Invited review for book ‘New Research on Food Habits’. Ed. F. Columbus.

Nova Science Publishers, New

York. Accepted March 2008.

What should we eat: contradictory researches and the confused consumer!

Poonam C. Mittal, Biochemistry Department, University of Allahabad, Allahabad – 211002, India.
e-mail: poonam_mittal@rediffmail.com.

Abstract:

Food habits are constantly being impacted by the perpetually increasing amount of
information available to every person. Print and electronic media is full of advise
regarding what should be eaten and why. Recommendations are often conflicting. Many
foods are declared detrimental to health on one day and beneficial the next. The average
consumer ends up being confused or even disillusioned by the scientific method.

A major reason for the confusion is that any food item is a heterogeneous mixture of
hundreds of compounds, and the effect of one isolated compound may be the opposite of
another compound in the same food. Taking the case of chocolate, if studied for the effect
of sugar, it is declared harmful, but when studied for its flavonoids, it is declared
desirable because it contributes to reduction in oxidative stress.

Confusion also arises due to the fact that findings regarding effect of various food
components on health are based on a variety of experimental and empirical models.
These include epidemiological and experimental studies on humans, animal models, and
in vitro studies of several types, involving tissue culture, cell isolates, and organelle
isolates. But what may work in a simple in vitro model may not be validated by a more
complex living system with more elaborate feedback controls.

Another reason may be that experimental studies generally employ much higher
concentrations of an isolated compound than is found in the natural food. Adding to the
confusion are questions regarding the absorbability of the active compound and the role
and feedback responses of the recipient system, which is ultimately the human body as a
whole. Further, differences may exist between short term and long-term feedback
responses from the recipient system, thus affecting the practical application of a
laboratory finding.

The present review will seek to identify controversies regarding the impact of some
common foods on health, study conflicting reports from existing research, and seek
reasons to explain them in the light of the foregoing. It may finally help us to conclude
that dogmas with regard to food habits are harmful, but may get reinforced if the average,
educated consumer gets a feeling that research is always conflicting and therefore not to
be relied upon.
What should we eat: contradictory researches and the confused consumer!

Poonam C. Mittal, Biochemistry Department, University of Allahabad, Allahabad – 211002, India.

e-mail: poonam_mittal@rediffmail.com.

Introduction

In the early stages of evolution, eating was a biological act, without any capacity
for thinking about what should be eaten and what should be avoided. Selection pressures
must have favored those who got to be programmed with better ability to eat desirable
foods and avoid undesirable ones.

At some stage, the capacity must have developed for observing the effects of
eating different things, memorizing, and passing the information so gained to others,
particularly to offspring. Natural selection would now have favored those who developed
this capacity better. It is no wonder that eating habits form an important component of all
cultures.

Probably in all cultures, food habits, though different, center around the
consumption of cereals and legumes together, making full use of the complementary
nature of their amino acid composition. The ancient seers must have deduced the
importance of various foods and their combinations by keen observation. Records of
some of these have been found, the Vedas being among the earliest records which
originated in the last years of the second millennium and the early years of the first
millennium BC. They contain much food related information as part of the indigenous
Indian system called ayurveda.

Nonetheless, both the biological and the cultural determinants of food habits are
inadequate in today’s world. There are several reasons for this.

Biologically, we are tuned to living in a food scarce world. By and large, cultural
habits have also evolved for such a world. However, for large numbers of people in
today’s world, any amount of food is available just for the asking, without any struggle,
without any realistic fear of having to go hungry the next day. Biological adjustment
cannot be expected to evolve in a short time. Cultural habits have also barely changed to
accommodate this new reality. As a result, there are more people suffering from an
excess of food and consequent obesity than from the problem of scarcity and hunger.

Another recent phenomenon is that the modern world has a huge food processing
industry which introduces artificial foods and molecules at a very rapid rate, compared to
the slow assimilation of food habits as an evolutionary process.

Then there is the pronounced intermingling of cultures due to movement of

people and food products. More and more people are enjoying foods of other cultures. It
is not clear what effects food from other cultures has, when taken by a body that has
evolved in a different niche.
 Clearly, in the modern world, neither biology (my taste buds must have evolved
to know what is best for me) nor conventional wisdom about eating practices (our society
has learnt the right eating practices over centuries best suited to our environment) can
provide adequate guidance.

Today science is an important component of human culture. The scientific method

is expected to provide objective, bias free and reliable guidance to modify our food habits
in order to live a healthier life.

Lavoisier who lived in the 18 th century is credited with having laid the
foundations of modern nutrition. Through the late 19th century till the early part of the
20th century, major developments took place in the area of nutrition, linking observations
of deficiency symptoms to possible causes, leading to discovery of many vitamins. This
was followed by more quantitative concepts to answer questions related to how much of
each nutrient is required, and nutrition became a more organized science. This involved
dependence on other sciences such as physics and technology for hitherto unknown
methodologies, and interaction with newly developing subjects such as biochemistry for
understanding the mechanisms by which food functioned in the body.

A large part of the 20th century had seen war, famine and strife. Naturally,
nutritional studies focused on food shortage and nutrient deficiencies. It was only towards
the seventh decade that the links between affluence, obesity and consequent harmful
effects on health became important issues requiring attention.

The advent of the internet and its easy availability for the common man in the last
two decades, has led to unprecedented access to information. Advice regarding what
should be eaten and why, occupies substantial space in print and electronic media. Food
habits are constantly being influenced by the perpetually increasing amount of
information. Despite this, or because of it, there are more dogmas and misconceptions
related to food and diet related issues than to most other areas of scientific study.
Recommendations are often conflicting, so that many foods are declared detrimental to
health on one day and beneficial the next. To inculcate rationality with regard to food
habits, and to ensure that the average consumer is not disillusioned by the scientific
method, it is desirable to examine some of the reasons for this trend.

Scientific analysis of food related questions can be divided into two approaches.
The first approach deals with analysis of different food items as chemical compounds and
understanding the mechanisms and processes that take place when these compounds enter
the body. The second approach is empirical. It tries to study the effects of a particular
food on a large experimental group, as compared to a control group using statistical
analysis. Of course, the two approaches interact with each other, knowledge gained from
one aiding the design and interpretation of the other.

Every food is a mixture of ‘healthy’ and ‘unhealthy’ compounds:

Conventional wisdom was mostly based on observation, propagated by folklore and

handed down generations. Much of folklore may not withstand the test of modern
scientific methods, for example, the classification of foods into hot and cold categories.
This concept is found in many cultures such as Indian, Chinese and Latin American, but
agreement with regard to details is lacking. For example, rice is considered cold in the
Indian ayurvedic system but neutral in the Chinese system [1]. On the other hand, the
utility of some traditional preparations like Chyawanprash, has been vindicated by
present methods of science. Chyawanprash, an ayurvedic preparation, said to have been
assembled by ancient sages two millenia ago, has the Indian gooseberry as its main
ingredient, now known to be one of the richest sources of ascorbic acid, and also contains
scores of herbs which contribute to the micronutrients required by the body. It has been
found to prevent steroid-induced cataracts [2], induce a greater beneficial effect on
glucose tolerance and lipoprotein profile as compared to vitamin C alone [3], and show
genoprotective efficacy on smokers [4].

Developments in chromatographic, colorimetric, spectroscopic, microbiological and

many other techniques have led to an understanding of biological phenomenon and
biochemical mechanisms. These have found widespread applications to study foods and
their effects on the body. They have also led to better methods of analysis and
characterization of large numbers of compounds in a food.

One of the major reasons for confusion with regard to benefits or harmful effects of
any food is that almost every food item is a heterogeneous mixture of hundreds of
compounds, and the effect of one isolated compound may be the opposite of another
compound in the same food. Therefore, the reported desirability of a particular food
would depend on which of its components has been investigated. Another investigation
based on a different component could lead to a contrary recommendation. Also, if a food
is a mixture of several compounds, and the experiment is designed to ensure that intake
of just one of these compounds is different, the effect attributed to that compound may
actually be because the compound under study interacts with some ingredient in the food
to produce the effect but may not function if taken alone as a supplement. Such
fluctuating reports can leave the general public confused.

Chocolate is just one example that can help explain this statement. Chocolates were
long condemned as junk foods, bad for the teeth, major causative for dental caries, an
unhealthy source of empty calories, full of sugar and saturated fat. Then it came to be
known that chocolates are a rich source of polyphenols such as (-)-epicatechin and (+)-
catechin, and their oligomers called procyanidins, all of which belong to the class of plant
compounds known as flavanols. So they started being investigated for their possible
healthy properties. This was because the health promoting properties of vegetables and
fruit were attributed to the various flavonoids in them [5]. Flavonoids belong to a class of
compounds known as polyphenolics. There are thousands of polyphenolics in the plant
kingdom, of which flavonoids are the most abundant [6]. Soon there was a surge of
studies to investigate the relationship between flavonoids in chocolate and health. The
early years of this decade reported their role in protection against cardiovascular diseases
[6], hypertension [7] and diabetes [8].
 Chocolates have also been recognized as ‘feel good’ food because of their
phenylethylamine content, which however, has also been implicated in the migraine
triggering effect of chocolate. According to a 1998 report [9] analysts have detected more
than 300 chemicals in chocolate. It is not clear whether this includes the polyphenols.

Chocolates also contain alkaloids caffeine and the chemically similar theobromine
[10]. Caffeine has been implicated as the reason of popularity of several drinks such as tea,
coffee and colas because it raises heart rate, blood pressure, and stimulates the brain by
raising dopamine. Theobromine has been implicated in the cough controlling function of
chocolate as well as in its anti-hypertensive function [11, 12]. Chocolate also contains
saturated fat and cholesterol, long considered harmful. However, it is now reported that
cocoa butter which is the saturated fat in chocolate, does not raise bad cholesterol and is
actually beneficial because it prevents the chocolate from sticking to teeth and causing
dental caries. Moreover, it has a melting point just below body temperature at 350 C,
which gives the melt-in-the-mouth quality to the final product.

Apart from the constituents of the cocoa bean, chocolate also contains milk solids and
sugar, so any attributes of chocolates have to be examined in the total product.
Chocolates, as marketed, are also said to promote pimples and acne because of the milk
fat that they contain, but since cocoa contains an abundance of antioxidant properties, the
overall effect on the skin remains an open question [13]. The high sugar content of
chocolates has also been a cause for concern due to a large number of detrimental effects
attributed to sugar [14, 15, 16].

However, the conclusions of a workshopError: Reference source not found [17] in

2003 on ‘Sugars and Health’ failed to establish health concerns for which a direct
association with sugar could be established. The only confirmatory evidence available at
reasonable intakes of sugar was its link to an increased risk and incidence of dental
caries. At high levels of intake of more than 125 g (the equivalent of 25 teaspoons) on a
2000 kcal diet, sugar was found to result in lowering of micronutrient intake (especially
calcium). The final verdict was that “negative energy balance (for example, sedentary
lifestyles) are more important in weight management, insulin sensitivity, and blood lipid
concentrations than is the inclusion or exclusion of any particular dietary component,
including sugars.” In this context, chocolate is also a calorie-rich food with a high fat
content, so daily intake of chocolate requires reducing caloric intake of other foods. It is
pertinent to note that the beneficial effects of chocolate on insulin sensitivity and blood
pressure, described by Grassi and associates [8], were on intakes of 100g of dark
chocolate, providing 480 kcal. The controversies continue, even as a recent study has
reported that older women who consumed chocolate daily had lower bone density and
strength [18].

These are just a few of the studies, based on some of the compounds in one food
product which demonstrate how research findings can confuse a consumer. The Indian
food table [19] lists about six hundred raw foods items, which can be combined in
thousands of ways. Each food, be it wheat, rice or one of hundreds of vegetables,
contains thousands of compounds, which can be studied for their individual properties,
and if the findings of each of these studies makes media headlines, the ensuing confusion
can well be imagined.

In this context, it is noteworthy that there is an explosion of research studies that are
easily accessible to scientists as well as others, due to the internet. More than 15 million
citations are available in MEDLINE, and 10 to 20 thousand are added every week [ 20].
These studies have varying methodology, are conducted on different populations, use
differing test conditions and interventions [21]. Even highly accepted studies are refuted
by subsequent work by the same and/or other investigators [22], so that experts are also
confused by different answers to the same question. Other reasons for complications with
regard to research findings that have been extensively discussed elsewhere include the
biases introduced due to financial and/or career interests of the researchers, the funding
agencies and the food industry [23, 24, 25, 26].

Varied models of research make similar eye-catching headlines:

Confusion also arises because findings regarding effect of various food components
on health are based on a variety of diverse experimental models and empirical studies.
Experimental models may involve in vitro techniques, where the experiment is performed
in a controlled environment, for example in cell isolates or organelle isolates, or where
the cell is made to grow outside the living organism in tissue culture. Since test
conditions may not correspond to those inside a living organism, results may not always
replicate in vivo conditions.

Studies may also be conducted in situ, which is intermediate between in vivo and in
vitro conditions, for example in a cell within the intact organ, but after the animal is
sacrificed, or ex vivo, where experimentation is done in or on living tissue in an artificial
environment outside the organism. Experiments are also conducted on non-human
animals such as rats, mice, dogs or guinea pigs, and results applied to humans, because
there is large physiological similarity between them. Such studies enable researchers to
perform experiments that are not ethical or feasible in humans. However, in all these
models, there is the element of extrapolation.

Studies involving humans are typically empirical, where research is based on

evidence and not just theory. These include limited experimental studies involving
intervention and clinical assessment, but are largely based on epidemiological models
such as cohort, case-control and observational studies. The variables in such studies are
enormous, so findings attributed to a particular event or phenomenon may actually be
caused by another reason. Varied applications of statistics to such studies can also often
yield diverse results for the same data.

We will seek to illustrate this conjecture, based on recent researches on flavonoids,

because in recent years, flavonoids are unparalleled in the media attention that they have
attracted as wonder molecules, and have been at the centre stage of investigation for their
role as anti-oxidants. The advice available to a consumer with regard to this group of
compounds has perhaps occupied more newsprint than any other, with people being
advised to consume large amounts of tea, chocolates, beer, wine, soybean, various
berries. The list of flavonoid-rich healthy foods can go on. Over 5000 naturally occurring
flavonoids, responsible for color and flavor, have been characterized from various plants,
and the USDA database [27] lists the content of several selected flavonoids in 225
commonly consumed foods.

As mentioned earlier, the beneficial effects of fruits, vegetables and beverages such as
tea, coffee, beer, wine and fruit drinks have been attributed more to flavonoids than to
traditional antioxidant vitamins. This is partly because foods contain quantitatively more
flavonoids than the antioxidant vitamins, ascorbic acid, the tocopherols, the carotenoids
[28, 29, 30] . Consequently, the dietary intakes of ascorbic acid, the tocopherols, the
carotenoids have been found to be one order of magnitude less (70, 7-10 and 2-3 mg
respectively) than that of total flavonoids. Early studies reported flavonoid intakes to be 1
and 1.1 g/day [31]. Later studies, using better analytic methodologies, corroborate that the
recommended nine daily servings of fruits and vegetables and moderate amounts of tea,
coffee, wine, beer, or chocolate can provide well over 1000 mg of total phenols per day
[32].

A large number of studies have demonstrated that ingestion of flavonoid-rich foods

raises the antioxidant levels of blood and tissues, which in turn is responsible for their
antiviral, anti-allergic, antiplatelet, anti-inflammatory, antitumor functions, and also for
prevention of cancer and cardiovascular diseases [33, 34]. These conclusions have been
derived from studies on animals [35, 36, 37], cell isolates [37, 38, 39], in vitro models of
human or animal tissues[40, 41]. Thus the strong antioxidant capacity and free-radical
scavenging activities of flavonoids in vitro seems indubitable and interest in the possible
health benefits of flavonoids has increased.

Studies on humans have mostly indicated a protective effect of flavonoids [42, 43, 44]
even though there are a few reports to the contrary [45]. Yet, epidemiologic studies
exploring the role of flavonoids in human health have been inconclusive. Some studies
support a protective effect of flavonoid consumption in cardiovascular disease and
cancer, other studies demonstrate no effect, and a few studies suggest potential harm.
Because there are many biological activities attributed to the flavonoids, some of which
could be beneficial or detrimental depending on specific circumstances, it has been
suggested that further studies in both the laboratory and with populations are required
[46].

Questions have also been raised with regard to their absorption, and the emerging
view is that their absorption is much lower and their half-life much shorter than that of
other dietary antioxidants such as ascorbic acid and tocopherols, suggestive of the
viewpoint that their capability to act as antioxidants in vivo is limited [47]. However, in
vivo studies have consistently shown an increase in total antioxidant capacity of plasma
on ingestion of flavonoids rich foods, leading to more questions regarding these
conflicting findings. It has now been postulated that the increase in total plasma
antioxidant capacity on ingestion of a wide variety of flavonoid rich fruits and vegetables
is a secondary phenomenon resulting from their extensive metabolism to urate in vivo,
and that the macro- and micronutrients present in fruits and vegetables may directly or
indirectly affect the total antioxidant capacity of plasma [32].

So the question regarding the in vivo efficacy of flavonoid-rich foods for their health
promoting functions remains wide open, even as, in addition to the natural sources, the
market is replete with nutritional supplements of specific flavonoids, such as quercetin,
isoflavones, catechins and various other bioflavonoids. In this context, the question arises
whether an isolated flavonoid will provide the same health benefit as it would, if it were
present in the whole food, and even whether the isolated compound may actually be
harmful [48].

Notwithstanding whether flavonoids have the beneficial effects attributed to them in

recent times or not, it is likely that if each of the studies on flavonoids made newspaper
headlines, as they have been found to do, dietary advise to the public to drink ‘n’ number
of cups of tea / coffee / wine / juice of any of several berries, and include ‘x’ bars of
chocolate and ‘y’ servings of tofu, yoghurt, salads, honey, fish, green chillies, olive oil
etc. etc. per day would add up to several kilograms of food. It is more likely than not that
this would translate into an excessive intake of energy, consequent obesity, and damage
to health!

Quantity matters! Physiological vs. pharmacological levels of intake:

Experimental studies generally employ much higher concentrations of an isolated

compound than is found in the natural food. When findings of such researches reach the
common man, he looks for ways to achieve them, and is led to non-dietary sources of the
nutrient. Thus nutritional supplements, called nutraceuticals, have become very popular.

The amounts of various nutrients, especially vitamins and minerals, recommended by

national and international agencies such as the Food and Nutrition Board (US) [49, 50] the
FAO [51] and the ICMR [52 ] are generally much lower than those supplied as
supplements. They are also not in the proportions as recommended, or as found in natural
foods. The question arises whether our metabolic machinery is designed to handle
amounts that are larger than can be obtained from diet. It is well known that fat-soluble
vitamins are absorbed readily, stored for prolonged periods in the liver but cannot be
readily excreted in urine. What is not clear is whether the large stores impact the
metabolism in the body. A recent meta-analysis of studies dealing with vitamin E
supplementation spanning from 1966 to 2004 revealed that high dosage >400 IU/d of
vitamin E supplements may increase all-cause mortality and should be avoided, even as
lower doses (< 150 IU/d )may have beneficial effects [53]. The amounts typically
available from dietary sources are approximately 14 IU/d) [54]. Similarly, high doses of
β -carotene supplementation have been reported to increase mortality rates [55].

Supplements of minerals are also common, but it is well accepted that minerals can
have negative interactions with other minerals, which can impact their intestinal
absorption, transport, utilization and storage.
 Calcium and iron are two minerals, the absorption of which are known to depend on
need of the body, because they are heavy metals, not easily excreted in the urine due to
low solubility of their salts.

Iron deficiency is the most common nutritional deficiency in the world, and has been
implicated in a wide range of problems, such as developmental delays, behavioral
disturbances and cognitive deficits in children and increased risk for a preterm delivery
and low-birth weight babies in women. Supplementation of iron has been prescribed,
almost universally, for pregnant women, adolescent girls, and newborns. However,
concerns have been raised with regard to its possible harmful effects [56], and maternal
complications consequent to oxidative stress have been reported when iron is given as a
prophylactic to pregnant women who do not have iron-deficiency anemia [57]. The
necessity of routine iron supplementation during pregnancy has been debated in
industrialized countries, but is still advocated in developing countries, because traditional
diets provide inadequate iron and where malaria and other infections causing increased
losses are endemic [58].

Ascorbic acid is routinely administered with inorganic iron supplements to keep the
ferrous salt in the reduced state. However, this has been reported to have adverse effects,
such as oxidative DNA damage and consequently cancer in well nourished adults [59] and
increased oxidative stress in the gastrointestinal tract [60]. High tissue iron concentrations
have been associated with the development and progression of several pathological
conditions, including certain cancers, liver and heart disease, diabetes, hormonal
abnormalities, and immune system dysfunctions, due to free radical-mediated tissue
damage, even as ingestion of antioxidant rich foods may prevent or delay primary and
secondary effects associated with iron overload-related diseases. [61].

Absorption of zinc is known to get reduced by non-heme iron supplementation,

although postabsorptive interactions between these nutrients are less clear [62]. Excessive
intake of zinc can reduce copper absorption, and excessive copper intake can result in
reduced absorption of manganese, zinc, and iron [63].

Among the macrominerals, calcium has attracted maximum attention as a desirable

supplement, especially for elderly women who are at risk for osteoporosis. Dietary
calcium deficiency has not been established as the major etiological factor for
osteoporosis, yet supplements are being taken by most postmenopausal women and even
elderly males as a prophylactic measure against osteoporosis. Apart from its benefits on
bone health, calcium supplementation has been linked to increase in high density
lipoprotein cholesterol and decrease in low density lipoprotein cholesterol [64]. However,
calcium supplementation has been reported to adversely affect vascular health [65] by
accelerating vascular calcification [66, 67, 68, 69]. More recently, calcium supplementation
in healthy postmenopausal women has been found to be associated with upward trends in
cardiovascular event rates [70]. It has also been implicated in development of brain lesions
[71].

Such examples, of positive and negative consequences of supplements providing

doses of known nutrients, which are nutritionally unattainable, abound in literature.
 Distinction is required between physiological and pharmacological doses. It is
therefore important that pharmacological doses should not be taken without sufficient
reason, that the consumer should be informed of possible harmful effects, that they
should be administered under supervision, and that their potentially detrimental effect
should be balanced against the likely benefits. Amounts obtained from a balanced diet
cannot be excessive, and therefore are unlikely to cause harm, so they should be
encouraged. Pharmacological doses of nutritional supplements should also be regulated
like other medicines, to be taken only under medical advice.

Short-term responses are different from long-term responses:

It is generally assumed that more is better, and this stands true for nutrients, growth
rates and parameters such as blood hemoglobin, even as it has been argued that functional
criteria are preferable [72]. However, calorie restriction is beneficial in the long run, and
promotes longevity [73, 74, 75 ], even though in the short run, growth is compromised. It is
possible that short-term responses to any deficiency are different from the long term
responses. So in case of energy deficits, for example, the immediate response is slowing
down of growth, but in the long run, as an adaptive measure, the organism is known to
reduce metabolic rates and achieve catch up. Life has evolved through shortages, and the
physiology of all organisms is designed to hold on to nutrients. Our digestive system is
designed to absorb several times the amounts of dietary fuels, carbohydrate, protein and
fat, normally present in the diet. However, it is unable to adapt to the relatively recent
phenomenon of availability of excessive food by reducing digestion and absorption. Our
adipose tissue is designed to hold on to the absorbed excess. Anyone who has lost weight
knows how difficult it is to keep it off. There is intricate metabolic regulation to conserve
body fat, and regain it even if availability of energy remains low, as in continued dieting.
It has been suggested that the human body is likely to have a genetically determined set-
point weight that is controlled by metabolic hormones and fat cell enzymes [76].

On the other hand, in countries such as India, poor children continue to grow on
dietary energy intakes barely above those required for basal metabolism, and the
gestation and lactation performance of poor women compares favorably in many respects
with that of upper class women [77]. In animal studies it has been shown that the
nutritional inadequacy of the diet is, to a large extent, compensated for, by better
utilization of dietary energy, protein and improved efficiency of nitrogen utilization [78, 79,
80 81
, ]. Also, rats switched from a low protein diet in early life to a moderate protein diet,
and back to a low protein diet were found to display a more efficient and rapid adaptation
to the switches, as judged by growth as well as rate of serum protein turnover, than those
switched from a high protein to a low protein diet after an intervening period on a
moderate protein diet [82]. This suggests that adaptive mechanisms that come into play
during periods of protein deprivation appear to persist later when the stress is reduced,
and may in fact be beneficial to the organism. Similarly, it is interesting to find that rats
fed carotene as the only source of vitamin A in early life were more efficient in
converting it to vitamin A later in life, than those fed preformed vitamin A throughout [ 83]
emphasizing the long term responses of the organism to nutrient utilization.
 The difference between short term and long-term feedback responses from the
recipient system are also likely to affect the practical application of laboratory findings,
because of elaborate feedback controls found in the complex living system.

The role of feedback responses of the recipient system: the complex human
body.

The recipient system of any nutrient can consist of a fairly homogenous system such
as found in a cell isolate or tissue culture, or it can be as heterogeneous and complex as
the human body. The human body is a conglomerate of about a hundred trillion cells
(which incidentally is four orders of magnitude more than all the Homo sapiens on earth!)
and they work in marvelous unison to maintain homeostasis, (or is it homeodynamics?).
To do this, there are intricate feedback mechanisms, so the effect of a particular
compound on the homogenous system may not get extrapolated to the complex system.
In biochemistry, it is generally found that if one set of chemicals drives the system in a
given direction, another set of chemicals will drive it in an opposing direction.

If one or both of these opposing influences are non-linear, an equilibrium point(s)

results to which the system gravitates. Any perturbation, generally in the form of a
chemical stimulus resulting from the influx of a nutrient, nutrient supplement, drug, or
even thought, can trigger a negative feedback system to reestablish the equilibrium. Since
ultimately the system moves to a predetermined equilibrium, there may be results of an
immediate response to an intervention, which are often not sustained in the long run, as
feedback systems come into play.

As a consequence of negative feedback mechanisms, many qualitatively different

stimuli oppose each other, and finally arrive at a point close to the equilibrium point. By
positive feedback, the system responds in the same direction as the perturbation, so the
signal is amplified. In biological systems, the controls are typically based on negative
feedback, through varied mechanisms. Many multienzyme pathways in metabolism are
regulated; the final product inhibits an early reaction in the pathway, after it is formed in
sufficient quantities, thus preventing excessive amounts of its own formation. Hormonal
regulation of blood sugar, blood pressure, body temperature and erythropoiesis are but a
few examples of well documented negative feedback controls.

Disruption of negative feedback can lead to undesirable results. Once the principle
that equilibrium is to be maintained in the long run is acknowledged, it is logical to
accept that all perturbations will eventually get attenuated, explaining why many drugs,
supplements and nutrients produce the expected result in short term studies, but in the
long run, the efficacy is lost.

Most traditional methodologies used in biological sciences have adopted the

reductionist approach to knowledge. However, as discussed in relation to feedback
approaches, the system is likely to behave differently when impacted by diverse stimuli,
than when it is in a controlled environment. One of the great current debates in biology
concerns whether the observed behavior of a system can be accounted for in terms of the
behaviors of its subcomponents, and it has been suggested that holistic approaches may
be more predictive and make for better understanding of the functioning of the body.
Systems biology develops these concepts and attempts to understand the integrated
function of complex, multicomponent biological systems ranging from interacting
proteins that carry out specific tasks to whole organisms [84]

Scientists dealing with varied aspects of food, nutrition and nutritional biochemistry
have more recently been involved in developing new approaches to accommodate the
complexities of the organism, leading to the development of new disciplines such as
nutrigenomics, which has the potential to prescribe tailored dietary regimens specific to
the individuals’ requirements.

Nutrigenomics links genomics, transcriptomics, proteomics and metabolomics to

human nutrition. Genomics allows the study of the genetic abilities of an individual to
metabolize nutrients based on its entire genome. It includes the study of genome-nutrient
interactions, including the role of nutrients and dietary components in regulation of
genome structure, expression and stability and the role of genetic variation on individual
nutrient requirements. Transcriptomics allows the global study of gene expression at the
RNA level, which will ultimately determine the extent of transcription of a gene.
Knowing the proteomics will help in determining whether the enzymes and other
required proteins have the desired conformation to perform optimal catalytic function as
required. Finally, it includes metabolomics, the development of which began in 1970, to
investigate the ideas of Linus Pauling with a view to studying relationships between
biological variability and wide ranges of nutritional requirements.

Metabolomics [85], specifically nutritional metabolomics, is concerned with

metabolic pathways and networks and includes regulation of metabolic pathways and
networks by nutrients and other food components. It summates all the metabolites in
body fluids, which are impacted by endogenous factors such as age, sex, body
composition, genetics, underlying pathologies, circadian rhythms and resting metabolic
rate and exogenous factors such as diet, including all known and hitherto unknown
nutrients as well as non-nutrients such as dietary fiber, additives, pollutants, drugs etc.,
and the large number of signals from the intestinal microflora. A very large number of
compounds make the metabolome, which can be likened to a metabolic fingerprint which
reflects the balance of an individual’s metabolism.

These compounds need to be identified, quantified and their relative proportions

analyzed and interpreted. This has led to the development of metabonomics which is
concerned with the quantitative measurement of the metabolome. [85]. Metabolomics
requires the establishment of analytical methods that can profile human serum and
urinary metabolites to assess nutritional imbalances and disease risk. Such analysis will
require the application of sensitive techniques such as nuclear magnetic resonance,
functional magnetic resonance imaging and high performance liquid chromatography,
and handling of a large amount of mathematical data.
 It should be clear from the foregoing, that the utilization of compounds from food
in the body is an extremely complex issue, with large individual variations due to
interactions between genetics and overall food habits. It is generally not true that every
laboratory finding is applicable to every one in the population. Therefore, categorical
statements about the benefits, or otherwise, of a food do not apply universally. Such
statements are inherently misleading and should be avoided.

Limitations of the methodology of science: The role and responsibility of the

media:

Statistics forms the basis for evaluating the results of modern scientific studies,
whether based on observation or on varied experimental designs. But even the basic
assumptions, on which statistical tests are based, are not devoid of controversy.

Usually, inferences are drawn, in a mechanical manner, using statistical techniques,

without appreciation of the assumptions on which these techniques are based. Very often
the techniques are based on assumptions of normal distribution, even for such data as are
not drawn from normal distribution.

If different samples are taken from a normal population, they will usually yield
different sample means. One of the most frequently used statistical techniques, the null
hypothesis testing, provides the probability that two samples have been drawn from the
same normal distribution. If this probability is less than an arbitrarily chosen number p
(usually 0.05, 0.01, or 0.001), the two samples (experimental group and the control
group) are regarded as drawn from different populations. It is usual to interpret this by the
statement that the treatment given to the experimental group has been effective at the
significance level p.

The null hypothesis testing is used in a very wide variety of experimental designs
because it allows a crisp decision – to accept or reject a hypothesis. It is used to compare
an experimental group with a control group, to determine whether they belong to the
same normal population, with regard to the parameter under investigation. It is assumed
that the populations under study are matched with regard to all other variables except the
one being investigated. Results are presented as dichotomous –the null hypothesis is
accepted or not, thereby concluding that the effect is / is not observed.

In the above procedure, the conventional choice of p = 0.05, 0.01 or 0.001 seems to
be arbitrary. The choice of p should depend on the nature of the question asked and an
assessment of the benefits and risks involved.

The practice of using the p value for testing the null hypothesis has been criticized for
arbitrarily dividing results into significant and not significant [86, 87]. Extremely small
differences can become statistically significant if the sample size is sufficiently large, and
find their way into scientific writings of repute. Null hypothesis significance testing has
been called a ‘research quality assurance test’ and is a requirement for experimental
research to be published. However, it is not always appreciated that this widely used
statistical methodology is not without its problems when extended to practical advice;
small statistically significant differences may not be of practical significance.

It is important that reports of scientific research are conveyed to the consumers in a

reasonably accurate manner. As discussed earlier, there is an extremely large, and often
conflicting, amount of scientific data being generated, and the consumer ends up being
confused.

It is important that the consumer is guided to be able to make informed choices with
regard to healthy food and desirable food habits. Media should desist from giving the
misleading impression that everyone who eats a particular food, or follows a particular
practice, will benefit from it. It is therefore suggested that media should report scientific
findings to the consumer only when reasonable consensus has been reached on a
particular issue. In the absence of such a consensus, the reports should include full details
of the study design and findings in a language that is comprehensible to the typical
educated consumer yet does not give misleading impressions. For example, they should
give the sample size and the extent of the effect observed, how many among the
experimental group/s exhibited a response in a direction opposite to the group average
and how many among the control group showed the response even without the
intervention. The consumer should also be made aware of the limitations of the scientific
methodology. The tendency to reduce a study to a simple statement expressing that ‘x’
food / habit causes ‘y’ effect needs to be restricted, both by scientists and media.

The common man should be encouraged to take his own decisions regarding
whether to try out a particular food for a particular condition. The media should facilitate
such decision making, by improving its information content, and seek to educate the
consumer to take informed decisions, rather than make categorical headlines about foods
as if they apply to all individuals in a society. For example instead of saying ‘Eat
chocolates for lowering blood pressure’, media should be encouraged to say that of ‘n’
persons who ate ‘m’ grams of chocolate everyday, ‘x’ persons showed a ‘y-z’ percent
reduction in blood pressure.

Headlines based on laboratory findings based on in vitro or animal studies can be

even more damaging. For example if a purified compound from a plant shows a health
benefit on rats, but makes media headlines that everyone who consumes a food
containing that compound will benefit from it, the result can sometimes even be harmful.
We need to have guidelines for media to refrain from making potentially irresponsible
headlines.

Conclusion

In the final analysis, we can conclude that there is a never-before quantum of

information available at the click of a button, but uncertainties and methodological
limitations are inherent in the scientific method. This is certainly not to denigrate research
per se, which, despite conflicting observations, or because of them, certainly promotes
understanding of food-physiological interactions and leads to wide applications in disease
amelioration and promotion of good health. Nevertheless, it is important to emphasize
that advice about food is a complex issue. In complicated medical conditions, food
choices can even make the difference between life and death.

Therefore, it is necessary to caution the media from creating euphoria over

viewpoints that may not extend to in vivo functionality based on superficial reports of
research findings. Education, in the modern world, must endeavor to make people
understand the strengths and limitations of the scientific method in general and nutritional
research in particular.

References:
. Bogumil, C. Humoral Theory In Cultural Food Beliefs. 2002 June10. Available from:
1

http://food.oregonstate.edu/ref/culture/humoral.html.
2
. Velpandian, T; Mathur, P; Sengupta, S; Gupta, SK. Preventive effect of Chyavanprash against steroid
induced cataract in the developing chick embryo. Phytotherapy Research, 1998. 12 (5): 320-323.

. Manjunatha, S; Jaryal, AK; Bijlani, RL; Sachdeva, U; Gupta, SK. Effect of Chyawanprash and
3

vitamin C on glucose tolerance and lipoprotein profile. Indian J Physiol Pharmacol. 2001. 45(1):71-9.

. Yadav, JS; Thakur, S; and P. Chadha. Chyawanprash Awaleha: A Genoprotective Agent for Bidi
4

Smokers. Int J Hum Genet. 2003. 3(1): 33-38.

5
. Hung, HC; Joshipura, KJ; Jiang, R; et al. Fruit and vegetable intake and risk of major chronic disease.
J Natl Cancer Inst. 2004. 96: 1577– 84.

. Fraga, CG. Cocoa, diabetes, and hypertension: should we eat more chocolate? Am J Clin Nutr. 2005;
6

81: 541–2.

. Keen, CL; Holt, RR; Oteiza, PI; Fraga, CG; Schmitz HH. Cocoa antioxidants and cardiovascular
7

health. Am J Clin Nutr. 2005. 80(suppl):1S– 6S.

8
. Grassi, D; Lippi, C; Necozione, S; Desideri, G; Ferri, C. Short-term administration of dark chocolate
is followed by a significant increase in insulin sensitivity and a decrease in blood pressure in healthy
persons. Am J Clin Nutr. 2004. 81: 611– 4
9
. Emsley, J. Molecules at an exhibition: the science of everyday life. Oxford University Press.1998.

. Usmani, OS; Belvisi, MG; Patel, HJ; Crispino, N; Birrell, MA; Korbonits, Korbonits, MD; Barnes,
10

PJ. Theobromine inhibits sensory nerve activation and cough. FASEB Journal. 2005. 19 (2): 231-233.

. Smit, HJ; Gaffan, EA; Rogers, PJ. Methylxanthines are the psychopharmacologically active
11

constituents of chocolate. Psychopharmacology (Berl). 2004. 176: 412–9.

. Kelly, CJ. Effects of theobromine should be considered in future studies. Am J Clin Nutr. 2005. 82:
12

483–9.

. Parker; M; Pond, D; Smith, W; Watson, A. A systematic review of the evidence for 'myths and
13

misconceptions' in acne management: diet, face-washing and sunlight. Family Practice. 2005. 22 (1):
62-70.
14
. Furth, A; Harding, J. Why Sugar Is Bad For You. New Scientist. Sep 23, 1989.
15
. Ceriello, A. Oxidative Stress and Glycemic Regulation. Metabolism. 2000. 49(2 Suppl 1):27-29.
16
. Yudkin, J; Kang, S; Bruckdorfer, K. Effects of High Dietary Sugar. Br J. Med. 1980; 1396.

. Lineback, JMJ; Rand D. Sugars and Health Workshop: summary and conclusions. Am J Clin Nutr.
17

2003. 78 (suppl):893S–7S.
 . Hodgson, JM; Devine, A; Burke, V; Dick, IM; Prince, RL. Chocolate consumption and bone density
18

in older women. Am J Clin Nutr. 2008. 87, 1, 175-180.

. Gopalan, C; Rama Sastri, BV; Balasubramanian, SC. Revised and updated by Narasinga Rao BS,
19

Deosthale YG, Pant KC. Nutritive value of Indian foods. National Institute of Nutrition, Indian Council
of Medical Research, Hyderabad, India. 1996.

. National Library of Medicine: Fact Sheet Medline.

20

http://www.nlm.nih.gov/pubs/factsheets/medline.html. Accessed 2007. November 15.

. Garg , AX; Hackam, D; Tonelli, M. Systematic Review and Meta-analysis: When One Study Is Just
21

not Enough. Clin J Am Soc Nephrol. 2008. 3: 253–260.

. Ioannidis, JP: Contradicted and initially stronger effects in highly cited clinical research. J Am Med
22

Assoc. 2005. 294: 218–228.

23
.Nestle, M. Eating made simple. Sci Am. 2007, Sept. 60-69.

. Nestle, M. Food Politics: How the Food Industry Influences Nutrition and Health. Revised edition.
24

University of California Press, 2007.

. Hilden, J; Jorgensen, AW; Gotzsche, PC. Cochrane reviews compared with industry supported meta-
25

analyses and other meta-analyses of the same drugs: Systematic review. Br Med J. 2006. 333: 782.

. Lesser, LI; Ebbeling, CB; Goozner, M; Wypij, D; Ludwig, DS. Relationship between Funding
26

Source and Conclusion among Nutrition-Related Scientific Articles. PLoS Medicine. 2007. 4, 1: e5,
41–46.

. Nutrient Data Laboratory; Beltsville Human Nutrition Research Center, USDA. Nutrient database
27

for standard reference, release. USDA Database; 2003. Available at

http://www.nal.usda.gov/fnic/foodcomp).

. Vinson, JA; Su, X; Zubik, L; Bose, P. Phenol antioxidant quantity and quality in foods: fruits. J.
28

Agric. Food Chem. 2001. 49:5315–5321.

29
.Lotito, SB; Frei, B. Relevance of apple polyphenols as antioxidants in human plasma: contrasting in
vitro and in vivo effects. Free Radic. Biol. Med. 36:201–211; 2004.

. Aaby, K; Skrede, G; Wrolstad, RE. Phenolic composition and antioxidant activities in flesh and
30

achenes of strawberries (Fragaria ananassa). J. Agric. Food Chem. 2005. 53:4032–4040.

. Kuhnau J. The flavonoids: a class of semi-essential food components: their role in human nutrition.
31

World Rev. Nutr. Diet. 1976. 24:117–91.

. Frei, B and Lotito, BS. Consumption of flavonoid-rich foods and increased plasma antioxidant
32

capacity in humans: Cause, consequence, or epiphenomenon? Free Radic Biol Med. 2006. 41: 1727–
1746.
 . Aviram, M; Fuhrman, B. Wine flavonoids protect against LDL oxidation and atherosclerosis. Ann.
33

N. Y. Acad. Sci. 2002. 957:146–161.

. Rietveld, A; Wiseman, S. Antioxidant effects of tea: evidence from human clinical trials. J. Nutr.
34

2003. 133:3285S–3292S.

. Goyarzu, P; Malin, DH; Lau, FC; et al. Blueberry supplemented diet: effects on object recognition
35

memory and nuclear factor-kappa B levels in aged rats. Nutr Neurosci. 2004. 7(2):75-83.

. Frei, B; Higdon, JV. Antioxidant activity of tea polyphenols in vivo: evidence from animal studies. J
36

Nutr. 2003. 133(10):3275S-3284S.

. Kavanagh, KT; Hafer, LJ; Kim, DW; et al. Green tea extracts decrease carcinogen-induced
37

mammary tumor burden in rats and rate of breast cancer cell proliferation in culture. J Cell Biochem.
2001. 82(3):387-398.

. Spencer, JP; Schroeter, H; Crossthwaithe, AJ; Kuhnle, G; Williams, RJ; Rice-Evans, C. Contrasting
38

influences of glucuronidation and O-methylation of epicatechin on hydrogen peroxide-induced cell

death in neurons and fibroblasts. Free Radic Biol Med. 2001. 31(9):1139-1146.

. Spencer, JP; Chen, JJ; Ye, ZQ; Koo, MW. Growth inhibition and cell cycle arrest effects of
39

epigallocatechin gallate in the NBT-II bladder tumour cell line. BJU Int. 2004. 93(7):1082-1086.

. Cheng, IF; Breen, K. On the ability of four flavonoids, baicilein, luteolin, naringenin, and quercetin,
40

to suppress the Fenton reaction of the iron-ATP complex. Biometals. 2000. 13(1):77-83.

. Polagruto, JA; Schramm, DD; Wang-Polagruto, JF; Lee, L; Keen, CL. Effects of flavonoid-rich
41

beverages on prostacyclin synthesis in humans and human aortic endothelial cells: association with ex
vivo platelet function. J Med Food. 2003. 6(4):301-308.

. Geleijnse, JM; Launer, LJ; Van der Kuip, DA; Hofman, A; Witteman, JC. Inverse association of tea
42

and flavonoid intakes with incident myocardial infarction: the Rotterdam Study. Am J Clin Nutr. 2002.
75(5):880-886.

. Yochum, L; Kushi, LH; Meyer, K; Folsom, AR. Dietary flavonoid intake and risk of cardiovascular
43

disease in postmenopausal women. Am J Epidemiol. 1999. 149(10):943-949.

. Liu, RH. Health benefits of fruit and vegetables are from additive and synergistic combinations of
44

phytochemicals. Am J Clin Nutr. 2003. 78(3 Suppl):517S-520S.

. Freese, R; Vaarala, O; Turpeinen, AM; Mutanen, M. No difference in platelet activation or

45

inflammation markers after diets rich or poor in vegetables, berries and apple in healthy subjects. Eur J
Nutr. 2004. 43(3):175-182.

. Ross, JA; Kasum, CM. Dietary flavonoids: bioavailability, metabolic effects, and safety. Annu Rev
46

Nutr. 2002. 22:19-34.

. Manach, C.; Williamson, G.; Morand, C.; Scalbert, A.; Remesy, C. Bioavailability and bioefficacy
47

of polyphenols in humans: I. Review of 97 bioavailability studies. Am. J. Clin. Nutr. 2005. 81:230S–
242S.
48
Lotito, SB; Frei, B. Relevance of apple polyphenols as antioxidants in human plasma: contrasting in
vitro and in vivo effects. Free Radic. Biol. Med. 2004. 36:201–211.

. Food and Nutrition Board. Dietary reference intakes for calcium, phosphorus, magnesium, vitamin D
49

and fluoride. Washington DC: National Academy Press, 1997.

. Food and Nutrition Board. Dietary reference intakes: Thiamine, riboflavin, niacin, vitamin B6, folate,
50

vitamin B12, pantothenic acid, biotin, and choline. Washington DC: National Academy Press, 1998.

. WHO/ FAO/ IAEA. Trace elements in human nutrition and health. Geneva: World Health
51

Organization. 1996.
52
. Indian Council of Medical Research. Nutrient requirements and recommended dietary allowances
for Indians, National Institute of Nutrition, Hyderabad, 1995.

. Miller, III ER; Pastor-Barriuso, R; Dalal, D; Riemersma, RA; Appel, LJ; Guallar, E. Meta-Analysis:
53

high-dosage vitamin E supplementation may increase all-cause mortality. Ann Intern Med. 2005;
142:37-46.

. Institute of Medicine. Dietary Reference Intakes for Vitamin C, Vitamin E, Selenium, and
54

Carotenoids. A report of the Panel on Dietary Antioxidants and Related Compounds, Subcommittees
on Upper Reference Levels of Nutrients and Interpretation and Uses of Dietary Reference Intakes, and
the Standing Committee on the Scientific Evaluation of Dietary Reference Intakes. Food Nutrition
Board. Washington, DC: National Academies Pr; 2000.

. Vivekananthan, DP; Penn, MS; Sapp, SK; Hsu, A; Topol, EJ. Use of antioxidant vitamins for the
55

prevention of cardiovascular disease: meta-analysis of randomized trials. Lancet. 2003; 361:2017-23.

. Scholl, TO; Reilly, T. Anemia, iron and pregnancy outcome. J Nutr. 2000.130(2S Suppl):443S-
56

447S.

Scholl,TO. Iron status during pregnancy: setting the stage for mother and infant Am J Clin Nutr.
57

2005. 81(5):1218S-1222S.
58
. Mungen, E. Iron supplementation in pregnancy. J Perinat Med. 2003. 31(5):420-6.

. Rehman, A; Collis, CS; Yang, M; Kelly, M; Diplock, AT; Halliwell, B; Rice-Evans, C. The Effects
59

of Iron and Vitamin C Co-supplementation on Oxidative Damage to DNA in Healthy Volunteers.

Biochem Biophys Res Comm. 1998. 246, 1: 293-298.

. Fisher, AEO; Naughton, DP. Iron supplements: the quick fix with long-term consequences. Nutr
60

Journal. 2004. 3:2.

61
. Fraga, CG and Oteiza, PI. Iron toxicity and antioxidant nutrients. Toxicology. 2002. 180: 1, 23-32.

. Donangelo, CM; Woodhouse, LR; King, SM; Viteri, FE; King, JC. Supplemental Zinc Lowers
62

Measures of Iron Status in Young Women with Low Iron Reserves. J. Nutr. 2002.132:1860-1864.
 . Anderson, JJB. Minerals. In Krause’s Food, Nutrition and Diet therapy. Mahan LK, Escott-Stump S
63

(Eds) WB Saunders Co. Philadelphia, 2000. p 111.

. Reid, IR; Mason, B; Horne, A; Ames, R; Clearwater, J; Bava, U; et al. Effects of calcium
64

supplementation on serum lipid concentrations in normal older women: a randomized controlled trial.
Am J Med. 2002.112:343-7.

. Reid, IR; Schooler, BA; Hannon, S; Ibbertson, HK. The acute biochemical effects of four proprietary
65

calcium supplements. Aust N Z J Med. 1986.16: 193-7.

. Pletcher, MJ; Tice, JA; Pignone, M; Browner, WS. Using the coronary artery calcium score to
66

predict coronary heart disease events: a systematic review and meta-analysis. Arch Intern Med. 2004.
164: 1285-92.

. Chertow, GM; Burke, SK; Raggi, P. Treat to Goal Working Group. Sevelamer attenuates the
67

progression of coronary and aortic calcification in hemodialysis patients. Kidney Int. 2002. 62: 245-52.

. Block, GA; Spiegel, DM; Ehrlich, J; Mehta, R; Lindbergh, J; Dreisbach, A; et al. Effects of
68

sevelamer and calcium on coronary artery calcification in patients new to hemodialysis. Kidney Int.
2005. 68:1815-24.

. Asmus, HG; Braun, J; Krause, R; Brunkhorst, R; Holzer, H; Schulz,W; et al. Two year comparison
69

of sevelamer and calciumcarbonate effects on cardiovascular calcification and bone density. Nephrol
Dialysis Transplant. 2005. 20:1653-61.

. Bolland, MJ; Barber, PA; Doughty, RN; Mason, B; Horne, A; Ames, R; Gamble, GD; Grey A and
70

Reid, IR. Vascular events in healthy older women receiving calcium supplementation: randomised
controlled trial. Br Med J. published online 15 Jan 2008.

. Payne, ME; Anderson, JJB; Steffens DC. Calcium and vitamin D intakes are positively associated
71

with brain lesions in depressed and nondepressed elders. FASEB J. 2007. 21:837.

. Beaton, GH. Iron needs during pregnancy: Do we need to rethink out targets? Am. J. Clin. Nutr.
72

2000. 72 (suppl):265S-71S.
73
. Weidruch, R. Calorie restriction and aging. Sc Am. January 1996, p32-38.
74
. Fontana, L. Excessive adiposity, calorie restriction and aging in humans. J Am Med Assoc. 2006.
293:13.
75
. Heilbron, LK;, de Jonge L; Frisard, MI; DeLany, JP; Enette, D; Meyer, L; Rood, J; Nguyen, T;
Martin, CK; Volaufova, J; Most, MM; Greenway, FL; Smith, SR; Williamson, DA; Deutsch, WA;
Ravussin, E. Effect of 6-month calorie restriction on biomarkers of aging, metabolic adaptation and
oxidative stress in overweight subjects. J Am Med Assoc. 2006. 293:13.

. Pi-Sunyer FX. Obesity. In Modern Nutrition and health and disease. Eds. Shils ME, Olson JA, Shike
76

M, Ross AC. Ninth edition. Williams and Wilkins, Baltimore. 1999. 1395-1418.
77
. Mittal PC. Pregnancy Outcome In Relation To Anemia, Nutrient Intakes and Socio-economic Status.
Abstract no. M16, p51. Micronutrient Forum Meeting, Istanbul, 16-18 April 2007.
78
. Mittal PC. Lactation performance, maternal behavior and activity of rats subjected to neonatal and/or
post weaning food restriction. Nutr Rep Int. 1984. 30, 305-310.

. Mittal, PC. Nitrogen balance during reproduction of rats fed diets simulating those consumed by
79

different population groups. Nutr Rep Int.1984. 30, 1355-65.

80
. Mittal, PC. Response of rats to lysine deficiency at different ages. Nutr Rep Int. 1985. 32, 453-461.
81
. Mittal, PC. Nitrogen balance during reproduction of second-generation rats fed diets typical of
different socio-economic strata. Nutr Rep Int. 1986. 33, 25-32.
82
. Mittal, PC. Response of rats to variations in dietary protein content. Nutr Rep Int. 1985. 31, 521-533.

. Mittal, PC. β - Carotene utilization in rats fed either Vitamin A or carotene in early life. Nutr Rep
83

Inter. 1983. 28, 181-188.

. Strange, K. The end of "naive reductionism": rise of systems biology or renaissance of physiology?
84

Am J Physiol Cell Physiol. 2005. 288: 968-974.

. Gibney, M J; Walsh, M; Brennan, L; Roche, HM; German, B; van Ommen, B. Metabolomics in

85

human nutrition: opportunities and challenges. Am J Clin Nutr. 2005. 82: 497-503.

. Sterne, JAC; Smith, GD. Sifting the evidence—what's wrong with significance tests? Br Med J.
86

2001. 322 – 27.

87
. Ioannidis, JPA. Why most published research findings are false. PLoS Med. 2005. 2(8): e124.