Food compositions for managing body weight

Abstract

Methods for managing the body mass index, the body fat percentage and/or the distribution of fat storage sites of humans are provided, and more particularly the use of food compositions for managing the body mass index, the body fat percentage and/or the distribution of fat storage sites by administering to a human in need of managing body mass index, body fat percentage and/or distribution of fat storage sites an effective amount of a food formulation comprised of an esterified pectin having a degree of esterification (DE) of less than 65%.

Claims

1. A method of managing visceral fat of a human, the method comprising administering to the human in need of managing visceral fat a food formulation comprised of a low methylated (LM) pectin having a degree of esterification (DE) of at least 1% to less than 50% to achieve an effective amount of the LM pectin of 0.01 g to 5 g per Kg of body weight of the human per day.

2. The method according to claim 1, wherein the LM pectin is not amidated.

3. The method according to claim 1, wherein the method further comprises obtaining the LM pectin by the steps comprising: (i) extracting a high methylated (HM) pectin having a DE of more than 50% from a pectin containing plant; (ii) controlling acidity, temperature and time of extraction according to step (i) to obtain the LM pectin; and (iii) including the LM pectin in the food formulation administered to the human in need of managing visceral fat.

Description

EXAMPLES

Example 1 Incorporation of LM Pectin in Diets Leads to More and Smaller Meals

(1) Rat Feeding Trial Set-Up

(2) 17 Male Wistar rats (weight±320 g; Harlan Netherlands BV, Horst, The Netherlands) were individually housed in TSE cages in a climate-controlled room (21 C±1) under a 12 h:12 h light-dark cycle (lights on at 10:00 AM). These specialized cages were equipped with food weighing sensor for continuous registration of food intake for multiple days (TSE Systems GmbH, Bad Homburg, Germany) to monitor circadian feeding patterns, meal sizes and meal numbers. Circadian food intake patterns were calculated as an average of the last two consecutive days, the first day was used for adaptation. These plexiglass cages (40×23×15 cm) consist of a sensitive weight balanced food station (stainless steel food container for standard size food pellets). Animals were maintained ad libitum on the diets. Water was available ad libitum throughout the study. Food intake and body weights were measured daily at 10 AM. For weighing a laboratory scale was used (sensitivity 0.1 gram). Experiments were approved by the Ethical Committee of Animal Experiments of the University of Groningen.

(3) All animals were instrumented with chronic heart catheters bilaterally in the jugular vein allowing stress free blood sampling during an intravenous glucose tolerance test (IVGTT). Surgeries were carried out under general isoflurane (2%) anesthesia. Animals had at least 10 days to recover before the start of the experiments. Cannulas were checked every week for patency.

(4) The full trial lasted 11 weeks: week1: meal pattern measurements (TSE) week3: Blood sampling during a single meal of 1.5 grams week4: meal pattern measurements (TSE) week6: Intravenous Glucose Tolerance Test week9: meal pattern measurements (TSE) week11: carcass analysis
Diets

(5) 9 rats were fed with a control diet, while 8 rats were fed with pectin enriched diet.

(6) The composition of the diets was as follows: 95% chow RMH-B meal (obtained from Arie Blok, Woerden, the Netherlands) and 5% Pectin (see Pectin sources). The diets were prepared by mixing all components (including 0.25% TiO2 as marker) with water to 600 ml/kilo in an industrial mixer until a homogeneous mixture/dough was obtained. After 20 minutes of mixing, the diets were pelleted using a pelleting machine (diameter 1.0 cm). The obtained pellets were dried for 48 hours using compressed air at room temperature.

(7) Pectin Sources

(8) Pectin with a DE 33 were isolated from citrus and obtained from Herbstreith & Fox (Neuenburg/Württingen, Germany).

(9) Feeding Pattern Analyses

(10) During the second meal pattern measurements the animals were observed and measurements were performed for 48 hours. The obtained data was averaged over all animals fed on a certain diet and is presented in the table below. Statistical analyses were performed using student T test.

(11) TABLE-US-00002 Control diet Pectin DE 33 diet Number of meals 20.56 ± 0.84  23.25 ± 0.82  Average meal size (g) 1.99 ± 0.07 1.75 ± 0.11 Average meal duration (min) 8.22 ± 0.54 9.22 ± 0.25 Average eating speed (g/min) 0.32 ± 0.04 0.21 ± 0.01

(12) Surprisingly, pectin fed animals consumed significantly more meals which are smaller in size. Still, the total amount eaten in the period for both diets is not significantly different (40.92 vs 40.69 grams), meaning that the reduced meal size is due to a satiating effect.

Example 2 Incorporation of LM Pectin in Diets Leads to Improved Homeostasis of Blood Glucose Levels

(13) The rat trial was as described in example 1.

(14) Intravenous Glucose Tolerance Test

(15) To assess the effects of pectin addition to the diet on glucose regulation, rats were subjected to an intravenous glucose tolerance test (IVGTT). The IVGTT was performed during week6. The IVGTT was performed during the third and fourth hour of the light phase. Food was removed at lights on and rats were connected to the blood sampling and infusion tubes at least one hour before the IVGTT. During the IVGTT, a 15% glucose solution was infused for 30 minutes at a rate of 0.1 ml/min. The start of the infusion was designated time point=0 min. Blood samples (0.2 ml) for determination of blood glucose levels were taken before, during, and after the infusion of glucose at time points=−10, −1, 5, 10, 15, 20, 25, 30, 35, 40, and 50 minutes. Note that the glucose infusion prevented any hypovolemic effect of the blood sampling. Blood samples were collected in EDTA (20 microliter/ml blood) containing tubes on ice. Blood was centrifuged at 2600 g for 10 minutes and plasma was stored at −20 C until analysis. Blood glucose levels were measured by Hoffman's ferrocyanide method (Hoffman, W. S. (1937). J. Biol Chem, 120, 51).

(16) Results

(17) Surprisingly, the data demonstrated that blood glucose levels after the 30-minutes intravenous infusion of glucose are lower (indicated by the grey boxes) in rats fed on the diet containing pectin with a DE of 33 as compared to the blood glucose levels of the rats fed on the control diet. Meaning that mammals fed on diets containing low DE pectin are better equipped to controlling their blood sugar levels, by levelling of peak concentrations, which is beneficial for preventing developing obesity, as well as managing the body mass index and/or the body fat percentage.

(18) TABLE-US-00003 Time points (min) Diet −11 −1 5 10 15 20 25 30 35 40 50 Control Glucose 5.56 5.71 7.37 8.55 9.49 10.05 10.34 10.61 8.73 7.16 5.86 (mmol) SEM 0.21 0.18 0.23 0.19 0.28 0.28 0.30 0.25 0.18 0.18 0.17 Pectin Glucose 5.65 5.62 7.55 8.68 9.50 9.36 9.52 9.60 7.83 6.49 5.72 DE 33 (mmol) SEM 0.11 0.12 0.23 0.28 0.51 0.32 0.35 0.32 0.24 0.33 0.27

Example 3 Incorporation of LM Pectin in Diets Leads to Improved Homeostasis of Plasma Insulin Levels

(19) The rat trial was as described in example 1.

(20) Blood Sampling During a Single Meal of 1.5 Grams

(21) The rats were fasted for 7 h (induce hungry animals), after which they were provided with 1.5 g of their diet. Blood samples (0.2 ml) for determination of plasma insulin levels were taken through the cannula at time points=−5, 0, 1, 2.5, 5, 7.5, 10, 15 and 20 minutes. Blood samples were collected in EDTA (20 microliter/ml blood) containing tubes on ice. Blood was centrifuged at 2600 g for 10 minutes and plasma was stored at −20 C until analysis. Plasma levels of insulin were measured by Millipore Rat Insulin Radioimmunoassay (Linco Research, St Charles, Mo., USA).

(22) Results

(23) Surprisingly, the data demonstrated that plasma levels of insulin after feeding were lower (indicated by the grey boxes) in rats fed on the diet containing pectin with a DE of 33 as compared to the plasma levels of insulin of the rats fed on the control diet. Meaning that mammals fed on diets containing low DE pectin are better equipped to controlling their plasma levels of insulin, thereby lowering the chances of so-called hyperinsulinemia, which is a risk factor for developing type 2 diabetes, and thereby acts positively towards managing the body mass index, the body fat percentage and/or the distribution of fat storage sites.

(24) TABLE-US-00004 Time points (min) Diet −5 0 1 2.5 5 7.5 10 15 20 Control Insulin 0.96 1.36 1.98 3.17 3.85 4.80 4.91 4.07 3.28 (ng/ml) SEM 0.08 0.10 0.37 0.43 0.41 0.38 0.34 0.39 0.49 Pectin Insulin 0.91 1.02 1.29 2.58 2.74 4.45 4.27 4.25 3.00 DE 33 (ng/ml) SEM 0.08 0.09 0.27 0.32 0.29 0.67 0.52 0.48 0.46

Example 4 Incorporation of LM Pectin in Diets Leads to Reduced Fat Deposition

(25) The rat trial was as described in example 1. After sacrifice a carcass analysis was performed to determine the amount of fat. Liver, stomach, gut (ilium to rectum), spleen, kidneys were removed and weighed. Retroperitoneal and epididymis fat was weighed as well. The fat content from the skin, carcass and gut was determined using a petroleum based Soxlet fat extractor. Visceral fat here was defined as the total of intestinal fat, epididymal fat and retroperitoneal fat.

(26) TABLE-US-00005 Control diet Pectin DE 33 diet Grams (or Relative Grams (or Relative g/g for vs body g/g for vs body the ratio) weight (%) the ratio) weight (%) Total fat 76.03 ± 3.26 19.5 67.74 ± 2.51 17.8 Visceral fat 21.66 ± 1.22 5.5 17.86 ± 1.21 4.7 Gut fat  4.46 ± 0.23 1.1  3.51 ± 0.31 0.9 Ratio fat:lean  0.54 ± 0.03 n.a  0.45 ± 0.03 n.a. mass

(27) Surprisingly, pectin fed animals have a reduced absolute as well as relative percentage of fat. This is also visible for the amount and relative percentage of visceral and gut fat.

Example 5 Incorporation of LM Pectin in Diets Leads to More and Smaller Meals

(28) Piglet Feeding Trial Set-Up

(29) Piglets were housed in weaner facilities pens with 7 (2.56×1.26 m) or 9 piglets per pen (1.3×2.85 m). These pens have been built according to practical regulations, resulting in respectively 0.44 and 0.40 m2 per piglet. A total of 27 pens (replicates) per treatment were used. Per treatment 4 of these replicates were housed in pens equipped with IVOG® feeding stations (Individual Feed Intake Recording in group housing; Isentec, Marknesse, The Netherlands) for weaning pigs. These pens contained 8 piglets per pen (1.75×3.00 m) resulting in 0.65 m2 per piglet. The housing conditions were typical for Dutch pig husbandry, operated according to the Dutch IKB farm standards. Each pen was equipped with a dry feeder and piglets were given ad libitum access to feed and water during the whole post-weaning period. The floor in the pens within a unit was a fully slatted, plastic floor. Enrichment (chain with a play ball) was provided as well. Environmental conditions during the trial (temperature and ventilation rate) were automatically controlled, and were adjusted to the age of the pigs. The start temperature in the weaner pens was 30 C and gradually decreased to 24 C over 35 days. The pigs had the following characteristics:

(30) Animals: 828 weaned piglets

(31) Origin animals: Laverdonk swine herd

(32) Breed: Pietrain×Topigs20

(33) Gender: Males and females

(34) Age at start: On average 26 days (at weaning)

(35) Age at end: On average 61 days

(36) Body weight at start: On average 7.4 kg (at weaning)

(37) Body weight at end: On average 18.2 kg

(38) Dietary Treatments and Feeding

(39) Four experimental treatments (=diets) were tested. The treatments were:

(40) TABLE-US-00006 Diet Description Code 1 Control 7% untreated SBM 2 Pectin soybean meal 7% processed SBM 3 Pectin DE33 3% pectin 33% DM 4 Pectin DE33 3% pectin 55% DM

(41) Treatment 1 was the control and contained 7% soybean meal (SBM) as a reference. In treatment 2, the SBM was pre-treated by autoclaving and 7% of the resulting modified SBM was included in the diets (exchanged for untreated SBM). Treatment 3 and 4, were similar to the control but here 3% lemon pectins were replacing 3% SBM. The experimental diets were produced by Research Diet Services (The Netherlands). Piglets had ad libitum access to feed and water.

(42) Number of treatments: 4

(43) Number of replicate pens: 27 per treatment

(44) Number of batches: 6

(45) Period 1: Weaner period, D0-D9

(46) Period 2: Nursery period, D9-D35

(47) Day 0: Day of weaning (Wednesday)

(48) Diets

(49) Feed Compositions (in g/kg):

(50) TABLE-US-00007 Weaner diet Piglet diet Diet 1 2 3 4 1 2 3 4 Moisture 119 112 117 118 121 112 115 115 Protein 174 181 161 161 164 169 147 147 Fibre 27 26 24 22 28 28 27 26 Fat 42 42 42 41 34 34 34 34 Starch 418 428 426 429 478 473 475 485 Phosphorus 5.41 5.39 5.12 5.14 5.64 5.5 5.12 4.89 Sodium 3.11 3.22 3.72 3.5 2.58 2.84 3.05 2.94
Pectin Sources

(51) Pectins with a DE 33 and 55 were isolated from citrus and obtained from Herbstreith & Fox (Neuenburg/Württingen, Germany). The Soy Bean Meal (SBM) was from South-American origin (mixture from Argentina, Brasil and/or Paraguay) and processed to extract the residing pectins by mixing the SBM at 33% dry matter with tapwater and autoclaving for 30 mins at 120 C. After cooling the obtained material was freeze dried and milled, and used as such in the diet.

(52) Experimental Design

(53) The experimental design was a complete randomized block design with four treatments.

(54) At weaning, both male and female piglets were assigned to one of the experimental treatments based on body weight, sex and ancestry. Boars and gilts were evenly distributed over treatments, based on the availability of the piglets.

(55) Two periods were distinguished. This was the so-called weaner period (day 0 to 9) and the nursery period (9 to 35 days).

(56) Piglet body weight was measured individually at day before weaning (D-1), D9 and D35. Feed intake was monitored over three periods (D0-D9, D9-35, D0-D35) continuously for the piglets housed in pens equipped with the feeding stations.

(57) Experimental Design

(58) TABLE-US-00008 Diet 2 1 Soy Bean 43 4 Period Control Pectin DE33 DE55 Number of meals D 0-D 9 8.9 10.0 11.7 11.4 (per day) D 9-D 35 13.8 16.1 18.2 15.8 Average meal D 0-D 9 18 19 19 19 size (g) D 9-D 35 43 41 34 34 Average meal D 0-D 9 4.7 4.4 4.4 3.7 duration (min) D 9-D 35 6.0 6.7 4.7 5.2 Average eating D 0-D 9 4.2 5.2 4.8 6.0 speed (g/min) D 9-D 35 7.8 8.2 7.5 6.8

(59) Surprisingly, pectin fed animals consumed significantly more meals which are in the majority phase of the tests also smaller in size.

Example 6 Incorporation of LM Pectin in Diets Leads to a Healthy Microbiota Composition

(60) Piglet Feeding Trial Set-Up

(61) The experimental farm for young piglets is located in Flanders (Belgium) and consists of 8 batteries, each containing 4 pens. The piglets under study are hybrids of Topigs Piétrains and are weaned at 21 days. The piglets are weighed individually at weaning and 2 and 4 weeks after weaning. Feed intake is registered per pen of 4 piglets at the moments of weighing. At arrival the piglets are earmarked with a new Sanitel-number. During the trial, a veterinarian and a Felasa D certified person supervise the performed piglet experiment according to the international guidelines described in law EC/86/609.

(62) Each pen (1.5 m×1.5 m) contains 4 piglets at the start of the trial. For each pen, one feeder (ad libitum) is installed for meal or pellets. One drinking nipple is installed per pen. The temperature at start is at 28±2° C. until 10 days after weaning. Afterwards, temperature is decreased to 25±2° C.

(63) Commercial non-medicated diets are given. Non-medicated means that the piglet doesn't receive any therapeutic antibiotics before and during the trial. The diets are given in the form of meal. All feed were analysed for their nutritional content.

(64) Four treatments were applied (diets A, B, C, D) on 7 replicates with 4 piglets per group. At the start of the trial, the piglets (around 7 kg body weight) are allocated to the different pens by weight. This allocation is made in order to have an equal average weight and an equal standard deviation around the average weight for each treatment and pen. For microbiological counting's and for taking biopsis, piglets receive an overdose of barbiturates (Nembutal) followed by sacrification. Afterwards, a section is performed on the piglets. Samples for microbial counts are immediately processed, while samples taken for histochemical experiments were fixed for later analysis. During the whole trial period the piglets are fed ad libitum, except for the period of microbiological countings. At that moment, three days before the microbiological countings are performed, the piglets are fed restricted. Piglets receive three times a day an amount of feed, which is carefully weighed and noted. The feed is given at 8.00, 13.00 and 18.00. When necessary, the sick piglets were treated individually (by injection). The following parameters were taken into account. (i) individual growth data, (ii) feed intake data per pen (corrected for eventual losses), (iii) feed conversion ratio during weaning, starter and whole trial period, (iv) fecal score and clinical score, (v) tight Junctions, (vi) microbial analysis, (vii) histochemical analysis.

(65) Diets

(66) Feed Compositions (in g/kg):

(67) TABLE-US-00009 Ingredient Feed A Feed B Feed C Feed D Corn 171.13 169.13 169.13 169.13 Grains (wheat and barley) 491.83 491.83 491.83 491.83 Protein sources (soy, potato) 227.72 227.72 227.72 227.72 Milk derivatives (whey) 52.65 52.65 52.65 52.65 Soy bean oil 14.19 14.19 14.19 14.19 Amino acids 10.80 10.80 10.80 10.80 Minerals & trace minerals 10.24 10.24 10.24 10.24 Limestone 10.63 10.63 10.63 10.63 Enzyme* 0.64 0.64 0.64 0.64 Premix** 10.17 10.17 10.17 10.17 Pectin DE33 — 2.00 — — Pectin DE55 — — 2.00 — Pectin soybean meal — — — 2.00 *Xylanase/beta-glucanase and phytase cocktail (BASF) ** Premix includes aroma's, extra trace minerals, vitamins (Vitamex N.V.)
Pectin Sources

(68) Pectins with a DE 33 and 55 were isolated from citrus and obtained from Herbstreith & Fox (Neuenburg/Württingen, Germany). The Soy Bean Meal (SBM) was from South-American origin (mixture from Argentina, Brasil and/or Paraguay) and processed to liberate the residing pectin. Pectins were extracted from SBM by mixing the SBM at 33% dry matter with tapwater and autoclaving for 30 mins at 120 C. After cooling the obtained material was freeze dried and milled, and used as such in the diet.

(69) Digesta Collection

(70) The pig fecal samples were collected on day 14 and 28 during the experimental diet feeding. After the fecal collection period, animals were anesthetized and euthanized. Digesta samples were collected from terminal ileum, proximal colon, mid colon and distal colon. Part of each digesta was stored in 1.5 mL Eppendorf tubes for analysis of microbiota composition and SCFA. These tubes were immediately frozen in liquid nitrogen and stored at −80° C. The remaining amount of digesta was immediately stored at −20° C. until further analysis.

(71) DNA Extraction and Microbiota Analysis

(72) Microbial DNA was extracted from 250 mg of digesta by using a fecal DNA extraction protocol (Salonen A, Nikkilä J, Jalanka-Tuovinen J, Immonen O, Rajilić-Stojanović M, Kekkonen R A, Palva A & de Vos W M. 2010. Comparative analysis of fecal DNA extraction methods with phylogenetic microarray: Effective recovery of bacterial and archaeal DNA using mechanical cell lysis. Journal of Microbiological Methods, 81: 127-134). The DNA is isolated by sequential precipitations and finally purified by using the QIAamp DNA Stool Mini Kit columns (Qiagen, Hilden, Germany) according to the manufacturer's recommendations. 16S rRNA gene was amplified and sequenced in paired-end mode by using the MiSeq platform (Illumina).

(73) Sequence Analysis

(74) Raw Illumina fastq files were de-multiplexed, quality-filtered and analysed using QIIME 1.9.0.

(75) TABLE-US-00010 Relative abundance of Bacteroidetes and Firmicutes species in the microbiota composition of experimental fed pigs Relative abundance* Ratio** Diet Pectin Fecal sample Bacteroidetes Firmicutes Bacteroidetes:firmicutes A control Terminal ileum 0 0.99 0 A control Proximal colon 0.07 0.93 0.07 A control Mid colon 0.12 0.88 0.14 A control Distal colon 0.10 0.89 0.11 B DE 33 Terminal ileum 0 1.00 0 B DE 33 Proximal colon 0.56 0.44 1.26 B DE 33 Mid colon 0.63 0.36 1.75 B DE 33 Distal colon 0.59 0.40 1.47 C DE 55 Terminal ileum 0 0.97 0 C DE 55 Proximal colon 0.56 0.44 1.25 C DE 55 Mid colon 0.48 0.51 0.94 C DE 55 Distal colon 0.52 0.48 1.07 D Soy Bean Terminal ileum 0.01 0.96 0.01 D Soy Bean Proximal colon 0.21 0.76 0.28 D Soy Bean Mid colon 0.23 0.75 0.31 D Soy Bean Distal colon 0.31 0.66 0.47 *The relative abundance is the % of 16S rRNA data of Bacteroidetes phylum (the summation of species from the families Porphyromonadaceae, Prevotellaceae, Rikenellaceae as well as species indicated by the codes RF16 and S24-7) and Firmicutes phylum (the summation of species from the families Aerococcaceae, Lactobacillaceae, Streptococcaceae, Christensenellaceae, Clostridiaceae, Lachnospiraceae, Peptostreptococcaceae, Ruminococcaceae, Veillonellaceae, Erysipelotrichaceae) in the total data set obtained through Illumina sequencing **The ratio is the relative abundance of Bacteroidetes divided by the relative abundance of Firmicutes.

(76) Surprisingly, addition of pectin to the diets leads to an increased ratio of species belonging to the phylum of the Bacteroidetes over species belonging to the phylum of Firmicutes, in the gut. Such a ratio is associated with a reduced prevalence of obesity, and will facilitate managing the body mass index and/or the body fat percentage.

Example 7 Incorporation of LM Pectin in Diets Leads to a Healthy Microbiota Composition

(77) The data was acquired and analysed as described in example 6.

(78) TABLE-US-00011 Prevotellaceae Ruminococcaceae Lactobacillaceae relative fold relative fold relative fold Diet Pectin Fecal sample abundance* increase** abundance* increase** abundance* increase** A control Terminal ileum 0 — 0.01 1 0.82 1 A control Proximal colon 0.06 1 0.06 1 0.71 1 A control Mid colon 0.12 1 0.08 1 0.64 1 A control Distal colon 0.10 1 0.09 1 0.57 1 B DE 33 Terminal ileum 0 — 0.01 0.95 0.78 0.95 B DE 33 Proximal colon 0.55 8.55 0.05 0.95 0.46 0.66 B DE 33 Mid colon 0.62 5.35 0.12 1.51 0.20 0.30 B DE 33 Distal colon 0.58 5.96 0.12 1.35 0.14 0.25 C DE 55 Terminal ileum 0 — 0 — 0.93 1.19 C DE 55 Proximal colon 0.54 8.40 0.15 2.59 0.03 0.05 C DE 55 Mid colon 0.47 4.06 0.16 2.05 0.01 0.02 C DE 55 Distal colon 0.50 5.15 0.15 1.70 0.01 0.02 D Soy Bean Terminal ileum 0 — 0 — 0.52 0.66 D Soy Bean Proximal colon 0.21 3.30 0.18 3.09 0.03 0.05 D Soy Bean Mid colon 0.23 1.97 0.15 1.94 0.03 0.04 D Soy Bean Distal colon 0.31 3.16 0.20 2.30 0.01 0.02 *The relative abundance is the % of 16S rRNA data of families of Prevotellaceae (the summation of species from the genus Prevotella), Ruminococcaceae (the summation of species from the genus Ruminococcus) and Lactobacillaceae (the summation of species from the genus Lactobacillus) in the total data set obtained through Illumina sequencing **The fold increase is the relative increase in % of specific 16S rRNA in the total data set, i.e. the relative abundance of determined for a specific pectin fed sample divided by the correspondingcontrol sample.

(79) Surprisingly, addition of pectin to the diets leads to an relative increase in presence of species belonging to the families of the Prevotellaceae and Ruminococcaceae, as well as a relative decrease in presence of species belonging family of Lactobacillaceae, in the gut. Such shifts in microbiota are associated with a reduced prevalence of obesity, and will facilitate managing the body mass index and/or the body fat percentage.

(80) All these examples clearly and surprisingly show that the specific pectins show improvements, which are of significant importance in managing the body mass index, the body fat percentage and/or the distribution of fat storage sites.