANESE
ACCUMULATION INDUCED BY IRON DEFICIENCY
Tore Syversen* [1], Stephanie J. Garcia [3], Kristin Gellein [2], Eiliv Steinnes [2], Trond P.
Flaten [2], Michael Aschner [3]
Department of Neuroscience [1] and Department of Chemistry [2], Norwegian University
of Science and Technology (NTNU), MTFS, N-7489 Trondheim, NORWAY; Depart-
ment of Physiology and Pharmacology, Wake Forest University School of Medicine,
Winston Salem, NC 27157, USA [3]
The adequate supply of trace metals is of great importance to the developing brain.
For many trace metals, the effects of nutritional deficiencies are well established.
However, such deficiencies may alter tissue levels of other trace elements. We have
used a high resolution inductively coupled plasma mass spectrometer (HR-ICP-MS) to
study the effects of iron deficiency (ID) on multi-element levels in neonatal rats. Since
iron (Fe) and manganese (Mn) share similar transport mechanisms, ID enhances Mn
accumulation even in the absence of excess dietary Mn (Erikson et al., 2002). Pregnant
Sprague-Dawley rats were fed either control (35 mg Fe/kg, 10 mg Mn/kg diet) or low
Fe (3 mg Fe/kg, 10 mg Mn/kg diet) semi-purified diets from gestational day (GD)7
until weaning at postnatal day (PN)21. Neonates were cross-fostered from control
dams to control or low Fe dams on PN4 and exposed to the diets via maternal milk
until PN21. Hematological parameters confirmed ID in dams and pups fed a low Fe
diet. In ID pups, Fe, chromium (Cr), and molybdenum (Mo) brain levels decreased
significantly while Mn and copper (Cu) levels increased. These data demonstrate that
ID increases brain Mn during early development and that ID is a risk factor for Mn as
well as Cr, Mo, and Cu alterations in the brain. Alterations in these essential trace
metals may be at least partially responsible for the consequences of ID during
neurodevelopment.
6.09
A MULTI-COUNTRY TRIAL ON IRON AND ZINC SUPPLEMENTATION IN
INFANTS: INTERACTIONS AND EFFECTS ON MICRONUTRIENT STATUS
AND GROWTH
SEAMTIZI study group: Frank T. Wieringa* [1], Jacques Berger [2,5], Marjoleine A.
Dijkhuizen [1], Adi Hidayat [4], Nguyen X. Ninh [5], Budi Utomo [3], Emorn Wasan-
twisut [6], and Pattanee Winichagoon [6]
SEAMTIZI: South-East Asia Multi-country Trial on Iron and Zinc supplementation in
Infants Study Group
LANDS [1]; Inst. Rech. Dev., Lab. Nutr. Trop., Montpellier, FRANCE [2] Univ.
Indonesia [3] and Univ. Trisakti [4], Jakarta, INDONESIA; Nat. Inst. Nutr., Hanoi,
VIETNAM [5]; Inst. Nutr., Mahidol Univ. Bangkok, THAILAND [6]
Pooled analysis of a double-blind placebo-controlled multi-country trial with 4 study sites
in Indonesia, Thailand and Vietnam. Infants (4-6 mo old, n=2603) were supplemented
with iron(10 mg) and/or zinc(10 mg) daily for 6 mo. Supplementation of iron improved
iron status (anemia reduced by 28%) and of zinc improved zinc status (zinc deficiency
reduced by 18%), but combined supplementation was less effective (reduction of 21% in
anemia and 10% in zinc deficiency). Iron reduced the effect of zinc supplementation
(P<0.01), but there was no significant interaction of zinc with iron supplementation.
Intriguingly, boys had a better response to iron supplementation than girls (effect size
12.0 g/L vs 6.8 g/L), as boys were more at risk for iron deficiency (P<0.01). Although
neither iron nor zinc supplementation prevented progressive growth faltering during the
first year of life, zinc supplementation improved linear growth in anemic infants
(P=0.01). However, in non-anemic infants, zinc supplementation may have a negative
effect on linear growth (P=0.05). (The SEAMTIZI study was financially supported by
UNICEF.)
Category 7: Trace Elements and Infectious and Immune Disorders
7.01
INHIBITORY ACTIVITY OF STAPHYLOCOCCUS AUREUS IN MILK FROM
COWS FED SLENIUM-SUPPLEMENTED AND SELENIUM DEFICIENT DIETS
Faik Atroshi*, Marge Malbe, Martti Attila, Tuomas Westermarck, Hannu Saloniemi
Dept. Vet. Clinical Science, Pharmacology & Toxicology, PL 57, 00014 Helsinki Uni-
versity, FINLAND
Nutritional support may improve the clinical outcome of the diseases and it is considered
a necessary strategy in the global treatment. Trace elements play an important role,
especially selenium, a key component of several enzymes e.g. GSHPx, that function as
antioxidant scavengers. Selenium availability for microbial growth, however, is not well-
recognized in host defence mechanisms. The role of Selenium supplementation and its
inhibitory activity in S. aureus growth in vivo and in vitro in cows’ milk was examined.
Selenium-supplemented cows received 0.2 ppm selenium yeast for 8 weeks. Non-sup-
plemented group received diet with no selenium. Cows were selected according to GSHPx
level in their blood as well as on low somatic cell count (<300,000 cells/ml) and negative
bacteriological findings in quarter milk samples during the experimental period. An a
association between high GSHPx activity in blood and growth inhibition in whey frac-
tions was found. In non-supplemented cows the growth of S.aureus showed no inhibition.
Comparing in vitro bacterial growth rate in whey from selenium supplemented cows
before and after supplementation period showed that whey became bacterial growth
inhibiting after supplementation. This trend was more clear in cow’s with GSHPx activity
in their blood above 4 _kat/g Hb. Cows with GSHPx activity below 4 kat/g Hb did not
show clear bacterial growth inhibition in their whey. We have shown that selenium
supplementation can provide protection against oxidative damage caused by mastitis.
7.02
ZINC PREVENTS CERAMIDE INDUCED JNK ACTIVATION, FAS LIGAND
EXPRESSION AND CASPASE-3 DEPENDENT APOPTOSIS IN CD4+ T
LYMPHOCYTES
S. Karandikar [1], S. Joshi-Barve [2], C. J. McClain [2] and S. Barve* [2]
University of Louisville, Department of Microbiology and Immunology [1] and
Department of Internal Medicine [2], Louisville, KY 40202
Zinc is a critical trace element required for the normal development and maintenance of
immune functions and hence was investigated for its ability to prevent ceramide-induced
apoptosis in human CD4+ T lymphocytes (Jurkats). Ceramide is a key component of
intracellular stress response pathways and mediates apoptosis induced by multiple agents
including inflammatory cytokines, irradiation, anticancer drugs and viral infections. In
this study we analyzed the effect of zinc (10 * 50 uM) on critical earlier mechanisms in the
ceramide mediated apoptotic cell death. Ceramide-induced apoptosis often involves the
SAPK/JNK signaling pathway. Hence, we evaluated the effect of zinc on JNK activation
induced by C-2 ceramide, a synthetic ceramide analogue. Pretreatment of Jurkat T
lymphocytes with zinc completely abolished ceramide induced JNK activation. Addi-
tionally, zinc completely inhibited ceramide induced FasL expression (mRNA and pro-
tein) and caspase 3 dependent apoptotic cell death. Taken together, these results
demonstrate that zinc can down-reugulate molecular mechanisms that are critical for the
apoptotic cell death and support the survival of CD4+ T lymphocytes. Importantly,
these data suggest that zinc has potential therapeutic applications in the prevention and
treatment of multiple immunosuppressive conditions that cause CD4+ T lymphocyte
depletion, including HIV infection and ageing.
7.03
THE EFFECTS OF ADJUNCTIVE ZINC AND MULTI-MICRONUTRIENT
SUPPLEMENTATION DURING TREATMENT OF PULMONARY TB:
A RANDOMISED, CONTROLLED TRIAL IN MWANZA, TANZANIA
Henrik Friis* [1], Nyagosya Range [2], John Changalucha [2], Pascal Magnussen [3],
Henrik Krarup [4], Aase Bengaard Andersen [5]
Dept Epidemiology, Inst Public Health, Univ Copenhagen, DENMARK [1]; Muhimbili
and Mwanza Medical Research Centres, NIMR, TANZANIA [2]; Danish Bilharziasis
Laboratory, DENMARK [3]; Dept Clinical Biochemistry, Aalborg Hospital, DEN-
MARK [4]; Dept Infectious Diseases, Rigshospitalet, DENMARK [5]
The effects of daily zinc (ZN, 45 mg) and multi-micronutrient (MMN, vitamins A, B, C
D, E, selenium, copper, 3-10 RDA) supplementation to pulmonary TB patients during 8
mo treatment were assessed in a randomized, controlled, two-by-two factorial trial in
Mwanza, Tanzania. Of 499 TB patients (age 35.4 (15-85) y, 41% women), 213 (42.7%)
had HIV. HIV load was determined at 2 mo in 184 (86.4%). The mean increase was 0.23
log10 (95% CI: 0.01, 0.46), but neither ZN (0.27; 95% CI: -0.18, 0.72) nor MMN (95% CI:
-0.22, -0.67, 0.23) had effects (interaction, p=0.37). Weight was measured after 7 mo in
389 (78.0%). ZN and MMN together increased weight gain by 2.37 kg (95% CI: 0.91,
3.83), whereas neither ZN (-0.21; 95% CI: -1.66, 1.25) nor MMN (0.30; 95% CI: -1.14,
1.73) alone had effects (interaction, p=0.03). According to preliminary data analyses, the
effect on weight gain was accompanied by effects on mortality, but only among those with
HIV infection. In conclusion, adjunctive supplementation with trace elements and vita-
mins during TB treatment seems to be beneficial.
7.04
THE EFFECTS OF ULTRASOUND ON SYNOVIAL FLUID ZINC LEVEL IN
PATIENTS WITH RHEUMATOID ARTHRITIS
Ethem Akçil [1], Bulent Seçkin [2], Ahmet Ergün [3], Rezan Hatungil* [4]
Dept.Pathophysiology, Faculty of Medicine, Ankara Üniversity, Ankara [1]; Dept.
Physical Medicine and Rehabilitation, Faculty of Medicine, Ankara University, Ankara
[2]; Dept. Physiology, Faculty of Medicine, Ankara University. Ankara [3]; Dept.phys-
iology, Faculty of Medicine, Mersin University,Mersin, [4],TURKEY
Recently, trace element levels such as serum zinc and copper in rheumatoid arthritis
patients became important. In some studies synovial fluid zinc levels of rheumatoid
arthritis patients were found to be increased. In this study, the effect of ultrasound on
synovial fluid zinc level and synovial fluid leukocyte count was investigated in classical
and/or definite rheumatoid arthritis patients, with the application of a dose of 2 watt/
centimeter2 for 5 minutes on the knee joints of twenty patients. Synovial fluid zinc levels
were estimated by atomic absorption spectrophotometer. Synovial fluid zinc levels
immediately after application of ultrasound was significantly decreased when compared
with that before ultrasound application (p<0.05). When the synovial fluid zinc level
immediately after ultrasound application was compared with that 24 hours after ultra-
sound application, the difference was found to be statistically significant (p<0.05). Sy-
novial fluid leukocyte count immediately after ultrasound application was statistically
significantly increased when compared with synovial fluid leukocyte count before ultra-
sound application (p<0.05). The decrease of synovial fluid zinc level and increase of
synovial fluid leukocyte count immediately after ultrasound application seems to be
related to the increase of inflammatory activity due to the ultrasound application.
7.05
LOCALIZATION OF TRACE METALS IN THE CEREBELLA OF SCRAPIE
INFECTED HAMSTERS
Christopher J. Johnson* [1,2], Valerie B. Hoefert [1,2], Bradley H. Frazer [3], Joel A.
Pedersen [4,5], Debbie McKenzie [2], Gelsomina De Stasio [3], Judd M. Aiken [1,2]
University of Wisconsin-Madison: Program in Cellular & Molecular Biology [1], De-
partment of Animal Health & Biomedical Sciences, 1656 Linden Dr, Madison WI 53705,
USA [2]; Synchrotron Radiation Center, Stoughton, WI 53589, USA [3]; Department of
Soil Science, Madison WI 53705, USA [4]; Program in Molecular and Environmental
Toxicology [5]
Bovine spongiform encephalopathy (mad cow disease), sheep scrapie, and human Cre-
utzfeldt-Jakob disease are members of a group of fatal, neurodegenerative diseases called
transmissible spongiform encephalopathies (TSEs). Pathology occurs in the cerebellum
and other brain regions. TSEs are caused by the misfolding of a host cellular protein
(PrPC) into a pathogenic isoform (PrPSc). The N-terminal regions of both PrP isoforms
contain metal-binding repeats and PrP misfolding is thought to disturb copper (Cu)
homeostasis in the brain. We hypothesize that TSE infection affects the distribution of Cu
and other metals in the brain. Using X-ray photoelectron emission spectromicroscopy, we
identified areas of high calcium concentration in the cerebella of infected, but not
uninfected animals. Several transition metals, including Cu, iron and cobalt are excluded
from these areas but are enriched on their periphery. Other metals, including aluminum
and chromium, are also excluded from calcified areas but without circumferential
enrichment. These data suggest novel, localized dysregulations of metals in TSE infec-
tion, which may contribute to the pathogenesis of disease.
7.06
REDINTEGRATION EFFECT IN NATURAL KILLER CELL ACITIVITY OF
ATHLETE WITH IMMUNOSUPRESSION BY TRACE ELEMENT
Mieko Kimura* and Takahisa Takeda
Takeda Research Institute of Life Science & Preventive Medicine, Kyoto Research Park
134 tyudoji-minamimati Simogyoku 600-8813, Kyoto, JAPAN
Intense physical exercise has been shown to be associated with immunosupression. The
notion that nutrients are involved in the regulation of immunological processes and in
the ability to cope with muscular and systemic exercise stress has been gaining atten-
tion. Particularly minerals have been shown to be related to cell mediated humoral
immunity such as natural killer (NK)-cell activity, T- and B-cell functions, and cytokine
release. On the other hand, magnesium has an important role as an enzyme cofactor in
energy metabolism. The effect on immune function of athletes through an assay of NK
cell activity of the peripheral blood lymphocytes was examined. For subjects, 33 young
men athletes and 19 non-athletes aged 19-22 were selected. NK cell activity was
determined by europium (Eu)-DTPA release assay based on measurement of time-
resolved fluorescence. This examination was performed in subjects obtained the in-
formed consent. To examine effect of magnesium on NK cell activity, magnesium, zinc,
manganese and copper was added in vitro. NK cell activity was lower in the athlete
group than in control group such as previous reports. NK cell activities in the athlete
subjects were significantly activated by addition of magnesium, zinc and manganese,
but negative effect was shown in NK cell activity by copper compared to no addition of
these minerals. These results suggest that athletic exercise induces to much exhaustion
of magnesium, zinc, manganese and consequently decline of immune function will be
caused. (Mieko Kimura serves on the Board of Director for Takeda Research Institute
of Life Science & Preventive Medicine.)
7.07
IRON DEFICIENCY REDUCES INTERLEUKIN-4 SECRETION IN MICE
Solo Kuvibidila*, Maria Velez, Ren#e Gardner, Lolie Yu, David Ode, Raj P Warrier
Pediatrics, LSU Health Sciences Center & the Research Institute for Children, New
Orleans, LA 70112, USA
Interleukin-4 (IL-4), a TH2 cytokine, regulates B cell proliferation and antibody secre-
tion. Iron deficiency, a public health problem for children and young women, impairs
interferon-gamma secretion and other T cell functions. Its effect on IL-4 secretion has not
been previously investigated. Considering that IL-4 down regulates interferon-gamma,
we hypothesize that iron deficiency increases IL-4 secretion. To test this hypothesis, we
measured IL-4 levels by ELISA in serum and supernatant of Con A- (2.5 lg/l) and anti-
CD3-antibody (50 ng/ml)-treated spleen cells from iron-deficient (ID), control, pairfed
mice (n=20-23), and iron-replete mice for 3, 7, and 14 days (n=9-13). Feeding the test
diet (5 ppm iron vs 50 ppm for control diet) for two months reduced hemoglobin,
hematocrit, and liver iron stores by 60-80% (p <0.001). Contrary to what we expected,
iron deficiency reduced IL-4 levels in serum and supernatant, and spleen cell proliferation
by 33% to 45% (p <0.05). Pairfeeding had no effect. Iron repletion increased all mea-
surements in a time dependent fashion. IL-4 levels positively correlated with indices of
iron status (r=0.568, p<0.05), but not body (r=0.222) and thymus (r=0.035) weights in
ID mice. Reduced IL-4 secretion may explain impaired antibody titers in immunized ID
rodents. Since IL-4 is required for IgE production, and IgE is involved in allergies and
asthma, our data suggest that iron deficiency may attenuate the severity of these diseases.
Whether IL-4 secretion is also altered in ID individuals and the mechanisms involved,
requires further investigation.
7.08
EFFECT OF ZINC DEFICIENCY ON NEUTROPHIL TRASMIGRATION AND
JUNCTION PROTEINS DISTRIBUTION IN CACO-2 CELLS
Alberto Finamore [1], Mara Massimi [2], Laura Conti Devirgiliis [2], Elena Mengheri* [1]
Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione, Roma, ITALY [1]; Dip.
Biologia di Base ed Applicata, Università. dell’Aquila, L’Aquila, ITALY [2]
Fundamental for the host defense is the capacity of neutrophils to migrate from blood
vessel to inflammatory sites. During the migration across epithelial monolayer the cellular
junctions are temporarily opened. Zinc deficiency can induce several alterations to im-
mune response. In this study we have investigated whether zinc deficiency affected neu-
trophil transmigration and the cellular junction proteins. We have used a human adeno-
carcinoma cell line (Caco-2) grown in complete medium (C cells), zinc depleted medium
(Zn- cells) or zinc repleted medium (ZnR cells). Neuthophil migration across inverted
monolayer of cells was measured by myeloperoxidase activity. Monolayer integrity was

checked by transepithelial electrical resistance (TEER) and paracellular [super
14]C mannitol flux. Tight and adherence junction proteins were analyzed by immuno
localization. The results showed a small decrease of TEER but not of mannitol passage in
Zn- cells, indicating no damages in cellular junctions. Zn- cells appeared morphologically
bigger and/or with irregular shape compared to C and ZnR cells. Different distribution of
Z0-1, occludin and -catenin on cellular membrane and a partial delocalization of E-
cadherin were seen in Zn- cells compared to C and ZnR cells. Neutrophil transmigration
occurred earlier in Zn- cells compared to C and ZnR cells, although the total amount of
neutrophil transmigrated were similar in all cells.
7.09
ATTENUATION OF BERYLLIUM INDUCED TOXICITY THROUGH TIFERRON
ALONG WITH PROPOLIS AS NUTRITONAL SUPPLEMENTATION IN RATS
Satendra Kumar Nirala* [1], R. Mathur [1], A. Mathur [2]
SOS in Zoology, Jiwaji University, Gwalior-474011, INDIA [1]; Naveen Girls College,
17-A, New Saket Nagar, Gwalior-474002, INDIA [2]
Laboratory based experimental studies and industrial experiences over the last five
decades have already established the toxic effects of beryllium in humans and experi-
mental animals and still there is no antidote concerning its toxicity. The study aims for
the first time to evaluate the therapeutic efficacy of Tiferron (Sodium 4,5-dihydroxy-
benzene 1,3-disulphonate) along with ‘‘Propolis’’ as nutritional supplement against
toxicological consequences of beryllium exposure in female albino rats of Sprague
Dawley strain. Animals were exposed to beryllium (as beryllium nitrate) at a dose of 1
mg/2ml/kg (ip) once a day for 28 consecutive days followed by therapy with Tiron (300
mg/2ml/kg, ip) and Propolis (200 mg/5ml/kg, po) individually and in combination for 5
consecutive days after toxicant administration. Data were statistically analyzed through
students‘t’ test followed by one way ANOVA and the values were considered to be
significant at P<0.05. Results of blood biochemistry revealed significant depletion in
blood sugar level, serum alkaline phosphatase, albumin and urea whereas non significant
fall in Hb and total serum protein contents was also noticed. Significant elevation was
noticed in AST, ALT, LDH, bilirubin, creatinine and G-GT. Tissue biochemistry re-
vealed severe alterations in liver and kidney. Significant fall in total protein and glycogen
contents, succinic dehydrogenase, alkaline phosphatase, adenosine triphosphatase and
glucose-6-phosphatase was noticed in both the organs. On the contrary, significant
elevation in acid phosphatase, triglycerides, total and esterified cholesterol level were also
recorded in both organs. Significant rise in lipid peroxidation and reduced glutathione in
both liver and kidney showed oxidative stress induced due to beryllium exposure.
Activities of microsomal drug metabolizing enzymes i.e. aniline hydroxylase and amid-
opyrene-N-demethylase were significantly declined due to beryllium toxicity. Although
Tiferron and Propolis exhibited therapeutic efficacy against beryllium induced toxicity
yet Tiferron in combination with Propolis exerted statistically more beneficial effects in
respect to control to reverse alterations in different variables altered due to beryllium
intoxication. Mobilization study of beryllium showed that combination of Tiferron and
Propolis significantly reduced beryllium burden in liver, kidney, blood, brain and bones
without any significant change in essential metals. Electron and photo micrographic
observations of liver and kidney also supported the data concluding that the combina-
tion of Tiferron and Propolis can be expected as better choice in treatment of beryllium
induced toxicity.
7.10
IMMUNE RESPONSE AND ZINC STATUS IN OLDER SUBJECTS
A.Finamore [1], D. Panno [1], V.C. Jewell [2], C. Hodkinson [2], J. M. O’ Connor [2], J.
Wallace [2], S. Strain* [2], E. Mengheri [1]
Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione, Via Ardeatina, 546, 00178
Roma, ITALY [1]; Northern Ireland Centre for Food and Health, University of Ulster,
Cromore Road, Coleraine, BT52 1SA, NORTHERN IRELAND [2]
Immune responses are well known to decline with aging and cell-mediated immunity
appears to be the most compromised. Zinc is fundamental for the activity of the immune
system and a suboptimal zinc status might be a frequent phenomenon in elderly popu-
lations. In this study we report baseline data on several markers of immune response of
96 male and female late middle-aged (55-70 years old) Northern Irish volunteers and 96
older (70-85 years old) Italian volunteers, both men and women. This study is a part of a
EU project, ZENITH, which aims to investigate the zinc status of older Europeans and
the effect of zinc supplementation (at 15 or 30 mg/day). In the old population, prolif-
erative response of lymphocytes to a common polyclonal mitogen was measured by,
[super 3]H-thymidine incorporation. Moreover, cytokine gene expression and secretion
by stimulated and non-stimulated lymphocytes were analyzed by RT-PCR and ELISA
assay repspectively. In the late middle-aged population, lymphocyte and leucocyte sub-
sets (Total T- and B-lymphocytes, T-helper lymphocytes numbers, natural killer cell
numbers, activated T-lymphocytes (early and late-activated)), intracellular cytokines in
lymphocytes and monocytes, phagocytosis and apoptosis (50 volunteers only) were
measured by flow cytometry. (ZENITH is supported by the European Commission
‘‘Quality of Life and Management of Living Resources’’ Fifth Framework Programme,
Contract No: QLK1-CT-2001-00168.)
7.11
REDUCED PRODUCTION OF IMMUNOREGULATORY CYTOKINES IN ZINC
AND VITAMIN A DEFICIENT INDONESIAN INFANTS
Frank T. Wieringa* [1], Marjoleine A. Dijkhuizen [1], Muhilal [2], Johanna vd Ven-
Jongekrijg [1], Clive E. West [3], Jos W.M. van der Meer [1]
LANDS [1]; Nutr Res Dev Center Bogor, INDONESIA [2]; Dept Human Nutr, Univ
Wageningen, NETHERLANDS [3]
Immunological assessment was done in 59 infants in Indonesia to determine the effect of
zinc, vitamin A and iron deficiency on the ability to produce immunoregulatory cyto-
kines. Production of T-helper cell type-1 cytokines interferon-[gamma] (IFNg) , inter-
leukin-12 (IL12), interleukin-18 (IL18) and T-helper cell type-2 cytokine interleukin-6
(IL6) were measured after stimulation with lipopolysaccharide and phytohaemagglutinin
in an ex vivo whole blood culture system. Circulating neopterin concentrations were
determined as an indicator of in vivo macrophage activity. Production of IFNg and IL12
were strongly correlated, IFNg and IL18 production were not. Zinc deficiency was
accompanied by significantly reduced white blood cell counts and reduced ex vivo pro-
duction of IL6. Vitamin A deficient infants had significantly reduced ex vivo production
of IFNg, but also significantly higher circulating neopterin concentrations. Iron status
was not related to cytokine production. This study shows that in vitamin A deficiency
there is Th1 dominance in a steady state, combined however with impairment of the Th1
response after stimulation, whereas in zinc deficiency, there is a decreased Th2 response.
Overall, vitamin A deficiency and zinc deficiency have marked albeit different effects on
the immunocompetence of infants, affecting both cell-mediated and humoral components
of the immune system.
Category 8: Trace Element Molecular Nutrition for the Clinician
8.01
FUMONISIN B1 DAMAGE DNA IN RAT LIVER. SELENIUM AND OTHER
ANTIOXIDANTS INHIBIT FUMONISIN TOXICITY
Faik Atroshi*, Aldo Rizzo, Terhi Ali-Vehmas, Tuomas Westermarck
Dept. Vet. Clinical Science, Pharmacology & Toxicology, PL 57, 00014 Helsinki Uni-
versity, FINLAND
Active oxygen radicals species are reported to cause organ damage. This study was
designed to determine whether oxidative stress contributed to the initiation or progres-
sion of hepatic cell DNA damage induced by fumonisin B1 (FB1) in rats. The protective
effects of selenium, vitamin E, coenzyme Q10, L-carnitine was studied. Fasted rats were
injected intravenously with a single dose of fumonisin B1 at 1.55 mg kg-1 body weight.
Treatment with FB1 led to hepatic DNA fragmentation in 85% of the tested animals.
DNA fragmentation was investigated as a critical event in toxic cell death by testing total
Ca 2+ in liver. FB1 administration caused total Ca2+ in liver to increase within 4 h ( 204%
of control) . Measurement of liver enzyme activities showed an increase in ASAT and
ALAT. Fumonisin B1 also markedly decreased hepatic glutathione (GSH) levels. Pre-
treatment with selenium (1 mg selenium as sodium selenite kg-1 diet), _-tocopherol (30 IU
vitamin E kg-1 diet) and CoQ10 (30 mg CoQ10 kg-1 diet) together with L-carnitine (2.8 mg
carnitine kg-1 diet), showed a change in DNA damage and decrease the activities of Ca2+,
ASAT and ALAT in the liver. The level of GSH was slightly increased. Oxidative damage
caused by FB1 may be one of the underlining mechanisms of FB1-induced cell injury and
DNA damage.
8.02
KISS AND RUN: THE ROUTE OF IRON FROM ENDOSOMES TO
MITOCHONDRIA CAN BYPASS THE CYTOSOL IN
HEMOGLOBIN-PRODUCING CELLS
Alex Sheftel [1], An-Sheng Zhang[1,2], Orian Shirihai [3], Prem Ponka* [1]
Dept. of Physiology and Medicine, McGill University, Montreal, QC, Canada [1]; Dept.
of Cellular and Developmental Biology, Oregon Health Sciences University, Portland,
OR, USA [2]; Biocurrents Research Center, Marine Biological Institute, Woods Hole,
MA, USA [3]
During differentiation, immature erythroid cells acquire vast amounts of iron at a
breakneck rate. Proper coordination of iron delivery and utilization in heme synthesis is
essential and disruption of this process likely underlies iron loading disorders such as
sideroblastic anemia and myelodysplastic syndrome with ringed sideroblasts. Iron is ta-
ken up by the cells via receptor mediated endocytosis, a process whereby diferric trans-
ferrin (Tf) binds to its cognate receptor (TfR) on the erythroid cell plasma membrane,
followed by internalization of the Tf-TfR complex. Subsequent to endocytosis, the en-
dosome is acidified by a H+-ATPase, allowing the release of iron from Tf. Through an
unknown mechanism, iron is targeted to the inner membrane of the mitochondria, where
the enzyme that inserts Fe into protoporphyrin IX, ferrochelatase, resides. Although it
has been demonstrated that the divalent metal transporter, DMT1, is responsible for the
egress of reduced Fe from the vesicle, the immediate fate of the iron atoms after their
transport across the vesicular membrane remains unknown. Because reduced iron is a
strong pro-oxidant, contributing to free radical formation through Fenton chemistry, it
has been predicted that an iron binding molecule shuttles Fe from the endosome to
mitochondria. However, this much sought iron binding intermediate, that would con-
stitute the labile iron pool (LIP), has yet to be identified. Thus, we hypothesize that, in
hemoglobin-producing cells, there is a direct relaying of Fe from the endosomal
machinery to that of the mitochondria. We have taken two strategies in examining this
supposition: 1) a biochemical approach by which the cytoplasm of cells was loaded with
an impermeant iron chelator, thus intercepting the delivery of Fe by the putative LIP
intermediate, and 2) a morphological approach employing time-lapse confocal micros-
copy which permits the tracking of iron-loaded endosomes and mitochondria with high
spatial and temporal resolutions. To examine whether iron delivered by Tf for heme
synthesis can bypass the cytosol, we have loaded reticulocytes with a high-molecular
weight version of desferrioxamine, hDFO, prior to incubation with 59Fe-Tf. The incor-
poration of transferrin iron into heme was unaffected by hDFO when compared to
controls. Importantly, iron delivered to these cells in a form that freely diffuses across the
membrane, iron-salicylaldehyde isonicotinoyl hydrazone (59FeSIH2), was significantly
prevented from being used for heme synthesis in hDFO-laden reticulocytes. Using con-
focal microscopy, as well as polarized light microscopy, we found that endosomes are
very mobile organelles. Immediately following budding from the plasma membrane, these
organelles continuously traverse the cytosol and touch a number of mitochondria mul-
tiple times. Experiments using various pharmacological agents indicate that these
movements are mediated by components of the cytoskeleton which are essential for
proper iron delivery for use in heme synthesis. Together, these data suggest that iron is
directly delivered to mitochondria by endosomes in a ‘‘kiss and run’’ paradigm. Our
current studies will examine the required components and regulation of this interaction
using the same experimental strategies as well as a cell free system consisting of isolated
organelles.
Category 9: Trace Elements and Neurological Diseases and Neuropsychological Impairment
9.01
EFFECT OF THERAPEUTIC IRON SUPPLEMENTATION ON
DEVELOPMENTAL INDICIES IN CHILDREN 1-3 YEARS OF AGE WITH
SEVERE ANEMIA
Sunil Sazawal* [1,2], Deepak Marwah [1], Priti Verma [1], Usha Dhingra [2], Saikat Deb
[1], Rakesh Juyal [1], Meena Osmany [1], Archana Sarkar [1], Pratibha Dhingra [1],
Jitender Kumar [1], Venugopal P Menon [1], Maureen Black [3], Robert E Black [2]
Center for Micronutrient Research, Annamalai University, India [1], Department of
International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore
USA [2], Department of Pediatrics, University of Maryland, School of Medicine, Balti-
more USA [3]
Although there have been some data relating iron deficiency to impaired development in
observational studies, the evidence from clinical trials of iron supplementation has been
inconclusive and sparse. Only two trials (Indonesia and Pemba) have reported impact of
iron therapy on development among children below 3 years. In a population where we
demonstrated a statistically significant association of severe anemia to lower develop-

mental scores we evaluated impact on these scores of therapeutic iron (3mg/kg/day)
supplementation for 3 months. In 94 children with baseline hemoglobin <=7 g/dl
Bayley’s Scales of Infant Development and Language scale were instituted at baseline and
after iron supplementation for 3 months at which time blood sample was also collected.
Hemoglobin values increased from baseline 6.19 ± 0.71 g/dl to 8.22 ± 1.83 g/dl; p<
0.001. There was also a significant increase in HCT 5.38 (95%CI 4.1-6.7) and decrease in
MCV 3.3; P=0.04 and RDW 3.04; P=0.005. Paired pre-post analysis indicated that
improvement in iron status resulted in significant improvement in PDI score 89.5±13.7
Vs 83.7±16.0 p=0.006), MDI Post 84.7±11.9 vs 81.1±10.4; p=0.03, Orientation
30.7±16.8 vs 24.1±15.8; p = 0.004, Emotional Regulation 29.8±11.3 vs 25.8±10.3; p
= 0.004) and Language scores 7.1±3.8 vs 4.3±3.9;p < 0.001. These data suggest that
developmental deficits associated with severe anemia are in fact caused by iron deficiency
and can be reversed by treatment.
9.02
THE NEW EVIDENCE ON THE ROLE OF ELEMENTAL HOMEOSTASIS IN
NEURO-DEGENERATIVE AND NEURO-PSYCHIATRIC DISORDERS
P. Shanmugavelu [1], S. S. Rao [2], M. L. Hegde [3], M. S. Mustak [3], R. B. Menon [1],
N.M. Shamasundar [2] and K. S. J. Rao [3]
B.A.R.C, Mumbai , INDIA [1]; J.S.S. Medical College, Mysore, INDIA [2]; Department
of Biochemistry and Nutrition, C.F.T.R.I, Mysore, INDIA [3]
Trace elements play pivotal role in structure and function of cell. Information on the level
of trace elements in different vital organs in control and pathological conditions is very
essential to interlink metal-biological functions. The study aims to provide new evidence
on the role of elemental homeostasis in neuro-disorders. Analysis of elements was carried
out using Inductively Coupled Plasma Atomic Emission Spectrometry in brain and
Cerebrospinal fluid (CSF) of Alzheimer’s disease (AD), serum samples of Parkinson’s
disease (PD) and Bipolar disorder (BD). The results showed imbalance in trace elemental
homeostasis and a definite pattern of inter-elemental relationships. In moderately affected
AD brain, monovalent elements were significantly reduced, while divalent elements (Zn,
Cu) were increased compared to control. In severely affected AD brains, trivalent ele-
ments Al and Fe, and S were predominantly present displacing monovalent and divalent
elements. An altered charge distribution pattern is established in AD. In case of AD CSF
samples, levels of K, P, and S were decreased. In both early and severe PD serum, Al and
S were decreased. In BD groups (I, II and V) Na, P, Cu, Al and Mn were elevated while S,
Fe and Zn were decreased. The observations are novel and significant, and lead to a new
hypothesis on role of trace elemental homeostasis in neuro-disorders.
9.03
ZENITH PROJECT: THE EFFECTS OF ZINC ON BEHAVIOURAL AND
PSYCHOLOGICAL PARAMETRES IN HEALTHY OLDER ADULTS:
PRELIMINARY FINDINGS
Liz Simpson* [1], Chris McConville [1], Gordon Rae [1], Barbara Stewart [1], Elizabeth
Maylor [2], Jenni Birrell [3], Victoria Jewell [1], Jacqueline O’Connor [1], Angela Polito
[4], Nathalie Meunier [5], Maud Andriollo [6], Charles Coudray [5]
School of Psychology, Niche University of Ulster, Cromore Road, Coleraine, County
Londonderry, NORTHERN IRELAND, BT52 1SA [1]; Department of Psychology,
University of Warwick, Coventry, ENGLAND CV4 7AL [2]; Cambridge Cognition

Limited Tunbridge Court Tunbridge Lane Bottisham Cambridge, ENGLAND CB5
9DU [3]; National Institute for Food and Nutrition Research Human Nutrition Unit, via
Ardeatina 546, 00178 Roma, ITALY [4]; Centre de Recherche en Nutrition Humaine
d’Auvergne, Unité Maladies Métaboliques et Micro-Nutriments, Inra, Rentre de
Recherche de Clermont-Ferrand/Theix, 63122 Saint Genès Champanelle, FRANCE [5];
Laboraroire de Biologie de Stress Fxydant, Faculté de Pharmacie, Domaine de la Merci,
38700 la Tronhe, FRANCE [6]
ZENITH is a large European multi-centre study looking at the effects of zinc on healthy
ageing, in older adults (n=440) aged 55-85 years, recruited through centres in France,
Italy and Northern Ireland. It is a random placebo controlled double blind intervention
study of zinc supplementation of either 15 or 30mg Zn/day or placebo lasting for six
months. Measures of psychological and behavioural parameters have been obtained at
baseline for health and lifestyle factors, personality, cognition, mood, taste acuity, stress
and coping. Centre and gender differences for stress, some coping styles, mood and
personality were found. The results will address confounding effects of age on all vari-
ables and the role of zinc supplementation.
9.04
ANEMIA IN CHILDREN 1-3 YEARS OF AGE IS ASSOCIATED WITH POORER
SCORES ON ALL DEVELOPMENTAL INDICIES
Priti Verma* [1], Sunil Sazawal [1,2], Usha Dhingra [2], Rakesh Juyal [1], Deepak
Marwah [1], Archana Sarkar [1], Pratibha Dhingra [1], Venugopal P Menon [1], Maureen
Black [3], Robert E Black [2]
Center for Micronutrient Research, Annamalai University, India [1], Department of
International Health, Johns Hopkins Bloomberg School of Public Health, Baltimore
USA [2], Department of Pediatrics, University of Maryland, School of Medicine, Balti-
more USA [3]
Approximately 25% of world’s children under 3 years of age have iron deficiency
anemia. In developing countries more that 60% of preschool children are anemic. Al-
though there are good animal data for association of iron deficiency and development,
data from human studies have been variable most of the studies have been small. We
evaluated association between anemia and developmental indices in a population with
high prevalence of anemia in peri urban New Delhi. As a part of baseline evaluation of
a randomized control trial we had assessments on the same day of hemoglobin status
and developmental indices (using Bayley’s II Scale and Language questionnaire) in 1242
children aged 1-3 years. Hemoglobin measured by coulter was < =7 g/dl in 129
children, 7.1- 10.0 g/dl in 793 children and >10 g/dl in 320 children. Using hemoglobin
> 10g/dl as reference category, children with hemoglobin 7.01-10.0 had significantly
lower scores on Psychomotor Development Index (PDI) (91.7 vs 94.41, p= 0.005),
Mental Development Index (MDI) (86.0 vs 87.9; p=0.03), Orientation (32.1 vs 35.1;
p=0.02), Motor Quality (MQ) (98.7 vs 99.0; p=0.03), Emotional Regulation (ER)
(29.1 vs 30.1; p= 0.04) and Language Scores (4.5 vs 5.5; p <0.001). Children with
Hemoglobin < =7 g% had higher deficits, PDI 85.0, MDI 81.2, Orientation 24.8, ER
25.9, Language Score 3.9, MQ 98.7 all with p<0.001 except for MQ, p=0.32). These
data confirm a negative association of anemia with development in children. The
gradation in response and similarity in socio-economic and home characteristics would
favor a causal relationship.
9.05
SELENIUM SUPPLEMENTATION IN JUVENILE NEURONAL
LIPOFUSCINOSIS PATIENTS. 30 YEARS EXPERIENCE
Tuomas Westermarck*, Faik Atroshi
Rinnekoti Research Center, 02980 Espoo; Dept. Pharmacology & Toxicology, University
of Helsinki, FINLAND
Juvenile neuronal ceroid lipofuscinosis (JNCL) is a recessively inherited progressive
neurodegenerative lysosomal storage disease leading to blindness and severe motor and
mental disability. The gene in chromosome 16 has been cloned, but its basic mechanism is
unknown. Excesses of free radicals have been found to be some secondary phenomenon
in JNCL. However, the use of antioxidants has shown to be beneficial. To determine
whether a nutritional supplement of selenium, combined with vitamin E and vitamins B-
complex will benefit JNCL patients. A total of 73 patients with a history of JNCL were
included. Oral administration of 0.05 mg kg -1 b.w. of selenium per day, %-tocopherol 10-
20 mg kg -1 b.w., vitamin B2 3-6 mg/day, vitamin B6 20-75 mg/day. The primary end
points for the trial were neurological-dysfunction of IQ, motor performance, balance,
coordination and speech. Selenium with vitamins E and B-complex treatment did show
beneficial effect. 10% of the selenium treated patients showed nausea or vomiting when
sodium selenite was administered at the same time with vitamin C rich food or beverage.
Selenium combined with vitamins E and B-complex treatment showed to have a bene-
ficial effect to JNCL patients which support the hypothesis that supplemental combined
antioxidants may help in reducing the incidence of motor performance, balance coor-
dination and speech in these. Recommendations regarding selenium and other antioxi-
dants supplementation for JNCL patients can be made.
9.06
PLASMA ZINC AND COPPER LEVELS IN AUTISTIC CHILDREN
Özgür Yorbik* [1], Cemal Akay [2], Ahmet Sayal [2], Adnan Cansever [3], Teoman
Söhmen [4], Ayhan O. Çavdar [5]
Child and Adolescent Psychiatry Department, GATA (Gülhane Military Medical
Academy) [1]; GATA-Toxicology Department [2]; GATA Psychiatry Department [3];
Chief, GATA Child and Adolescent Psychiatry Department, Etlik, Ankara [4], Chief
Trace Element Research Group of TÜBA (Turkish Academy of Sciences), UNESCO
Satellite Center in Ankara, TURKEY [5]
The transition metal zinc is an essential trace element for normal development and
function of biological systems including the CNS. Zinc deficiency during early brain
development may cause malformations, while during the later phase impairs neuronal
growth, synaptogenesis and may cause behavioral sequel. The aim of this study was to
investigate blood (plasma and erythrocyte) zinc and plasma copper levels in children with
autistic disorder. The plasma, erythrocyte zinc levels and plasma copper concentrations
were measured by atomic absorption spectrophotometry in the 45 autistic children. Forty
one healthy children of the same age group and sex were used as controls. Plasma and
erythrocyte zinc levels in autistic children were found to be significantly lower (13.0 + 1.7
lmol/L and 1.51 + 1.6 lmol/L against 16.8 + 1.6 and 1.95+ 1.9 lmol/L in normals)
while plasma copper levels (153 + 10.8 lg/dl ) were significantly higher (p<0,05) in
autistic children as compared to controls (122 + 10 lg/dl). These results may indicate
that either chronic zinc deficiency (low plasma and erythrocyte zinc levels) or a metabolic
abnormality related to zinc may exist in autistic children. Interestingly, in our previous
study plasma and erythrocyte levels of antioxidant enzymes, namely GSH-Px and SOD
were also found to be significantly low in autistic children. Further studies are required to
analyze blood selenium, manganese and ceruloplasmin levels in addition to that zinc and
copper to clarify underlying mechanism in this disorder.
Category 10: Trace Elements and Osteoporosis and Other Bone Disorders
10.01
UROV KASHIN-BECK DISEASE: BIOGEOCHEMICAL ASPECTS
Vadim V. Ermakov
V.I. Vernadsky Institute of Geochemistry and Analytical Chemistry of RASci., Kosigin
Str., 19, 119991 Moscow, RUSSIA
In connection with the supposition of significance of geochemical factors in etiology of
Urov Kashin-Beck disease endemic deforming osteoarthrous, widespread mainly in
China and East Zabaikalia (Russia), between 1998-2003 are carried out complex ecology-
biogeochemical research of Urov provinces of a polymetallic zone of Chita district (basin
of the rivers Shilka, Unda, Gazimur, Urumkan and Urov). The basic attention is given to
distribution and behaviour of biologically active chemical elements in soils, waters and
plants of endemic and control territories. Enrichment of soils of Urov biogeochemical
provinces by manganese, phosphorus, iron and to a lesser degree strontium that reflects
landscape-geochemical contrast of an examined territories and features of migration of
macro-and trace elements in conditions of a permafrost and bogging of territories is
discovered. Concentration of calcium in soils of endemic villages in some cases surpass,
and in most cases do not exceed a level of its content in soils of control sites. The fact of
sharp enrichment of soil by strontium in flood-lands of the river Urov is obvious, that is
connected to structure of soil-forming of rocks. The most part of endemic villages is dated
to gold-fields to displays and characterized by higher concentration of arsenic in flood-
lands landscapes. Coefficient of biological absorption of calcium and strontium in system
the plant - soil grows in endemic areas, and phosphorus - decreases. Manifestation of
Urov bone-articulate pathology in East Zabaikalia occurs on a background of deficiency
of selenium in all parts of a biogeochemical food chain, a low level of sulfates in natural
waters with weak mineralization and fungous invasion of grain crops. Nevertheless, on a
number of parameters the areas of spreading of Urov Kashin Beck disease in Russia
differ from those in China, except for the common deficiency of selenium. (Supported by
grant of RFFR 03-05-64597 and grant of MSTC 2046.)
10.02
EFFECT OF METALLOTHIONEIN ON BONE FORMATION PROCESSES IN
RAT MARROW MESENCHYMAL STEM CELLS
Yoshiko Dohi [1], Hideki Shimaoka [1,2], Masako Ikeuchi [2,3], Hajime Ohgushi [3],
Kunio Yonemasu [1], and Takeshi Minami* [4]
Dept Public Health [1], Oral and Maxillofacial Surg [2], Nara Med Univ, Kashihara 634-
8521, JAPAN; Tissue Engineer Res Center, Natl Inst Advanced Indust Sci & Technol.,
Amagasaki 661-0974, JAPAN [3]; Dept Life Sci, School Sci & Technol, Kinki Univ, 3-4-1
Kowakae, Higashi-osaka 577-8502, JAPAN [4]
Metallothionein (MT) is a ubiquitous low-molecular-mass protein. There are four
major isoforms, and MT-1 and MT-2 are known to be acute phase proteins. In bone
tissue culture, it is suggested that the MTs might be involved in zinc homeostasis, but
the details are still not clear. We have established an experimental model in which
osteoblasts with the capability of mineralization are induced in mesenchymal stem cells
(MSCs) cultured in the presence of dexamethasone, ß-glycerophosphate, and ascorbic
acid. However, the processes underlying osteoblastic differentiation remain to be de-
fined in detail. Therefore, the present study is conducted with two aims: to determine
whether the expression of MT isoform genes changed temporary in dexamethasone-
treated MSCs and to assess whether MTs have an important role in osteoblastic dif-
ferentiation. Subculture of rat bone marrow MSCs was achieved with ß-glycerophos-
phate, and ascorbic acid phosphate, and with or without dexamethasone for 15 days.
Levels of MT-1, MT-2, MT-3, osteocalcin, ALP, and GAPDH mRNAs were measured
by real-time quantitative RT-PCR. Osteoblastic differentiation after the addition of
sense or antisense oligonucleotide for MT-1 and MT-25 was compared with reference
to Ca content and ALP activity at day 14. Both MT-1 and MT-2 mRNA expression in
the MSCs with dexamethasone showed maximum levels at day 1, and the expression
levels of MT-2 mRNA was 9.4 fold higher than that of MT-1 mRNA. Osteoblast
differentiation and mineralization of MSCs at day 14 was inhibited by the addition of a
common antisense oligonucleotide for both MT-1 and -2 in the culture medium during
the first 4 days. The present study suggests that the large amounts of MT-2 are pro-
duced in the early stage of subculture of MSCs and this may regulate their differen-
tiation.
10.03
LOW-LEVEL CADMIUM INTAKE TOGETHER WITH LACTATION MODULATE
BONE METABOLISM AND RENAL FUNCTION IN MOTHER RATS
Hisayoshi Ohta*, Yasuhiro Nakamura, Yohei Ohkawa, Yoshifumi Ohmori, Hiroshi
Sano
Dept. of Environ. Occup. Health and Toxicol., Kitasato University, 1-15-1 Kitasato,
Sagamihara, Kanagawa 228-8555 JAPAN
Female Wistar rats (6 weeks old) were given Cd (as CdCl2) at a dose of 1, 2, and 5
mgCd/kg/day by gastric tube daily for 6 consecutive days a week during the period of
pairing with male rat, of pregnancy and of nursing to newborn rats. Renal Cd con-
centration, urinary excretion of enzymes (NAG, AAP, GST, ALP), beta2-MG, amino
acids, calcium, pyridinoline (Pyr), deoxypyridinoline (Dpyr), and intact-osteocalcin
(BGP) and vitamin D activity in plasma were determined to evaluate bone metabolism.
Bone mineral density and bone strength, and osteoid volume (%) of the femur tissue
were also determined. Cd accumulation in kidney and bone increased dose-dependently
according to the Cd administration. Prominent renal dysfunction was observed after
gestation and lactation, as an increased urinary excretion of beta2-MG, NAG, and
protein. The prominent decrease of bone mineral density was also found in the mother
rats of groups of 2 and 5 mgCd/kg after lactating to newborn rats. An increase of
urinary excretion of Pyr and Dpyr by lactation was observed in both mother rats of the
Cd exposed group and of the non Cd exposed group. The activity of vitamin D and
BGP in plasma were not changed significantly except the decreased 1alpha-25(OH)-D
of the 5 mg Cd/kg group. These results suggest that low-level cadmium intake affected
more adversely to renal function and bone metabolism together with the effect of
lactation, although the renal dysfunction and the bone metabolism disorder of mother
rats were caused by gestation-lactation.
10.04
OSTEOARTHRITIS AND POST MENOPAUSAL OSTEOPOROSIS RATES IN A
BORATE AREA IN TURKEY
Ugur Sayli* [1], Bekır Sıtkı Saylı [2], Tanju Baskan [3], Aslı Koktener [4],Gulcın Dılmen
[4], Mehmet Korkmaz [2], Ayse Saylı [5]
Dept. of Orthopaedıcs and Traumatology, Fatıh Unıversıty, Ankara , TURKEY [1];
Boron Research Center, Balıkesır Unıversıty, Balıkesır, TURKEY [2]; Orthopaedıcs and
Traumatology Clınıc, Labor’s Hospıtal, Balıkesır, TURKEY [3]; Dept. of Radıology,
Fatıh Unıversıty, Ankara, TURKEY [4]
In order to reveal ıf a seemıngly ıncreased amount of boron exposure envıronmentally
affects osteoporosıs prevalance rate ın postmenopausal women, we compared bone mı-
neral densıty (BMD) measurements of natıves from a borate town wıth a group from
elsewhere. Nınety-eıght females aged 49 to 75 from the town Iskele mıght be consıdered
a ‘natural human boron laboratory’, had BMD measures. Accordıng to WHO crıterıa
27.6% were found osteopenıc and 22.4% were osteoporotıc wıth equıvocal results con-
cernıng 208 controls. Sıxty sıx males between 38 and 84 years of age from the same area ,
eıther actıve (54) or retıred (12), were evaluated by physıcal examınatıon and radı-
ologıcally for sıgns of knee osteoarthrıtıs and results were compared wıth the fındıngs of a
control group from elsewhere, wıth sımılar demographıc data. The fındıngs would ımply
that a yet-undefıned factor mıght be requıred for preventıon of knee osteoarthrıtıs and
osteoporosıs ın addıtıon to boron supplementatıon.
Category 11: Toxicity of Trace Elements
11.01
FIELD-FLOW FRACTIONATION-INDUCTIVELY COUPLED PLASMA MASS
SPECTROMETRY: A NEW APPROACH TO STUDY METAL-PROTEIN
INTERACTION
Atitaya Siripinyanond [1,2], and Ramon M. Barnes* [1]
University Research Institute for Analytical Chemistry, 85 N. Whitney Street, Amherst,
MA 01002-1869, USA [1]; Department of Chemistry, Mahidol University, Rama VI Rd.,
Bangkok 10400, THAILAND [2]
Field-flow fractionation (FFF) is a gentle size separation technique applicable to both
macromolecules and particles. The technique provides unique species information.
Coupling FFF with inductively coupled plasma mass spectrometry (ICP-MS) offers the
possibility of size based elemental speciation. In the nutritional viewpoint, FFF-ICP-
MS provides information about biodistribution of essential and toxic metals in pro-
teins. Binding of metals to metal transport and storage protein, i.e., serum albumin,
transferrin, and ferritin was examined by FFF-ICP-MS. Binding of U and La to
albumin monomer (67 kDa) was observed. Lanthanum appears to show preferential
binding to the large molecular weight components (MW 280 kDa) at extended contact
times. Cobalt, La, Tl, and U were found to bind with ferritin (MW 440 kDa).
Moreover, binding capability of Cr(III), Cr(VI), and Cr(picolinate) to human apo-
transferrin was studied. Only Cr(III) binds to transferrin. Chromium (VI) and Cr(pic)
do not bind to transferrin. The capability of FFF-ICP-MS to differentiate binding
ability of various species of Cr with transferrin provides clear evidence that the tech-
nique is a useful tool for nutritional study.
11.02
EXTREME HIGH BLOOD SELENIUM CONCENTRATIONS IN POLAR
ESKIMOS
Bente Deutch*, Henning Sloth Pedersen, and Jens C Hansen
Centre for Arctic Environmental Medicine, University of Aarhus, Vennelyst Boulevard 6,
Aarhus DK-8000, DENMARK
In Greenland the human intake of selenium has always been high and is closely connected
to intake of the traditional food of marine origin. Analyses of historic and present day
human and animal hair samples have indicated that the selenium level in the marine
environment has been constant over time, while the levels in humans have declined
corresponding to a decrease in intake of traditional food. The Inuit population in
Greenland is in dietary transition where western-style food will increasingly dominate. As
a consequence the ample supply of selenium may not be sustained in the future. We
report here the selenium status in the Polar Eskimo population living in the northernmost
part of Greenland. Mean whole blood concentrations 585 lg/L (range: 73-1910 lg/L, the
geometric mean was 483 lg/l). 33 or 41% of the 79 participants had a blood concentration
above 560 lg/L corresponding to a daily intake of 400 ı̀g regarded as the individual daily
maximum safe dietary intake. Selenium blood levels were highly significantly correlated
with long chain marine fatty acids in plasma. Dietary survey and food composition data
showed that whale skin, muktuk, is the main source of Se followed by birds, seal meat
and organs, and fish. Mercury blood concentrations were also high with a mean of 70 lg/
L (range: 2.3 240 lg/L) 99% (all but one person) had blood concentrations above 4.4
lg/L corresponding to a daily intake at the U.S. EPA RfD. The mean Se/Hg molar ratio
was 24 with a minimum of 4.5 indicating surplus of selenium, which might counteract
potential effects of mercury.
11.03
GENETIC VARIATION IN ATP7B PROMOTOR AND 5’ UTR IN HAN CHINESE
PATIENTS WITH WILSON’S DISEASE
Yan-Hong Gu* [1], Si-Lian Du [2], Hiroko Kodama [1], Qing-Jiu Gu [3], Han-Jun Sun [4]
Dept. Pediatrics, Teikyo University School of Medicine, Tokyo, 173-8605 JAPAN [1];
Dept. Pediatrics, The Affiliated Hospital of Qingdao Medical College, Qingdao, 266000
CHINA [2]; Dept Surgery, TieJian Hospital of China Railway Construction Corpora-
tion, Beijing, 100855 CHINA [3]; Dept Otolaryngology, Chinese Armed Police General
Hospital, Beijing, 100039 CHINA [4]
Wilson’s disease (WD) is an autosomal recessive disorder of copper transport charac-
terized by the accumulation of intracellular copper. The WD gene (ATP7B) encodes a
copper transporting P-type ATPase with 50-70% similarity to the Menkes disease (MNK)
protein, and is primarily expressed in the liver, while the MNK gene (ATP7A) is almost
ubiquitously expressed in a variety of tissues, but not in the liver. The ATP7B gene, but
not ATP7A, contains several metal response elements (MRE) and MRE-like sequences
(MLS) in the promotor region that seem to play important roles during the ATP7B gene
expression. Considering the previous studies on ATP7B mutations, it is possible that
disease-caused mutations may exist in the promotor, 5’ untranslated region, or introns.
So far, only two mutations have been reported in the promotor region of the ATP7B
gene. In the present study we identified a new mutation and a new polymorphism, and
there were totally eight genetic variations in the promotor region of the ATP7B gene in
Chinese population. The identified mutation -215 A to C, occurred close to MREd,
between MREd and the E-box. The results of the present study suggest that mutations in
the regulatory elements of the ATP7B gene are rare in Chinese patients.
11.04
EFFECT OF HIGH DOSE MANGANESE EXPOSURE ON KIDNEY TISSUE
M. Cengiz Güven* [1], Nezahat Zaloğlu [2], Gülseli Yıldırım [2], Metin Baştuğ [2], Hakan
Fıçıcılar [2], Emine Koç [2], Yüksel Saran [1]
Ankara University, School of Medicine, Department of Histology-Embryology [2] and
Physiology [2], Ankara, TURKEY
Manganese (Mn) is an essential trace element. It is known as a free oxygen radical
scavenger and necessary for the activity of some important enzymes. The beneficial effects
of Mn can be seen when it is in low concentrations. On the other hand exposure of high
concentrations via diet or environment it may accumulate in some tissues and can be
toxic. Manganese is one of the widely used elements in the world. The use of Mn and its
compounds, in industry, generates adverse health effects in workers who are exposed to
Manganese. In the present study we aimed to investigate the effects of high dose and
chronic Mn application on kidney tissue. In the experiments, 20 wistar rats were used.
Ten of the rats were injected (i.p.) manganese chloride as a dosage of 30 mg/kg/day for 50
days (group II-experimental). The remainder of the 10 rats was injected serum physio-
logic at the same time and same amount (group I-control). At the end of experimental
period the kidneys were removed under anesthesia for the histologic investigation. The
light microscopic examination of control group kidneys was normal in appearance. Renal
(Malpighian) corpuscles appeared as rounded structures, the glomeruli, surrounded by
narrow Bowman’s spaces. The flattened epithelial lining of Bowman’s capsule was easily
seen. The proximal and distal convoluted tubules making up the bulk of the parenchyma
between the renal corpuscles were normal aspect.. In the experimental group kidneys
most glomerular and tubular structures of cortex and medulla appeared normal but some
histopathologically striking lesions were observed. Some glomeruli showed capillary
congestion, seemed as occlusion of the capillary lumen with erythrocytes. Besides nor-
mocellular glomeruli, hypercellular glomeruli probably due to inflammatory cells, pro-
liferation of mesangial and endothelial cells were also seen. Focal segmental necrotizing
glomerular lesions were present. Cortex contained dilated distal tubules lined by flattened
cells. Some of these cells had no nuclei, others have closely juxtaposed nuclei which may
indicates that having formed by mitosissuch signs of regeneration. Some of the proximal
tubules had no nuclei and their cytoplasm appeared pale stained. Medullary tubules
contained granular casts derived from necrotic cells of tubules. Interstitial spaces con-
tained inflammatory infiltration cells. The results suggested that high doses of manganese
chloride may have no beneficial effects but effected some histopathological changes in
kidney tissues of rats.
11.05
INFLUENCE OF ACUTE CADMIUM TREATMENT ON THE LIPID
PEROXIDATION AND ANTIOXIDANT ENZYMES IN KIDNEY AND LIVER OF
OVARIECTOMIZED RATS
Rezzan Hatungil* [1], Serap Yalin [2], Nefise Ozlen Sahin [3], Ulku Comelekoglu [4],
Selda Bagıs [5]
Mersin University Medical School, Department of Physiology [1]; Faculty of Pharmacy,
Department of Biochemistry [2]; Faculty of Pharmacy Pharmaceutical Sciences
Department of Pharmaceutics [3]; Medical School, Department of Biophysics [4];

Department of Physical Treatment and Rehabilitation [5], Yenisehir Campus, Mersin
33169, TURKEY
We aimed to investigate the acute effect of cadmium (Cd) treatment on the levels of
lipid peroxidation and antioxidant enzymes in liver and kidney of rats following
ovariectomy operation. 14 of Wistar albino female rats in the age of 10 weeks were
operated to mimic menopause period in women. Next, animals were maintained under
controlled conditions for recovery from the ovariectomy operation. Twelve weeks
later, the animals in the sham-operated group (Ovx, n=7) were provided free access
to water. On the other hand, cadmium was administered i.p. at a single dose of 5 mg/
kg (isotonic parenteral solution) to the ovariectomized-cadmium group (Ovx-Cd,
n=7). 24 hours after the injection, rats were sacrificed. Then, liver and kidney tissues
were removed. Superoxide dismutase (SOD) and catalase (CAT) activities and the
levels of malondialdehyde (MDA) in the liver and kidney of ovariectomized rats were
measured employing the biochemical methods. SOD activity significantly decreased
(p<0.05) in both organs. Although CAT activity showed significant decrease
(p<0.05) in liver, no change of that was obtained in kidney. MDA concentrations
were determined to be higher in both organs than those of the control group. Based
on the data, it can be stated that cadmium impairs the antioxidant defense system and
induce the oxidative stress.
11.06
XENOBIOTIC-METABOLIZING ENZYMES IN IODINE and/or
SELENIUM DEFICIENT RATS THE EFFECTS OF FENVALERATE
EXPOSURE
Pinar Erkekoglu, Aydan Caglayan, Belma Giray, Filiz Hincal*
University of Hacettepe, Faculty of Pharmacy, Department of Toxicology, Ankara,
06100, TURKEY
This study was undertaken to examine the effects of iodine and/or selenium deficiency
on hepatic and extrahepatic enzyme systems and activities responsible for detoxifica-
tion or bioactivation of drugs and other xenobiotics on a rat model. Three weeks-old,
male Wistar rats were used, and feeding period was 7 weeks. Selenium deficiency was
induced by feeding the animals with a diet containing < 0.005 mg/kg selenium, and
iodine deficiency was introduced by sodium perchlorate containing drinking water.
The levels of total microsomal cytochrome P450 and cytochrome b5 (CYP b5); ac-
tivities of microsomal NADPH- cytochrome P450 reductase (P450R), CYP2E1, and
cytosolic GST were determined. Total P450 levels did not change in iodine deficiency
(ID) and combined iodine/selenium deficiency (ISeD), whereas 100% enhancement
was observed in selenium-deficient (SeD) rats along with an increase of 80% in the
activity of CYP2E1. ID and ISeD caused >50% decreases in P450R, CYP2E1, and
GST activities; and CYP b5 decreased markedly (60%) in ISeD rats. Further
experiments examining the effects of the widely-used pyrethroid insecticide, fenvalerate
exposure on the same parameters of the same groups of animals are undergoing.
Preliminary results showed that fenvalerate exposure elevates both hepatic and cere-
bral GST activities in control animals, and causes a relative enhancement of 160% in
hepatic GST activity in SeD, and 100% elevation in cerebral GST activity in ISeD
animals.
11.07
URINARY DEOXYPYRIDINOLINE EXCRETION IN ENVIRONMENTAL
CADMIUM EXPOSURE
Ryumon Honda*, Ikiko Tsuritani, Yuka Noborisaka, Hisa Suzuki, Masao Ishizaki and
Yuichi Yamada
Dept. Social and Environmental Medicine, Kanazawa Medical University, Uchinada,
Ishikawa 920-0293, JAPAN
Six hundred fifteen women aged 40-84 years living in a non-Cd-polluted area in Japan
were analyzed for urinary cadmium (Cd), deoxypyridinoline (DPD), calcium (Ca), N-
acetyl-ß-D-glucosaminidase (NAG) activity, ß2-microglobulin (B2MG) concentration,
and for the stiffness index (STIFF) of calcaneal bone using an ultrasound method. The
urinary Cd in the subjects, with a mean and range of 2.86 and 0.25-11.4 lg/g creatinine,
respectively, showed a significant positive correlation with DPD. The association re-
mained significant after adjusting for age in the postmenopausal subjects. DPD showed
significant positive correlation with NAG but not with B2MG and Ca. STIFF was
significantly inversely correlated with Cd and DPD in the premenopausal subjects. These
results suggest that bone resorption is elevated by Cd exposure in the general Japanese
population.
11.08
HAIR INVESTIGATION OF CHILDREN FROM TECHNOGENE PROVINCE
Shahista Ishniyazova* [1], Abdumalik Khudayberdiev [2], Khurshida. Avazova [1],
Zuhra.Saidmuratova [1], Ahmadjon Tursunov [1]
Samarkand State Agricultural Institute, Mirzo Ulugbek Street, 77, Samarkand, UZBE-
KISTAN [1]; Samarkand State Economics and Service Institute, Amir Temur, 9, Sa-
markand. UZBEKISTAN [2]
The microelement composition of hair of children that live in the vicinity to chemical
plant was studied. Hair samples were taken from different age groups of girls and boys
that ranged from 2 to 7 years old. Detection of arsenic (As), mercury (Hg), antimony (Sb)
and selenium (Se) were carried out by atomic absorption method and neutron activation
analysis. Results of analysis showed that the concentration of arsenic (As) and antimony
(Sb) is higher than the average level of these elements in adult population up to 15 and 3
times, respectively. While the composition of mercury (Hg) in hair is lower than in adult
population up to 2 times. It was demonstrated that hair analysis is a valuable diagnostic
method for indicating metal composition in the organism and for estimating metal poi-
soning.
11.09
CADMIUM LEVELS IN MATERNAL BLOOD, FETAL CORD BLOOD, AND
PLACENTAL TISSUES IN JAPANESE PREGNANT WOMEN WHO SMOKE OR
NOT SMOKE
Satomi Kameo*, Kunihiko Nakai, Keita Suzuki, Tomoko Oka, Norio Sugawara,
Naoyuki Kurokawa, Hiroshi Satoh
Environmental Health Sciences, Tohoku University Graduate School of Medicine,
Sendai, 980-8575, JAPAN
A cohort study was designed to examine the influences of perinatal exposures to heavy
metals and other chemicals in Japanese children. In this study, we focused to examine the
maternal and fatal cadmium (Cd) levels, in relation to mother’s habit of smoking and
fetal development. One hundred eight pregnant (non-smoker group: n=54, sum of
smoker and ex-smoker groups: n=54) women were participated in this study with their
written informed consent. Rice consumption and ages of participants were matched with
both groups. Maternal peripheral blood, cord blood and placenta samples were collected
for Cd analysis. Cd concentrations were determined with GFAAS. Mean Cd concen-
trations of non-smokers were 1.29 ng/ml in maternal blood and 0.48 ng/ml in cord blood,
these of smokers A (Brinkman index (B-index) 1-200) were 1.31 ng/ml in maternal blood
and 0.43 ng/ml in cord blood, and these of smokers B (B-index 200<) were 1.52 ng/ml in
maternal blood and 0.44 ng/ml in cord blood. Cd was significantly higher in maternal
than in cord blood among each groups. Cd in maternal blood of smokers B tended to be
higher than non-smokers. In a linear regression analysis, the birth weights of infant
decreased with increased smoking (B-index), weights of placenta decreased with Cd in
placenta and cord blood in the smoking groups (smokers A+ smokers B). It was sug-
gested that the birth weights of infant might be associated with placental and fatal Cd
levels and long time smoking habit not only during pregnant period.
11.10
DILTIAZEM AND ATROPIN ENHANCE SELENIUM-INDUCED CONTRACTILE
RESPONSE IN ISOLATED RAT ILEUM
Emine Koc* [1], Belma Turan [2]
Ankara University School of Medicine, Departments of Physiology [1], Biophysics [2],
Ankara, TURKEY
Selenium is known to play an important role in the physiology of many cell types and
extracellular application of high concentration (1 mM and higher) of sodium selenite
causes cellular dysfunction in different types of mammalian tissues such as heart. In our
previous study, we have shown that in rat ventricles, this amount of sodium selenite
caused an increase in the resting tension and a decrease in contractile force, in a time-
dependent manner. In the present study, we aimed to investigate the effects of sodium
selenite (1 mM) application on the contractile activity of isolated rat ileum. We also
investigated the effect of diltiazem and atropin application on these contractions. Adult
Wistar rats (200-250 g) were used. Isolated ileum contractility was measured by using
convential organ bath system with Tyrode perfusion solution. All contractions were given
as percentage changes. Sodium selenite application (15 min) caused a very significant
(p<0.001) incrase (230±55%) in the ileal contraction. Both diltiazem and atropin en-
hanced this contraction significantly (p<0.001) in the same order (280±34% and
275±49%, respectively). On the other hand, acetylcholine did inhibit this sodium sele-
nite-induced contraction in the order of 43±14%. Selenium, to be a multifunctional trace
element, can show different types of effects mostly due to tissue, dose, or time de-
pendency. This hypothesis is supported with many studies even including our studies. We
have published that the same amount of sodium selenite application caused a biphasic
(both positive and negative inotropic responses) contractile responses in the heart pre-
parations but in smooth muscle preparations such as ileum, it caused only enhancement
of contractile response. On the other hand, enhancement mechanisms of the ileal sodium
selenite-induced responses with both diltiazem and atropin are not clear yet and this
subject should be under further investigation.
11.11
EFFECT OF COMBINATION THERAPY WITH COPPER AND A CHELATOR IN
THE MACULAR MOUSE, AN ANIMAL MODEL OF MENKES DISEASE
Hiroko Kodama*, Emi Sato, Yan-Hong Gu, Katsuaki Shiga
Dept. Pediatrics, Teikyo University School of Medicine, Tokyo, 173-8605 JAPAN
Menkes disease is a disorder of copper transport; copper accumulates in the intestine,
kidney, and blood brain barrier (BBB) while the brain remains in a state of copper
deficiency and cannot be transported to neurons. The copper deficiency in the brain
cannot be improved by copper injections. We investigated the effect of combination
therapy with administrations of copper and sodium diethyldithiocarbamate (DEDTC), a
lypophylic chelator, in the macular mouse. Four-week old macular mice that had been
treated with 50 ug of CuCl2 on the 7th day after birth were used. The mice were separated
into three groups. Mice in group A were treated with a subcutaneous injection of CuCl2
(4 ug) and an intraperitoneal injections of DEDTC (0.2 mg/g) twice a week for 4 weeks
and then sacrificed. Mice in group B were treated with only CuCl2, and group C were
treated with saline control in the same manner as group A. The brain, kidney and
intestine were then dissected, and copper concentration and cytochrome C oxidase (CCO)
activity were analyzed. The copper concentration in the brain of group A was higher than
that in group B and C. The CCO activity in the brain was highest in group A. Although
the copper concentrations in the kidneys and intestines of group A were also higher than
group B and C, no adverse effect of the combination therapy was found. These results
suggest that the chelator enables administered copper to pass through BBB, and then
improves copper deficiency and CCO activity in the brain.
11.12
SERUM CONCENTRATION OF TRACE ELEMENTS IN CHOLERA
TOXIN-TREATED ANIMALS
F. Marni* [1], A. M. S. Alam [2], G. H. Rabbani [1], S. K. Saha [1], M. Akhtar [1]
ICDDR, B: Centre for Health and Population Research, GPO Box 128, Dhaka 1000 [1];
Department of Chemistry, University of Dhaka, Ramna, Dhaka 1000, BANGLADESH
[2]
The 1-10 microg cholera toxin (CT) was directly injected to a segment of intestine of
rabbit (n=10), rat (n=10) and guinea pig (n=10) to induce intestinal secretion and the
venous blood samples were taken 5 hours after incubation. Blood samples were also
collected from equal number of animals which were served as control. The trace elements
were analyzed by atomic absorption spectrophotometry, and ceruloplasmin by UV-vis-
ible spectrophotometry. It was found that the mean ± SD serum concentrations
(micromole/l) of copper (Cu) were significantly lower in the CT-treated animals than in
the control (rabbit:16.9±5.16 vs. 21.6±6.22, rat: 26.6±5.95 vs. 31.8±4.98 , guinea pig:
9.8±2.92 vs. 17.0±2.52 ). Serum concentration of copper oxidase ceruloplasmin was also
lower in the CT-treated animals than in the control (rabbit:11.9±3.22 vs. 21.7±4.77,
rat:19.6±1.70 vs. 23.2±1.93 , guinea pig: 5.3±2.39 vs. 12.4±3.90 millig/deciL). Similar
changes were found in the serum concentration of zinc (Zn). However, serum concen-
tration of iron (Fe) was higher in the CT-treated animals than that in the control.
Therefore, CT-induced secretion of intestinal fluid may contain large amount of trace
elements, ultimately reducing serum concentration. A decrease in absorption may also
reduce serum concentration of trace elements. Although there is a species variation in the
mean concentrations, the CT-induced changes in trace elements and ceruloplasmin were
similar in all species.
11.13
NEPHROPATHY INDUCED BY CHRONIC EXPOSURE TO DIETARY
CADMIUM IN DIABETIC AND NON-DIABETIC SUBJECTS
Melissa Haswell [1], Soisungwan Satarug [2], Victor McGrath [1], Maria Walmby [1],
Jack Ng [2] and Michael R Moore*[2,3]
Centre for Indigenous Health, The University of Queensland [1], National Research
Centre for Environmental Toxicology [2], Queensland Health Scientific Services, AUS-
TRALIA [3]
Cadmium (Cd) is a nephrotoxicant that is absorbed by the body from dietary sources,
drinking water, cigarette smoking and polluted air. Safe levels of dietary intake, the
Provisional Tolerable Weekly Intake (PTWI) for Cd is 7 lg/ kg bw, equivalent to
70lg daily for a 70-kg individual. We conducted investigations in two populations to
derive dietary Cd exposure estimates and identification of sources of exposure and
renal toxicity. The first was of 197 healthy Thai men and women aged between 19 and
47 yr. The second group was 60 female Indigenous Australians, aged 30 to 50 yr. This
group included 13 subjects with diabetes. Cd burden of women in group 1 showed an
inverse correlation with serum ferritin (P < 0.001). Women with low iron stores
showed a 3.4-fold greater Cd body burden than did those with normal iron store
status (P = 0.003). In group 2, 32% excreted urine Cd of greater than 2 lg/g crea-
tinine and their Cd burden showed a positive association with consumption of liver
and kidney of dugong and turtle, clams, coconut and peanuts. A trend toward higher
risk of abnormal urinary excretion of albumin was seen in diabetic subjects (P =
0.06) although there were no differences in Cd burden between diabetic and non-
diabetic subjects. This result suggests that toxicity occurs at lower renal Cd con-
centrations in diabetic than in non-diabetic subjects. (Michael R. Moore is the Di-
rector of EnTOX and QHSS.)
11.14
POPULATION EXPOSURES TO CADMIUM AND IMPACTS ON RENAL
FUNCTION
Michael R Moore*, Soisungwan Satarug
National Research Centre for Environmental Toxicology, and Queensland Health Sci-
entific Services, Brisbane, AUSTRALIA
In evaluation of impacts of metals on health we conventionally focus on the impacts of
toxic elements. More recently the concept of the acceptable range of intake (AROI) has
allowed us to recognize in addition to toxicity of some elements there is the associated
effect of deficiency of essential elements. Metals such as copper or zinc have relatively
narrow margins of safety associated with optimal intake to provide nutritional sufficiency
but sufficiently low to avoid toxicity. The concurrent effects of toxic and non-toxic ele-
ments are critical in the understanding of optimal dietary intake and likely toxicological
consequences. The capacity to do this has been greatly assisted by increased analytical
capabilities through the use of liquid chromatography linked to inductively coupled
plasma mass spectrometry to establish the presence of different species of elements.
Population studies in different parts of the world have helped to give further guidance on
the additional impacts of variations in genotype and gender on metabolism. A good
example of this comes from studies in Thailand and in the Torres Strait showing that
women are more susceptible to the nephrotoxic effects of Cadmium than men and that as
a consequence of this there is increased excretion of protein in the urine. Other forms of
metabolic change such as development of diabetes can further compound these effects.
These impacts are not restricted to the kidney but are found in other organs as well,
particularly the liver where variations in genotype for the monoxygenases can have
downstream effects on other organ systems. (Michael R. Moore is the Director of En-
TOX and QHSS.)
11.15
DETERMINATION OF THE TOTAL NON-DIETARY ARSENIC IN URINE BY
INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY (ICP-MS)
Manoch Muantongchin*, Ela Bakowska, and Judy Vinosky
National Medical Services, Willow Grove, PA, USA
This new method was used for the quantitation of arsenic in urine to evaluate
occupational exposure or poisoning. The sum of concentrations of trivalent arsenic
(arsenite As+3), pentavalent arsenic (arsenate As+5), monomethyl arsonic acid
(MMAA), and dimethylarsinic acid (DMAA) is reported as Total Inorganic Arsenic.
The non-toxic seafood derived arsenobetaine and arsenocholine are not reported by
this method. The advantages of this method are: a combination of utilization of the
specialized extraction protocol, employment of state-of-the-art analytical instrumen-
tation and optimization of the instruments’ performance, which included a novel
approach to the data acquisition. The various forms of arsenic bind to iodine under
acidic conditions and are then extracted into toluene. A portion of the organic phase
is back extracted into 2% nitric acid. This aqueous phase, which contains non-dietary
forms of arsenic is mixed with Yttrium (internal standard). The employment of the
Inductively Coupled Plasma Mass Spectrometer with a Dynamic Reaction Cell (ICP-
DRC-MS) and utilizing oxygen as the reaction gas eliminated the polyatomic inter-
ference from chloride. The use of internal standard is a requirement for the analysis of
biological materials by ICP-MS. The choice of the optimal internal standard could be
a critical parameter in achieving the best accuracy of the results. In this study
(involving approximately 2,000 urine samples) the arsenic was converted to arsenic
oxide (AsO) and measured at mass 91, instead of mass 75. The operational param-
eters of the instrument were optimized in order to completely convert yttrium (Y) to
yttrium oxide (YO), which was measured at mass 105. This approach allowed us to
measure the non-dietary arsenic at the level of 1 mcg/L. For the duration of 25-week
long study we conducted extensive measurements of control materials certified for
Total Arsenic and for Total Non-dietary Arsenic. With each analytical run we were
also verifying that the new approach did not produce false positive results. This
verification was achieved by employing a certified reference material containing dietary
(seafood) arsenic and negligible levels of non-dietary arsenic. The determination of
total non-dietary Arsenic by ICP-DRC-MS measured as AsO and calculated against
YO demonstrated improvement in lowering the reporting limit and avoiding false
positive results. The results obtained from the new method were compared with the
results obtained with Hydride Generation Atomic Absorption Spectroscopy (HGAAS)
and standard ICP-MS methods.
11.16
DIFFERENT BINDING AFFINITY OF ALUMINUM FOR ASIALO-TRANSFERRIN
COMPARED WITH SIALO-TRANSFERRIN STUDIED BY HPLC/HIGH
RESOLUTION-ICP-MS
Megumi Hamano Nagaoka*, Hiroshi Akiyama, and Tamio Maitani
National Institute of Health Sciences, Kamiyoga 1-18-1, Setagaya, Tokyo 158-8501,
JAPAN
Although originally considered non-toxic, aluminum (Al) neurotoxicity has had a long
and controversial history in neuroscience since the discovery of dialysis encephalopathy.
The Al in the blood is bound to transferrin (Tf). Tf is N-glycosylated proteins generally
containing four molecules of sialic acids in the C-lobe site, and has two metal binding
sites, the N-lobe site and the C-lobe site. Carbohydrate-deficient Tfs and sialic acid-
deficient Tfs are observed in several diseases such as coronary heart diseases and alcohol
abuse. In this study, focusing on the fact that Tfs are glycoproteins, the binding affinity of
both Al and Fe for asialo-Tf was compared with that for native (sialo)-Tf by on-line
connected HPLC/high resolution-inductively coupled plasma-mass spectrometry (HR-
ICP-MS). Three major peaks of Fe-binding human-Tf (hTf) and one apo-hTf peak were
detected in Fe-supplemented apo-hTf and assigned to FeC-hTf, metal2-hTf, apo-hTf, and
FeN-hTf in sequence. The binding affinity of Fe was similar between native- and asialo-
hTfs. Al added as both Al-citrate and Al-NTA selectively bound to the N-lobe site, and
its binding affinity to asialo-hTf was higher than to native-hTf. After the addition of Al as
Al-oxalate, the affinity to the N-lobe site of asialo-hTf increased further. In the absence of
bicarbonate, Al-oxalate showed preference for the C-lobe site in native-hTf and com-
parable affinity to both lobes of asialo-hTf. Thus, the lack of sialic acid in the glycans and
the presence of oxalate enhanced the binding affinity of Al to hTf. Therefore, it may
suggest that the binding affinity of Al to Tf is enhanced in patients with Carbohydrate-
deficient Tfs. The possibility of adverse effects of Al may be increased in several diseases.
(M.H.N. is the recipient of a Grant-in-Aid for Scientific Research from the Ministry of
Education, Culture, Sports, Science and Technology of Japan.)
11.17
ADVERSE EFFECT OF LOW-LEVEL CADMIUM INTAKE AND THE ROLE OF
METALLOTHIONEIN ON CADMIUM TRANSPORT FROM MOTHER RATS TO
NEWBORN RATS
Yasuhiro Nakamura*, Youhei Ohkawa, Hisayoshi Ohta
Dpt. Environ. Occup. Health and Toxicol., Graduate school of Medical Sciences,
Kitasato University, 1-15-1 Kitasato, Sagamihara , Kanagawa 228-8555 JAPAN
Female Wistar rats were given Cd (as CdCl2) at a dose of 0, 1, 2, and 5 mgCd/kg/day by
gastric tube daily for 6 consecutive days a week. The concentration of Cd, Zn, and Cu of
kidney and liver, and metallothionein (MT) of were determined at the first day and at the
4th week after the birth. Furthermore, gene expression of each iso-MT (I, II, and III) was
measured by RT-PCR for uterus. Cd concentration in liver of newborn rats was higher
than that in kidney. But Cd in kidney of 4 weeks old rats was significantly higher than
that of liver. Zn in liver and Cu in kidney of newborn rats decreased according to the Cd
accumulation in uterus of 5mgCd/kg group. The birth rate of newborn rats was ob-
structed, and the growth of newborn rats was decreased in the offspring of 5mg Cd/kg
group. MT concentration increased in uterus and in intestine, and in stomach milk of
offspring rats. It was considered that more Cd was accumulated into liver rather than
kidney of fetus as the form of non-MT-Cd from mother rats. Also it was speculated that
MT in uterus does not participate positively the Cd transport through the placenta from
uterus to fetus. But in rats (4 weeks old), it was considered that more Cd was transported
into kidney as the form of Cd-MT combined by MT induced in the intestine. An in-
creased gene expression of iso-MT (I, II, and III) was found in uterus of 5mgCd/kg,
especially MT-III was prominently. But MT protein induction of amount of according to
gene expression of iso-MT-III was not found in uterus.
11.18
RENAL TUBULAR DYSFUNCTION AND MORTALITY IN RESIDENTS OF A
CADMIUM POLLUTED AREA IN JAPAN, A 20-YEAR FOLLOW-UP STUDY
Muneko Nishijo* [1], Hideaki Nakagawa [1], Yuko Morikawa [1], Katsuyuki Miura [1],
Kenji Tawara [1], Teruhiko Kido [2], Etsuko Kobayashi [3] , Koji Nogawa [3]
Dpt. Public Health, Kanazawa Medical University, Uchinada, JAPAN [1]; Dpt. Com-
munity Health Nursing, Kanazawa University, Kanazawa, JAPAN [2]; Dpt. Occupa-
tional and Environmental Medicine, Chiba University, Chiba, JAPAN [3]
We have conducted a 20-year follow-up study of the subjects (1424 men and 1754 wo-
men) aged >=50 years old of a health impact survey carried out in 1981-2 living in the
cadmium (Cd) polluted Kakehashi River basin. On the basis of the survival data during
observation period extended to February, 2003, we analyzed mortality risk ratios of the
subjects with renal tubular dysfunction due to Cd exposure using Cox’s Proportional
Hazard Model. In the present study, mortality risk ratios of urinary beta2-microglobulin
(b2-MG) positive (> = 1000ug/gCr), protein positive (> = 10mg/dl), glucose positive
(> = 20mg/dl), and amino acids positive (> = 300ug/gCr) subjects were increased in
both sexes. As compared with < 300ug/gCr of urinary b2-MG group, mortality risk ratio
of the 300-1000 group was 1.16 in men and 1.67 in women, that of 1000-10000 group
1.41in men and 1.90 in women, and that of the > = 10000 group 2.28 in men and 2.26 in
women. Also, mortality risk ratio of the 5-10mg/dl of urinary protein group was 1.14 in
men and 1.46 in women, and that of the 10-20 group was 2.10 in men and 2.14 in women,
and that of the > = 20 group was 2.95 in men and 1.78 in women, as compared with < 5
group. These results suggest that the life prognosis of the subjects with renal tubular
dysfunction is unfavorable, and that the mortality was increased in proportion to the
degree of renal tubular dysfunction.
11.19
LIFE PROGNOSIS OF RENAL TUBULER DYSFUNCTION IN RESIDENTS OF
AN ENDEMIC AREA OF ITAI-ITAI DISEASE IN JAPAN: TENTATIVE ANALYSIS
Muneko Nishijo* [1], Hideaki Nakagaw [1], Masumi Minowa [2], Toshiharu Fujita [2],
Jun-ichi Kuriwaki [1], Hidetoyo Teranishi [3], Etuko Kobayashi [4], Koji Nogawa [4]
Dpt. Public Health, Kanazawa Medical University, Uchinada, JAPAN [1]; Dpt. Epide-
miology, National Public Health Institute, Wako, JAPAN [2]; Dpt. Public Health,
Toyama Medical Pharmaceutical University, Toyama, JAPAN [3]; Dpt. Occupational
and Environmental Medicine, Chiba University, Chiba, JAPAN [4]
We have conducted a 20-year follow-up study of the subjects examined in 1979-1984
living in a cadmium (Cd) polluted Jinzu River basin in Toyama, an endemic area of itai-
itai disease. At present, the survey on life status was cover Cd polluted Fuchu town (1519
men and 1715 women) and a control area of Ohyama town (247men and 291 women).
This study is intended to investigate relevant factors to the increase of mortality in these
subjects with Cox’s Proportional Hazards Model. The result reveals that the mortality
risk ratio of residents of a Cd polluted area was 1.51 in men, 1.32 in women as compared
with controls. Among the subjects in a polluted Fuchu town, the mortality rate of con-
current urinary protein and glucose positive subjects was significantly increased, with the
risk ratio of 1.39 in men and 2.18 in women, as compared with the subjects without this
urinary finding. Also, mortality risk ratio of the of urinary protein positive (> = 10mg/
dl) group was 1.31 in men and 1.97 in women. These results suggested that the life
prognosis of the subjects living in a Cd polluted area is unfavorable, with high mortality
risk of the subjects with renal tubular dysfunction. We continue to conduct a follow-up
study cover all subjects in polluted and control areas.
11.20
THE EFFECTS OF CHRONIC MANGANESE CHLORIDE APPLICATION ON
LEARNING AND MEMORY OF RATS
Metin Bastug, Nezahat Zaloglu, Hakan Ficicilar, Gökhan Karaorman, Ahmet Aydın,
A. Banu Ocakcioglu*
Dept. of Physiology, School of Medicine, Ankara University, Ankara, TURKEY
Manganese (Mn) is an essential trace element at low concentrations. When it is present in
exess quantities in the tissues it may be toxic. As neurotoxic effects are seen in brain and
changes in central nervous system are considered as a critical health effect we aimed to
investigate over dose (30mg/kg/day) effects of Mn application on learning and memory in
rats.30 Wistar rats grouped as experimental (MnCl2 injected for 50 days), sham operated
(saline injected) and control group. The rats were trained as 25 trials/day for three
consecutive days by using two ways active avoidance shuttle box to test the learning and
memory on the 31st day. Ten days after the last acquisition test, the retention test was
performed and conditioned avoidance responses (CAR) of the rats were evaluated. At the
end of learning and memory tests, plasma and erythrocyte Mn levels in blood samples
and Mn and malonildialdehyde (MDA) levels in brain samples were measured and
evaluated. As a result, Mn levels increased significantly and there were not considerable
differences between the brain MDA levels. The CAR of all rats from three groups showed
a significant increase in three consecutive days while the differences observed in CAR of
same sessions were not significantly different among three groups. The memory process of
these rats also was not affected significantly. According to our results we suggest that the
learning and memory mechanisms were not affected by the chronic MnCl2 application
due to antioxidative adaptation and other possible regulatory mechanisms.
11.21
RENAL METAL LOAD AND CYP4F2: IMPLICATION IN RENAL EICOSANOID
SYNTHESIS AND HIGH BLOOD PRESSURE DEVELOPMENT
Soisungwan Satarug* [1], Jason R Baker [2], Aedah Abu-Bakar [1] Rob J. Edwards [3]
Jerome M. Lasker [4] and Michael R. Moore [1,5]
National Research Centre for Environmental Toxicology, University of Queensland [1];
Flinders University, AUSTRALIA [2]; Imperial College, UK [3], Hackensack University,
USA [4], Queensland Health Scientific Services [5]
Cadmium (Cd) is one of the nephrotoxicants that is absorbed into the body from dietary
sources and cigarette smoking. It accumulates most extensively in the kidney cortex.
Chronic feeding of small amounts of Cd has been shown to induce high blood pressure in
rodents. Studies of genetic rat models of hypertension and the CYP4A14 knockout mice
revealed a key role for renal cytochrome P450 4A (CYP4A) subfamily in blood pressure
regulation. The CYP4A enzymes in rats and CYP4F2 in humans catalyze x-hydroxyl-
ation of the fatty acid arachidonate and generate the second messenger molecule 20-
hydroxyeicosatetraenoic acid (20-HETE), involved in renal salt re-absorption and
excretion, vascular tone and volume homeostasis. We hypothesize that Cd promotes the
pathogenesis of hypertension in susceptible individuals through its induction of renal
expression of CYP4F2. We used ICPMS in conjunction with immuno blotting to
determine levels of Cd, Zn, Cu, CYP4F2 and CYP2E1 protein abundance in post-mor-
tem liver, and kidney cortex samples from 30 Australian subjects who had not occupa-
tionally been exposed to metals. Liver CYP2E1 protein levels did not correlate with liver
Cd content, but correlated positively with CYP4F2 protein. In contrast, no expression of
CYP2E1 was detected in any of the kidney cortex samples tested, but kidney CYP4F2
protein levels showed a positive correlation with kidney Cd content.
11.22
INFLUENCE OF CHRONIC EXPOSURE TO TOXIC METALS ON ZINC STATUS,
RENAL FUNCTION BIOMARKERS AND BLOOD PRESSURE
Soisungwan Satarug* [1], Pailin Ujjin [3], Yuvaree Vanavanitkun [3] and Michael R
Moore [1,2]
National Research Centre for Environmental Toxicology, University of Queensland,
AUSTRALIA [1]; Queensland Health Scientific Services [2]; Dept. of Laboratory Med-
icine, Chulalongkorn University, Bangkok, THAILAND [3]
Cadmium (Cd) and lead (Pb) are toxic elements that are absorbed into the body from
dietary sources, drinking water, cigarette smoking and polluted air. Cd accumulates in
soft tissues notably the liver and kidney while Pb accumulates largely in bone. Cd in liver
and Pb in bone are mobilizable, providing an opportunity for renal toxicity to occur with
no additional exposure. The manifestations of Cd nephrotoxicity notably proteinuria,
calciuria, aminoaciduria are detectable at renal Cd concentrations that are well below the
previous estimate of 180-200 lg/g wet kidney cortex weight. In the present study, the
relationships between blood pressure, zinc status, nephropathy and exposure to Cd and
Pb were analysed in a group of 118 healthy, non-smoking Thai men and women (21-57
yrs). Their urinary Cd excretion ranged from 0.05 to 2.36 lg/g creatinine while their
urinary Pb excretion ranged from 0.1 to 12 lg/g creatinine. Average age and Cd burden
of men and women did not differ. Women, however, showed a 46 % higher average
urinary Pb excretion (P < 0.001) and lower Zn status (lower average serum Zn and
urinary Zn excretion) than men. Cd-linked nephropathy was detected in both men and
women. Pb-linked nephropathy was seen only in women, probably due to higher Pb
burden coupled with lower protective factors notably Zn (P < 0.001) in women.
11.23
COMBINED EFFECTS OF LONG-TERM LOW-DOSE EXPOSURE TO
METHYLMERCURY AND SELENIUM DEFICIENCY ON NEUROBEHAVIORAL
EXAMINATIONS IN MOUSE OFFSPRING
Kunihiko Nakai [1], Satomi Kameo [1], Norio Sugawara [1], Naoyuki Kurokawa [1],
Komyo Eto [2], Hiroshi Satoh* [1]
Environmental Health Sciences, Tohoku University Graduate School of Medicine,
Sendai, 980-8575 JAPAN [1]; National Institute for Minamata Disease, 867-0008, Min-
amata, JAPAN [2]
Methylmercury (MeHg) has been shown to be a neurobehavioral toxicant to the

developing nervous system of human fetuses. In addition, nutritional factors may have
a potent impact on long-term low-dose MeHg exposure. The main purpose of this
study is to examine the combined effects of long-term low-dose MeHg exposure and
Se deficiency using an animal model. For this purpose, female C57BL/6Cr mice at 8
weeks old were fed either Se-deficient or control diet containing 0 or 1 ppm (as Hg)
MeHg for 4 weeks before mating, and during pregnancy and lactation. The male
offspring obtained were successively given the same diet. Neurobehavioral examina-
tions were performed with open field test when the offspring were at 12 and 67 weeks
old. MeHg exposure did not affect the results of the open field test, although several
effects of aging and Se deficiency were observed. Some mice showed abnormal hind
limb walking when they were at 43 weeks old and the incidence was increased with
aging. Since this phenomenon was remarkable in the aged mice of Se-deficient and
MeHg-exposed group, it is considered Se deficiency enhanced the toxicity of MeHg.
Although the mechanism by which the combination of Se deficiency and long-term
MeHg exposure caused the hind limb abnormal walking is still unknown, these
findings suggest that Se plays an essential role in the defensive system against the
long-term low-dose MeHg exposure.
11.24
TRACE ELEMENT MEASUREMENTS IN BLOOD AND HAIR OF EXPOSED
YOUNG POPULATION AND IN THEIR ALIMENTATION IN MARRAKECH
REGION, MOROCCO
A. Sedki*, and N. Lekouch
Cadi Ayyad University; Faculty of Sciences Department of Biology; Laboratory of
Ecotoxicology. Marrakech, MOROCCO
To determine lead and cadmium content in the most frequently consumed foods of plant
origin grown in different regions of Marrakech, assess lead and cadmium content in the
biological materials (blood and hair) from Marrakech young population, and verify the
hypothesis that the trace elements pollution of the environment varies across regions in
Marrakech. We analysed lead and cadmium concentration in 185 composite samples of
foods of plant origin, and determined the trace elements burden in the population by
analysing lead and cadmium content in whole blood and hair samples 439 randomly
selected healthy children aged 1-15 years. There were significant differences in lead and
cadmium content in the analysed foods with respect to the regions in Marrakech where
were the foods of plant animal origin were produced (p<0.001) especially in the area
polluted by solid and liquid waste. The risk to humans through consumption of polluted
vegetable and animal products is apparent. In fact, the means lead and cadmium content
in 439 blood of children’s was 98.5±31lg/l; 0.9±0.4 lg/l in 245 children’s hair was
12.5±4.5 lg/g and 3.4±1.3 lg/g. Lead and Cadmium content in the blood and hair
samples from study participants varied significantly across regions in Marrakech (p <
0.001 and p < 0.003,respectively). Cadmium content in foods and cadmium burden in
the blood showed only a weak correlation (Spearman’s r = 0.13), whereas the correlation
between cadmium content in the foods and hair was much stronger (Spearman’s r =
0.55). Our research confirmed the hypothesis of regionally different environmental pol-
lution with lead and cadmium, and clearly showed the connection between toxic metals
burden in foods and in population in especially in the area exposed to pollution by the
solid and liquid.
11.25
BENEFICIAL CO TREATMENTS OF TIRON AND SUPPLEMENTS AGAINST
VANADIUM INTOXICATION
Sadhana Shrivastava*, and R. Mathur
UGC-SAP School of Studies in Zoology, Jiwaji University, Gwalior 474011 (M.P.)
_
INDIA
Vanadium, the 21st most abundant element in the earth’s crust, is present in varying
concentrations in almost all mammalian cells. It is now considered an essential nutritional
trace element & has therapeutic value in pharmacological doses. It is an essential element
for normal growth and function of animals (12.4-28ug/day). Its salts have been used
medicinally as antiseptic, spirochetocide, antituberculotic & antianemic agents & as general
tonic. It is used in the hardening of steel & in insecticides. It causes irritation of respiratory
tracts, nasal catarrh and hemorrhage from nasal mucosa. Other symptoms include rhinitis,
bronchitis due to broncho spasm, and conjunctivitis. It compounds inhibit a large number
of cellular activities, with profound effects on enzymes of plasma membranes. Vanadate
exerts oxidative damage affecting mainly the mitochondria. In the present study an attempt
has been made to evaluate the effectiveness of Tiron along with zinc, selenium and vitamin
E. Tiron (4,5-dihydroxy-1, 3-benzene disulphonic acid disodium salt) is a phenolic com-
pound, which is widely used in analytical chemistry and has been reported to be effective in
preliminary study for reducing the concentration of vanadium. Zinc, selenium and vitamin
E are powerful antioxidants. Female albino rats (body wt 130+10g) were randomly di-
vided into 6 groups of five animals each. Group-1 was treated as normal control, group 2-6
were administered vanadyl sulphate (18.5 mg/kg oral), group 2 served as experimental
control. Group 3 was administered Tiron per se (606 mg/kg) Group 4-6 were administered
Tiron along with zinc (4mg/kg), selenium (0.5mg/kg) and +vitamin E (50 mg/kg) .All
animals were sacrificed 24 hrs after last treatment. Vanadium induced a decrease in serum
albumin and triglycerides while serum LDH, serum urea showed increase values. It caused
elevation in glycogen and protein content of liver, kidney uterus and ovary. There was
increased enzymatic activity of acid phosphatases, adenosine tri phosphatase whereas
alkaline phosphatase showed inhibition, Lipid peroxidation showed elevation and gluta-
thione was inhibition. Administration of vanadyl sulphate caused degenerative changes in
the ultrastructure of liver. Numerous vacuoles noted in the cytoplasm. Mitochondria were
swollen and damaged. The collagen bundles were clearly seen. At some places glycogen
bodies were arranged around the vacuoles. Rough and smooth endoplasmic reticulum were
observed associated with the mitochondria and the nuclear wall after treatment of Tir-
on+Se. Therapy with Tiron, Tiron +zinc, Tiron+selenium and Tiron+vitamin E was
effective in restoring the blood and tissue biochemistry. In comparison Selenium along with
Tiron was found significantly effective. Histopathological changes were also recouped
significantly with the treatment of Tiron+Se in liver and kidney.
11.26
EFFICACY OF CHELATING AGENT AND SELENIUM SUPPLEMENTATION
FOLLOWING BERYLLIUM INTOXICATION: A BIOCHEMICAL APPROACH
Sangeeta Shukla*, and Sonia Johri
UGC-SAP School of Studies in Zoology, Jiwaji University,Gwalior-474011 M.P.,
_
INDIA
The present study aims in evaluating the detoxification of beryllium with chelating
agents in combination of Se, an antioxidant. Beryllium was administered subchroni-
cally at a dose of 1mg/kg followed by therapy with chelating agents for 5 consecutive
days. Ascorbic Acid, NTA, DMSA, GSH, DMPS and DPA were screened against
beryllium toxicity. Distribution of beryllium (AAS) in various tissues after toxicant
exposure was further substantiated by blood biochemical variables, light and electron
microscopic studies. Results revealed that DPA+Se proved to be the most efficacious
combination in reducing the beryllium body burden in liver, kidney, lungs and uterus.
There was a rise in transaminases pattern whereas a fall was observed in blood sugar
level, serum alkaline phosphatases and serum proteins with beryllium exposure.
DPA+Se helped in the recovery of these variables. EM studies revealed deformed
nucleus with collagen fibres, however, with DPA+Se the mitochondria were well
formed in close proximity with ER. Thus the study was narrowed down to pregnant
animals where beryllium was exposed at a bolus dose of 50 mg/kg on day15th, pc
followed b administration of DPA+Se for 5 consecutive days. Results reveal a sig-
nificant fall in the blood sugar level, Serum ALP, and proteins. A significant rise in
transaminases pattern is indicative of leakage of enzymes resulting alteration in per-
meability of the cells. A rise in hepatic lipid peroxidation is a direct indication of
oxidative damage. The distribution studies showed an increase in concentration of
beryllium in maternal and fetal liver, kidney, lungs and uterus. Biochemical and
distribution studies confirm that DPA+Se can be said as a better therapeutic agent
against beryllium toxicity.
11.27
EFFECTS OF COPPER IONS ON THE FREE RADICAL-SCAVENGING
PROPERTIES OF THIOL-CONTAINING AMINOACIDS: IN VITRO STUDIES
Hernán Speisky*[1,2], Catalina Carrasco [1], Claudia Rocco [1] and Claudio Olea-Azar
[2]
Micronutrients Unit, INTA [1], Faculty of Chemical & Pharmaceutical Sciences [2],
University of Chile, Santiago, CHILE
Homocysteine (HCys) and cysteine (Cys) have been implicated in various oxidative-stress
related diseases. Depending on whether Cu ions occur concomitantly, these thiol-ami-
noacids could either prevent or promote free radical generation. Here we have addressed
the consequences of the in vitro interaction between HCys, Cys and Cu2+ ions, on the
free radical-scavenging properties of the former. The bleaching of ABTSÆ+, a color-stable
free radical, was employed to assess the free radical-scavenging capacity. Both amino-
acids bleached ABTSÆ+ concentration-dependently. In the presence of increasing Cu
concentrations, only Cys lost its BC completely. HCys, in turn, retained a substantial part
of its BC, and its thio-reactivity, even in the presence of a large molar Cu excess. Under
such conditions, increasing concentrations of a fixed ratio of excess Cu plus HCys led to a
linear and concentration-dependent increase in BC. Addition of TRIEN, EDTA or
histidine to a pre-incubated Cu plus HCys mixture failed to affect the BC. Yet, the
incubation of Cu with TRIEN or EDTA (but not histidine) prior to HCys addition,
totally prevented the loss of BC induced by Cu. EPR, bathocuprein (which binds Cu+)
and ascorbate-oxidation studies strongly indicate that the reduction of Cu2+ ions in-
duced by HCys (but not Cys) is associated with a redox stabilization of the metal under
the form a Cu-aminoacid complex. Thus, only HCys reduced Cu2+ without promoting
the O2Æ--dependent Cyt c reduction. Data support the contention that Cu interacts with
HCys forming a complex which partially retains the free-radical scavenging properties of
HCys. (Supported by Fondecyt # 1040736.)
11.28
DIAGNOSTICS OF CHRONIC HYPERMICROELEMENTOSES ON THE
CHEMICAL ELEMENTAL COMPOSITION OF HAIR (CECH)
Svetlana I. Usenko* [1], Vadim V. Ermakov [2]
Russian Federal Nuclear Center All-Russian Scientific Research Institute of Experi-
mental Physics (RFNC-VNIIEF), Sarov [1]; V.I. Vernadsky Institute of Geochemistry
and Analytical Chemistry of RASci., Kosigin Str., 19, 119991 Moscow, RUSSIA
At interpretation of an element composition of a hair is underlined, that the hair as the
bioindicator can be used in ecological analysis and medicine. The analysis of a hair can
appear useful for detection of disturbance in a mineral feed of man, for estimation of the
status one or several trace elements depending on feeding habits and biogeochemical situ-
ation. The problem of CECH as applied to diagnosis of microelementoses is in the focus of
keen attention of scientists and active exploration of optimal solutions. Considerable ana-
lytical difficulties are associated with the choice of biomaterial, its preparation for analysis
and interpretation of data obtained. Conditions of washing up of the hair from exogenous
pollution by distilled water are investigated. The estimation of the necessary size of frag-
ments of particles of hair for reception representative mass of sample in weight of 0.5 g is
carried out. Conditions of a microwave mineralization of hair of DC are picked up. Using
unified hair analysis methods and correlating elemental chemical composition of hair, the
clinical and subclinical manifestation forms of chronic microelementoses in Pb and Mo
biogeochemical provinces of Russia were discovered. (Supported by grant of MSTC 2046.)
11.29
IS THE TRACE ELEMENTS IN HUMANS A SUBSTANTIAL SCIENTIFIC
DISCIPLINE?
Vladimir Zaichick* [1,3], Anatoly Skalny [2,3]
Medical Radiological Research Center, Russian Academy of Medical Sciences, Obninsk
249020, RUSSIA [1]; Institute of Bioelementology, Orenburg State University, Orenburg,
RUSSIA [2]; Russian Society of Trace Elements in Medicine, Moscow, RUSSIA [3]
All legitimate scientific disciplines are characterized by: the clear definition of subjects of
the study, some postulates, set of methods, specific terminology and determinations, and
a clear-cut name. The examination of the trace elements in humans has been made from
these positions. Many inaccuracies and uncertainties have been found throughout the
study beginning with the name of the new scientific discipline and ending with extensively
used terms. The inaccuracies and uncertainties are discussed and some corresponding
statements are made. Another and no less important problem with the trace elements in
humans as a ‘‘scientific discipline’’ is the critical unsatisfactory reproducibility of data.
The complex arrangements required for the harmonization of data acquired for the trace
elements in humans study are offered, including the unified protocols of sampling, sample
preparations, quality control of measurements and statistical treatment of results.
Category 12: ISTERH 2004 Invited Speakers
12.01
THE MECHANISM AND REGULATION OF INTESTINAL IRON ABSORPTION
Gregory J. Anderson* and David M. Frazer
Iron Metabolism Laboratory, Queensland Institute of Medical Research, PO Royal
Brisbane Hospital, Brisbane Queensland 4029, AUSTRALIA
In recent years there has been enormous progress in understanding how iron moves
across the epithelial cells lining the small intestine and how this process responds to
changes in body iron requirements. Most iron in the diet is present in the Fe3+ or ferric
form but it must first be reduced to Fe2+ before it can be utilized by the body. This
reduction is probably catalyzed by the brush border membrane protein Dcytb. The
resultant ferrous iron is then moved across the apical membrane by the iron import
protein DMT1. The precise form of iron within the enterocyte has yet to be defined, but
any iron that does not move across the epithelium can be incorporated into the iron
storage protein ferritin, later to be lost when the enterocytes are exfoliated. Passage of
iron out of the enterocytes into the circulation requires at least two components. One of
these is the basolaterally located ferrous iron efflux protein Ireg1 (or ferroportin1). The
other is the membrane-bound ferrous iron oxidase hephaestin. The regulation of iron
absorption appears to be complex, but strong evidence indicates that the liver-derived
peptide hepcidin plays a major role and acts as a repressor of absorption. Hepcidin
expression is under the control of HFE (the protein mutated in the iron overload disease
hemochromatosis) and likely hemojuvelin (mutated in juvenile hemochromatosis) and
transferrin receptor 2. This pathway may respond to variations in body iron need by
monitoring levels of diferric transferrin in the plasma, which in turn reflect the iron
supply to the tissues.
12.02
SELENIUM AND PROGRESSIVE CHAGAS DISEASE CARDIOMYOPATHY
Tania C. Araújo-Jorge* [1], Andréa P. de Souza [1], Solange L. de Castro [1], Baiyu Tang
[2], Herbertz B. Tanowitz [3], Maria Teresa Rivera [1], and Linda A. Jelicks [2]
Lab. de Biologia Celular, Dept. de Ultra-estrutura e Biologia Celular, Instituto Oswaldo
Cruz, Fundação Oswaldo Cruz, Av. Brasil 4365, Manguinhos, 21045-900, Rio de Ja-
neiro, RJ, BRAZIL [1]; Department of Physiology & Biophysics, Albert Einstein College
of Medicine, Bronx, 10461, NY, USA [2], Department of Pathology (Division of Tropical
Medicine and Parasitology) [3]
Selenium (Se) is an essential trace element required in human nutrition. Se deficiency has
been implicated in development of cardiovascular problems, frequent consequences of the
infection by Trypanosoma cruzi. In our recent work we found that a significant per-
centage of patients with progressive chronic chagasic cardiomyopathy present low Se
levels that positively correlate with heart insufficiency. Besides, mice fed with a Se defi-
cient chow are more susceptible to T. cruzi infection and oral Se supplementation at low
doses contributes to alleviation of heart damage in T. cruzi-infected mice. We investi-
gated the effect of Se supplementation (2 or 4 ppm) for 120 days in mice chronically
infected by T. cruzi [5 x 104 (Brazil strain)]. The right ventricular inner dimension (RVID)
determined by magnetic resonance imaging of infected non-supplemented mice was 45%
larger than the controls throughout infection. Long-term Se supplementation prevented
the increase of RVID in infected mice. A study on the effects of Se supplementation as a
treatment to reduce an already established hypertrophy is warranted. (T. C Araújo-Jorge
serves on the Board of Lab. de Biologia Celular (Instituto Oswaldo Cruz).
12.03
ROLE OF MAGNESIUM IN INSULIN ACTION, DIABETES AND
CARDIOMETABOLIC SYNDROME X
Mario Barbagallo* and Ligia J. Dominguez
Geriatric Unit, Dept. of Internal Medicine, University of Palermo, 90100 Palermo,

ITALY
Magnesium (Mg) is one of the most abundant ions present in living cells and its plasma
concentration is remarkably constant in healthy subjects. Plasma and intracellular Mg
(Mgi) concentrations are tightly regulated by several factors. Among them, insulin is one
of the most important. In vitro and in vivo studies have demonstrated that insulin
modulate the shift of Mg from extra- to intracellular space. Mgi concentrations modulate
insulin action (mainly oxidative glucose metabolism), offset calcium-related excita-
tion contraction coupling, and decrease smooth cell responsiveness to depolarizing
stimuli. A poor Mgi concentration, as found in noninsulin-dependent diabetes mellitus
(NIDDM) and in hypertension, may result in a defective tyrosine-kinase activity at the
insulin receptor level and exaggerated cellular calcium levels, ending up in an impairment
in insulin action and a worsening of insulin resistance in NIDDM and hypertensive
patients. By contrast, in NIDDM patients Mg administration, restoring a more appro-
priate Mgi concentration, contributes to improve insulin-mediated glucose uptake. The
benefits deriving from daily Mg supplementation in NIDDM patients are supported by
studies showing that high daily Mg intake are predictive of a lower incidence of NIDDM.
In conclusion, a growing body of studies suggest that Mgi may play a key role in
modulating insulin-mediated glucose uptake and vascular tone. We further suggest that a
reduced Mgi concentration might be the missing link helping to explain the epidemio-
logical association between NIDDM, hypertension, and cardiometabolic Syndrome X.
12.04
SELENIUM AND VIRAL INFECTION
Melinda A. Beck
Depts. Pediatrics and Nutrition, University of North Carolina at Chapel Hill, Chapel
Hill, NC 27599, USA
The emergence of new infectious diseases and of old diseases with new pathogenic
properties continues to be a problem worldwide. What factors contribute to the devel-
opment of new infectious diseases? Various hypotheses have been suggested, including
changes in agricultural practices, climate changes, rainforest clearing and air travel.
However, until recently, the nutritional status of the host has not been considered as a
factor for the emergence of new viral disease. Work from our laboratory and others have
demonstrated that a host deficiency in selenium can not only affect the immune status of
the host resulting in an increased susceptibility to viral infection, but can alter the viral
genome as well. Two very different viruses, coxsackievirus and influenza virus, develop
increased virulence when replicating in a selenium-deficient host. This increase in viru-
lence is due to specific viral genome changes. Once these genome changes occur, even
hosts’ with normal selenium status are susceptible to the newly virulent viruses. Fur-
thermore, data from a population in Cuba suggests that the effect of host nutritional
status on a viral genome is not limited to experimental laboratory conditions, but can
occur in human populations as well. This work represents a new paradigm for the
interaction between host nutritional status and infectious disease.
12.05
ZINC CHILDHOOD INFECTIOUS DISEASES
Robert E. Black
Dept. International Health, Johns Hopkins Bloomberg School of Public Health, 615 N.
Wolfe Street, Baltimore, MD 21205, USA
Zinc deficiency, which is highly prevalent in children in developing countries, results in

an increased incidence and severity of infectious diseases. Randomized controlled zinc
supplementation trials provide the best evidence for a causal role of zinc deficiency.
Daily supplementation with zinc has been found in a pooled analysis on 9 studies
done in developing countries to result in 20% lower incidence and 25% lower prev-
alence of diarrhea. Five studies found a pooled reduction of 34% in the incidence of
pneumonia. Two of three trials suggest a benefit of zinc supplements for preventing
clinical malaria, but a multicenter trial using zinc as an adjunct to anti-malarial
therapy found no benefit. A meta-analysis of 12 trials of zinc in the therapy of acute
diarrhea found a significant reduction in mean duration and lower stool frequency or
stool output. WHO now recommends that zinc be used to treat diarrhea along with
oral rehydration therapy. Two trials indicate a benefit of zinc as an adjunct to
antibiotic therapy of pneumonia. Finally, zinc supplementation has been found to
reduce total mortality by 2/3 in small for gestational age infants given daily supple-
ments in India and by 50% in children given zinc during and for a short time after
acute diarrhea in Bangladesh. Provision of additional zinc to children in developing
countries would result in a substantial reduction in the burden of infectious disease
morbidity and mortality.
12.06
THE DIAGNOSIS AND CLINICAL FEATURES OF THE METABOLIC
SYNDROME
George A. Bray
Pennington Center, Baton Rouge, LA, USA
The metabolic syndrome describes a collection of classic clinical signs and symptoms
including central adiposity, impaired fasting glucose, low HDL-chol, elevated trigly-
cerides and elevated blood pressure. The early view was that this syndrome reflected
insulin resistance with hyperinsulinemia. As our understanding of the syndrome has
grown it has become clear that it involves alterations in vascular function, inflam-
mation and disturbances in coagulation. The concept of the ‘‘metabolic syndrome’’
arose from studies of Professor Vague and his colleagues in France, of Professor
Bjorntorp and his associates in Sweden and Professor Kissebah and associates in the
United States. These groups of investigators found that some of the complications of
obesity were more clearly related to the central distribution of fat than to overall
obesity. This and a variety of other pieces of information led Reaven in 1988 to
propose ‘‘Syndrome X’’ as a syndrome associated with insulin resistance. Over the
years since its first description, it is clear that the metabolic syndrome encompasses
more than just insulin resistance and its associated consequences. Because insulin
resistance is difficult to measure and not clinically available, the collection of physical
symptoms and signs have been used to identify this syndrome (Abdominal obesity
(waist circumference: >102 cm (>40 in) (men), >88 cm (> 35 in) (women); HDL-
cholesterol: <40 mg/dL (men), <50 mg/dL (women); Triglycerides: >150 mg/dL;
fasting glucose: >110 mg/dL; Blood pressure (SBP/DBP): >130/>85 mmHg). The
syndrome appears to reflect an increased accumulation of visceral adipose tissue that
secretes a variety of factors, including free fatty acids and cytokines that may be
responsible for the syndrome. When the factors in this table were examined in the
U.S. population, 24% of adults qualified for the metabolic syndrome with an
increasing percentage with each decade.
12.07
LOWERING COPPER AVAILABILITY REDUCES SIGNALING FROM KEY
CTYOKINES: USE FOR ANTIANGIOGENESIS, ANTIFIBROSIS AND
ANTIINFLAMMATION
George J. Brewer
University of Michigan, Ann Arbor, MI 48109, USA
Tetrathiomolybdate (TM), originally under development for the initial treatment of
Wilson’s disease, is now being used to lower copper levels in non-Wilson’s patients to
inhibit dysregulated cytokines contributing to a variety of diseases. Many angiogenic
cytokines become less active if copper levels are lowered to a midrange, using cerulo-
plasmin levels as a marker of copper status. In this midrange, enzyme requirements for
copper are met, and copper deficiency is avoided. TM therapy has given positive results in
five rodent tumor models, and produced encouraging results in spontaneous advanced
cancers in pet dogs. Finally, a clinical study of 42 patients with advanced cancers pro-
duced very encouraging results. Phase II trials of TM therapy of specific cancers are now
underway. Recently we have used TM to inhibit dysregulated cytokines that contribute to
fibrotic and inflammatory disease. In the bleomycin mouse model of pulmonary damage,
TM prevents both inflammation and fibrosis while inhibiting the inflammatory cytokine,
tumor necrosis factor alpha (TNFß) and the fibrotic cytokine tissue growth factor ß
(TGFß) in the lungs. TM also inhibits the hepatitis and cirrhosis produced in the mouse
by concanavalin A, or carbon tetrachloride, while inhibiting TGF?. Finally, TM inhibits
the hepatitis produced in the mouse by acetaminophen (Tylenol) while inhibiting an-
other inflammatory cytokine, interleukin one ß (IL-1). (The University of Michigan has
licensed the antiangiogenic uses of TM to Attenuon LLC, and the author has equity in
and is a paid consultant of Attenuon.)
12.08
RECOVERY FROM COPPER-INDUCED BRAIN DAMAGE IN WILSON’S DIS-
EASE USING ZINC THERAPY
George J. Brewer
University of Michigan, Ann Arbor, MI 48109, USA
Wilson’s disease is a rare autosomal recessively inherited disease of copper accumulation
and toxicity, due to mutations in the ATP7b gene. Clinical presentation can be primarily
hepatic, neurologic, or psychiatric. The neurologic presentation is that of a movement
disorder often involving speech and swallowing problems, tremor, dystonia, and inco-
ordination. Often behavioral problems occur coincident with or before the neurologic
symptoms. Initial treatment is best carried out with tetrathiomolybdate (TM). Penicil-
lamine causes neurologic worsening in these patients very frequently, and many patients
do not recover to their prepenicillamine baseline. Trientine also has the risk of causing
neurologic worsening, although less frequently than penicillamine. Zinc, by itself, is too
slow acting for these patients. TM protects neurologic function while controlling copper
toxicity quickly. After TM treatment for 8-16 weeks, these patients are placed on zinc
maintenance therapy. Neurologic recovery in most of these patients is very gratifying.
Recovery begins at about 5-6 months after therapy initiation as measured by quantitative
neurologic and speech testing, and brain magnetic resonance imaging, and continues for
another 18 months, then plateaus. For the most part, disability that remains after 24
months will be permanent. We presume the combination of TM and zinc reduces the
‘‘free copper’’ of the brain from toxic to non-toxic levels. (The University of Michigan has
licensed the antiangiogenic uses of TM to Attenuon LLC, and the author has equity in
and is a paid consultant of Attenuon.)
12.09
OXIDATION, BIOMETALS AND ALZHEIMER’S DISEASE
Ashley I. Bush
Laboratory for Oxidation Biology, Genetics and Aging Research Unit, Harvard Medical
School, Massachusetts General Hospital, Boston, MA, USA; Oxidation Disorders
Laboratory, Mental Health Research Institute of Victoria and The University of Mel-
bourne, AUSTRALIA
Aß is the principal component of the plaque pathology which is the hallmark of Alz-
heimer’s disease (AD). We have characterized Aß as a ubiquitous metalloprotein, with
selective high-affinity binding sites for zinc (Ka 100 nM) and copper (Ka 10 attoM).
Several studies have reported that there is an elevation of zinc and copper in the plaques
in Alzheimer’s disease. Furthermore, genetic ablation of the ZnT3 transporter, which
loads zinc into the glutamatergic synapse, abolishes amyloid deposition in the Tg2576
mouse model for AD. ZnT3 activity at the synapse contributes to a pool of Zn2+ in the
cebral blood vessel wall that fosters congophilic angiopathy at that site in the Tg2576
mouse model. Aß binds Cu and Zn through a site that involves oligomeric peptide
assembly, and resembles the structure of SOD1. When synthetic Aß binds copper in the
absence of zinc, it is highly redox active. Biological reducing agents such as dopamine,
cholesterol and vitamin C can act as a reservoir of electron donation, so that the Aß/Cu
complex acts as a catalyst generating H2O2 (Km= 5 lM, Vmax= 30 nM/min). The
toxicity of Aß species is proportional to the peptide’s ability to reduce Cu or Fe and
generate H2O2 (Aß42> Aß40 >rat Aß). H2O2 is a freely permeable pro-oxidant and is
the chemical source of much of the oxidation damage that is abundantly evident in AD
brain. Where cholesterol is the substrate for H2O2 production, it is specifically oxidized
by Aß:Cu complexes into 4-cholesten-3-one, a pro-apoptotic toxin that is markedly in-
creased in AD post-mortem brain tissue compared to age-matched normal and neuro-
logical disease control tissue. Chelators of copper and zinc both disaggreagate Aß
deposits from post-mortem human brain and also inhibit redox activity and H2O2 pro-
duction. We have found that one such orally bioavailable chelator, clioquinol, markedly
inhibits brain amyloid pathology in Tg2576 mice. This compound is a retired antibiotic
and has recently completed a successful pilot phase 2 clinical trial in Alzheimer’s disease,
where plasma Aß levels were lowered and cognitive decline significantly arrested. (Dr.
Bush is a paid consultant, scientific advisory board member, and shareholder of Prana
Biotechnology Ltd.)
12.10
CHROMIUM AND METABOLIC SYNDROME
William T. Cefalu
Biomedical Research Center Louisiana State University Baton Rouge, Louisiana, USA
Considerable controversy exists regarding the effect of chromium on insulin action in
vivo in clinical states of insulin resistance. This controversy exists because despite
widespread use and antedotal reports, there is a paucity of data in humans in regard to
the effect of chromium to improve measures of insulin action in vivo or on cellular aspects
of insulin action. However, emerging clinical and epidemiologic evidence suggests that
chromium may play a role in carbohydrate metabolism. Specifically, several lines of
evidence in both in vitro and in vivo models suggest that chromium may modulate
intracellular pathways of glucose metabolism. Further, evidence suggests the existence of
chromodulin, a chromium binding protein, that may participate in insulin signaling. On a
clinical level, studies in individuals with and without diabetes have demonstrated that
chromium supplementation, in certain populations, may significantly enhance glucose,
insulin, and lipid metabolism. In general, the results appear to be more consistent for
subjects with diabetes than among healthy subjects. However, support for chromium
supplementation is currently limited by the highly varied results obtained in generally
small-scale, short-term clinical studies. A more consistent clinical response is observed
with daily supplementation of chromium >200 lg/day for a duration of at least 2
months.
12.11
LONG-TERM ARSENIC EXPOSURE AND CARDIOVASCULAR DISEASES
Chien-Jen Chen* [1], Chih-Hao Wang [2], Chi-Ling Chen [1], Lin-I Hsu [1], Hung-Yi
Chiou [3], Yu-Mei Hsueh [3], Shu-Yuan Chen [4], Meei-Maan Wu [3]
Graduate Institute of Epidemiology, National Taiwan University, Taipei, TAIWAN [1];
Department of Cardiology, Cardinal Tien Hospital, Taipei, TAIWAN [2]; School of
Public Health, Taipei Medical University, Taipei, TAIWAN [3]; Division of Biostatistics,
National Health Research Institutes, Taipei, TAIWAN [4]
The pleiotropism of arsenic-induced cardiovascular effects may be characterized by its
associations with peripheral vascular disease, ischemic heart disease, cerebral infarction,
carotid atherosclerosis, as well as prolongation and increased dispersion of QTc on
electrocardiogram in a dose-response relationship. Ingested inorganic arsenic has been
documented to cause blackfoot disease, a unique endemic peripheral vascular disease in
southwestern Taiwan. Clinically, the disease ends with dry gangrene and spontaneous
amputation of distal parts of affected extremities. Pathologically, the disease is com-
patible with thromboangiitis obliterans (30%) and arteriosclerosis obliterans (70%) with a
fundamental change of an unduly developed severe systemic arteriosclerosis. During the
period of 1988-1992, we enrolled 1,563 subjects from the southwestern area of endemic
arseniasis in Taiwan and 23,942 subjects from seven non-endemic areas in Taiwan and
Penghu archipelago. The arsenic-exposed cohort had a significantly higher cardiovascular
mortality than the unexposed cohort, showing the multivariate-adjusted relative risks of
2.4 for ischemic heart disease and 2.2 for stroke mortality (P < 0.01). There was a
significant dose-response relationship between the cardiovascular mortality and the ar-
senic level in drinking water (P < 0.0001 trend test).
12.12
CONSIDERING CHEMICAL SPECIATION IN ASSESSING TRACE ELEMENT
BIOAVAILABILITY: THE CASE OF SELENIUM
Gerald F. Combs Jr.
Grand Forks Human Nutrition Research Center, USDA-ARS, Grand Forks, ND,
58201, USA
The biological activities of selenium (Se) are those of its various covalent compounds and
not of the element per se. Accordingly, the dominant forms of Se in foods, selenome-
thionine (SeMet) and selenocysteine (SeCys) are not direct-acting; each must be metab-
olized to exert either nutritional function via a fairly small number of selenoproteins, or
anticardinogenic activity, probably via methylselenides such as methylselenol (CH3SeH).
In general, the absorption of the Se-amino acids and other organic Se-species in foods
(e.g., methylSeMet, selenobetaine) is 70-95%, but can be highly variable, depending on
the digestibility of food proteins and the efficiency of metabolism of the resulting Se-
amino acids. For example, SeMet enters the general protein pool as a mimic for Met; it is
thus well retained but may not be efficacious in supporting SeCys-enzyme expression or
CH3SeH production. In contrast, SeCys, which cannot be incorporated directly into
proteins but is instead catabolized to selenide (H2Se), is not well retained but is well
utilized for both the SeCys-enzymes and CH3SeH. Inorganic Se-compounds (selenite,
selenate) feed the H2Se-pool by reduction by glutathione and/or NADPH. Thus, selenite,
selenate, SeMet, SeCys and Se in most plant materials have reasonably good bioavaila-
bilities; Se in many animal products has low-to-moderate bioavailability; and the reduced
(and insoluble) inorganic forms (selenides, elemental Se) are virtually unavailable.
Methods are needed to determine the Se-species of Se in foods for the assessment of the
nutritional and cancer-prevention potential.
12.13
TRACE ELEMENTS AND TUBERCULOSIS
Carlton A. Evans
Wellcome Centre for Clinical Tropical Medicine, Dept Infectious Diseases, Imperial
College London, Hammersmith Hospital Campus, London W12 0NN, UNITED
KINGDOM
Tuberculosis kills more people than any other infection. Tuberculosis is a disease of
poverty and malnutrition, characterized by such severe wasting that tuberculosis was
traditionally called ‘consumption’. Tuberculosis pathogenesis involves a chronic balance
between mycobacterial replication and the host immune defense. This immune defense
against tuberculosis is modulated by nutritional factors that influence the rapidity of
tuberculosis treatment response and susceptibility to latent infection; symptomatic dis-
ease; and death. Patients with tuberculosis are often zinc deficient and analysis of zinc
levels in hair demonstrated that deficiency preceded the disease. In an animal model, zinc
deficiency impaired antimycobacterial immunity and in humans, the administration of
both oral and topical zinc augmented delayed-type hypersensitivity immune reactions to
tuberculosis antigens. In a blinded, randomized placebo-controlled trial in humans, zinc
given with vitamin A accelerated the response to tuberculosis treatment. Intervention
trials are investigating whether micronutrient supplementation also protects individuals
from tuberculosis. Selenium, copper and especially iron levels have also been linked with
tuberculosis susceptibility. In addition to these trace elements, deficiency and/or sup-
plementation of dietary protein, vitamin A and D also modulate tuberculosis suscepti-
bility and it is likely that deficiencies of these micronutrients interact to increase the risk
of tuberculosis in malnourished individuals. These findings have considerable potential to
improve public health.
12.14
THE ROLE OF ZINC IN DNA DAMAGES AND APOPTOTIC CELL DEATH
Alain Favier
LAN, University and Nuclear Research Center of Grenoble, CENG, 17 Avenue des
Martyrs, 38056 Grenoble, FRANCE
DNA damages are an important disturbance in cell that can lead as well to mutations and
cancerogenesis as to cell death by apoptosis. In such a process, zinc can act differently
according to cell type and to zinc concentration. First zinc can protect DNA from
oxidative stress by two mechanisms: preventing direct binding of redox metals or pre-
venting binding of redox metal to transcription factors. In the two cases it prevents in situ
DNA oxidation. Such a binding is regulated by nuclear metallothionein. But zinc is also
an essential cofactor of DNA repair system. The decrease in intracellular zinc concen-
tration induces a characteristic apoptosis with apoptotic bodies formation and nuclear
DNA condensation and fragmentation. Zinc addition in cell culture medium prevents the
apparition of morphological and biochemical signs induced by intracellular zinc chela-
tion, but also by other inducers like etoposide or tumour necrosis factor alpha. The
antiapototic action of zinc is not only related to DNA protection but also to inhibition by
zinc of various apoptotic enzymes. We described a strong inhibitory effect of zinc on the
main killer enzymes the caspases. Conversely on some cell type as neurons, excess of zinc
is a proapoptotic agent. These cells are certainly more sensitive as they use zinc as a
neuromediator by excreting zinc-rich vesicles and so excess of extracellular zinc can
trigger some transduction signal of death. But large zinc excess is toxic for all cells as by
necrosis as by apoptosis. Clinical consequences are important and apoptosis may be
related to a zinc deficiency, i.e. the depletion in T lymphocytes in elderly, or to a zinc
excess, i.e. the decrease in memory neurons occurring in Alzheimer disease. According to
the biphasic effect of zinc, we have to be very careful and well documented on molecular
mechanisms before to supplement patients.
12.15
THE ESSENTIALITY OF ZINC FOR REPAIR OF BRAIN DNA
Christopher Frederickson
NeuroBioTex, Inc., Galveston Island, TX, USA
Free zinc ions are released into the extracellular fluids of the brain from at least two
sources. The first is the well-known synaptic release of zinc ions from the stores in the
presynaptic vesicles of certain neurons, which, in the mammalian cortex, are all gluta-
matergic neurons. This synaptically - released zinc is an important modulator of NMDA
receptor activity, and of certain forms of synaptic plasticity. The second source of free
zinc ions occurs in the so-called ‘‘excitotoxic’’ injury mode and (potentially) during other
types of neuronal injury as well. This latter, ‘‘somatic’’ release occurs when nitric oxide
rises to pathological levels and mobilizes zinc off the thiol ligands of MT3. In this
scenario, neuronal somata fill up with free zinc and this zinc spills into the surrounding
extracellular fluids. Whether the latter, somatic release, also occurs in sub-clinical ni-
trosative stress remains open.
12.16
TRACE ELEMENTS AND HIV INFECTION
Henrik Friis
Dept Epidemiology, Inst. Public Health, Univ. Copenhagen, DENMARK
Trace elements are likely to be important in HIV as in other infections. Trace elements
may affect HIV transmission and progression, as well as morbidity from common or
opportunistic infections, and efficacy of antiretroviral drugs. However, current data are
inadequate and their interpretation complicated by co-existence of multiple deficiencies
and interactions between micronutrients. This is unfortunate, since micronutrient sup-
plementation is a potentially beneficial, feasible and affordable intervention among
people with HIV infection. Selenium may affect HIV replication and virulence. Low
serum selenium was a predictor of mortality in observational studies, but selenium
supplementation trials have not been able to show that selenium supplementation reduces
HIV load, and no data on the effect of clinical outcomes have been reported. Data from
in vitro and observational studies suggest that a high iron status or intake increase HIV
replication and progression. However, a small trial found no effect of low dose, twice
weekly iron on viral load, but the effect of usual iron regimens given to pregnant or
anemic HIV infected people needs to be assessed in larger randomized, controlled trials.
Given the importance of zinc for immunity and host defense to diarrhea and pneumonia
in children in developing countries, the effect of zinc supplementation on HIV progres-
sion and morbidity should be assessed. Although observational data from the US, sug-
gesting that a high zinc intake increases HIV progression, have deferred the needed trials,
data from intervention trials are now emerging.
12.17
BREEDING PLANTS WITH BIOAVAILABLE TRACE ELEMENTS
Robin D. Graham
University of Adelaide, South AUSTRALIA
For some of the nutritional traits of interest, sufficient genetic variation exists for breeders
to select new, more nutrient-dense varieties. This has been demonstrated for the major
cereals, beans and cassava for iron, zinc and carotenoids. However, for iron and zinc the
trait appears to be quite complex with several to many genes involved. For carotenoids,
only two genes appear to control most of the variation among genotypes so breeding for
carotenoids in toto or for pro-vitamin A carotenoids in particular appears relatively easy
where highly pigmented donors are available as is the case for maize, sweet potato and
cassava, for example. Good donor parents are not currently available for the major
cereals, rice and wheat, although a little genetic variation is known. For iodine and
selenium, the evidence for genetic variation is not strong, and where we have looked, it is
further complicated by relatively much greater variation in soil so that identifying
superior germplasm is problematical. For these elements, and also for zinc where the
breeding is also complicated genetically, fertilizer strategies offer a viable alternative
strategy for biofortify staple foods. Iron remains the enigma: on a planet staggeringly rich
in iron, its bioavailability to the biosphere is greatly limited by plentiful free oxygen,
genetics are a limitation for breeders, and contamination is common, all considerations
conspiring against biofortification, though it is clearly not impossible. A better alternative
for iron is suggested: exploiting interactions in the gut between iron and carotenoids and
zinc to increase the bioavailability of iron already in the diet. These various strategies are
illustrated and discussed.
12.18
NUTRITIONAL IRON DEFICIENCY EARLY PERSPECTIVE AND PRESENT
APPLICATIONS (RAULIN AWARD LECTURE)
Leif Hallberg
Institute of Internal Medicine, Dept of Clinical Nutrition, Univ. of Göteborg, Göteborg,
SWEDEN; Sahlgrenska University Hospital, SE 413 45 Göteborg, SWEDEN
Nutritional iron deficiency is caused by an intake of dietary iron insufficient to cover
physiological iron requirements. Present problems with iron deficiency being the most
frequent deficiency disorder are related to nonbiological changes in our societies over
the most recent 10,000 years. Menstrual iron losses are constant in the single woman
but varies by a genetic control in the same way in different populations making dif-
ferences in dietary iron the main source of variation between iron status in different
populations. Pregnancy is a main source of variation in iron requirements between
women. Early, two thirds of pregnancy are associated with reduced iron requirements
but with increased ones in the last trimester making access of iron stores a necessity.
The newborn infant has a hemoglobin with an increased affinity for oxygen requiring
higher Hb to deliver oxygen to tissues. At delivery the fetus thus has increased amounts
of iron to be sufficient for the first 4-6 months. The iron requirements during the
following weaning period up to 2 years are the highest. Iron is transported by the hard-
binding transferrin within the body. The iron bound to transferrin is then released to
tissues needing iron by a special protein, the transferrin receptor, which control the
delivery of iron to iron-needing tissues. The extrinsic tag technique to measure nonheme
iron absorption and the labeling of heme iron with a suitable heme iron tracer has
made it possible to measure dietary iron absorption and to identify factors that
influence iron absorption.
12.19
DIETARY BORON AS A FACTOR IN GLUCOSE AND INSULIN
METABOLISM
Curtiss D. Hunt
USDA, Agric. Res. Ser., Grand Forks Human Nutrition Research Center, PO Box 9034,
Grand Forks, ND 58202, USA
Insulin resistance is a component of the Metabolic Syndrome, a condition that affects
approximately 22% of the American population (47 million persons). The Syndrome
is an important risk factor for cardiovascular disease, the leading cause of death for
the obese. There is emerging evidence that boron is one of several dietary factors
important in insulin and glucose metabolism. Boron deprivation perturbs energy
substrate metabolism in humans and animal models, particularly when other nutrients
are provided in suboptimal amounts. For example, boron deprivation can increase
fasting serum glucose concentrations in volunteers fed a low magnesium diet (115
mg/d) and can induce hyperinsulinemia in the vitamin D-deficient rat. In the chick,
boron deprivation exacerbates the rise in plasma glucose concentrations induced by
marginal vitamin D deficiency. Moreover, newer findings indicate that boron reduces
insulin resistance independent of other nutritional stressors. For example, in the
growing rat, boron can independently reduce plasma insulin while maintaining glu-
cose concentrations. In the chick model, boron can independently decrease in situ
peak pancreatic insulin release. Because boron reduces circulating insulin concentra-
tions while maintaining serum glucose concentrations within the normal range, die-
tary boron might help prevent pancreatic exhaustion caused by chronic
hyperinsulinemia. The findings to date are probably relevant to human health because
the amounts of dietary boron that improve glucose and insulin metabolism are
similar to those found in diets that include ample amounts of fruits, nuts, vegetables,
and legumes.
12.20
COMBATING IRON DEFICIENCY - SUPPLEMENTATION, FORTIFICATION,
AND DIETARY TACTICS
Janet R. Hunt
USDA-ARS, Grand Forks Human Nutrition Research Center, PO Box 9034, Grand
Forks, ND 58202, USA
Deficiency of iron and the associated anemia, which adversely influence cognitive and
immune function and diminish work capacity, affect more than 3.5 billion people
worldwide. Strategies for the improvement of iron nutrition include supplementation,
fortification, and dietary approaches. Iron status indices have improved with the quan-
titative assessment of body iron, based on the ratio of serum transferrin receptor to serum
ferritin. Challenges of iron supplementation include effective delivery, acceptance, and
gastrointestinal tolerance. Daily iron supplementation reduces the intestinal absorptive
efficiency, and is not clearly superior to weekly iron supplementation. Effective fortifi-
cation of sauces in rice-based diets has been demonstrated with NaFeEDTA, a form
efficiently absorbed even from diets high in phytic acid. Other promising fortificants
include iron chelated with amino acids, and iron micronized and encapsulated to main-
tain a readily soluble particle size. The effectiveness of elemental iron powders (carbonyl,
electrolytic, reduced) varies considerably with large differences in solubility and surface
area, dependent on commercial production methods. Diets with high, but not low iron
bioavailability enable women to absorb iron in inverse proportion to their body iron, but
the high impact of dietary bioavailability in short-term absorption studies is not readily
observed in longer trials to improve iron status. Addressing iron deficiency requires a
multifactorial approach with supplementation, fortification, and, ultimately, accessible
and varied diets.
12.21
ADVANCES IN NUTRITIONAL METROLOGY
Venkatesh Iyengar
IAEA, A-1400 Vienna, Austria; pesent address: 6202 Maiden Lane, Bethesda, MD
20817, USA
Harmonization of chemical measurements is an essential requirement to ensure compa-
rability of analytical data. Application of principles of metrology (the science of mea-
surements) enables such reliable comparisons. For more than 3 decades the International
Atomic Energy Agency (IAEA) has supported projects using nuclear and isotopic tech-
niques in several areas such as food, nutrition and environment with the objective of
strengthening the analytical capabilities in the IAEA Members States. Over time such
efforts have led to the development of proper study designs, harmonization of sampling
protocols, adequate contamination control and evaluation of the suitability of competing
analytical techniques for specific analyte determinations. These in turn have contributed
to the emergence of coherent and practical analytical approaches resulting in measurable
improvements in the quality of analytical data generated by the IAEA Member States.
These positive impacts are demonstrated by (i) the development of many natural matrix
reference materials (RMs), (ii) enhancement of the capacity of participants for upgrading
the existing RMs for establishing reference values for project specific analytes, and (iii)
improvement in biomonitoring techniques in the areas of occupational exposure to heavy
elements and airborne particulate matter (APM) sampling. Over time these efforts were
also extended to improve measurement processes in the human physiology area resulting
in further progress documented under IAEA supported activities (i) development of the
characteristics of a Reference Asian Man to strengthen radiological safety approaches in
the Asian region; (ii) establishment of reference values for human milk and dietary intake
of minor and trace elements under supported initatives, and (iii) contribution of the
IAEA Co-ordinated Research Projects and Technical Co-operation Projects to
strengthen the new FAO/WHO/UNU global recommendations on human energy intake
and expenditure. Collectively, these consistent IAEA initiatives nurtured over a period of
several years have promoted harmonization of chemical measurements to a significant
degree, thus infusing a measure of metrological awareness at the level of field projects.
This has been possible since over the years, the IAEA has gone through a process of
continuous refinement, modelled on accumulating practical experience. Nuclear and
isotopic techniques have played an important role in reaching these goals.
12.22
HEALTH EFFECTS OF CADMIUM-EXPOSED WORKERS RESIDING IN
ENVIRONMENTAL CADMIUM-POLLUTED AREA IN CHINA
T. Jin*[1,2], T. Ye [3], Q. Kong [4], X Wu [1], G. Zhu [5], H. Wang [5], X. Zeng [1], 1X.
Jiang [1], and G. F. Nordberg [2]
Department of Occupational Health, School of Public Health, Fudan University, 200032
Shanghai, PR CHINA [1]; Unit of Environmental Medicine, Umeå University, S-901 87
Umeå, Sweden [2]; Department of Preventive Medicine, School of Public Health, School
of Public Health, Fudan University, 200032 Shanghai, PR CHINA [3]; Institute of
Environmental Health, Zhejiang Academy of Medical Science, Hangzhou, CHINA [4];
Department of Bone Metabolism, Institute of Radiation Medicine, Fudan University,
Shanghai, CHINA [5]
The present interest is to find whether combining occupational with environmental
exposure to cadmium might result in a different severity of health effects, i.e. kidney,
bone and prostate, compared with solely environmental or occupational exposure in
China. The smelter started to produce in 1961 and discharged cadmium polluted
wastewater into the nearby river, which was used for irrigation of the rice fields. The
historical data from the local industrial health inspector showed that the cadmium
concentration of air in workplace was 0.655 mg/m3 (0.026 to 3.623 mg/m3). Cadmium
concentration of rice was 3.7 mg/kg. Rice is the main food of local residents. Group A
(44 persons) was from those who were the residents in polluted area and worked in the
smelter. Group B and C were 88 persons for each group and age and sex matched with
Group A. The persons from Group B and C lived in Cd-polluted and non Cd- polluted
area, respectively. Blood cadmium was 9.66, 7.82 and 1.53 lg/l and urinary cadmium
11.86, 9.51 and 1.81 lg/g creatinine in group A, B and C, respectively. There was an
increasing prevalence of hyper-b2 microglobulinuria, NAGuria and albuminuria in
both Group A and Group B. BMD was 0.754, 0.752 and 0.747 and the prevalence of
osteoporosis was 9.09%, 4.55% and 2.27% in group A, B and C respectively. The mean
value of prostate specific antigen (PSA) was 1.19, 1.32 and 2.09, and the prevalence of
hyper PSA was 12.12%, 9.09% and 4.55%, respectively. In the present study, the
prevalence of renal dysfunction, bone and prostate damage in group A were higher
than those in group B. It was suggested that exposure to cadmium both occupationally
and environmentally resulted in higher prevalence of renal dysfunction, bone and
prostate damage in these workers, compared with persons who exposed to cadmium
only in the general environment. (This project was granted by European Commission
and SIDA.)
12.23
COPPER AND ZINC DYNAMIC CHANGES IN PROGRESSION OF
CARDIOMYOPATHY
Y. James Kang
Dept. Medicine, Pharmacol. Toxicol., Univ. Louisville, Louisville, KY 40202, USA
Many clinical studies have shown that patients with hypertrophic cardiomyopathy dis-
play depressed serum zinc and increased serum copper concentrations and those with
dilated cardiomyopathy show serum zinc depletion along with increased zinc urinary
excretion. Animal studies have duplicated the clinical observation and shown that during
the compensatory phase (hypertrophy) of cardiac response to pathological challenges,
zinc concentrations increase and copper decreases in the heart, whereas at the end stage of
dilated cardiomyopathy, zinc decreases and more often copper increases. Studies using
isolated hearts have shown that perfusion with zinc increases copper efflux from the heart
and inhibits ischemia/reperfusion-induced cardiac injury. Transgenic mice overexpressing
metallothionein (MT) specifically in the heart have displayed a significant increase in zinc
and a slight increase in copper concentrations in the heart, which is highly resistant to
ischemia/reperfusion injury. MT depression along with decreased zinc concentrations in
the heart occur in the progression from compensated hypertrophy to decompensated
dilated cardiomyopathy (heart failure) in animal models. This phase corresponds to the
clinical observation of serum zinc depletion and zincuria. Although high concentrations
of copper in the heart occur with cardiomyopathy, copper deficiency with or without zinc
elevation in the heart is also detrimental. Therefore, zinc repression along with copper
elevation is often associated with cardiomyopathy, but an appropriate balance between
zinc and copper is required for prevention of cardiomyopathy. (Supported in part by
NIH grants HL63760 and HL59225.)
12.24
CONTROL OF MICRONUTRIENT MALNUTRITION IN VIETNAM: CURRENT
SITUATION AND STRATEGIES IN COMING YEARS
Nguyen Cong Khan* [1], Nguyen Xuan Ninh [1], Ha Huy Khoi [1], Luong Ngoc Khue
[2]
National Institute of Nutrition, Hanoi, VIETNAM [1]; Ministry of Health, Hanoi,
VIETNAM [2]
In Vietnam, vitamin A deficiency (VAD), iron deficiency anemia, and iodine deficiency
disorders have been reduced during the last decade but still remain prevalent. In the early
1990’s, a nationwide strategy for VAD control that included nutrition education, uni-
versal distribution of vitamin A capsules and promotion of household production and
consumption of vitamin A-rich foods has been implemented. Presently, prevalence of
clinical exophthalmia is lower than the cut-off point established by the WHO but sub-
clinical VAD remains high. Iron deficiency anemia is wide-spread in Vietnam. The cur-
rent prevention program has focused in 1,282 out of more than 10,000 communes in the
country with two components: supplementing women with iron and folic acid and pro-
viding them with nutrition education together with the prevention of intestinal parasites,
especially hookworm. Results of studies on the efficacy of FeEDTA-fortified fish sauce
are very promising such that there are plans to scale-up a fish sauce fortification program.
Control of goiter has been implemented since the early 1970s. The universal salt iodi-
zation program initiated in 1999 by government legislation resulted in 61% of Vietnamese
households using iodized salt by 2001 but with large variation among regions. As out-
lined by the National Nutrition Strategy which was ratified by the Government and seeks
to overcome micronutrient deficiencies in Vietnam, we are proceeding to combine dif-
ferent nutritional strategies that maintain success and provide for further sustainable
achievements.
12.25
EMERGING ROLE FOR ENDOGENOUS ZINC IN NEUROLOGICAL DISEASES
Jae-Young Koh
Department of Neurology, CRI Center for the Study of CNS Zinc, University of Ulsan,
College of Medicine, Seoul, South Korea
Zinc is one of the most abundant transition metals in the brain. A substantial fraction of
brain zinc is located inside presynaptic vesicles of certain glutamatergic terminals in a free
or loosely bound state. This vesicle zinc is released with neuronal activity or depolar-
ization, probably serving physiologic functions. However, with excess release, as occurs in
a variety of pathologic conditions, zinc in presynaptic terminals translocates to post-
synaptic neurons. Another mechanism of the rise in intracellular free zinc concentrations
is the intracellular release from zinc-binding proteins such as metallothioneins. Regard-
less of the routes, increases in intracellular free zinc levels can induce death of neurons
and glial cells. Intracellular mechanisms of zinc neurotoxicity may include disturbances in
energy metabolism, increases in oxidative stress, and activation of apoptosis cascades.
Zinc inhibits glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and depletes nico-
tinamide adenine dinucleotide (NAD+) and adenosine triphosphate (ATP). On the other
hand, zinc activates protein kinase C (PKC) and extracellular signal-regulated kinase
(Erk-1/2), and induces NADPH oxidase; these events result in oxidative neuronal injury.
Zinc can also trigger caspase activation and apoptosis via the p75NTR pathway. Inter-
estingly, the converse, depletion of intracellular zinc, also induces neuronal death, but in
this case, exclusively via classical apoptosis. In addition to the neurotoxic effect, zinc may
contribute to the pathogenesis of chronic neurodegenerative diseases. Zinc promotes
aggregation of amyloid. Consistently, in Alzheimer’s disease, mature amyloid plaques,
but not preamyloid deposits, are found to contain high levels of zinc, suggesting the role
of zinc in the process of plaque maturation. Further insights into roles of zinc in brain
diseases may help set a new direction toward the development of effective treatments.
(Supported by National Creative Research Initiatives of Korean Ministry of Science and
Technology.)
12.26
THE LINK BETWEEN ZINC AND REDOX METABOLISM
Wolfgang Maret
Depts. Preventive Medicine and Community Health & Anesthesiology, University of
Texas Medical Branch, Galveston, TX 77555, USA
Coordination environments of zinc with sulfur ligands of cysteines are redox-active and
couple the redox-inert zinc ion to redox metabolism. Thus, oxidation of sulfur ligands
mobilizes zinc while reduction of the oxidized ligands enhances zinc binding. In this way,
changes in the redox state modulate the availability of zinc. Under redox stress, reactive
species release zinc from proteins and transfer zinc to sites on other proteins, thereby
compromising signal transduction, generation of metabolic energy, and maybe even ge-
netic programs. In this regard, the relation between nitric oxide signaling and cellular zinc
homeostasis is receiving particular attention. Both decreases and increases of cellular zinc
can elicit oxidative stress. Yet, zinc also has antioxidant effects. Such an action is indirect
and includes, e.g., induction of thionein, a strong reductant. Jointly, these functions
indicate that zinc has been selected to control some aspects of redox metabolism. In order
to utilize this ion for biological regulation and to balance its antioxidant and prooxidant
potentials, tight control of cellular zinc ions is required. The link between zinc and redox
metabolism is implicated in the pathogenesis of a variety of chronic diseases. While acute
oxidative stress increases cellular zinc, chronic oxidative stress may result in ‘‘local zinc
deficiencies’’ that drastically alter the physiology of affected tissues. It is expected that in
nutritional interventions simultaneous administration of both zinc and antioxidants will
be beneficial.
12.27
MULTIPLE MICRONUTRIENT SUPPLEMENTATION TO ADDRESS
CONCURRENT MICRONUTRIENT DEFICIENCIES IN INFANTS AND
PREGNANT WOMEN
Siti Muslimatun
SEAMEO TROPMED Regional Center for Community Nutrition, University of Indo-
nesia, PO Box 3852, Jakarta 10038, INDONESIA
Multiple micronutrient deficiencies are significant problems among women in developing
countries, including Southeast Asia. Moderate to severe deficiencies of iron, folic acid and
zinc are associated with increased risk of low birth weight and pregnancy complications.
Until dietary improvements take place, supplementation with multiple micronutrients is
one alternative solution to reduce maternal deficiencies, improve reproductive outcomes,
and child survival and growth. Studies in Indonesia found that prenatal supplementation
of vitamin A together with iron/folic acid reduce the risk of anemia during pregnancy,
maintain maternal vitamin A status up to lactation period, and improve infant vitamin A
status. Adding zinc and ß-carotene to iron/folic acid supplementation during pregnancy
has beneficial effects on birth weight and vitamin A status compared to iron/folic acid
only. The benefits of prenatal multi micronutrient supplementation over iron/folic acid
supplementation on pregnancy outcomes and child survival and growth need to be been
tested in large trials in a Southeast Asian setting. Future multiple micronutrient sup-
plementation research should take into account the limiting nutrient in the population
and nutrient-nutrient interactions.
12.28
ESSENTIALITY OF COPPER, ZINC, MAGNESIUM, BORON AND SILICON IN
BONE DEVELOPMENT AND FUNCTION
Forrest H. Nielsen
USDA, Agric. Res. Ser., Grand Forks Human Nutrition Research Center, PO Box 9034,
Grand Forks, ND 58202, USA
Among the nutritional factors that affect bone development and function are copper
(Cu), zinc (Zn), magnesium (Mg), boron (B) and silicon (Si); this is confirmed by findings
obtained in my laboratory. B deprivation (0.07 mg/kg diet) decreased femur strength in
rats with the effect most marked in females fed a diet high in polyunsaturated fatty acids.
B deprivation also altered femur shape and decreased tibia calcium (Ca) and phosphorus
concentrations. B apparently has a function that affects bone development and is influ-
enced by the fatty acid composition of the diet. Si deprivation (2 mg/kg diet) decreased
plasma osteopontin, increased urinary helical peptide excretion, and altered femur shape
in rats. Si deprivation did not affect femur Ca concentration, but decreased potassium
and Zn, minerals associated with the organic matrix. Si may have a function that affects
bone growth prior to bone crystal formation. In humans, a combination of low dietary
Zn and Cu, or low dietary Zn alone, may be factors affecting bone health. In post
menopausal women fed low Cu (1 mg/d), plasma osteocalcin was higher when dietary Zn
was low (3 mg/day) than when it was high (53 mg/day). Regardless whether dietary Cu
was low or luxuriant (3 mg/d), high compared to low dietary Zn increased the urinary
excretion of N-telopeptides and decreased serum calcitonin. Mg deprivation (118 mg/d)
decreased urinary Ca excretion, and increased Ca balance and intracellular Ca in post
menopausal women. Mg deprivation may affect bone health through changing cellular
Ca such that inflammatory or oxidative processes result.
12.29
EFFECTS OF CADMIUM EXPOSURE ON BONE HEALTH IN THE CHINESE
POPULATION
Gunnar Nordberg* [1], Guoying Zhu [2], Hongfu Wang [2], and Taiyi Jin [3]
Environmental Medicine, Umea University, SE-90187 Umea, SWEDEN [1]; Department
of Bone Metabolism, Institute of Radiation Medicine, Fudan University, Shanghai,
CHINA [2]; Department of Occupational Health, School of Public Health, Fudan
University, Shanghai, CHINA [3]
Cadmium exposure in the working or the general environment has, for long been asso-
ciated with osteoporosis and osteomalacia in Japan, France and the UK. Cadmium
intake via rice has been shown in Japan to be associated with osteoporosis and renal
tubular dysfunction in several population groups, but until we performed our recently
reported studies, no such effects had been recognised in other countries in Asia. The
World Health organisation (WHO) has defined osteoporosis as bone mineral density
(BMD) 2.5 SD lower than the average in normal young adults. BMD among 790 persons
in three areas located at various distances from a non-ferrous metal smelter in South
Eastern China was negatively correlated with urinary cadmium in both males and females
(p < 0.01). The prevalence of osteoporosis among women above age 50 increased from
34 % in the control area to 52% in the heavily polluted area near the smelter. The
prevalence of fracture was higher in the most polluted area compared with controls and
there was renal dysfunction clearly related to urinary and blood cadmium. Worldwide
osteoporosis is a main cause of fractures in the elderly. The association between cadmium
and osteoporosis in China, the most populous country in the world, and in Belgium and
Sweden at much lower exposure levels, underlines the need for further studies aiming at
defining the importance of such a relationship for preventive measures.
12.30
ANTI-OXIDANT EFFECT OF ZINC IN HUMANS
Ananda S. Prasad
Department of Medicine, Division of Hematology-Oncology, Wayne State University
School of Medicine, Barbara Ann Karmanos Cancer Institute, University Health Center,
4201 St. Antoine, Detroit, MI 48201, USA
Oxidative stress is an important contributing factor in several chronic diseases. In this
study we have tested the hypothesis that zinc is an effective antioxidant in vivo. We used
in vitro activation of NFjB by TNFa in peripheral blood mononuclear cells (MNC),
direct measurement of lipid peroxidation and modified DNA as biomarkers of oxidative
stress in humans. We assayed plasma zinc, NFjB activation, lipid peroxidation and DNA
oxidation products, gene expression of TNFa and IL-1ß (known to generate ROS) by
RT-PCR, in LPS stimulated MNC before and after either zinc or placebo supplemen-
tation in healthy human volunteers. Zinc supplementation in vivo protected MNC
against oxidative stress. The gene expression of cytotoxic cytokines TNFa and IL-1ß,
lipid peroxidation and DNA oxidation products were decreased in the zinc supplemented
subjects in comparison to the placebo group. In HL-60 cells, the gene expression of A20 a
zinc-finger protein known to inhibit the gene expression of TNFa and IL-1ß, were pos-
itively regulated by zinc, suggesting that the effect of zinc on negative regulation of gene
expression of TNFa and IL-1ß may be via A20 molecule. A therapeutic trial of zinc either
alone or in conjunction with other modalities in many chronic diseases, in which oxi-
dative stress is known to play an important role, may prove to be beneficial.
12.31
ZINC NUTRITION AND RESPIRATORY INFECTION IN THAILAND
Sakda Pruenglampoo
Research Institute for Health Sciences, Chiang Mai University, Chiang Mai 50202,
THAILAND
Zinc is present throughout all biologic systems and it is required for catalytic function of
more than 100 enzymes. The zinc content of the adult human body ranges from 1.5 to 2.5
g, with higher average values in men compared to women. Dietary intake of zinc cor-
relates with the protein content of food. Thus, good sources of zinc are lean meat,
seafoods and dairy products. Zinc absorption is concentration-dependent and takes place
throughout the small intestine. Calcium and iron may reduce zinc uptake. Other inhib-
itors of zinc uptake include phytate, oxalic acid, polyphenols, and folic acid. The ab-
sorbed zinc is carried directly via the portal system to the liver, where it is taken up
rapidly and released into the systematic circulation for delivery to other tissues. Excretion
of zinc occurs mostly via the feces through secretion into the digestive tract. Small
amounts are lost in the urine and in skin cells. The recommended dietary zinc intake to
meet normative physiologic requirements depends on the population group, age and
estimated zinc bioavailability in the diet. Acute zinc toxicity may result in symptoms,
such as nausea, vomiting, epigastric pain, diarrhea, lethargy and fatigue. With zinc
deficiency, several biochemical abnormalities and clinical signs may occur. Some studies
report changes in immune function that are clinically important, and decreased rates of
infection (diarrhea, respiratory infections and malaria) have been observed following zinc
supplementation in population-based studies. In 2001, the Thai Ministry of Public Health
showed that the reported cases of pneumonia per 100,000 population during 1992-2001
varied from 153.00 to 218.65. We are interested in investigating whether there is rela-
tionship between zinc dietary intake and the incidence of pneumonia, especially in chil-
dren under 5.
12.32
ZINC IN METABOLIC SYNDROME
M. Roussel
Laboratoire NVMC, J. Fourier University, 38700 Grenoble, FRANCE
In the metabolic syndrome, the inability of cells to respond to physiological levels of
insulin results in impaired insulin function. Zn is involved in glucose homeostasis and
insulin function. It acts in stabilizing insulin structure, increasing insulin receptor
expression and activating insulin receptor binding. Zn also functions as a biological

antioxidant, protects endothelial cells and induces antioxidant metallothionein synthesis.
Several biological signs of metabolic syndrome including increased insulin resistance,
elevated triglycerides, hypertension and increased oxidative stress are also found in zinc
deficient patients. Zinc deficiency is associated with alterations of adipocyte metabolism,
visceral obesity, hyperleptinemia, increased lipoperoxidation and increased risk of dia-
betes. All these symptoms are also observed in the metabolic syndrome. Impaired zinc
status in obese people and in people with diabetes are well documented (increased urinary
losses, modified tissue pools and lowered plasma zinc). In cell models, we have observed
the protective effects of Zn against glucose-induced oxidative damage. In rats receiving a
fructose fed diet, we have shown improved insulin sensitivity when supplemented by Zn. In
humans, data are still controversial. In our group, we have reported the beneficial effects of
Zn supplementation as 30 mg/d of Zn as Zn gluconate in people either type I or II diabetes
against lipoperoxidation. Therefore, given the numerous possible effects of Zn nutriture in
patients at risk of metabolic syndrome, zinc should be taken into account among nutri-
tional factors acting in the prevention of the metabolic syndrome and diabetes.
12.33
ZINC AND IRON NUTRITURE OF PREMENOPAUSAL WOMEN AND
ESSENTIALITY FOR COGNITION
Harold Sandstead* [1], Katsuhiko Yokoi [2], James Penland [3], Norman Egger [1], VMS
Ramanujam [1], Nancy Alcock [1], Hari Dayal [1]
U Texas Medical Branch, Galveston, TX 77555, USA [1], Seitoku U, Matsudo, Chiba
271-8555, JAPAN [2], and USDA ARS Human Nutrition Research Center, Grand
Forks, ND 58201, USA [3]
Zn & Fe are essential for brain function. Zn and Fe associations were assessed by food
frequency, Fe status and Zn kinetics in 50 premenopausal women; and effects of Zn, Fe,
and other micronutrients treatments on neuropsychological functions were measured in
92 through 16-week double blind randomized trial. Food frequency correlated with Zn
kinetics and serum(s) ferritin in subjects not taking oral contraceptives. In 30 subjects
with pZn > 700 lg/L sferritin, sFe, and %-saturation of Fe binding protein were higher
(p< 0.03). Serum ferritin correlated significantly with the central Zn pool (Q1, repre-
senting the extracellular Zn) / fat free mass (FFM), the lesser peripheral Zn pool (Q2,
representing rapidly turning over tissue Zn) / FFM, & the greater peripheral Zn pool (Q3,
representing slower turning over tissue Zn) / FFM. Increase in sferritin above 17.8 lg/L
was proportionate to Q2. Treatment affected perception, visual memory, reasoning and
neuromotor function: ZnM > FeM for memory and > M for reasoning; FeM > M for
perception; ZnM & FeM > M for a neuromotor task. Thus Fe & Zn status are highly
related and ZnM and FeM treatments improved some neuropsychological functions.
(Support: DAMD 17-95-C-5112 & USPHS M01-RR-00073.)
12.34
GENETIC DEEFECTS OF COPPER TRANSPORT: WILSON AND MENKES
DISEASES
Bibudhendra Sarkar
The Hospital for Sick Children and The University of Toronto, Toronto, Ontario M5G
1X8, CANADA
Copper is an essential trace element which is a necessary component of many
developmentally important enzymes. Excess copper can lead to extensive pathophys-
iological problems. The two major inherited disorders of copper metabolism are
Wilson and Menkes diseases. Wilson disease is characterized by a defect in copper
excretion which results in the accumulation of copper in vital organs causing neuro-
logical and liver diseases. In contrast, Menkes disease involves a defect in intestinal
copper absorption which then results in global deficiency of copper-dependent en-
zymes and death in early childhood. Wilson and Menkes disease genes encode copper-
transporting P-type ATPases. These ATPases perform their roles by interacting with
many cellular components in order to transport copper. There are several pharma-
cologic regimens and therapeutic modalities available for the treatment of Wilson
disease. However, the only treatment available currently for Menkes disease is the
administration of copper-histidine soon after birth. The detection of copper-histidine
in human blood led us to use this form of treatment for Menkes disease patients.
Although death usually occurs by 3 years of age due to severe neurodegeneration, our
first patient showed nearly normal development. We have completed a long-term
clinical follow-up of copper-histidine treated patients over a period of 10-20 years.
Results strengthen our earlier findings that the treatment has to start soon after birth
before the neurological abnormalities develop. Patients may still develop connective
tissue disorders. Further improvement of the treatment of Menkes disease is currently
underway.
12.35
IRON SUPPLEMENTATION AND RISK OF INFECTIOUS DISEASE
Sunil Sazawal*, Girish S. Hiremath, and Robert E. Black
Department of International Health, Johns Hopkins Bloomberg School of Public Health,
Baltimore, MD, USA
Iron deficiency is the most common nutritional deficiency disorder in world, for which
iron supplementation is most commonly employed intervention. There are at least 4
possible pathways through which excess iron could be harmful, competitive inhibition
in the uptake of nutrients like zinc, enhanced growth of enteric pathogens, enhanced
growth of pathogens in tissues and oxidative damage. The other side of the coin,
however, is that reduced lymphocyte function, particularly T-cell proliferation has
been shown in iron deficiency, increase in transferrin saturation is likely to benefit
immune response. Finally clinical studies indicate impairment of neutrophil function
(which plays key role in combating any infectious process) in iron deficiency. Overall
data from clinical trials has also been conflicting, some studies suggesting an increased
susceptibility to malaria with iron consumption, while others have not found such
association. We present results of a meta analysis evaluating effect of routine iron
supplementation in preschool children in developing countries on morbidity and also
newer data from large clinical trial of iron supplementation. The results of meta
analysis suggests that there is lack of data evaluating effect of iron supplementation
on severe morbidity in preschool children, and data from trials when iron is given a
preventive strategy rather than therapy. Results from a large study 30,000 children in
Pemba Zanzibar found 10% increase in adverse events (hospitalizations or mortality),
95% CI 0-20%, p=0.04. This increase was primarily contributed by malaria related
causes 12% increase, p=0.06 and infection related causes 29% increase, p=0.004. In
view of these results routine preventive supplementation of children in malaria holo-
endemic area’s needs to be reviewed both in terms of potential risk/benefit and also
alternative strategies for delivery of iron.
12.36
ZINC AND CELL CYCLE CONTROL IN THE CANCER CELL
Paul J. Smith
Department of Pathology, University of Wales College of Medicine, Heath Park, Cardiff,
CF14 4XN, UNITED KINGDOM
The transition metal zinc is a physiologically important divalent cation in biological
systems and its availability can affect various macromolecular processes including DNA
synthesis, microtubule polymerization, apoptosis, and gene expression. In most cells the
free intracellular concentrations of Zn2+ are extremely low, the majority being bound to
proteins at various subcellular locations. Two-photon-excitation fluorescence microscopy
was used to track the intracellular location of free Zn2+ complexes to cytoplasmic ves-
icles. Therapy-related characteristics of cancer cells include the loss of appropriate pro-
liferation controls under stress and a propensity for metastasis. Changes in cell-cell and
cell-matrix adhesion accompany the transition from benign tumours to invasive, malig-
nant cancer and the subsequent metastatic dissemination of tumour cells. It is an in vitro
characteristic of classical forms of human small cell lung carcinoma (SCLC) cell line to
grow in an anchorage-independent manner as micro-spheroids, enacting autocrine loops
and demonstrating characteristics associated with elevated metastatic potential. Gene
expression array analysis (of 22K genes) and immunofluorescence studies of cell surface
markers for adherent isogenic variants of SCLC cells revealed changes consistent with the
acquisition of a reduced metastatic potential. However, no significant changes could be
found in the expression of 195 genes encoding proteins with zinc ion binding capacity, or
4 genes (including LIV-1) encoding metal ion transporters. Thus significant shifts in zinc-
requiring proteins and transporters do not appear to be a feature of modified metastatic
potential. However, the availability of zinc can affect macromolecular processes including
stress response pathways that engage cell cycle checkpoints. The presentation will de-
scribe the mapping of the availability of free Zn2+ throughout the cell cycle of tumour
cells using novel live cell reporters and the analysis of the reactive changes in intracellular
free Zn2+ in the cell cycle under genomic stress. Flow cytometry was used to detect an
early but limited increase in the normally low levels of Zn2+ (0.1 fmol/cell) in lymphoma
cells after exposure to agents that induce arrest of cells at the damage-sensitive checkpoint
in G2. The zinc transients were found to collapse as competent cells engaged caspase-
driven apoptosis. The data are consistent with the collapse being a feature of cells des-
tined to undergo DNA fragmentation within the apoptotic response. These findings and
additional studies using the zinc-chelator TPEN suggest that the threshold for DNA
damage-induced apoptosis in cancer cells can be reduced by endogenous-driven changes
in Zn2+ availability. (Professor PJ Smith is a director of Biostatus Ltd, UK.)
12.37
MAGNESIUM AND ZINC DEFICIENCY AND CARDIOVASCULAR
COMPLICATIONS IN SUDDEN UNEXPLAINED DEATH SYNDROME (SUDS)
IN THAILAND
Piyaratana Tosukhowong
Department of Biochemistry, Faculty of medicine, Chulalongkorn University, Bangkok
10330, THAILAND
Sudden Unexplained Death Syndrome (SUDS) disproportionately affects the rural
population of Northeast (NE) Thailand and Southeast Asia. Other common metabolic
diseases found in NE of Thailand are hypokaliuric, hypomagnesiuric, and hypocitraturia
associated renal stone disease, hypokalemic periodic muscular paralysis, and renal
tubular acidosis. Among these four metabolic diseases, intracellular magnesium, potas-
sium, sodium, and zinc play the key roles in pathophysiological consequences. Moreover,
studies of Thai men have demonstarated that SUDS-victims show a characteristic elec-
trocardiographic pattern of right bundle-branch block (RBBB) and ST elevation, as
described in the patients with Brugada Syndrome. Since magnesium, potassium and zinc
deficiency have been shown to contribute significantly to several cardiovascular com-
plications, our researcher group from Bangkok and NE Thailand were interested to
assess mangesium, potassium, and zinc status of patients with sudden respiratory distress
and cardiac arrested who had survived resuscitation attempts or a near-SUDS episode
(N-SUDS). All subjects were free of structural heart disease. Plasma, RBC, WBC and 24
Hr-Urine magnesium and zinc were assessed. The mean levels of magnesium in the RBC,
WBC, and 24-hour urine of N-SUDS patients and rural villagers were significantly lower
than those urban control groups while the plasma levels did not show any differences.
When comparing the Zn-status of N-SUDS with that of the urban groups, the plasma,
RBC, and WBC levels were found to be significantly lower in N-SUDS, while the 24-hour
urine levels was higher. The prevalence of hypomagnesuria, hypozincemia, and hyper-
zincuria was higher in the N-SUDS group. These findings suggest that the homeostasis of
both magnesium and zinc is altered in N-SUDS patients. Our study also found that
potassium depletion is common among N-SUDS patients. We suggest that magnesium
and zinc deficiency might be precipitating factors that reinforce intracellular potassium
deficiency in cardiomyocytes, thereby inducing cardiac arrhythmia or SUDS in people
from NE Thailand who have a positive Brugada sign. From these associations, supple-
mentary potassium, magnesium, and zinc for high risk groups may prevent cardiac
arrhythmia and SUDS events.
12.38
SELENIUM RESTORES INCREASED INTRACELLULAR FREE ZINC
CONCENTRATION IN STREPTOZOTOCIN-INDUCED DIABETIC RAT
CARDIOMYOCYTES
Belma Turan
Department of Biophysics, School of Medicine, Ankara University, 06100, Sihhiye,
Ankara, TURKEY
Evidence from animal models suggests that reactive oxygen species (ROS) play an
important role in the development of diabetic cardiomyopathy. The antioxidants that
have been used to reduce cardiomyopathy lower circulating lipids, which can explain
their beneficial effects in diabetic cardiomyopathy. Since free zinc ion is capable of
altering the function of numerous cellular proteins, its mobilization by ROS in diabetic
animal heart can be likely to cause significant effects. Although the regulation of free
calcium ion concentration in the myocardium is a critical determinant of contractile
performance, current controversies are present regarding the specific alterations in cal-
cium ion signaling pathways to contribute to these cardiac defects during diabetes
mellitus. From the multiple functions of zinc, it is highly likely that any mechanism that
alters the concentration and distribution of intracellular free zinc concentration (IFZnC)
in cardiomyocytes will cause profound functional effects. In our previous study, we have
demonstrated that oxidants increased IFZnC in cardiomyocytes, and could contribute to
oxidant-induced alterations of excitation-contraction coupling. Administration of oral
and/or injectable trace elements and some antioxidants have been proposed as a thera-
peutic adjuvant in the treatment of diabetes. Therefore, considering possible involvement
of oxidative stress-induced alterations in cardiac function, in the present study, we fo-
cused first on the quantification of STZ-induced alterations of IFZnC in cardiomyocytes

and the metallothionein (MT) level of heart tissue, and secondly on the beneficial effects
of selenium on these parameters. We showed that diabetes caused a significant increase in
IFZnC of cardiomyocytes in oxidized glutathione of the heart tissue while decreasing the
MT and reduced glutathione significantly. We also found that sodium selenite treatment
of the diabetic rats markedly restored these altered parameters. Our data indicate that an
oxidant shift of cellular thiolic pools can modulate intracellular zinc metabolism in
diabetic heart. (This work has been supported by Ankara University Research Fund No.
2003-08-0998.)
12.39
BIOAVAILABILITY: AN OVERVIEW OF FACTORS AFFECTING THE
UTILIZATION OF ESSENTIAL TRACE ELEMENTS
Ross M. Welch
USDA-ARS, U.S. Plant, Soil & Nutrition Laboratory, Tower Road, Cornell University,
Ithaca, NY 14853-2901, USA
Essential trace element malnutrition afflicts over 50% of the world’s population especially
among resource-poor families in the Global South resulting in increased morbidity and
mortality rates, reduced labor productivity, lower intellectual attainment, meager family
incomes which greatly contributes to sluggish national development efforts and poor
health and livelihoods for those affected. One of the leading contributing factors to this
global health crisis is the low bioavailability of trace elements in diets comprised of
common staple food crops (e.g., rice, wheat, maize, and beans) that feed the poorest of
the poor. Most of these crops, as commonly eaten, can contain high levels of antinutritive
substances (i.e., phytate, polyphenols, certain fibers, etc.) that greatly reduce the bio-
availability of trace elements in ordinary diets. What sustainable approaches can be used
to alleviate the poor bioavailability of these nutrients in these diets? This review will
discuss potential solutions to this problem focusing on sustainable food system ap-
proaches. The roles that plant breeding, agronomy, and biotechnology can play in
improving the output of bioavailable trace elements in food systems will be discussed as
will means to increase the levels of those factors [e.g., ascorbate, cysteine, the "meat
factor(s)", etc.] that promote the bioavailability of trace elements from diets based on
staple plant foods. The possible negative effects of reducing certain antinutrients (e.g.,
phytate) on both crop productivity and consumer health will be presented. The potential
importance of the hindgut, and associated microorganisms, in determining trace element
bioavailability will also be explored. (R. Welch serves on the planning committee of
HarvestPlus, the CGIAR Biofortification project.)
12.40
IMPROVEMENT OF TRACE ELEMENT NUTRITION IN CHINA
Yiming Xia
Dept. of Trace Element Nutrition, Institute of Nutrition and Food Safety, Chinese
Center for Disease Control and Prevention, 29 Nan Wei Road, Beijing 100050, CHINA
Low or excess intake of trace elements impact trace element nutrition status directly, and
the trace element related diseases would gradually occur. Some of them are recognized as
public health problems due to high prevalence or incidence and wide regional distribu-
tion. In China, supplementation of selenium has tremendously reduced the incidence of
Keshan disease (KD) - a selenium related disease. The national salt monopoly ensures
more than 90% people receiving adequate quantities of iodized salt and the criteria for
goal of elimination of iodine deficiency disorders (IDD) has been almost reached.
Altering drinking water source and improvement of stove design have been used for
preventing endemic fluorosis and arsenism. Extended trial on using NaFeEDTA fortified
soy sauce to control iron deficiency anemia (IDA) is undergoing.
12.41
ASSOCIATION OF LEAD TOXICITY WITH d-AMINOLEVULINIC ACID
DEHYDRATASE (ALAD) AND VITAMIN D RECEPTOR (VDR) GENE
POLYMORPHISMS
Xibiao Ye*, Hua Fu, Weimin Ni
School of Public Health, Fudan University, 200032 Shanghai, PR CHINA
Lead has been a known toxicant for thousands of years, and it remains a persistent
environmental health threat. Toxic effects to nervous, hematologic, renal and repro-
ductive systems have been studies extensively and are well documented. d-Aminolevulinic
acid dehydratase (ALAD) is a principal erythrocytic binding site for lead and is encoded
by a gene localized to human chromosome 9q34. Expression of the two common alleles,
ALAD1 and ALAD2, results in a polymorphic enzyme system with three distinct charge
isozymes, designated 11,12, and 22. There are several polymorphisms of vitamin D
receptor (VDR) gene with the Bsm I polymorphism (VDRB and VDRb alleles) studied
most often. The BB genotype has been associated with lower bone mineral densities,
particularly in women, and may play a role in the development of osteoporosis and
rheumatoid arthritis. The findings suggested that VDR Bsm I polymorphism may modify
calcium metabolism. Because of their similar biochemical nature, lead and calcium can
modify each other’s absorption. It is expected that VDR Bsm I might change lead
metabolism and body lead burden, which may influence lead toxicity. Several groups
have studied the effects of ALAD and VDR polymorphisms on blood lead levels and
related toxicity. However, the study results were inconsistent with each other. None of the
former studies assessed the subjects’ cumulative exposure doses. Therefore, these studies
failed to present effects of gene polymorphisms on dose-response of lead toxicity. The
principle aim of our study was to explore the effects of ALAD and VDR gene poly-
morphisms on dose-response of lead toxicity. There were 216 lead-exposed workers in the
present study. Blood lead (BPb) levels were measured by flameless atomic absorption
spectrophotometry. Blood were measured 3 times, after taking 5 minutes rest. Meta-
analysis showed that ALAD gene polymorphism could play a role in elevation of blood
lead (BPb) level for population with blood lead level more than 25 ug/dl. For subjects
from our study, BPb level were 26.94 and 40.29 ug/dl (P>0.05) for ALAD11 and
ALAD12 groups respectively, 26.64 and 37.94ug/dl(P>0.05) for VDRbb and VDRBb/BB
groups respectively. Compared to ALAD11 group, ALAD12 group had larger elevations
in kidney damage with increasing BPb level. Workers with VDRB allele had larger ele-
vations in systolic blood pressures with increasing BPb level than VDRbb genotypes.
ALAD12 group had higher level of LH than ALAD11 at the significant level of 0.05, 15.55
mIU/ml and 8.19 mIU/ml respectively. However, workers with VDRB allele had lower
level of LH than VDRbb genotypes, 3.26 mIU/ml and 9.38 mIU/ml respectively
(P<0.05). No differences were found for testosterone and follicle-stimulating hormone.
Our analysis and study suggested that ALAD and VDR polymorphisms might change
the pharmacokinetics and lead toxicity of kidney, blood pressure, and reproductive
system.
12.42
INVESTIGATION AND ANALYSIS FOR ARSENICOSIS DUE TO COAL
BURNING IN GUIZHOU PROVINCE, CHINA
A. H. Zhang* [1], X. X. Huang [2], G. H. Yang [1], F. Hong [1], P. Luo [1], X. L. Pan [1]
Dept. Toxicology, Guiyang Medical College, Guizhou 550004, CHINA [1]; 44th Hospital
of PLA, Guizhou 550009, CHINA [2]
Severe arsenic poisoning affects about 3000 people in Guizhou province in southwest
China. The primary cause is the mining and use of arsenic-rich coals, which resulted in
food and air pollution. This pollution and the less developed economic and culture
constituted the main reasons of arsenicosis. In order to better prevent and treat the
disease, many methods were taken to determine and analyze it, such as clinical, bio-
chemical, genetic, toxicologic, immunochemical and molecular pathological ones (in situ
hybridization, PCR-SSCP, PCR product cloning, sequencing, and PCR-based methyla-
tion assay), on the basis of epidemiological investigation. Besides the typical features of
arsenicosis (abnormal dermal pigmentation and hyperkeratosis), liver, kidney, nerve and
eye damage, and irritation of the respiratory tract were also noticed. Dermal cancer and
liver cirrhosis were major sequels causing death. Experimental data showed that arsenic
could influence DNA synthesis and repair, and severely damage DNA. Moreover, it
could induce abnormal expression of oncogene and tumor suppressor, as well as disor-
ders of cell proliferation and apoptosis. Further research found that arsenic caused gene
p53 to G:C fi A:T transition and P16 hypermethylation, resulting in the carcinogenicity.
Arsenicosis is a poly-system damage disease and its mechanism is very difficult to
understand. Only an expeditiously developed local economy, popular health education,
improved living environment, and health protection consciousness can eradicate the
disease successfully. (A. Zhang serves as the Director of Public Health School, Guiyang
Medical College.)

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The Journal of Trace Elements in Experimental Medicine 17:181–272 (2004)

‘‘TRACE ELEMENT NUTRITION AND HUMAN DISEASE’’

Host of ISTERH 2004

ISTERH 2004 Planning Committee Chair

Plenary Speakers and Symposia Committee

Abstracts, Platform and Poster Sessions Committee

Student Travel Award Committee

2004 Wiley-Liss, Inc.

Category 1: Bioavailability of Trace Elements and Dietary Patterns

foods is yet to be clariﬁed. Carnosine (b-alanyl-L-histidine) is found in millimolar

availability was developed. The method is based on a simulated gastrointestinal digestion

dietetic source of the element in food. The selenomethionine proceeds of vegetables

Category 2: Trace Elements and Cancer

Category 3: Trace Elements and Cardiovascular and Pulmonary Diseases

CuD cardiomyopathy as demonstrated by the reversal of depressed cardiac hemody-

2207.5±899.1 lg/g vs. 1665.5±722.2lg/g, respectively, with p<0.05; copper,

levels in STZ-induced diabetic rats. Diabetes was induced by a single intraperitoneal

Category 5: Trace Elements and Disorders of Aging

Category 6: Trace Elements and Fetal Development and Pediatric Disorders

Category 7: Trace Elements and Infectious and Immune Disorders

monolayer of cells was measured by myeloperoxidase activity. Monolayer integrity was

Category 8: Trace Element Molecular Nutrition for the Clinician

Category 9: Trace Elements and Neurological Diseases and Neuropsychological Impairment

demonstrated a statistically signiﬁcant association of severe anemia to lower develop-

University of Warwick, Coventry, ENGLAND CV4 7AL [2]; Cambridge Cognition

Category 11: Toxicity of Trace Elements

Department of Pharmaceutics [3]; Medical School, Department of Biophysics [4];

Methylmercury (MeHg) has been shown to be a neurobehavioral toxicant to the

Category 12: ISTERH 2004 Invited Speakers

Geriatric Unit, Dept. of Internal Medicine, University of Palermo, 90100 Palermo,

Zinc deﬁciency, which is highly prevalent in children in developing countries, results in

expression and activating insulin receptor binding. Zn also functions as a biological

cused ﬁrst on the quantiﬁcation of STZ-induced alterations of IFZnC in cardiomyocytes

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