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Background: Legumes are an excellent plant source of the limiting indispensable amino acid (IAA) lysine in vegetarian, cereal-based diets. However, their digestibility is poor largely because of their antiprotease content. Extrusion can enhance digestibility by inactivating trypsin inhibitors and thus potentially improve the protein quality of legumes.Objective: We measured the digestibility of extruded chickpea and yellow pea protein with use of a dual stable isotope method in moderately stunted South Indian primary school children.Methods: Twenty-eight moderately stunted children (height-for-age z scores <-2.0 SD and >-3.0 SD) aged 6-11 y from low to middle socioeconomic status were randomly assigned to receive a test protein (extruded intrinsically [2H]-labeled chickpea or yellow pea) along with a standard of U-[13C]-spirulina protein to measure amino acid (AA) digestibility with use of a dual stable isotope method. Individual AA digestibility in the test protein was calculated by the ratios of AA enrichments in the test protein to the standard protein in the food and their appearance in blood plasma collected at 6 and 6.5 h during the experiment, representing a plateau state.Results: The mean AA digestibility of extruded chickpea and yellow pea protein in moderately stunted children (HAZ; -2.86 to -1.2) was high and similar in both extruded test proteins (89.0% and 88.0%, respectively, P = 0.83). However, lysine and proline digestibilities were higher in extruded chickpea than yellow pea (79.2% compared with 76.5% and 75.0% compared with 72.0%, respectively, P < 0.02).Conclusion: Extruded chickpea and yellow pea protein had good IAA digestibility in moderately stunted children, which was 20% higher than an earlier report of their digestibility when pressure-cooked, measured by the same method in adults. Higher digestibility of lysine and proline highlights better retention of these AA in chickpea during extrusion-based processing. Extrusion might be useful for developing high-quality protein foods from legumes. This trial was registered at www.ctri.nic.in as CTRI/2018/03/012439. [ABSTRACT FROM AUTHOR]

Severe acute malnutrition (SAM) in children under five years is an important public health problem due to associated high mortality and long-term health consequences. Research on the dietary causes of SAM, especially the role and relative importance of dietary protein, in the aetiology of oedematous malnutrition, has led to considerable debates and controversies. The present article revisits some of the debates in this field, where the researchers at the National Institute of Nutrition (NIN), Hyderabad, India, with their pioneering work, have contributed to the global literature on the various facets of the disease. Highlighting the importance of energy as a bigger problem than protein malnutrition is a noteworthy contribution of NIN's research. It is, however, important to examine the protein quality of the diets in light of the new information on the lysine requirements. The article argues that the currently dominating hypothesis of free radical theory requires a critical review of the supporting evidence. Over the past few decades, the research has focused on low-cost diets using locally available foods. The article also argues that solutions based on local foods, being acceptable and sustainable, need to be strengthened for their effective delivery through the existing nutrition programmes. Recent evidence shows that the use of ready-to-use therapeutic foods (RUTF) with high micronutrient density may be linked with higher mortality possibly due to the high iron content, which could be counterproductive. There are several unaddressed concerns regarding the potential long-term impact of consumption of RUTF in children with SAM. More evidence and a cautious approach are, therefore, needed before implementing these solutions., Competing Interests: None

Background: Lysine requirements of well-nourished children from developing regions have been found to be similar to those of children from developed regions (33.5 mg · kg⁻¹ · d⁻¹). However, intestinal parasites have been shown to increase lysine requirements in undernourished adults, and it is not known if a similar phenomenon occurs in undernourished children from poor and unsanitary environments., Objective: Our objective was to measure the lysine requirement of moderately undernourished school-aged Indian children by the indicator amino acid oxidation technique before and after successful treatment for intestinal parasites., Methods: Twenty-one undernourished school-aged children (∼8 y of age) with z scores between -2 SD and -3 SD for height-for-age or weight-for-age, who tested positive for intestinal parasites, were studied before and after successful antiparasite treatment. Children were fed any 2 of 7 levels of lysine intakes (5, 15, 25, 35, 50, 65, and 80 mg · kg⁻¹ · d⁻¹) in random order. The lysine requirement was determined by applying a 2-phase linear regression crossover analysis on the fractional oxidation rate of the tracer L-[1-¹³C] phenylalanine in response to the graded lysine intakes., Results: The lysine requirement of undernourished children with intestinal parasite infestations was determined to be 42.8 mg · kg⁻¹ · d⁻¹ (95% CI: 32.6, 53.1 mg · kg⁻¹ · d⁻¹), and after successful antiparasitic treatment it was determined to be 35.5 mg · kg⁻¹ · d⁻¹ (95% CI: 25.5, 45.5 mg · kg⁻¹ · d⁻¹). The results were significantly different (P < 0.05), although the 95% CIs overlapped., Conclusions: The lysine requirement in undernourished children is similar to that of well-nourished children, and intestinal parasitic infestation increased the lysine requirement by ∼20%., (© 2015 American Society for Nutrition.)

Cereals and legumes are staple foods in India and are limiting in lysine and sulphur amino acids, respectively. Available lysine loss, due to Maillard-type reactions that may occur during food preparation, exacerbates the problem of lysine deficiency particularly in cereals. Consequently, determining the contents of digestible essential amino acids, particularly lysine, is important. True ileal digestibilities of most amino acids (including total and reactive lysine) were determined for ten food ingredients and eleven foods commonly consumed in India. Semi-synthetic diets each containing either an ingredient or the prepared food as the sole protein source were formulated to contain 100 g kg− 1 protein (75 g kg− 1 for rice-based diets) and fed to growing rats. Titanium dioxide was included as an indigestible marker. Digesta were collected and the amino acid content (including reactive lysine) of diets and ileal digesta determined. Available (digestible reactive) lysine content ranged from 1·9–15·4 g kg− 1 and 1·8–12·7 g kg− 1 across the ingredients and prepared foods respectively. True ileal amino acid digestibility varied widely both across ingredients and prepared foods for each amino acid (on average 60–92 %) and across amino acids within each ingredient and prepared food (overall digestibility 31–96 %). Amino acid digestibility was low for many of the ingredients and prepared foods and consequently digestibility must be considered when assessing the protein quality of poorer quality foods. Given commonly encountered daily energy intakes for members of the Indian population, it is estimated that lysine is limiting for adults in many Indian diets. [ABSTRACT FROM PUBLISHER]

Co-fermentation of finger millet with horse gram was carried out to produce inexpensive protein-rich food (dosa-an Indian breakfast food). Natural fermentation of finger millet-horse gram flour blend in different proportions (2:1, 3:1, 4:1 and 5:1) was performed for 24 h. Biochemical analysis showed reasonable drop in pH (6.6-4.2) and starch content (25.52%) with considerable augment in titratable acidity (0.168-1.046%), soluble proteins (1.1-fold) and free amino acids (2.6-fold) at 16 h. Lactic acid bacteria dominated yeast counts throughout the fermentation accompanied by a decrease in total soluble and reducing sugars. Total essential amino acids increased 1.1-fold at 16-h fermentation with protein containing 48.68% of essential amino acids over total amino acids. Lysine increased from 5.87 to 6.73 g of amino acid/100 g of total amino acids. Dosa, prepared from 16-h fermented batter, showed better sensory attributes for 4:1 ratio. The formulated new product might be used to overcome the protein-energy malnutrition problems.

We recently reported the lysine requirement of school-aged children living in Canada consuming a mixed diet to be 35 mg.kg(-1).d(-1). Because the majority of children in the world live on cereal-based diets in developing countries, we measured the daily lysine requirement in healthy children living in India and consuming a mostly cereal-based diet. Our objective in this study was to determine the lysine requirement in healthy, school-aged children in the developing world by using the indicator amino acid oxidation method with l-[1-(13)C] phenylalanine. Six healthy, school-aged children consumed 7 levels of lysine (5, 15, 25, 35, 50, 65, and 80 mg.kg(-1).d(-1)) each in a random order along with an amino acid mixture providing energy and protein intakes of 1.7 x resting energy expenditure and 1.5 g.kg(-1).d(-1), respectively. The mean lysine requirement was determined by applying a 2-phase linear regression crossover analysis on tracer oxidation (F(13)CO(2)) data, which identified a breakpoint (requirement) at minimal F(13)CO(2) in response to the graded lysine intakes. The mean lysine requirements with the upper 95% CI for children were determined to be 33.5 and 46.6 mg.kg(-1).d(-1), respectively, by breakpoint analysis of the F(13)CO(2) data. The mean lysine requirements of Indian children were almost identical to that of Canadian children (35 mg.kg(-1).d(-1)). There is no evidence for any adaptation in lysine requirements in children from developing countries such as India.

Outbreaks of highly pathogenic avian influenza (HPAI) H5N1 virus were reported for the first time in India during February 2006. Herein, we have sequenced and analyzed the PB2 genes of five influenza virus isolates obtained from three affected states (Gujarat, Madhya Pradesh and Maharashtra) in India during the outbreaks. In the phylogenetic analysis, the Indian isolates were grouped in the mixed-migratory bird sub-lineage of the Eurasian lineage. From the phylogenetic tree, it is evident that viruses were probably introduced to India from China via Europe because they share a direct ancestral relationship with the Indian isolates. The virus might have spread through migratory waterfowls that survived the HPAI H5N1 infection. These viruses were able to replicate in cultured cells of avian and mammalian hosts and posses lysine at position 627 of the PB2 protein, indicating that they might be able to cross the host barrier to infect mammals.

Background: We previously used the 24-h indicator amino acid balance method to show that the lysine requirement in undernourished Indian men from low socioeconomic and unsanitary environments is approximately 50% higher than the mean requirement of 30 mg lysine. kg(-1). d(-1) in well-nourished men., Objective: It is possible that this higher lysine requirement in persons with chronic undernutrition is due to environmental influences, including the presence of intestinal parasites. We assessed this possibility by using 24-h indicator amino acid balance (with leucine) at both the "normal" requirement for lysine intake and the higher requirement, before and after successful treatment to eradicate intestinal parasites in affected, undernourished men., Design: Fourteen chronically undernourished men were studied before and after treatment for intestinal parasites, during each of two 7-d (6-d dietary adaptation plus 1-d tracer experiment) diet periods supplying either 30 (n = 7) or 45 (n = 7) mg lysine. kg(-1). d(-1) from an L-amino acid diet. Twenty-four-hour indicator amino acid balance was estimated on day 6 by [(13)C]leucine tracer infusion., Results: Before the parasite treatment, subjects were in neutral 24-h leucine balance at both lysine intakes. After the eradication of intestinal parasites, there was a significant (P < 0.001) improvement in 24-h leucine balances, which were positive at both lysine intakes., Conclusions: On the basis of the 24-h indicator amino acid balance approach, it appears that intestinal infestation with parasites increases the requirement for lysine and that this may be one factor responsible for the higher lysine requirement observed in persons with chronic undernutrition.

Twenty soil communities from the northeastern forests (Assam) and the Western Ghats (Maharashtra) were screened for the presence of the lysine aminotransferase (lat) gene from Nocardia. Hybridization probes and primers were synthesized in accordance with the reported sequence of the Nocardia lat gene from GenBank (number: G1 49355). Seven positives were obtained from the 20 soils. Six of the seven positive were from the Western Ghats and one from the northeast Assam forests. Eighteen actinomycete isolates from the 7 positive soils showed the presence of the lat gene. Only 9 isolates actually produced an antibiotic. These results are discussed.

Background: In earlier studies with well-nourished subjects that used a 24-h indicator amino acid oxidation or balance approach, we concluded that the 1985 FAO/WHO/UNU requirement for lysine (12 mg x kg(-1) x d(-1)) was inadequate for healthy South Asian subjects and proposed a tentative requirement of 30 mg x kg(-1) x d(-1)., Objective: We assessed whether chronic undernutrition, with low habitual dietary protein and lysine intakes, leads to changed lysine requirements., Design: Twenty-seven otherwise clinically healthy, chronically undernourished Indian men were studied during 2 randomly assigned 7-d diet periods supplying 12 and 30, 18 and 36, or 24 and 42 mg lysine x kg(-1) x d(-1), based on an L-amino acid diet. The subjects' leucine intake was 40 mg x kg(-1) x d(-1). At 1800 on day 6, a 24-h intravenous [(13)C]leucine tracer-infusion protocol was conducted to assess leucine oxidation and daily leucine balance at each test lysine intake., Results: A breakpoint was not identified in the lysine intake-leucine oxidation or balance response over the range of intakes studied. Mixed-models linear regression analysis indicated a mean requirement of 44 mg lysine x kg(-1) x d(-1) (95% CI: 36, 63) for the lysine intake-leucine balance relation., Conclusions: The mean lysine requirement in chronically undernourished men is estimated to be higher than the value of 30 mg x kg(-1) x d(-1) proposed for well-nourished individuals. This may be related to body-composition differences. It also suggests that these subjects have not elicited a metabolic adaptation in response to their habitually low lysine intakes by substantially improving their efficiency of dietary lysine utilization.

Background: The mean lysine requirement of healthy Indian subjects was estimated from short-term experimental diet periods to be 29 mg x kg(-1) x d(-1), which is higher than the 1985 FAO/WHO/UNU upper requirement of 12 mg x kg(-1) x d(-1)., Objective: Our objective was to confirm our proposed requirement of 29 mg x kg(-1) x d(-1) by extending the diet period to 21 d and by using 4 test lysine intakes (12, 20, 28, and 36 mg x kg(-1) x d(-1)) and a 24-h indicator amino acid oxidation and balance approach., Design: During two 21-d diet periods, 18 healthy Indian men were randomly assigned to receive 12 and 28 or 20 and 36 mg lysine x kg(-1) x d(-1) as part of an L-amino acid diet. At 1800 on days 6 and 20, [(13)C]leucine was infused over 24 h to assess leucine oxidation and daily leucine balance at each test intake., Results: Leucine oxidation, balance, and flux did not differ significantly between days 7 and 21. Twenty-four-hour leucine oxidation was lower at lysine intakes of 28 and 36 mg than at 12 and 20 mg. Leucine balances at lysine intakes of 12 and 20 mg were negative and significantly less than equilibrium (P < 0.01) and lower (P < 0.02) than balances at 28 and 36 mg lysine. Two-phase regression analysis indicated a breakpoint at 31 mg lysine x kg(-1) x d(-1) in the relation between lysine intake and 24-h leucine oxidation and balance., Conclusions: Full adaptation to a low lysine intake occurs within 7 d. The previously proposed tentative mean lysine requirement for Western subjects of 30 mg x kg(-1) x d(-1) is confirmed for healthy Indian adults.

Background: In an earlier study, using a modification of the indicator amino acid oxidation approach, we concluded that the 1985 FAO/WHO/UNU-proposed lysine requirement of 12 mg x kg(-1) x d(-1) is likely inadequate to maintain body amino acid homeostasis in apparently healthy south Asian subjects and that our proposed requirement of 30 mg x kg(-1) x d(-1) is more appropriate., Objective: We assessed the lysine requirement in a similar population by using 4 test lysine intakes (12, 20, 28, and 36 mg x kg(-1) x d(-1)) with an indicator amino acid balance approach., Design: Sixteen healthy male Indians were studied during each of 2 randomly assigned 8-d L-amino acid diets that supplied either 12 and 28 or 20 and 36 mg lysine. At 1800 on day 8, a 24-h intravenous [(13)C]leucine tracer-infusion protocol was conducted to assess leucine oxidation and daily leucine balance at each lysine intake., Results: Mean 24-h leucine oxidation rates decreased significantly (P = 0.005) across different lysine intakes and were 104.1, 97.8, 87.3, and 87.3 mg x kg(-1) x d(-1) at intakes of 12, 20, 28, and 36 mg x kg(-1) x d(-1), respectively; mean 24-h leucine balances were 3.3, 9.1, 19.7, and 20.7 mg x kg(-1) x d(-1), respectively (P = 0.015, mixed-model analysis of variance). Oxidation and balances differed significantly between the lower and higher lysine intakes but were not significantly different between the 12- and 20-mg and 28- and 36-mg test intakes. Two-phase regression analysis indicated a mean breakpoint at 29 mg lysine x kg(-1) x d(-1) in the relation between lysine intake and leucine oxidation or balance., Conclusion: We propose a mean lysine requirement of 30 mg x kg(-1) x d(-1) for healthy Indian adults, which is the same amount we proposed previously for Western populations.

Since germination has been found to increase the nutritive qualities of grains, proso millet, a minor millet, was allowed to germinate for 1-7 days and analysed for protein fractions, free amino acids, lysine, tryptophan, methionine, non-protein nitrogen, starch, and sugars. Overnight soaking and germination up to 7 days significantly increased the free amino acids and total sugars while the content of dry weight and starch decreased. Increases in albumin and globulin and large decreases in prolamin accompanied sprouting. The percent protein in germinated grains was higher than in the initial grain as a result of dry matter loss during germination. Further, there was an increase in lysine, tryptophan and non-protein nitrogen contents during germination. No change was noticed in methionine content.