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Protein and Amino Acids


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Title: Protein and Amino Acids

Protein and Amino Acids
  • Leo Galland, M.D., F.A.C.P., F.A.C.N.
  • Foundation for Integrated Medicine

How much protein does a person really need? Does
the source of protein matter?
  • __________________________________
  • Few issues in nutritional science have aroused
    such long-standing and deep-seated controversies
    as protein and amino acid requirements.
  • Millward, J Nutr 1997

The Elements of Life
  • Carbon
  • Oxygen
  • Hydrogen
  • Nitrogen
  • Sulfur
  • Phosphorus
  • Minerals Ca, Mg, Na, K, Fe, Zn, Cu, Se, Mn,
    Cr, Mo, Vn, Ni...

Essential Amino Acids
  • Isoleucine
  • Leucine
  • Lysine
  • Methionine
  • Phenylalanine
  • Threonine
  • Tryptophan
  • Valine
  • Arginine
  • Histidine

Major Non-essential Amino Acids
  • Alanine
  • Asparagine
  • Aspartate
  • Cysteine
  • Glutamate
  • Glutamine
  • Glycine
  • Proline
  • Serine
  • Tyrosine

The Fate of Dietary Protein
  • Digestion and absorption (efficiency varies by
    25, depending upon protein source)
  • Maintenance of body protein stores
  • Net protein synthesis (less than 10 of minimum
    requirements, even in children)
  • Synthesis of non-protein compounds
  • Oxidative deamination

Essential Non-Protein Nitrogen-Requiring
  • Nucleic acid synthesis, including adenosine for
    ATP (glutamate, aspartate, glycine, glutamine)
  • Nervous system Neurotransmitter/neuropeptide
    synthesis (phenylalanine, tryptophan,
  • Immune system Cytokine production,Anti-oxidant
    protection (taurine glutathione cysteine,
    glutamate, glycine)
  • Skeletal muscle Creatine (methionine, arginine,
  • Cardiovascular system Nitric oxide synthesis
  • GI tract Mucin secretion (threonine, cysteine,
    serine, proline)

How dietary protein is recorded
  • grams/day 50-135
  • mg/kg body weight 800-1500
  • of kcalories consumed 10-20

Minimum Protein Requirements
  • Obligatory Nitrogen Loss (metabolic ward)
  • 36 mg/kg/day urinary (85)
  • 12 mg/ fecal (13)
  • 8 mg/kg/day sweat, skin, saliva, hair (2)
  • 54 mg of nitrogen/kg/day total
  • Replacement requires 340 mg protein/kg/day
  • 70 kg person 24 gm/day
  • WHO safe minimum 750 mg/kg/day, of which 84
    mg/kg should be essential amino acids
  • 70 kg person 52 gm/day
  • 60 kg person 46 gm/day

Minimum Safe Protein Requirements for Children
  • Newborns 1.85 mg/kg/day
  • Infants 1.5-1.65 gm/kg/day
  • Toddlers 1.1-1.2 mg/kg/day
  • School children 1.0 mg/kg/day
  • Adolescents 0.9-0.95 mg/kg/day
  • N-balance depends upon caloric intake. Fewer
    calories increase protein needs. Profound
    inter-individual differences

US Adult Protein RDA's
  • female male
  • Calories 1900-2200 2300-2900
  • Protein 46 gm 71 gm
  • about 10 of calories
  • typical US diet 15 of calories
  • popular weight loss diets 30 of calories
  • excess protein is oxidized by deamination

(No Transcript)
Ketogenic Amino Acids
  • Exclusively ketogenic
  • Leucine
  • Lysine
  • Ketogenic and glucogenic
  • Threonine
  • Isoleucine
  • Phenylalanine
  • Tyrosine
  • Tryptophan

Hunter-Gatherer Diets
  • Compared to typical U.S. diet
  • high dietary protein (19-35 of calories)
  • relatively low level of dietary carbohydrate
    (22-40 of calories)
  • variable fat intake (28-58 of calories)

The paradoxical nature of hunter-gatherer diets
meat-based, yet non-atherogenic L Cordain, S B
Eaton, et al Eur J Clin Nutr 2002 56, Suppl 1,
  • 13 known quantitative dietary studies of HG
    demonstrate that animal food provided the
    dominant (65) energy source, while gathered
    plant foods comprised the remainder (35).
  • Comprehensive review of 229 HG societies mean
    subsistence dependence upon gathered plant foods
    was 32, whereas it was 68 for animal foods.

Changes in nutritional status and patterns of
morbidity among free-living elderly persons a
10-year study. Vellas et al, Nutr 1997 13
  • 304 healthy French elderly (median age 72 in
    1980) 97 (34.2) remained healthy74 (26.5)
    became frail or sick 54 (19.1) died, 57 (20.1)
    dropped out. Median protein intake was 0.8-1.2
    g/kg of body weight in 1980-81.
  • Protein intake 1.20-1.76 g/kg associated with
    fewer health problems over the next 10 years vs
    protein intakes lt 0.8 g/kg

High-protein low fat diets are effective for
weight loss and favorably affect biomarkers in
healthy adults. Johnson et al. J Nutr 2003 134
586-91. 2003
  • 20 adults, low fat (lt30 kcal) diets, consuming
    either 10 or 30 protein
  • Weight loss (-6 over 6 weeks) and fat loss (-9
    to -11) not affected by protein
  • Cholesterol (-10 to -12), insulin (-25), uric
    acid (-22 to -30) similar changes
  • Hunger and satiety better with high protein
  • Ca-balance unaffected, N-balance better

A Reduced Ratio of Dietary Carbohydrate to
Protein Improves Body Composition and Blood Lipid
Profiles during Weight Loss in Adult Women.
Layman DK. J.Nutr. 2003 133411-417
  • 24 women age 45 to 56, BMI gt26 kg/m2
  • Isocaloric, matched-fat diets, 68 vs 125 g
    protein/day for 10 weeks.
  • HP loss of fat/lean (6.3 1.2 g/g) vs LP (3.8
  • Cholesterol down 10 for both, TGs down only for
    HP (21)
  • LP had higher insulin responses to meals and
    postprandial hypoglycemia
  • HP reported greater satiety.
  • Increasing the proportion of protein to CHO in
    the diet of adult women has positive effects on
    body composition, blood lipids, glucose
    homeostasis and satiety during weight loss.

Randomized trial on protein vs carbohydrate in ad
libitum fat reduced diet for the treatment of
obesity. Skov et al. Int J Obes 1999 50418-30
  • 50 subjects, protein of 12 vs 25 of kcal, mean
    BMI of 30.8 and 30.0.
  • 27 week weight loss high protein 8.9 kg, high
    CHO 5.1 kg
  • 27 week fat loss high protein 7.6 kg, high CHO
    4.3 kg (84-85 of total weight loss)
  • Kcal consumed 2139 high protein vs 2605 high CHO.

Dietary protein and risk of ischemic heart
disease in women. Hu et al, Amer J Clin Nutr,
1999. 70 221-7
  • 80,082 women aged 3459 y
  • Median protein 14.7 of kcal to 24.0 of kcal by
    quintile. Higher protein associated with less
    tobacco and alcohol use, more exercise and
    multivitamin and vit E supplements, more fat,
    saturated fat, cholesterol, folate, meat, dark
    bread, fruits and vegetables, and less
    carbohydrate (white bread, potatoes, sweets and
  • High protein intakes reduced risk of ischemic
    heart disease RR 0.74 (95). Both animal and
    vegetable protein contributed to the lower risk,
    but most of the protein came from animal sources.
    This inverse association was similar in women
    with low- or high-fat diets.
  • Conclusion Increased dietary protein does not
    increase and may reduce risk of ischemic heart

Short-term effects of substituting protein for
carbohydrate in the diets of moderately
hypercholesterolemic human subjects. Wolfe BM,
Giovannetti PM. Metabolism 1991 Apr40(4)338-43.
  • Hypercholesterolemic human subjects (four men,
    six women)
  • Diets 23 v 11 of energy as protein, 24 as
    fat, 53 v 65 as carbohydrate. Isocaloric
    substitution of protein for CHO during low fat
  • High protein HDL-C higher by 12 (plt.01),
    total cholesterol (TC) lower by 6.5 (plt .001),
    LDL-C lower by 6.4 (plt.02), TG lower by 23
    (plt.02). LDL-C/HDL-C lower by 17 (plt.001).

Cardiovascular Disease and Diabetes Inverse
relationship between urinary markers of animal
protein intake and blood pressure in Chinese
results from the WHO Cardiovascular Diseases and
Alimentary Comparison (CARDIAC) Study.Liua L et
al, Int J Epidemiology 200231227-233
  • Urinary 3-methylhistidine (3MH) excretion (a
    biological marker of animal protein intake) and
    BP in 11 Chinese population samples
  • 3MH and 3MH/creatinine were inversely associated
    with BP and hypertension, even after adjustment
    for age, sex, Na/K ratio, BMI, calcium and
  • The results provide strong evidence that animal
    protein intake is associated inversely with BP in
    Chinese populations.

Impact of diet on blood pressure and age-related
changes in blood pressure in the US population.
Hajjar et al. Arch Intern Med. 2001161589-93.
  • All individuals gt20 yrs old (n 17,030) surveyed
  • Systolic BP and pulse pressure positively
    associated with higher Na, alcohol, and protein
    intakes (Plt.05)
  • Age-related increase in systolic BP was
    attenuated by higher Ca and protein intakes.

An increase in dietary protein improves blood
glucose response in persons with type 2 diabetes.
Gannon et al. Am J Clin Nutr 78 734-41.
  • 10 men, 2 women, age 39-72, BMI 22-37
  • 2250 kcal 55 CHO, 15 protein, 30 fat vs 40
    CHO, 30 protein, 30 fat
  • Glycemic response to high protein diet (AUC) was
    40 lower.
  • Glycated Hb decreased 0.8 at 30 protein vs 0.3
    (plt.05) over 5 weeks.

Meat and dairy food consumption and breast
cancer a pooled analysis of cohort studies.
Missmera et al. Int J Epidemiol 20023178-85
  • Eight prospective cohort studies 351 041 women,
    7379 diagnosed with invasive breast cancer during
    15 years of follow-up.
  • No significant association between intakes of
    total meat, red meat, white meat, total dairy
    fluids, or total dairy solids and breast cancer
  • J-shaped association for egg consumption where,
    compared to women who did not eat eggs, breast
    cancer risk was slightly decreased among women
    who consumed lt2 eggs per week but slightly
    increased among women who consumed 1 egg per day.

Dietary factors and the survival of women with
breast carcinoma. Holmes MD, et al. Cancer
  • 1,982 female registered nurses with breast cancer
    diagnosed between 1976-1990 who completed a food
    frequency questionnaire.
  • The main outcome measure was time to death from
    any cause.
  • The relative risk (95 confidence interval) of
    mortality comparing highest with lowest quintile
    of protein intake was 0.65 (0.47-0.88).

Meat, cancer and dietary advice to the public.
Hill, Eur J Clin Nutr 200256 Suppl 1S36-41
  • Epidemiological data are consistent with a
  • protective role for fruit, vegetables and whole
    grain cereals in colon cancer with no role for
    meat consumption as a risk factor.
  • Meat may play a protective role in gastric
  • Data from Europe are not consistent with those
    from the US because of the different contexts
    (meal composition) within which meat is consumed
    in different countries

Meat consumption and colorectal cancer a review
of epidemiologic evidence. Noral Riboli,
Nutrition Reviews 200159 37-47
  • Review of 36 studies, most show no significant
    effect for any kind of meat
  • Relative risk ranged from 0.41-2.87
  • Studies with significant adverse association
  • RR 1.33-2.87 for total meat in 5/32 studies
  • RR 1.04-2.35 for red meat in 8/26 studies
  • RR 1.18-2.87 for processed meat in 13/32 studies

Protein consumption and bone mineral density in
the elderly the Rancho Bernardo Study.
Promislow et al. Am J Epidemiol 2002155636-44
  • 572 women and 388 men aged 55-92 years.
  • Positive association between animal protein
    consumption, assessed by FFQs in 1988-1992, and
    BMD measured 4 years later.
  • Negative association between vegetable protein
    and BMD was observed in both sexes.
  • Increasing protein consumption appearing to be
    more beneficial for women with lower calcium
    intakes, but evidence for this interaction was
    not consistently strong.

Effect of Protein Intake on Bone Mineralization
during Weight Loss A 6-Month Trial. Skov et al.
Obesity Research 10432-8 (2002)
  • 65 overweight adults, 6-month of high protein
    (HP 107.8 g/d)vs low protein (LP 70.4 g/d)
    matched fat diets.
  • BMC loss, adjusted for differences in fat loss,
    was greater in the LP group than in the HP group
    difference in LP vs. HP, 44.8 g (95 confidence
    interval, 16 to 73.8 g) p lt 0.05.
  • Independent of change in body weight and
    composition during the intervention, high protein
    intake was associated with a diminished loss of
    BMC (p lt 0.01).

Protein supplements increase serum IGF-1 levels
and attenuate proximal femur bone loss in
patients with recent hip fracture. Schurch et
al. Ann Int med 1998 128 801-9.
  • 82 patients, mean age 80 years, mean dietary
    protein 45-51 g/day, recent femoral fracture, vit
    D3 200,000 IU given
  • Supplement 250 kcal, 20 g protein/35 gm CHO/3 g
    fat vs 54 g CHO, Ca/Mg/P/A/K1
  • Protein improved femoral healing at 1 y,
    increased IGF-1 and pre-albumen

A high dairy protein, high-calcium diet minimizes
bone turnover in overweight adults during weight
loss. Bowen et al. J Nutr 2004 134 568-573
  • 60 subjects (BMI 27-40) lost mean of 9.7 kg over
    12 weeks on diets of 34 protein, 41 CHO, 24
  • Diet 1 dairy-based, 2400 mg Ca/day. Diet 2,
    meat-based 500 mg Ca/day
  • Bone resorption markers increased more with
    meat-based diet and osteocalcin increased only
    with meat-based diet

Meta-analysis of the effects of soy protein
intake on serum lipids.Anderson et al NEJM.
  • Meta-analysis of 38 controlled clinical trials
  • Soy protein intake averaged 47 g/day.
  • Compared with animal protein, soy effect
  • Total-C decreased 23.2 mg/dl (9.3, plt.05)
  • LDL-C decreased 21.7 mg/dl (12.9 , plt.05)
  • TG decreased 13.3 mg/dl (10.5 , plt0.05)
  • HDL-C increased 2.4 (NS).
  • TC and LDL-C Changes directly related to the
    initial serum cholesterol (P lt 0.001)

A Dietary Portfolio Approach to Cholesterol
Reduction. Jenkins et al. Metabolism 2002 51
  • 7 men, 6 women age 43-84, BMI 21-31, on diets low
    in saturated fat and cholesterol
  • Per 1000 kcal
  • Soy protein 22.7 g
  • Viscous fiber 8.2 g (oats, barley, psyllium,
    okra, eggplant)
  • Plant sterols 1 g (margarine)
  • Raw unblanched almonds 2.9 g
  • LDL-C decreased 29 (plt.001), LDL/HDL decreased
    26.5 (plt.001)

Legume consumption and risk of coronary heart
disease in US men and women. Bazzano et al. Arch
Intern Med. 20011612573-8
  • 9632 men and women who participated in NHANES 1
    and were free of CVD
  • Frequency of legume intake was estimated using a
    3-month food frequency questionnaire,
  • 19 years of follow-up, 1802 incident cases of CHD
    and 3680 incident cases of CVD were documented.
  • Legume consumption was significantly and
    inversely associated with risk of CHD (P .002
    for trend) and CVD (P .02 for trend) after
    adjustment for established CVD risk factors.
  • Legume consumption 4 times or more per week
    compared with less than once a week was
    associated with a 22 lower risk of CHD (relative
    risk, 0.78 95 confidence interval, 0.68-0.90)
    and an 11 lower risk of CVD (relative risk,
    0.89 95 confidence interval, 0.80-0.98).

The Impact of protein on renal function decline
in women with normal renal function or mild renal
insufficiency.Knight et al, Ann Int med 2003
138 460-7
  • 1624 nurses age 42-68, followed for 11 years,
    protein intake measured by FFQ (mean 76 g/d)
  • GFR estimated from serum creatinine, age, height,
    weight by 2 formulas
  • No effect of protein on GFR in women with
    baseline GFRgt80 ml/min/1.73 m-squared.
  • Initial GFR of 55-80 ml/min (S-creatinine
    .77-1.1mg/dl, mean .88) showed a decline in GFR
    of 7.72 ml/min/10 g increase in protein. Meat
    proteingtdairy protein. No effect of vegetable

Purine-Rich foods, Dairy and Protein Intake, and
the Risk of Gout in Men. Choi et al, NEJM 2004
350 1093-1103
  • 47,150 men followed for 12 years
  • Incidence of gout increased with increasing
    intake of protein from meat or seafood and
    decreased with increasing intake of dairy
    protein. No effect of total protein
  • Relative risk
  • 1.41 for 2.5 vs 0.5 meat servings/day
  • 1.51 for 0.8 vs 0.04 seafood servings/day
  • 0.56 for 4.2 vs 0.5 dairy servings/day

Protein Consumption May Affect Mineral
  • High meat diets increase zinc bioavailability in
    elderly women (Hunt et al, Am J Clin Nutr 1995
    62 621-32) and iron and zinc utilization in
    young women, with no effect above 3 oz lean
    beef/d. (Johnson Walker, J Am Diet Assoc 1992
    92 1474-8).
  • Replacement of 25 meat protein by soy protein
    decreased zinc absorption and protein
    digestibility (Sandstrom et al, J Nutr 1986 116

Large Neutral Amino Acids Share Common
  • The ratio of tryptophan to leucine, isoleucine,
    valine, phenylalanine and tyrosine (5LNAA)
    influences brain serotonin concentration
  • Exercise and CHO loading raise this ratio
  • High protein diets lower this ratio
  • Control of brain serotonin by the diet. Wurtman
    RJ, Fernstrom JD. Adv Neurol 1974519-29

Implications of Competitive Transport of LNAA
into Brain
  • Tryptophan/5LNAA is lower in major depression,
    correlates with severity of depression and with
    plasma cortisol level following dexamethasone
  • Tryptophan/5LNAA is higher after strenuous
    exercise and in patients with fibromyalgia,
    correlates with intensity of post-exertional
    fatigue, reversible with BCAA feeding

References on LNAA Transport
  • Decreased plasma tryptophan concentration in
    major depression relationship to melancholia and
    weight loss. Anderson IM, et al. J Affect Disord
  • Relationship between the dexamethasone
    suppression test and the L-tryptophan/competing
    amino acids ratio in depression. Maes M, et al.
    Psychiatry Res 198721323-35
  • Plasma tryptophan and five other amino acids in
    depressed and normal subjects. DeMyer MK, et al
    Arch Gen Psychiatry 198138642-6
  • Serotonergic markers and lowered plasma
    branched-chain-amino acid concentrations in
    fibromyalgia. Maes M, et al Psychiatry Res 2000
  • Amino acids and central fatigue. Blomstrand E.
    Amino Acids 200120(1)25-34

Glutathione (GSH) Levels Depend Upon Dietary
Sulfur Amino Acids
  • GSH/GSSG is the major human redox pair
  • Malnutrition, fasting and alcoholism deplete
    hepatic and WBC GSH levels
  • Depletion of GSH impairs immune function and
    detoxification of substrates like acetaminophen
  • Loading with sulfur AA reverses these effects

  • High protein intake (up 30 of kcal) is not
    detrimental and may produce improved health
  • Dairy and vegetable protein may have different
    effects than meat protein.
  • Elderly patients often consume too little protein
    and should be encouraged to increase intake to gt1
    gm/kg/day (pulse feeding of 0.8 g/kg at one meal
    may work best Arnal et al, Am J Clin Nutr 1999
    69 1202-8)