Intake of bilberries (Vaccinium myrtillus L.) reduced cardiovascular diseases

risk factors through positive influences in lipoprotein profiles

Marta Habanova, Jorge A. Saraiva, Miroslav Haban, Marianna Schwar-

zova, Peter Chlebo, Lenka Predna, Jan Gažo, Joanna Wyka

PII: S0271-5317(16)30693-5
DOI: doi: 10.1016/j.nutres.2016.11.010
Reference: NTR 7700

To appear in: Nutrition Research

Received date: 3 June 2016

Revised date: 16 November 2016
Accepted date: 18 November 2016

Please cite this article as: Habanova Marta, Saraiva Jorge A., Haban Miroslav,
Schwarzova Marianna, Chlebo Peter, Predna Lenka, Gažo Jan, Wyka Joanna, In-
take of bilberries (Vaccinium myrtillus L.) reduced cardiovascular diseases risk factors
through positive influences in lipoprotein profiles, Nutrition Research (2016), doi:
10.1016/j.nutres.2016.11.010

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 ACCEPTED MANUSCRIPT

Intake of bilberries (Vaccinium myrtillus L.) reduced cardiovascular diseases risk

factors through positive influences in lipoprotein profiles

Marta Habanova a,*, Jorge A. Saraiva b,*, Miroslav Haban c, Marianna Schwarzova a, Peter

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Chlebo a, Lenka Predna a, Jan Gažo d, Joanna Wyka e

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a
Department of Human Nutrition, Slovak University of Agriculture in Nitra, 94976, Slovakia

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b
Department of Chemistry, University of Aveiro, Campus Universitário de Santiago, 3810-
193 Portugal

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c
Department of Pharmacognosy and Botany, Faculty of Pharmacy, Comenius University in
Bratislava, 83232, Slovakia
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d
Department of Genetics and Plant Breeding, Slovak University of Agriculture in Nitra,
94976, Slovakia
e
Department of Human Nutrition, Wrocław University of Environmental and Life Sciences,
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51630, Poland

*Corresponding authors at: Jorge Manuel Alexandre Saraiva, Universidade de Aveiro,

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Campus Universitário de Santiago, 3810-193 Aveiro, Portugal, Tel.: +351 234401513, fax:
+351 234370084 (email: jorgesaraiva@ua.pt) and Marta Habanova, Department of Human
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Nutrition, Slovak University of Agriculture in Nitra, Trieda Andreja Hlinku 2, 94976, Nitra
Slovakia, Tel.: +421 37 641 4467, fax: +421 37 6415599 (email addresses:
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marta.habanova@uniag.sk, marta.habanova1@gmail.com).

E-mail addresses: marta.habanova@uniag.sk (M. Habanova), jorgesaraiva@ua.pt (J.

Saraiva), miroslav.haban@gmail.com (M. Haban), Marianna.schwarzova@uniag.sk (M.
Schwarzova), peter.chlebo@hotmail.com (P. Chlebo), lenkapredna@gmail.com (L. Predna),
jan.gazo@uniag.sk (J. Gažo), joanna.wyka@up.wroc.pl (J. Wyka)

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Abbreviations: CVD, cardiovascular diseases; T-C, total cholesterol; LDL-C, low-density

lipoprotein cholesterol; HDL-C, high-density lipoprotein cholesterol; TG, triglycerides; ALT,
alanine aminotransferase; AST, aspartate aminotransferase; GMT, gamma-
glutamyltransferase; ALP, alkaline phosphatase; TAS, total antioxidant status; AA,

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antiradical activity

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ABSTRACT

Cardiovascular diseases (CVDs) are the leading causes of death, and lifestyle modification,
including dietary changes, is recommended to improve this condition. In this study, regular

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consumption of bilberries was hypothesized to have beneficial effects on CVD risk reduction,

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by changes in human health indicators such as decreasing low-density lipoprotein cholesterol

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(LDL-C), and triglycerides (TG) and increasing high-density lipoprotein cholesterol (HDL-
C). The research involved women (n = 25) and men (n = 11) who consumed 150 g of frozen

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stored bilberries three times a week for 6 weeks. Anthropometric parameters, blood pressure,
lipid profile, glucose, liver enzymes, creatinine, albumin, magnesium, and antiradical activity

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were measured. Except for the body mass index (BMI) of women (P = 0.019), no significant
changes were found for anthropometric indicators. The consumption of bilberries led to a
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decrease in the following parameters: total cholesterol (T-C, P = 0.017), LDL-C (P =
0.0347), TG (P = 0.001), glucose (P = 0.005), albumin (P = 0.001), gamma-
glutamyltransferase (GMT, P = 0.046), and a positive increase in HDL-C (P = 0.044). In
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men, additionally favorable changes were observed in T-C (P = 0.004), glucose (P = 0.015),
albumin (P = 0.028), aspartate aminotransferase (AST, P = 0.012), GMT (P = 0.013), and in
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HDL-C (P = 0.009; in this group LDL-C increased [P = 0.007]). Changes in other parameters
were not significant, for both women and men. Thus, the regular intake of bilberries can be
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important to reduce CVDs risk, by decreasing LDL-C/TG and increasing HDL-C.

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Keywords: bilberries, lipid profile, cardiovascular diseases risk, biochemical parameters,

normal subjects

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1. Introduction

Bilberries (Vaccinium myrtillus L.) are recognized as a good source of anthocyanins 1,
chlorogenic acid, flavonoids, alpha-linolenic acid, pterostilbene, and vitamins 2, 3. Recent

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studies demonstrated the benefit of consuming bilberries and blueberries to prevent age-

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related chronic diseases such as cardiovascular diseases (CVDs) 4-6, cancer 7-11, diabetes

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12-14, hyperlipidemia 15, hypertension 16, obesity 17, metabolic syndrome 18], and
antiinflammation 19-21]. Bilberries have a high content of total polyphenols and

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anthocyanins, and their phenolic composition has been often the subject of various studies
22-27. The detailed analysis of phenolics in fresh bilberries and blueberries confirmed that

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anthocyanins are the main phenolics in both bilberries and blueberries, as previously reported
in two studies 28-29.
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While limited epidemiological data inversely associate the consumption of berries with
inflammation and CVD, these conclusions need to be strengthened. Clinical studies in healthy
human subjects with diabetes mellitus, dyslipidemia, metabolic syndrome, hypertension, or
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smokers show a significant decrease in CVD risk factors, especially glucose, lipids and lipid
peroxidation, and systolic blood pressure, after the consumption of berries. The principal
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mechanisms of action underlying the potential cardio-protective effects of berries include

counteracting free radical generation, attenuating inflammatory gene expression,
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downregulating foam cell formation, and upregulating endothelial nitric oxide synthase
(eNOS) expression. Through these effects, the progression of atherosclerosis is slowed, and
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normal vascular function and blood pressure are preserved. Moreover, in light of the possible
decrease in nutritional and functional value that occurs during food processing, including
drying and pasteurization, the consumption of fresh or minimally processed foods, such as
frozen whole berries as part of a regular diet, may be better than the intake of juices or
extracts that need more elaborate processing for consumption 5.
To test our hypothesis, the present study examined the effects of regular consumption of
bilberries on CVD risk reduction. Specifically, bilberries cause positive changes in human
health indicators, such as decreasing low-density lipoprotein cholesterol [LDL-C] and
triglycerides [TG] and increasing high-density lipoprotein cholesterol [HDL-C]). Volunteers
(apparently healthy women and men) ingested whole frozen preserved bilberries during 6
weeks and anthropometric (weight, height, and volume of the body), blood pressure, and
several blood health indicators (lipid profile/(T-C, LDL-C and HDL-C, and TG), as well as

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alanine aminotransferase [ALT], aspartate aminotransferase [AST], gamma-

glutamyltransferase [GMT], alkaline phosphatase [ALP], creatinine, albumin, glucose,
magnesium [Mg], and antiradical activity [AA]). These parameters were monitored at the
beginning and after 3 and 6 weeks of the initiation of the ingestion of bilberries.

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2. Methods and materials

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2.1. Participants and study design

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The objective of the present study was to evaluate the effect of regular consumption of

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bilberries on parameters related to energy and blood lipids, antioxidant status (antiradical
activity), and other human health indicators, based on the possibility that the intake of
bilberries reduce CVD risk factors. This was a pre–post intervention study 30, involving a
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total of 65 subjects from the staff of the Slovak Agriculture University who were screened for
eligibility to participate in the study. Twenty two subjects did not satisfy the inclusion
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criteria and seven subjects dropped out (Figure). Twenty-five women (n = 25) with a mean
age (± SD) 47.7±5.23 and 11 men (n = 11) with a mean age (± SD) 49.00±6.48, all of whom
were apparently healthy, were included in the study. The inclusion criteria were as follows:
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volunteers could not have been treated for cardiovascular, renal, and hepatic diseases, could
have neither high cholesterol nor high blood pressure for the last 6 months, and could not
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have intake of any nutritional supplements, namely (vitamins, minerals, antioxidants, and
flavonoids).
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Volunteers were asked not to change neither their eating habits nor their lifestyle during
the study. Anthropometric parameters (weight, height, and volume of the body parameters)
and blood pressure were observed and recorded. The following were monitored during the
study: blood profiles (T-C, LDL-C, HDL-C, TG) and ALT, AST, GMT, ALP, creatinine,
albumin, glucose, Mg, and AA. The values obtained before (pre) the study were used as
control.
This study was conducted according to the guidelines of the Declaration of Helsinki, and
approved by the Slovak Agriculture University in Nitra, Department of Human Nutrition,
Slovakia, and by the Ethical Committee of the Specialized Hospital of St. Svorad Zobor in
Nitra, Slovakia, study No. 02/0906/2015. All subjects gave written, informed consent.

2.2 Intervention

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To test the biological efficiency of bilberries (Vaccinium myrtillus L.), wild bilberries
originating from the region Kysuce, and from the northwestern part of Slovakia were used.
The volunteers were instructed to consume 150 g of frozen bilberries three times per week

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(Monday, Wednesday, and Friday, during the day, e.g. as a snack with natural yogurt) for 6

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weeks. The amount (per dry weight) of some important bioactive compounds (total

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polyphenols and total anthocynins, quercetin, chlorogenic acid, and pterostilbene) as well as
the AA of the bilberries were quantified. The total phenolic content was determined

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according to the Folin-Ciocalteu method 40, the total anthocyanin content was analyzed as
described by Giusti and Wrolstad 41, and the quantification procedures for quercetin and

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chlorogenic acid were adapted from the work of Justesen and Knuthsen 42. The AA of the
bilberries extract was measured using the stable radical 2.2-diphenyl-1-picrylhydrazyl DPPH
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33, 34, and the content of pterostilbene was quantified according to Remsberg et al. 45.

2.3 Anthropometric measurements

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Anthropometric measurements were performed by trained personnel before (pre) at the

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beginning of the consumption of the bilberries (week 0) and at the end (post) of the study
(week 6). Body weight (kg) and height (m) were measured on the outpatient electronic
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medical scales TANITA WB-3000. The body mass index (BMI) was calculated by dividing
the body weight in kg by the square of the height in meters. Body fat (%) was determined
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using a bioelectric impedance analyzer Bodystat Quadscan 4000 apparatus, and waist
circumference was quantified using Tanita ViScan 140. Systolic and diastolic blood pressure
were measured twice in mmHg using a Sphygmomanometer DM-3000 while participants
were seated at screening visits, and the means of measurements were used for analysis.
Subjects rested for at least 15 minutes before the measurements.

2.4 Clinical analyses

Blood samples were obtained as follows: the first samples (before the start of bilberry
consumption) were used as control, the second blood sampling was performed after 3 weeks
of bilberry consumption, and the third blood sampling was performed after 6 weeks of
bilberry consumption (at the end of the study). Blood was collected in the morning after 8

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hours of fasting in a standard manner using 2.5 mL of a ethylenediaminetetraacetic acid

(EDTA) solution and in a 7.5 mL serum gel tube. After the separation of blood serum
samples, they were stored at -80°C until the analyses were carried out. The lipid profile in
blood serum (T-C, LDL-C, HDL-C, and TG), ALT, AST, GMT, ALP, creatinine, albumin,

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glucose, Mg, and TAS were determined from thawed plasma samples using a biochemical

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analyzer LISA 200, with commercially available kits Spinreact (Spain) from Ecomed

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Slovakia and Randox (UK). Methods for the determination of biochemical parameters were
programmed using the application protocol, provided by the supplier of reagents (Ecomed,

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Slovakia) and adapted when necessary. The lipid profile was compared with the reference
values of The National Cholesterol Education Program: Adult Treatment Panel III (NCEP

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ATP III) [46]. To convert the concentration to mmol/L, values in mg/dL were multiplied by
0.02586 for T-C, HDL, and LDL and multiplied by 0.01129 for TG. [47].
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2.5 Statistical analyses
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Data are expressed as means ± standard deviation (SD) (Tables 1 and 2) and as median and
interquartile ranges (Table 3). A power analysis was conducted in G*Power to determine a
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sufficient sample size using an alpha value of 0.05, a power of 0.80, and a large effect size
48. The data were analyzed by t-test and the Wilcoxon signed-rank test, depending on the
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data under analysis, using Statistica CZ10 software. Differences were considered statistically
significant at P < 0.05.
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3. Results

The amount of relevant bioactive compounds (total polyphenols and total anthocynins,
quercetin, chlorogenic acid, and pterostilbene), as well as the AA of billberries is shown in
Table 1 together with comparison with values reported in the literature 8, 31-39. Generally
the results obtained in the present work were in the range of the results reported by others 8,
31-39.
The analyzed anthropometric parameters and blood pressure of the participants before
(pre) and after (post) the consumption of bilberries are presented in Table 2. No significant
differences were observed for all monitored anthropometric indicators at the start and at the
end of the study, except for a slight increase in the BMI of women (P = 0.019).

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Concerning the parameters quantified in the blood samples, the results are presented in
Table 3, where it can be seen that T-C levels decreased from 5.63 mmol/L at the beginning of
the research to 5.29 mmol/L (P = 0.017) after bilberries intervention. Similar observed
beneficial effects were found for LDL-C (3.39 to 3.14; P = 0.035), TG (1.46 to 1.06; P =

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0.001), glucose (5.54 to 5.14; P = 0.005), albumin (47.90 to 45.60; P = 0.001), and GMT

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(0.54 to 0.49; P = 0.046), and there was a favorable increase in HDL-C (1.57 to 1.67; P =

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0.044). Similar advantageous changes were observed for men for T-C levels that decreased
from 5.38 mmol/L to 4.96 mmol/L (P = 0.004), glucose (5.95 to 5.24; P = 0.012), albumin

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(48.77 to 46.64; P = 0.028), AST (0.47 to 0.37; P = 0.012), and GMT (0.71 to 0.58; P =
0.013), and there was a beneficial increase in HDL-C (1.28 to 1.49; P = 0.009). In men, LDL-

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C levels increased (3.26 to 3.78; P = 0.007). The changes observed for the other parameters
were not statistically significant.
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The percentage change for the lipid profiles in the number of subjects (T-C, LDL-C,
HDL-C, and TG) in plasma before (pre) and after, following a 3-week and 6-week (post)
intake of bilberries according to NCEP ATP III, is presented in Table 4. Generally, the lipid
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profiles for the number of volunteers improved. The number of volunteers with optimal
values, near or above the optimal and borderline high values, increased and the number of
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volunteers with high and very high values decreased. This is evaluated as positive in terms of
reducing CVD risk factors.
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4. Discussion
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Results of the present study show that the short-term inclusion of bilberries in the diet can
contribute to improved blood plasma lipid profiles of women and men although the LDL-C
levels increased in men they were still below the considered risk values (> 4 mmol/L). Other
studies also pointed to the benefits of bioactive substances of bilberries for health 22-24, 49-
51. Wild bilberries (Vaccinium myrtillus L.) are currently not commercially available on the
market, but only cultivated species of blueberries can be purchased (Vaccinium corymbosum
L.). The composition of bioactive compounds of bilberries and the comparison with that of
blueberries was studied by Giovanelli and Buratti 52 and Habanova et al. 27. Their results
indicate a high concentration of important bioactive substances in bilberries, contributing to
increasing references to bilberries as a natural functional food 53, with potential health
benefits.

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The amount of bilberries consumed by volunteers was selected to be comparable to that of

other studies, taking into account the processed form used in other studies such as
fresh/frozen/dried, juice, smoothie, beverages, or capsules 54. The anthropometric
parameters and blood pressure of volunteers before (pre) and after (post) bilberries

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consumption that were obtained show that the consumption of bilberries led to a significant

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change in the BMI of women (but still less than 1% increase) and had no significant effect on

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the other studied parameters (body weight, waist circumference, body fat, body mass, and
blood pressure). Basu et al. 55 have previously reported some health benefits of blueberries

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consumption. In the study by these authors, 48 participants with metabolic syndrome
consumed freeze-dried blueberries, and the decrease in plasma oxidized LDL and serum

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malondialdehyde and hydroxynonenal concentrations was higher in the group ingesting
blueberries (28 and 17%, respectively), than in the control group (9 and 9%; P = 0.01,
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respectively).

In our study, the positive effects of the consumption of bilberries on CVD risk factors
were also observed. Basu et al. 55 and Erlund et al. 56 reported similar results, but our
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findings are not directly comparable with the results of these authors, because they used
different doses of berries or a different time for consumption, and additionally, the study was
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carried out with volunteers with CVD risk (obese men and the metabolic syndrome in
women). The results of the two teams of previously cited authors 55-56 showed significant
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reductions of CVD risk factors linked to the consumption of berries. The intake of bilberries
in the present work was linked to the normalization/improvement of several parameters in
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blood related to CVD risk factors, similarly to results presented by other authors 57-59.
Stamler et al. 60 evaluated the relationship between serum cholesterol and the risk of
premature death from coronary heart diseases in 356,222 men aged 35 to 57 years. This was
the largest cohort with standardized serum cholesterol measurements and long-term mortality
follow-up. Of all CVD deaths, 46% were estimated to be deaths attributable to serum
cholesterol excess levels of 180 mg/dL or greater (≥4.65 mmol/L). Approximately 1 mmol/L
or lower T-C is associated with approximately a one-half lowering of CHD mortality in both
sexes at ages 40-49, 50-69, and 70-89 years.

Moreover, beneficial changes (%) in the lipid profile of volunteers before (pre) compared
to a 3-week and 6-week (post) intervention of bilberries were found in the present work,
according to NCEP ATP III 46. The results show that there was a favorable shift of
volunteers with risk values into more favorable lipid profile categories. Results from
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intervention studies aimed at monitoring the impact of fruits rich in polyphenols to improve
health status were reported by several authors 56-57, 61-65, indicating that regular
consumption of small fruits plays an important role in preventing CVD. Clinical trials that
use diet or drugs to lower serum cholesterol levels have consistently shown a 2% reduction in

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the incidence of coronary heart disease for every 1% reduction in total serum cholesterol

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level 66.

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Halliwell 67 reported that despite the results of several intervention studies, some cause-
effect relationships are still unclear, because the relationships and mechanisms are complex.

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For example, it is known that free radicals are detrimental, because they contribute to aging
and certain age-related diseases, especially cancer and neurodegenerative diseases, and

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therefore, antioxidants mechanism are difficult to interpret. However, the intervention trials
with antioxidants often failed to demonstrate benefits in humans.
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In addition, different results reported in the literature may arise from the diversity of study
designs, insufficient control, relatively brief intervention periods, low doses of active
substances, and as a result of low amounts of consumption of the tested foods, like berries.
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Also, some studies do not indicate details on the content of the polyphenols of consumed
berries, making it difficult to compare data. To confirm the observations verified in the
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present study, it is recommended that well-organized intervention studies be conducted to

investigate a wider range of fruits and the long-term impact of their consumption on the body
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68-69.
In conclusion, the present study shows that even a short period of regular consumption of
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whole wild bilberries, preserved through minimal processing (frozen), is associated with an
improvement of the lipid profile in humans. The results obtained in the present work support
the hypothesis designed to investigate the study that the regular consumption of bilberries can
contribute to beneficial effects on CVD risk reduction, such as decreasing LDL-C and TG
and increasing HDL-C. Nonetheless, one has to be aware that the current study was limited in
a number of ways that deserve careful attention: first, the study population was relatively
small, and this could be the main reason why significant between-group differences for many
of the biochemical parameters assessed in this study were not detected; second, the current
study was subject to potential bias because of the lack of blinding and placebo controlling;
third, the intake period in the current trial was 6 weeks, which may not have been sufficient
to detect significant differences for some of the parameters studied; and lastly, it would be

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ideal to standardize the intake of berries based on the content of phytochemicals, such as
anthocyanins and flavonoids, rather than on crude fruit weight.

Acknowledgments

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This work was supported by the Scientific Grant Agency of the Ministry of Education,

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Science, Research and Sport of the Slovak Republic no. 1/0127/14. The determination of the
capacity of total polyphenols and antioxidants of the plant sources of natural and

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agroecological conditions of the Slovak Republic and their utilization in improving
population health; project ITEBIO “Support and innovations of a special and organic

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products Technologies for human healthy nutrition” ITMS 26220220115, implemented under
Operational Program Research and Development and by FCT/MEC to the QOPNA research
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unit (FCT UID/QUI/00062/2013), through national funds and where applicable co-financed
by the FEDER, within the PT2020 Partnership Agreement. Authors also acknowledge
editorial assistance to improve the manuscript. The authors have no conflict of interest to
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disclose.
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FIGURE

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Volunteers screened for eligibility

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(n = 65)
Excluded (n = 22)

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Not meeting
inclusion/exclusion

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criteria (e.g.,
choleserol)
medications)

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Women (n = 30) Men (n = 13) Pre
Pre (before) bilberries (before) bilberries
intervention analyses intervention analyses
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(control) (control)
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Drop out Drop out

(n = 5) (n = 2)
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Post (after) bilberries Post (after) bilberries

intervention intervention
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Data analyzed in Data analyzed in men

women group (n = 25) group (n = 11)
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Figure - Flowchart of subject selection in the pre-post intervention and those that completed
the study.

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TABLES

Table 1 - Amount (in dry weight) of bioactive substances in frozen bilberries used in

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the present study and comparison with literature valuesa
Parameter Units Values of the Values of References

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present study

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Total polyphenols mg/kg 20752.50±848.66 868.3-2950.9 [31]
Total anthocyanins mg/kg 3038.25±74.44 2651±781 [32]

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2878.0±607.0 [33]
2050.0±566.0 [34]
3985.0±1290.0 [35]
2619.0-9296.2 [36]

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Quercetin µg/g 6.10±0.23 29-30 [37]
Chlorogenic acid µg/g 25.86±0.48 23.1±1.0 [38]
Pterostilbene ng/g 190.00±22.5 99-520 [8]
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Antiradical activity, inhibition % 76.937±1.474 73.54±1.94 [39]
of DPPH
a
The data are expressed as means ± standard deviation (for each sample, n = 4).
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Table 2 - Anthropometric parameters and blood pressure measurements of

volunteers before (pre) and after (post) bilberries interventiona, b
Parameter Women (n=25) Men (n=11)

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Units Pre Post Pre Post

P
Body weight kg 73.23±13.45 73.68±13.61 90.67±15.68 90.95±15.78

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(0.062) (0.556)
Waist

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circumference cm 87.24±12.19 88.24±12.56 99.00±11.84 98.91±11.37
(0.100) (0.922)
Body fat % 34.06±6.39 33.31±7.60 23.64±5.58 22.98±5.58

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(0.722) (0.746)
Body fat mass kg 25.51±9.04 25.67±10.03 21.88±7.83 22.21±8.43
(0.215) (0.217)
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BMI kg/m2 26.3±4.73 26.5±4.78 29.04±3.94 29.15±4.03
(0.019) (0.545)
Systolic blood
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pressure mmHg 131.4±15.86 133.56±17.31 134.0±14.52 134.45±12.69

(0.430) (0.881)
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Diastolic blood
pressure mmHg 76.4±12.09 78.24±11.59 85.55±8.64 82.91±7.38
(0.250) (0.355)
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a
n = number of subjects; pre = measurements taken before (pre) of the study; post =
measurements taken at the end of the study; data are expressed as means ± standard
deviation; t-test; P < 0.05.
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b
Values in parentheses show the P-value.

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Table 3 - Change in blood plasma parameters of volunteers before (pre) and after 6-weeks
(post) bilberries interventiona, b

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Women (n=25) Men (n=11)

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Units Pre Post P-value Pre Post P-value
T-C mmol/L 5.82 (4.74- 5.19 (4.85- 0.017 5.36 (4.77- 4.94 (4.47- 0.004

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6.39)1 5.74) 6.07) 5.43)
HDL-C mmol/L 1.52 (1.38- 1.70 (1.44- 0.044 1.22 (0.98- 1.49 (1.21- 0.009

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1.81) 1.88) 1.52) 1.68)
LDL-C mmol/L 3.42 (2.60- 2.95 (2.75- 0.035 3,05 (2.95- 2.79 (2.50- 0.007
4.15) 3.68) 3.78) 3.12)

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TG mmol/L 1.56 (0.83- 0.99 (0.84- 0.001 1.69 (0.92- 1.39 (0.66- 0.061
2.02) 1.24) 2.82) 2.06)
TAS mmol/L 1.69 (1.66- 1.72 (1.63- 0.553 1.85 (1.71- 1.89 (1.66- 0.789
1.75) 1.80) 1.98) 1.94)
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Glucose mmol/L 5.43 (5.02- 4.92 (4.57- 0.005 5.97 (4.83- 5.05 (4.61- 0.012
5.62) 5.37) 6.44) 6.25)
Mg mmol/L 0.89 (0.82- 0.84 (0.82- 0.103 0.82 (0.77- 0.82 (0.75- 0.307
0.94) 0.92) 0.96) 0.87)
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Creatinin mol.l-1 76.2 (71.8- 72.5 (66.7- 0.353 88.6 (79.1- 84.9 (74.7- 0.062
9.90) 79.1) 103.2) 98.1)
Albumin g.l-1 46.8 (45.4- 46.0 (44.5- 0.001 49.2 (47.5- 46.9 (45.0- 0.028
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49.6) 47.2) 50.0) 48.6)

ALT kat.l-1 0.15 (0.09- 0.16 (0.12- 0.976 0.29 (0.19- 0.25 (0.17- 0.059
0.28) 0.25) 0,52) 0.39)
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AST kat.l-1 0.34 (0.29- 0.35 (29- 0.236 0.34 (0.30- 0.33 (0.27- 0.012
0.44) 0.39) 0.59) 0.49)
GMT kat.l-1 0.39 (0.29- 0.34 (0.25- 0.046 0.47 (0.31- 0.43 (0.32- 0.013
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0.68) 0.48) 0.98) 0.83)

ALP kat.l-1 2.19 (1.88- 2.10 (1.85- 0.989 2.13 (1.50- 1.78 (1.53- 0.894
2.64) 2.47) 2.62) 2.79)
a
n = number of subjects, pre = blood samples taken before (pre) the study, post = blood samples taken
at the end of the study, T-C = total cholesterol, LDL-C = low-density lipoprotein cholesterol, HDL-C =
high-density lipoprotein cholesterol, TG = triglycerides, TAS = total antioxidant status, Mg =
magnesium, ALT = alanine aminotransferase, AST = aspartate aminotransferase, GMT = gamma-
glutamyltransferase, ALP = alkaline phosphatase.
b
Data are expressed as medians and interquartile ranges in parentheses. Statistical significant results
differences were verified by the Wilcoxon test, P < 0.05.

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Table 4 - Distribution percentage changes (%) of blood lipids in subjects before (pre),
following a 3-week intervention of bilberries, and after a 6-wk (post) intervention of
bilberries according to NCEP ATP III.
Volunteers (%)

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Reference values of NCEP ATP III Women (n=25) Men (n=11)

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Baseline 3-wk 6-wk Baseline 3-wk 6-wk

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T-C
desirable ( 5.18 mmol/L) 38.9 50.0 50.0 42.8 42.8 85.7
higher borderline (5.2 – 6.19 mmol/L) 33.3 27.7 33.3 42.8 28.6 14.3

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high ( 6.2 mmol/L) 27.8 22.3 16.5 14.4 28.6 0
LDL-C
optimal ( 2.6 mmol/L) 0 11.2 0 0 0 14.3
near or above optimal (2.6-3.3 mmol/L) 27.8 22.2 22.2 28.6 57.1 28.6

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higher borderline (3.4-4.1 mmol/L) 22.2 38.9 55.4 28.6 14.3 42.8
high (4.2-4.9 mmol/L) 27.8 22.2 11.2 42.8 28.6 14.3
very high (> 4.9 mmol/L) 22.2 5.5 11.2 0 0 0
HDL-C
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high ( 1.55 mmol/L) 66.7 77.8 83.3 42.8 57.2 57.2
higher borderline (1.04-1.54 mmol/L) 27.8 22.2 16.7 57.2 42.8 42.8
low ( 1.03 mmol/L) 5.5 0 0 0 0 0
TG
optimal ( 1.7 mmol/L) 66.8 77.9 83.4 42.8 42.8 57.1
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higher borderline (1.70-2.25 mmol/L) 27.7 16.6 11.1 14.3 28.6 28.6
high (2.26-5.64 mmol/L) 5.5 5.5 5.5 28.6 28.6 14.3
very high ( 5.65 mmol/L) 0 0 0 14.3 0 0
Baseline, 3-week, and 6-week = number of participants in % in each category before
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intervention, following a 3-week intervention, and after 6-week (post) intervention. T-C
=total cholesterol, HDL-C and LDL-C = high-density and low-density lipoprotein
cholesterol, TG = triglycerides, compared with the reference values of The National
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Cholesterol Education Program: Adult Treatment Panel III (NCEP ATP III) [46].
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