Protein Dietary Supplementation Research Articles

Whey Protein Dietary Supplements: Metal Exposure Assessment and Risk Characterization.

Protein supplements (PS) are trendy foods, especially those made from whey. In addition to providing protein, these products are a source of metals, providing essential elements (Na, K, Mg, Ca, Mo, Mn, Fe, Co, Cu, and Zn) and other potentially toxic elements (Al, B, Sr, V Ba, and Ni). In this study, 47 whey PS samples were analyzed for mineral elements by ICP-OES, and their dietary exposures were assessed for three consumption scenarios. Elements found in higher concentrations were K (4689.10 mg/kg) and Ca (3811.27 mg/kg). The intake of 30 g PS (average recommended amount/day) provides about 20% of the established reference value (NRI) for Cr (18.30% for men and 25.63% for women) and Mo (26.99%). In a high daily consumption scenario (100 g PS/day) and when the maximum concentrations are considered, Cr, Zn, Fe, Mo, and Mg dietary intakes of these metals exceed the daily recommended intakes and could pose a risk. The daily intake of 30, 60, and 100 g of whey PS for 25 years does not pose a health risk since the hazard index (HI) is less than one in these consumption scenarios, and the essential elements contributing most to HI are Co, followed by Mo and Cr. It is recommended to improve the information to the consumers of these new products. Furthermore, to help in the management and prevention of these potential health risks, it would be advisable to improve the regulation of these dietary supplements and their labeling.

Cronkhite-Canada Syndrome Presenting with life-threatening protein-losing enteropathy: a case report

Background: Cronkhite Canada Syndrome (CCS) is a rare gastrointestinal polyposis syndrome. The diagnosis of CCS is made by clinical, endoscopic and histopathological findings. Chronic diarrhea, malnutrition, alopecia, skin hyperpigmentation, onychodystrophy, hypogeusia, protein loss due to chronic inflammatory changes in the intestinal mucosa are common findings.Nutritional support, high protein diet, antibiotics, correction of electrolyte imbalance, and corticosteroids are frequently used in CCS treatment. Aim of the study: Here we present a newly diagnosed Cronkhite Canada Syndrome patient who has suffered life-threatening protein loss. Material and methods: Medical documentation was used. Case report: A 62-year-old male patient presented with bloodless diarrhea, happening 8-10 times a day for 4 months, and general condition disorder. On admission, arterial blood pressure was 80/50 mm/hg, pulse was 110 per minute and body temperature was 38.8°C. In laboratory tests, total protein: 38 mg/dl and albumin: 20 g/L.Upper and lower gastrointestinal system (GIS) endoscopy revealed 2-20 mm polyps in the stomach, duodenum, colon, and a small amount in the distal esophagus. Pathological examination of polypectomy materials revealed edematous and inflamed lamina propria consisting of plasma cells, neutrophils and eosinophils.The patient benefited from total parenteral nutrition, high protein dietary supplementation, antibiotic therapy and was followed with upper and lower GIS endoscopy. Conclusions: CCS is a rare disease that can cause life-threatening hypoalbuminemia and requires close follow-up.

Effect Of Protein Supplementation On Dietary Intake, Muscle Function And Soreness During Military Field Exercise

PURPOSE: Military training can result in negative energy balance, low energy availability (≤ 30 kcal·kg FFM-1·d-1), negative protein balance and muscle damage. This study investigated the effect of a high protein dietary supplement on ad libitum energy and macronutrient intake, muscle function and muscle soreness, during an arduous military field exercise. METHODS: Thirty male British Army Officer Cadets were randomly assigned to a control (CON; n = 15, mean ± SD; age 23 ± 2 y, body mass 80.8 ± 6.6 kg) or supplementation group (SUP; n = 15, age 25 ± 3 y, body mass 84.4 ± 12.5 kg). Both groups consumed food ad libitum during a 2-day field exercise and a 4-day post-exercise recovery period. SUP were also provided two protein-rich bars (434 kcal, 46.6 g protein, 27.2 g carbohydrate and 16.4 g fat) per day during field exercise and recovery. Energy expenditure was estimated using a tri-axial accelerometer and energy intake was estimated from food diaries and food wrappers. Isometric mid-thigh pull force, isokinetic knee extension/flexion peak torque, and muscle soreness and perceived fatigue were measured at baseline, and after field exercise and recovery. RESULTS: For energy balance and energy availability there was a main effect of time, but no group or group x time interaction, where participants were in greater negative energy balance (CON: −3717 ± 687, SUP: −3638 ± 1194 kcal·d-1) and lower energy availability (CON: −30 ± 8, SUP: −21 ± 16 kcal·kg FFM-1·d-1) during field exercise. Protein intake was greater in SUP compared to CON during field exercise (CON: 56 ± 22, SUP: 105 ± 30 g·d-1; p < 0.001) and recovery (CON: 125 ± 37 vs. SUP: 161 ± 35 g·d-1; p < 0.05) as desired. Isometric knee extension at a velocity of 180 °/s was 6% greater during recovery than during FEX only, but with no difference between groups. There was no significant difference in any other muscle function measures between conditions or group (all p > 0.05). Muscular soreness and fatigue were increased after FEX (p < 0.05), irrespective of group (p > 0.05). CONCLUSIONS: The provision of a high protein dietary supplement during a military field exercise increased protein intake, but did not attenuate the energy deficit or improve muscle function or soreness.

Whey Protein Powder Analysis by Mid-Infrared Spectroscopy.

There is an ever-expanding number of high protein dietary supplements marketed as beneficial to athletes, body builders, infant formulas, elder care, and animal feed. Consumers will pay more for products with high protein per serving data on their nutritional labels, making the accurate reporting of protein content critical to customer confidence. The Kjeldahl method (KM) is the industry standard to quantitate dairy proteins, but the result is based on nitrogen content, which is an approximation of nitrogen attributable to protein in milk. Product tampering by third-party manufacturers is an issue, due to the lack of United States Food and Drug Administration regulation of nutraceutical products, permitting formulators to add low-cost nitrogen-containing components to artificially inflate the KM approximated protein content in products. Optical spectroscopy is commonly used for quality control measurements and has been identified as having the potential to complement the KM as a more nuanced testing measure of dairy protein. Mid-infrared (MIR) spectroscopy spectra of eight protein standards provided qualitative characterization of each protein by amide I and amide II peak absorbance wavenumber. Protein doping experiments revealed that as protein amounts were increased, the amide I/II peak shape changed from the broad protein powder peaks to the narrower peaks characteristic of the individual protein. Amino acid doping experiments with lysine, glutamic acid, and glycine, determined the limit of detection by MIR spectroscopy as 25%, suggesting that MIR spectroscopy can provide product quality assurance complementary to dairy protein measurement by the KM.

Effects of Pre-Sleep Whey vs. Plant-Based Protein Consumption on Muscle Recovery Following Damaging Morning Exercise.

Pre-sleep whey protein intake has been shown to improve overnight muscle protein synthesis, muscle size and strength, and muscle recovery. Despite a growing interest in alternative protein sources, such as plant-based protein, there is no evidence regarding the efficacy of plant-based proteins consumed pre-sleep. Therefore, we aimed to compare whey vs. plant-based pre-sleep protein dietary supplementation on muscle recovery in middle-aged men. Twenty-seven recreationally active, middle-aged men performed 5 sets of 15 repetitions of maximal eccentric voluntary contractions (ECC) for the knee extensors (ext) and flexors (flex), respectively, in the morning. Participants consumed 40 g of either whey hydrolysate (WH, n = 9), whey isolate (WI, n = 6), rice and pea combination (RP, n = 6), or placebo (PL, n = 6) 30 min pre-sleep on the day of ECC and the following two nights. Catered meals (15% PRO, 55% CHO, 30% Fat) were provided to participants for 5 days to standardize nutrition. Plasma creatine kinase (CK), interleukin-6 (IL-6), and interleukin-10 (IL-10) were measured at pre, immediately post (+0), +4, +6, +24, +48, and +72 h post-ECC. Isometric (ISOM) and isokinetic (ISOK) maximal voluntary contraction force were measured at pre, immediately post (+0), +24, +48, and +72 h post-ECC. Muscle soreness, thigh circumference, and HOMA-IR were measured at pre, +24, +48, and +72 h post-ECC. CK was increased at +4 h post-ECC, remained elevated at all time points compared to baseline (p < 0.001), and was significantly greater at +72 h compared to all other time points (p < 0.001). IL-6 was increased at +6 h (p = 0.002) with no other time differing from baseline. ISOMext was reduced after ECC (p = 0.001) and remained reduced until returning to baseline at +72 h. ISOMflex, ISOKext, and ISOKflex were reduced after ECC and remained reduced at +72 h (p < 0.001). Muscle soreness increased post-ECC (p < 0.001) and did not return to baseline. Thigh circumference (p = 0.456) and HOMA-IR (p = 0.396) did not change post-ECC. There were no significant differences between groups for any outcome measure. These data suggest that middle-aged men consuming 1.08 ± 0.02 g/kg/day PRO did not recover from damaging eccentric exercise at +72 h and that pre-sleep protein ingestion, regardless of protein source, did not aid in muscle recovery when damaging eccentric exercise was performed in the morning.

Cysteine/cystine-rich undenatured whey protein supplement in patients' pressure ulcers outcomes: an open label study

The prevalence and costs associated with treating pressure ulcers (PU) are at high levels. Frequently, PUs heal slowly or not at all, which may be due to the patient's catabolic state which may include protein energy malnutrition. The objective of this open label clinical trial was to improve healing rates by providing patients with a patented, high-quality protein containing all essential amino acids to ensure positive nitrogen balance. An additional benefit of this protein is the delivery of bioavailable cysteine (cystine) to promote glutathione (GSH) synthesis which supports immune function and heightens antioxidant defences. Patients with category II, III and IV PUs were fed 20g BID whey protein dietary supplement for 16-120 days, without change in ongoing 'best practice' PU management and their progress recorded. A total of 10 patients were recruited, with an average age of 77 years. Most had shown no improvement in healing for ≥2 months before treatment and usually had other complications including chronic obstructive pulmonary disease (COPD), diabetes and various cardiovascular diseases. There were a total of 23 PUs, with some patients having more than one. Of these, 44% (n=10) showed complete resolution 83% (n=19) had better than 75% resolution over the observation period. Healing rates ranged from 16.9-0.2cm2/month (healed PUs) and 60.0-1.6cm2/month for resolving PUs. By providing the necessary amino acids to rebuild tissues and bioactive cysteine (cystine) to promote synthesis of intracellular GSH and positive nitrogen balance, improvement in PUs healing was achieved.

Nutrition Supplementation and Education May Increase the Healing Rate in Diabetic Patients with Foot Ulcers (P19-005-19)

ObjectivesThis study examined the effects of high protein dietary supplementation and nutrition education on the healing of diabetic foot ulcers. MethodsTwenty-nine adults with diabetic foot ulcers were recruited from the Tallahassee Memorial Hospital Wound Care Clinic. Participants were randomly assigned to treatment (n = 15) or control (n = 14) group. The treatment group received two carbohydrate controlled high protein shakes to consume daily, and was educated regarding adequate intake of low-fat protein sources, fruits, vegetables, complex carbohydrates and minimizing simple carbohydrates. The control group did not receive any intervention. Daily supplementation provided a total of 500 kilocalories, 28 grams of high-quality protein, and approximately 50% of micronutrient requirements. Anthropometric measurements (weight, height, BMI) and wound planimetry were examined at baseline and every four weeks until the wound healed or up to 12 weeks. ResultsThere was no significant difference between the groups at baseline for BMI (mean = 33.8 ± 7 kg/m2), age (mean = 53.34 ± 11.14 years), duration of diabetes (mean = 13 years), duration of wounds (mean 10.8 months) or wound area (mean = 450 mm2). Wound healing rate (decrease in the area of the wound/week) for the treatment group was three times faster than the control group (151.1 mm2/week vs. 45.2 mm2/week respectively). ConclusionsImproved nutritional intake using nutritional supplementation and patient education has a strong positive effect on the wound healing rate of diabetic foot ulcers. Faster wound healing improves quality of life and functionality of patients with diabetic foot ulcers. Funding SourcesDissertation award.

Análise da adequação da rotulagem de suplementos à base de proteínas

O presente estudo teve como objetivo analisar a conformidade com a legislação vigente, dos rótulos de suplementos proteicos comercializados em um município do Maranhão. A análise foi realizada com suplementos alimentares proteicos, a venda em lojas especializadas e drogarias comercializados no referido município. Os rótulos foram analisados conforme a legislação vigente para praticantes de atividades físicas, RDC nº18/2010, de acordo com parâmetros apresentados pela ANVISA desacordo com a legislação. Dentre as irregularidades encontradas citam-se a não apresentação do número de registro do produto (70,6%), não poder ser adicionado de fibras alimentares e não nutrientes (37,3%), designação do produto de 1/3 do tamanho da fonte da marca (23,5%), mínimo. Foram analisados 51 rótulos de suplementos produzidos por dezesseis fabricantes, observou-se que 90,2% das amostras apresentaram algum item em 10g de proteínas na porção (23,5%), 50% do VET provenientes de proteínas (17,6%), entre outras. Os resultados evidenciam a necessidade de uma fiscalização contínua e rigorosa dos rótulos de suplementos proteicos, com vistas à garantia da orientação adequada do consumidor na aquisição de produtos.

Relationships between use of dietary supplements, caffeine and sensation seeking among college students

Objective: Caffeine and dietary supplement (DS) use by college students is not well-documented. Given reported associations between energy drink consumption and sensation seeking, we used the Sensation Seeking Scale Form V (SSS-V) to assess relationships between sensation-seeking, caffeine, and DS use. Participants: Data from 1,248 college students from five US institutions were collected from 2009 to 2011. Methods: Linear regression was used to examine relationships between scores on the SSS-V and caffeine and DS use, demographic, and lifestyle characteristics. Results: Male sex, nonHispanic race-ethnicity, higher family income, tobacco use, consuming caffeinated beverages, more than 400 mg caffeine per day, and energy drinks with alcohol at least 50% of the time, were significantly associated with higher total SSS-V scores (P < 0.001). Those using protein DSs had higher total, disinhibition, and boredom susceptibility SSS-V scores (Ps < 0.001). Conclusions: Results demonstrate a positive correlation between sensation-seeking attitudes and habitual caffeine, energy drink, and DS consumption.

Effects of sheep/goat whey protein dietary supplementation on the redox status of rats.

The purpose of the present study is to estimate the effects of sheep/goat whey protein dietary supplementation on the redox status of blood and tissues of rats. Twelve male Wistar rats were divided into the control group (standard commercial diet) and whey group [standard commercial diet + sheep/goat whey protein (1 g kg b.w/day)] (6 rats/group). The animals were maintainted on their respective diet for 28 days. At the end of the experimental period, reduced glutathione, catalase activity, total antioxidant capacity, thiobarbituric reactive substances, protein carbonyls and the decomposition rate of H2O2 were measured in blood and tissues of rats. According to the results, the rats fed with the sheep/goat whey protein exhibited improved antioxidant status and decreased free radical-induced toxic effects on lipids and proteins. Specifically, in blood, GSH and CAT levels were significantly increased while TBARS and protein carbonyl levels were significantly decreased compared to the control group. Regarding the effects on tissues, it was observed that GSH levels were significantly increased in small intestine, quadriceps muscle, pancreas and lung tissue compared to the control group. The decomposition rate of H2O2 was significantly decreased in liver, brain and quadriceps muscle, but was significantly increased in spleen tissue compared to the control group. TBARS levels were significantly decreased in liver, brain, quadriceps muscle, pancreas, lung and spleen tissue compared to the control group. Finally, protein carbonyl levels were significantly decreased in brain, small intestine, kidney, pancreas and spleen tissue compared to the control group. Thus, the present findings show the beneficial effects of sheep/goat whey protein, a by-product of cheese manufacturing, on the redox status in an in vivo model.

Effects of biscuit-type feeding supplementation on the neurocognitive outcomes of HIV-affected school-age children: a randomized, double-blind, controlled intervention trial in Kenya

Objective To determine if meat or soy protein dietary supplementation will enhance the neurocognitive performance of HIV-affected children at-risk of malnutrition and food insecurity. Methods A randomized, double-blind, controlled intervention trial evaluated the effect of nutritional supplementation on the neurocognitive outcomes of 49 HIV-affected school-age children in western Kenya. The intervention consisted in providing the mother, target child, and siblings with one of three isocaloric biscuit-type supplements – soy, wheat, or beef – on 5 days per week for 18 months. Neurocognitive outcomes of the target children were assessed by a battery of eight measures and followed up longitudinally for up to 24 months. Results Mixed effects modeling demonstrated significant differences in the rates of increase over time among all three groups (F test degrees of freedom of 2, P Conclusion HIV-affected school-age children provided with soy protein supplementation showed greater improvement in nonverbal cognitive (fluid intelligence) performance compared with peers who received isocaloric beef or wheat biscuits. Soy nutrients may have an enhancing effect on neurocognitive skills in HIV-affected school-age children.

The effects of protein dietary supplementation on fecal egg counts and hematological parameters in goat kids with subclinical nematodosis.

Aim:The aim of the present study was to assess the effect of dietary supplementation with different levels of protein on fecal egg counts and hematological parameters in goat kids with subclinical nematodosis under semi-intensive condition.Materials and Methods:20 goat kids (3-5 months old with an average body weight of 8.90 kg) were randomly allocated to four groups: T1, served as a negative control, without receiving concentrate feed, and T2, T3, and T4 that received concentrate feed containing 16, 20, and 24% digestible crude protein, respectively. The experiment was carried out for 60 days.Results:In this study, protein supplementation had a significant (p<0.05) effect on fecal egg counts even after 15 days; hemoglobin (Hb) (g/dl) after 45 days; total leukocyte count (103/mm3) and total erythrocyte count (106/mm3) after 30 days; packed cell volume (%), lymphocyte (%), and eosinophil (%) after 15 days of supplementation, whereas monocyte (%) and neutrophil (%) values were not significantly influenced by protein supplementation effect during the entire experiment. The values of mean corpuscular volume (fl) were affected significantly (p<0.05, p<0.01) due to protein supplementation after 30 days, mean corpuscular Hb (MCH) (pg) after 45 days, but MCH concentration (g/dl) was not significantly different among the experimental groups during the entire experiment.Conclusion:The dietary supplementation with different levels of protein significantly improved the hematological profiles and inhibited the nematodosis infection in the experimental goat kids.

Effect of L-arginine dietary supplementation on salivary urea concentration and pH in physically active individuals

The aim of this study was to assess if the consumption of 3g of a commercially available L-arginine dietary supplement causes a postabsorptive rise in urea concentration or pH of unstimulated saliva in a group of physically active individuals. Salivary urea and pH were determined for 117 participants in a randomized double-blinded placebo-controlled study. Samples were collected by 'spitting' method in fasting conditions. One hour prior to their second visit, participants consumed three tablets of L-arginine or placebo. Urea concentration was significantly lower at second measurement for both the study and control group. The magnitude of the change was not significant between the groups. pH was higher for both groups at second measurement, but only significant for the study group. The magnitude of the change was significant between the groups. Participants who intermittently ingested protein dietary supplements and those with a Body Mass Index (BMI) higher than 25 had significantly higher basal urea concentration. The results of this study did not confirm the hypothesis. Further studies are needed to determine the effects of different doses of L-arginine supplements on the biochemical composition of saliva and the influence of their long-term consumption on the risk of developing dental diseases.

Rhabdomyolysis: When Exercising Becomes a Risk

www.jpedhc.org was his first spin class, and although he tired, he kept going in order to keep up with the rest of the class. During the prior month, he had started conditioning workouts, which consisted of lifting weights, squats, and treadmill running, and he experienced no significant muscle soreness or weakness. The adolescent claimed to be well hydrated while working out and used a 100% whey protein dietary supplement after exercising. After his workout 3 days ago, he admitted drinking alcohol but denied the use of other drugs. He does not routinely take other medications or use dietary supplements, such as creatine.

The effects of soy protein containing phytoestrogens on menopausal symptoms in postmenopausal women

Objective To analyze the impact of soy protein dietary supplements containing phytoestrogens on menopausal symptoms in healthy postmenopausal women.Methods A double-blind, placebo-controlled trial was conducted in 94 healthy postmenopausal women aged 50–75 years, with 44 randomized to soy supplements containing 118 mg of isoflavones (daidzein, genistein, glycitein and their respective glycosides), and 50 to an identically presented casein placebo. A validated questionnaire on menopausal symptoms was administered at baseline and at 3 months of treatment. Compliance was assessed by high-performance liquid chromatography assay of urinary phytoestrogens. Statistical analysis was completed using non-parametric statistical methods and multivariate analysis.Results At baseline 80% of women recruited were experiencing menopausal symptoms, although symptom severity was mild. Those consuming phytoestrogen supplements had 13- and 17-fold increases in urinary excretion of genistein and daidzein, respectively, with no change in the placebo group. Active soy supplements did not significantly alter either individual symptoms or specific symptom category scores when compared to placebo. Within-group comparisons revealed that the active group reported a significant improvement in vaginal dryness (p = 0.01), libido (p =0.009), facial hair (p = 0.04) and dry skin (p =0.027). However, similarly, those on placebo reported an improvement in libido (p =0.015), facial hair (p = 0.014) and dry skin (p = 0.011) but not vaginal dryness.Conclusions In this group of 94 older postmenopausal women with a high frequency of mild menopausal symptoms, 3 months of soy supplements containing phytoestrogens did not provide symptomatic relief compared with placebo.

Effect of Fish Protein Supplementation on Aminoacid Profile and Nutritional Status in Haemodialysis Patients

Plasma concentrations of aminoacids, albumin, prealbumin, transferrin, C3 and C4 complement, and anthropometric measurements were determined before and after oral administration of a new preparation of fish protein in 19 haemodialysis patients participating in a randomised, double-blind, placebo-controlled crossover study lasting 6 months. Furthermore, plasma concentrations of aminoacids were measured in 24 control subjects. Before fish protein dietary supplementation, serine, threonine, tyrosine, valine, methionine, isoleucine, phenylalanine, leucine, and lysine were lower than in control subjects. Serine, histidine, valine, tyrosine, methionine, isoleucine, leucine, and lysine increased after treatment, and phenylalanine tended to increase. The ratios of essential: non-essential aminoacids and alanine:branched-chain aminoacids (BCAA) became normal, whereas valine: lycine and serine:glycine increased significantly and tyrosine:phenylalanine was unchanged. Plasma protein concentrations were unchanged after treatment. Serum transferrin, triceps and subscapular skinfolds, and bodyweight index were slightly reduced in the patients, whereas arm muscle circumference was normal. Bodyweight, weight index, and arm muscle circumference increased after active treatment. In conclusion, the increase in body weight and the return to normal of the plasma aminoacid profile and aminoacid ratios seem to indicate that the treatment with this new fish protein preparation may have beneficial effects on some of the nutrition-related abnormalities in haemodialysis patients.

The synthesis of amino acid derivatives by catalytic phase-transfer alkylations

The current widespread interest in preparation and utilization of amino acids,2 the building blocks of life, is based on their use in many diversified areas. For example, amino acids are employed as protein dietary supplements,3 as drugs4 and as starting materials for synthesis of biologically active natural products.5 New and general synthetic routes to amino acids are, therefore, of interest for both large-scale production of known amino acids as well as preparation of rare and/or unknown amino acids. Phase-transfer reactions6 provide a potentially attractive synthetic route for preparation of higher amino acids from derivatives of simplest amino acid, glycine. We recently reported7 first general synthesis of amino acid derivatives using phase-transfer alkylations.B3g The Schiff base l7 is an active methylene compound in which both carboxylate and

Protein dietary supplementation and physical performance

ABSTRACTHalf of a Physical Training Platoon at Officer Candidate School, Quantico, Virginia, were given protein supplements, while the other half received a placebo. Both groups significantly improved their physical performance scores as a result of the training program, but there was no significant

Effect of a Protein Dietary Supplement on Muscular Strength and Hypertrophy

Effect of a Protein Dietary Supplement on Muscular Strength and Hypertrophy

Aspects of nutritional anaemia in pregnancy in Gambia

1. (1) Eleven patients with anaemias of pregnancy were studied over a period of 4 months. Six with megaloblastic, one with normocytic, and one with microcytic anaemia were given a high protein dietary supplement, but no haematological response was observed. 2. (2) A study of eight patients with tropical macrocytic anaemia of pregnancy and bone marrows containing “transitional megaloblasts” leads to the proposition that two factors are present in the aetiology of this anaemia as found in the Gambia: 1. i. a haemolytic factor, which ceases directly on delivery and, in the production of which past malarial infection can be suspected of playing a part. 2. ii. a failure of utilization of haemopoietic substances in pregnancy which lasts for at least a week after delivery. The findings are thought to provide evidence that, in the Gambia, dietary deficiency of haemopoietic substances, or direct competition for them between foetus and mother, are not major factors in the production of this anaemia.