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    Can anyone grab the pdf file for this study?

    Combined Effects of Caloric Restriction and Branched-Chain Amino Acid Supplementation on Body Composition and Exercise Performance in Elite Wrestlers

    A. Mourier1, 2, A. X. Bigard1, E. de Kerviler3, B. Roger4, H. Legrand3, C. Y. Guezennec3
    1 Centre d'?tudes et de Recherches de M?decine A?rospatiale, D?partement de Physiologie Syst?mique, Br?tigny-sur-orge, France
    2 Service d'Endocrinologie, H?pital Saint-Louis, Paris, France
    3 Service de Radioltogie, H?pital Saint-Louis, Paris, France
    4 Service de Radiologie, H?pital Piti?-Salp?tri?re, Paris, France

    Twenty-five competitive wrestlers restricted their caloric intake (28 kcal kg-1 ? day-1) for 19 days, using a hypocaloiric control (hC, n = 6), hypocaloric high-protein (hHP, n = 7), hypocaloric high-branched-chain amino acid (hBCAA, n = 6), hypocaloric low-protein (hLP, n = 6) diet to determine the effects of caloric restriction on body composition and performances versus control diet (C. n = 6). Anthropometric parameters (weight, percent body fat) and adipose tissue (AT) distribution measured by magnetic resonance imaging (MRI) obtained before and after diet, were compared. A significant highest body weight loss (-4 kg, p<0.05) and decrease in the percent of body fat (-17.3 %, p< 0.05) were observed for subjects of the hBCAA group. Subjects of the hBCAA group exhibited a significant reduction (- 34.4 %, p<0.05) in abdominal visceral adipose tissue (VAT). There was no change in aerobic (VO2max) (p>0.75) and anaerobic capacities (Wingate test) (p > 0.81), and in muscular strength (p :> 0.82). We conclude that under our experimental conditions, the combination of moderate energy restriction and BCAA supplementation induced significant and preferential losses of VAT, and allowed maintainance of a high level of performance
    PMID: 9059905 [PubMed - indexed for MEDLINE]
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    Introduction
    Prior to competing, all wrestlers are required to attain aspedfic
    body weight (weight class). The majority of wrestlers attempt
    to lose body weight, maximise the amount of lean tissue and
    minimise the amount of body fat.
    Numerous methods are employed to reduce body weight for
    wrestling competition. Wrestlers can reduce body weight
    either rapidly or gradually. Reduction of body weight in less
    than 1 week is defined by Fogelholm (9) as rapid body weight
    reduction. Reduction over a longer period (>7 days) is defined
    as gradual body weight reduction. The main difference is that
    gradual body weight reduction is accomplished by negative
    energy balance, whereas active or passive dehydration (fluid
    loss) is a necessary part of rapid body weight loss. The primary
    goal of restriction of energy intake is the restriction in body
    fat. But previous studies in moderately active men have shown
    that substantial reductions in body weight include loss of both
    lean and fat tissues (22). Therefore, wrestlers cutting weight
    through the restriction of energy intake can expect to lose
    muscle mass. Results of the effects of gradual body weight reduction
    on aerobic performance in athletes are variable. It has
    been shown that during gradual body weight reduction, rnaximal
    oxygen intake (W0,rnax) might deteriorate (16.38). remain
    unchanged (26) or might improve (24). When reducing
    body weight gradually with a normal carbohydrate diet (50%
    of energy intake, 2.5 g kg-'. day1), anaerobic performance was
    impaired (28). Conversely, anaerobic performance was not affected
    after body weight loss with a higher intake of carbohydrates
    (4.1 g . k g ' . d a y l ) (28). After gradual body weight reduction,
    the isometric endurance capacity of the skeletal
    muscle was reduced (13,36) while the endurance capacity
    during isokinetic exercise was unaffected (36).
    Steen and McKinney (34) found that college wrestlers from
    several universities consumed diets composed of approximately
    15 % protein. 33 to 37 % fat and 43 to 47% carbohydrate
    with the composition being somewhat dependent on the time
    of the season (pre-season, mid-season, etc.). During periods of
    weight reduction, the authors found that 37% of the wrestlers
    studied consumed less than two-thirds of the Recommended
    Dietary Allowances (RDA) for energy. At the reduced level of
    energy intake, all macronutrients were significantly lower, although
    theaverage protein intake remained at 0.9g. kg1. day1
    (14), which was almost the RDA for these subjeas (Committee
    on Dietary Allowances 1989). Based on nitrogen balance. Walberg
    et al. (36) suggested that a protein intake of 0.8 g- kg1.
    day1 was insufficient to maintain nitrogen balance during
    body weight reduction, whereas 1.6g. kg1 .day1 was enough.
    It has been shown that in catabolic states such as starvation.
    the plasma concentrations of branched-chain amino acids
    (BCAA), i.e. leucine, isoleucine and valine, were selectively increased
    (33). It has become widely accepted that BCAA and
    leucine in particular can directly stimulate protein synthesis.


    C group hC group hHP group h0CAA group hLP group composition1.
    Variable (n - 6) (n = 6) (n = 7) (n = 6) (n = 6)
    Carbohydrates (%)2 55 55 60 60 60
    Proteins (%) 12 12 25 20 15
    Fats (%) 33 33 15 20 2 5
    Energy intake (kcal . kg-' . day-') 40 28 28 28 28
    ' C= normocaloric control diet: hC = hypocaloric diet: hHP = high-protein diet; hBCAA = high-branched-chain amino acids diet;
    hLP = low-protein diet
    percentage of calorie intake


    Louard et al. (25) observed that in normal man. BCAA infusion
    suppresses skeletal muscle proteolysis independently of any
    rise of plasma insulin. Furthermore, Schena et al. (31) concluded
    that the administration of a suitable amount of BCAA
    can prevent the ubiquitous muscle loss observed during high
    altitude exposure. BCAA administration potentiated the release
    of some anabolic hormones, mainly human growth hormone
    (GH) (17), known to accelerate amino acid uptake and protein
    synthesis in skeletal muscle (32). It may thus be suggested that
    under conditions of increased protein catabolism (hypercaloric
    diet), BCAA will help minimise muscle wasting.
    The purpose of this study was to investigate the effects of 19-
    day qualitatively and quantitatively varied energy intakes on
    body composition and exercise performance in elite male
    wrestlers.



    Material and Methods
    Subjects


    Thirty-one male wrestlers from the French National Institute
    of Sports volunteered to serve as subjects in this study. Testing
    was conducted mid-season. The study was approved by Cochin-
    Port Royal Hospital Ethics Committee. All subjects were
    informed of risks and signed consent fonns. All were highly
    trained in prolonged exercise and had been wrestling for many
    years. None reported ever using anabolic steroids. The subjects
    were randomly assigned to one of five groups and consumed
    diets whichvaried in their energy and macronutrient composition
    (four levels of dietary protein intake, detailed in Table 1):
    .N ormocaloric control (n = 6): C Hypocaloric control (n =x6): hC
    Hypocaloric high-protein (n = 7): hHP
    .H ypocaloric high-branched-chain amino acid (n = 6): hBCAA Hypocaloric low-protein (n = 6): hLP

    Anthropometric measurements
    Height, weight and BMI

    Height of the barefoot subject was measured to the nearest
    0.1 cm. Before and immediately after the dietary period, body
    weight (BW)of subjects wearing same shorts was measured
    on the same standard medical balance with an accuracy of
    + 100g. Body Mass Index (kg. m-2) was then calculated (5).

    Skinfold thickness
    The anthropometric assessment consisted of three skinfold
    sites: chest, quadriceps and abdomen, which were measured
    with a Harpenden skinfold calliper (18). A minimum of two
    measurements were made at each skinfold site by the same
    experienced investigator at each time of measurement. The
    values were averaged and used to estimate body fat percentage
    using an equation developed by Jackson and Pollock (18).


    MRI
    Magnetic resonance images were taken at 0.5 Tesla (General
    Electric. Milwaukee. WIS. U.S.A.). Before and after the dietary
    period, subjects were positioned in the magnet in a supine feetfirst-
    position. A multislice spin-echo sequence (TR = 500 ms.
    TE = 25 rns) was used to obtain TI-weighted images with a reasonable
    contrast between adipose tissue (AT) and lean tissue.
    The magnetic resonance examination consisted of two series
    of 7 mm thick (3 mm gap) axial images. One series was taken
    at midthigh. and two other series were taken in the abdomen
    at the level of the umbilicus.
    As discussed by Ross et al. (30). magnetic resonance imaging
    (MRI) pixel intensity value for a given tissue may not be consistent
    from slice to slice or between individuals. To set a common
    dynamic range for all images, a 1.5-cm diameter reference
    tube was filled to capacity with a solution ofGadolioium-DOTA
    (0.5 mmol. I-') and placed on either of the sides of the subjects
    during data acquisition.The end points ofthe signal range were
    0 for air and 4095 (maximum value for the 12 bit integers returned
    by the MRI system) for the reference tube. All the images
    were transferred onto personal Iris Computer (Silicon
    graphics) for further analysis.

    Segmentation of Adipose Tissue (AT)
    Two large regions of interest (ROI) were drawn manually. One
    ROI included the largest cross-sectional area of subcutaneous
    adipose tissue (SAT) and the other R01 included the largest
    cross-sectional area of visceral adipose tissue (VAT). The next
    step consisted of highlighting and counting the pixels which
    correspond to AT in each ROI. The reference tube allows determination
    of the threshold of AT. The threshold selected for AT
    was based on the analysis of a sample of typical images and
    their respective gray level histograms. The optimal threshold
    for AT was 180 (on a scale of 256); above this value, pixels were
    considered as representing AT. The next step involved
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    ...

    highlighting the pixels that were counted as AT in response to
    the selected threshold. Each slice was reviewed using an interactive
    slice editor program that allowed verification and. when
    necessary, correction of the segmentation result. The operation
    was facilitated by superimposing the original gray level image
    on the binary segmented image using a transparency mode.
    Visceral and subcutaneous adipose tissue regions on each
    abdominal slice were then assigned different colour codes.
    Similar operations were performed on each midthigh slice to
    identify adipose tissue and muscle tissue (MT).

    Calculation of areas
    The areas of the respective AT regions in each slice were computed
    automatically by summing AT pixels and multiplying by
    the pixel surface area. A previous investigation (30) verified
    that the error in the spatial dimensions of MR images using
    this system ranged from 0.1 to 1.5 %. Thus the AT area on the
    MR images is fairly accurate.

    Diet analysis
    Wrestlers recorded their daily weighed-meals intake over
    seven successive days in notebooks supplied by the investigators.
    One training session was held to show the subjects how
    to keep records. During the weeks of the recording, the investigators
    met with the subjects and reminded them to record
    the amount and type of all foods consumed. The complete records
    of food intake of all subjects were analysed using Dietetique
    1 software (copyright S.D. Crun for SDG '1986-1987,
    Database by Mouton-Pastre-Peres for SPEFS "1985-1986-
    1987) before the beginning of the study. Total daily intake of
    energy, protein, carbohydrate and fat were estimated daily and
    averaged for one week.

    Dietary protocol
    During the study, all the meals of the wrestlers were eaten in
    the same refectory. Estimates of the nutrient compositions of
    food ingested were made by weighing foods and recording the
    energy content of each portion. All wrestlers generally ate from
    a common menu. A dietician gave instructions on what to
    choose to obtain the desired diet. according to the specifications
    of the diet group.
    The 19day diet period began immediately following the period
    of nutrient composition food intake analysis. In all groups, diets
    exceeded RDA for protein intake (0.8 g . kg' . day'). Subjects on
    hypocaloric diets consumed the same food and received a commercial
    supplement depending on the group they belonged to.
    The experimental diets were isocaioric and the daily energy
    intake of each subject (except C subjects) was estimated to be
    28 kcal . kg1. The commercial dietary supplements used in the
    ptesent study represented 3.6 kcal. kg1 .day1.
    Subjects of the C group consumed ad libitum (normocaloric
    diet), while subjects of the hC group consumed a 28 ltcal. kg1
    body weight .day-' hypocaloric diet without any supplementation.
    The subjects of the hHP group consumed a hypocaloric diet
    (24.4 kcal - kg1. day') and a dietary supplement which consisted
    of a mixture of soja protein (0.9g. k g ' - day') so that
    protein intake was increased to 2 g . kg' . day': 25% of the total
    energy intake (6.1 g BW/100g protein: 46% leucine. 22%
    isoleucine and 32 % valine) with 60 %carbohydrate and 15 % fat.
    The hBGQA diet consisted of a hypocaloric diet (24.4 kcal. kg1
    . day-') and a protein dietary supplement enriched with 0.9 g -
    kg1. day'' branched-chain amino acids (51.9g BCAA/lOOg protein:
    76% leucine. 19% isoleucine and 5% valine), approximately
    20% protein. 60% carbohydrate and 20% fat.
    The hLP diet consisted of a hypocaloric diet (24.4 kcal . kg'.
    day1) and a dietary supplement of 0.9g glucose. kg1. day1
    (3.6 kcal .kg1. day'), approximately 15% protein, 60% carbohydrate
    and 25 % fat.

    Exercise testing
    During the two days prior to the dietary program, the maximal
    oxygen uptake (W0,max) was determined using an open system
    to analyse the expired gases (Sensor Medics 2900. Sensor
    Medics corporation. Yorba Linda. CA, USA). On the morning of
    the first day. the subjects exercised on a treadmill according
    to an incremental protocol for V0,max determination (W0,max
    T). The exercise session of W0,max T consisted of a five min
    warm-up at 12 km. h-I followed by an incremental test during
    which successive speeds of 1 km. h-I were reached every two
    minutes. On the second morning, they exercised on an arm
    ergometer (881 E, Monark-Crescent AB., Valberg, Sweden) for
    V0,max determination (V02max U). The exercise session of
    V0,rnax U consisted of a two min warm-up at 36.8 W followed
    by incremental work during which successive loads of 18.4 W
    were reached every two minutes. A pedalling frequency of
    75 rpm was selected until 110 W was obtained and then upwards
    to 85 rpm. The test protocol was adjusted to ensure attainment
    of maximal effort within 15 to 20 min. Oxygen intake.
    minute ventilation. respiratory exchange ratio (R) and heart
    rate (HR) were continuously monitored. V0,max was the value
    obtained over 60s when two of the following criteria were retained:
    1 ) an increase in exercise intensity produced no further
    increase in V02, 2) a respiratory exchange ratio > 1.1, and 3)
    visuals signs of tiredness and distress on the treadmill or with
    the crankam ergometer.
    The first afternoon, right isometric knee extensions were performed
    using an isokineticdynamometer (Cybex 11. Lumex Inc.,
    N.Y., U.S.A.). The subjects were in a sitting position, securely
    strapped into the muscle-test chair. The seated posture met
    the following specifications: hip angle IOU, knee angle 80'. The
    experimental procedure comprised the following steps: 1 )subjects
    were asked to perform maximal isometric contractions
    of short duration (2-2s) of the knee extensor muscles. The
    maximal force was measured, and the best performance after
    4 trials was selected as maximal voluntary isometric contraction
    (MVC). Approximately 2-3 minutes elapsed between each
    of the 4 trials. 2) following a sufficient recovery period lasting
    between 15 and 17 minutes, subjects maintained a prolonged
    isometric contraction of the knee extensor muscles at 50% MV
    as long as possible. The required force of contraction was
    shown by an oscilloscope placed in front of the subject. The
    endurance time for this submaximal muscle contraction was
    measured until the subject was unable to maintain the required
    tension.
    The second afternoon, the anaerobic capacity was determined
    using the Wingate Anaerobic Capacity Test (2). Subjects
    completed a 30-sec Wingate Anaerobic Capacity Test on a
    Monark cycle ergometer (Monark-Crescent A.B., Valberg,
    Sweden) with two clips and adapted with an electronic reed
    switch to measure speed (revolutions per minute, rprn). The
    resistance for each subject (0.098. kg' body weight) was determined
    according to Evans and Quinney (7). Five sec revolutions
    were counted For 30sec using an electronic rpm counter.
    From these measurements, peak power. total power and mean
    power were calculated (2).
    All physical performance tests were conducted two days prior
    to the initiation of the study and the two last days of the dietary
    period.

    Blood sample analyses
    Blood samples of approximately 10ml were collected from an
    antecubital vein at rest, and at the end of the incremental
    treadmill exercise. Blood samples were collected before and
    after the dietary program, and plasma was immediately obtained
    by centribgation and frozen until assays were performed
    within 30 days. At rest, glucose, lactate and glycerol
    were assayed in neutralised blood samples using an enzymatic
    method (3). Free fatty acids (FFA) were assayed according to
    the method of Ho (12). Because low caloric diets are known to
    affect the regulation of peripheral thyroid hormones (27) and
    because protein and amino acid intakes promote a rise in
    growth hormone serum levels (19.27). the endocrine responses
    to diets were tested. Hormone determinations were
    made in duplicate using commercially available radioimmunoassay
    kjts (CIS-bio International. Gif-sur-Yvette. France):
    insulin (SB-INSI-5), growth hormone (GH) (SB-GH)an d triiodothyronine
    (RIA-gnost T3).

    Statistical analysis
    All data are given as means &standard error of the mean (SEM).
    Significant time (pre- and post-diet measurements) and diet
    effects (C, hC, hHP, hBCAA. hLP) were determined by a two-way
    repeated measures analysis of variance (ANOVA). If the F-value
    indicated significance between group variance. comparisons
    between pre- and post-values in each experimental group (C,
    hC. hHP, hBCAA and hLP) were determined using a paired Student's
    t-test p<0.05 after adjustment using the Bonferroni
    correction was considered to indicate a significant difference
    between groups. The percentage differences between measurements
    obtained pre- and post-dietary period in each group
    were compared with an unpaired Student's t-test. A level of
    p < 0.05 was selected to indicate significant differences between
    mean values.
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    Results

    Body weight
    Analysis of variance (ANOVA) showed that there is a global effect
    of both time and diet on BW changes (p <0.05, Table 2).
    BW losses recorded in all hypoenergy groups were significantly
    higher than those observed in control subjects (Fig. 1, p < 0.05).
    Moreover, the highest BW loss was observed for subjects of
    the hBCAA group. who reduced significantly their BW (4kg.
    - 5.4%, Fig. 1).

    Body composition
    Body fat estimated by skinfold thickness


    There was a global effect of both time and diet on body fat
    changes (Table 2. p < 0.05). The decrease in the percentage of
    body fat (BF) of hBCAA subjects was significantly higher than
    that recorded in the other hypoenergy groups (p<0.05). Subjects
    receiving BCAA supplementation lost an average of 17.3 %
    of their estimated % BF using skinfold measures (Fig. 2).

    Body fat estimated by MRI
    Adipose tissue and lean tissue were clearly identified due to
    large differences in signal on magnetic resonance images
    (MRI).cated a significant effect of diets on SAT area changes
    (p < 0.05). The percent changes in SAT were not significantly
    different between hypoenergy groups (p = 0.12. Fig. 5). Analysis
    of variance showed a global effect of both time and diet on VAT
    area changes (p < 0.05). The VAT losses (- 34.4 %) observed in
    the hBCAA group were significantly higher than those recorded
    in other groups (p < 0.05, Fig. 5).
    As shown in Fig.4. AT at midthigh were clearly distinguished
    from muscle tissue. Analysis of variance showed a significant
    effect of both time and diet on thigh muscle AT(p < 0.05). After
    the 19-day hBCAA diet, losses of AT were significantly higher
    (- 23.4 %) than those observed in other hypoenergy groups
    (p dc0.05, Fig. 6). However, muscle mass loss was not significantly
    affected by the diets in hypoenergy groups.

    Exercise capocity
    The 19-day diet program did not significantly affect V@max
    measured either on the treadmill or using the crankann
    ergometer in any group (Table 3).
    Nineteen days of caloric restriction did not reduce peak power
    output in any group. Dieting did not significantly alter the subject's
    capacity to perform anaerobic work.
    The MVC of the knee extensor muscle was unaffected by the
    dietary protocol (Table 3) as well as the endurance time of the
    knee extensor muscle at 50 % MVC.

    Blood metabolic changes
    Plasma concentrations of glucose, lactate and insulin were unchanged
    after the 19day dietary regimen (Table 4). However,
    the nitrogen-enriched (N-enriched) diets induced lower blood
    concentrations of FFA (- 25 %, - 35.7 %, in the hBCAA and hHP
    groups, respectively) and higher blood concentrations of
    glycerol (62.9 %, 58.7 %, in hBCAA and hHP groups, respectively,
    p < 0.05).
    Plasma T3 concentration was significantly decreased after 19-
    day N-enriched diets (p i 0.05). Prior to dietary restriction, resting
    GH concentrations were similar for all groups. The 19-day
    diet affected all groups equally, resulting in a significant rise
    at rest (p< 0.05). In conditions of pre- or post-diet, exercise increased
    the GH concentrations for all groups. However, the Nenriched
    diets induced a significant increase in post-exercise
    plasma GH concentrations (p < 0.05).
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    Originally Posted by Elliptical Envy View Post
    Thanks a million
    you are welcome. For legal reasons, I cannot post the entire pdf, but the most important parts (Methods & Results) are covered.
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