scholarly journals Ileal digestibility of intrinsically labeled hen's egg and meat protein determined with the dual stable isotope tracer method in Indian adults

2018 ◽  
Vol 108 (5) ◽  
pp. 980-987 ◽  
Author(s):  
Sindhu Kashyap ◽  
Nirupama Shivakumar ◽  
Aneesia Varkey ◽  
Rajendran Duraisamy ◽  
Tinku Thomas ◽  
...  
Keyword(s):  
Amino Acid ◽  
Tracer Method ◽  
Dual Isotope ◽  
Ileal Digestibility ◽  
Egg White Protein ◽  
Meat Protein ◽  
Isotope Tracer ◽  
Hen’S Egg ◽  
The Mean ◽  
Cooked Meat

ABSTRACTBackgroundProtein quality assessment through the Digestible Indispensable Amino Acid Score requires accurate measurements of true ileal protein and amino acid digestibility, for which a dual isotope technique was recently developed. However, the ileal digestibility of indispensable amino acids (IAA) in humans from high-quality proteins is not well known.ObjectiveThe aim of this study was to intrinsically label hen's egg and meat protein by the use of uniformly 2H-labeled amino acids, and to measure their true ileal indispensable amino acid (IAA) digestibility via the dual isotope method in humans.Design2H-labeled lyophilized boiled egg white protein, whole boiled egg, and cooked meat were obtained from layer hens (BV-300) administered a uniformly 2H-labeled amino acid mix orally for 35 d with their daily feed. The ileal IAA digestibility of these proteins was determined with reference to digestibility of previously characterized [U-13C]spirulina in a dual tracer method in healthy Indian subjects whose intestinal health was measured by the plasma kynurenine-to-tryptophan (KT) ratio.ResultsAll subjects had normal KT ratios. The mean ± SD true ileal IAA digestibility of 2H-labeled egg white protein, whole boiled egg, and cooked meat was 86.3% ± 4.6%, 89.4% ± 4.5%, and 92.0% ± 2.8%, respectively. Leucine digestibility correlated with the KT ratio (r = −0.772; P = 0.009).ConclusionsUniformly 2H-labeled hen's egg and meat protein can be used to measure ileal IAA digestibility by the dual isotope tracer approach in humans. The mean IAA digestibility values for these high-quality proteins in the healthy Indians studied were similar to values obtained in earlier human and animal experiments. Leucine digestibility in these meal matrices correlated with the KT ratio, but this aspect needs further evaluation. This trial was registered at the Clinical Trials Registry of India (http://ctri.nic.in) as CTRI/2018/03/012265.

scholarly journals True ileal digestibility of legumes determined by dual-isotope tracer method in Indian adults

2019 ◽  
Vol 110 (4) ◽  
pp. 873-882 ◽  
Author(s):  
Sindhu Kashyap ◽  
Aneesia Varkey ◽  
Nirupama Shivakumar ◽  
Sarita Devi ◽  
Rajashekar Reddy B H ◽  
...  
Keyword(s):  
Mung Bean ◽  
Deuterium Oxide ◽  
Protein Digestibility ◽  
Plant Protein ◽  
Tracer Technique ◽  
Feeding Method ◽  
Dual Isotope ◽  
Ileal Digestibility ◽  
Isotope Tracer ◽  
Plant Protein Sources

ABSTRACTBackgroundGood-quality plant protein sources are important for protein adequacy in a balanced diet. Legumes are known to be a source of good quality plant protein, but the true ileal digestibility of indispensable amino acids (IAAs) of commonly consumed legumes is not known in humans.ObjectivesIn this study we measured the true ileal IAA digestibility of 2H-intrinsically labeled chickpea, yellow pea, and mung bean (hulled and dehulled) protein, using the dual-isotope tracer technique referenced to a standard protein ([U-13C] spirulina). The study also aimed to validate the use of [U-13C] spirulina as a reference protein in this method.Methods2H-intrinsically labeled legumes, obtained by watering plants with deuterium oxide (2H2O), were administered in a plateau feeding method to healthy Indian adults to measure their true ileal IAA digestibility with the dual-isotope tracer technique, using [U-13C] spirulina protein or a 13C-algal IAA mixture as the standard.ResultThe true ileal IAA digestibilities (mean ± SD) of chickpea, yellow pea, and mung bean were 74.6 ± 0.8%, 71.6 ± 1.3%, and 63.2 ± 1.5%, respectively. The true mean ileal IAA digestibility of mung bean when referenced to [U-13C] spirulina protein or a 13C-algal IAA mixture did not differ significantly (63.2 ± 1.5% versus 64.0 ± 2.4%, P > 0.05). The true ileal IAA digestibility of mung bean improved to 70.9 ± 2.1% after dehulling.ConclusionsThe true mean ileal IAA digestibility of legumes in healthy Indian adults was lower than expected. Traditional processing techniques such as dehulling improve protein digestibility by about 8%. This study was registered in the Clinical Trials Registry of India (CTRI): CTRI/2017/11/010468 (http://ctri.nic.in, accessed on 28/03/2019).

scholarly journals Bioefficacy of a mono-component protease in the diets of pigs and poultry: a meta-analysis of effect on ileal amino acid digestibility

2013 ◽  
Vol 2 ◽  
Author(s):  
Aaron J. Cowieson ◽  
Franz F. Roos
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Meta Analysis ◽  
Control Diet ◽  
Feed Ingredients ◽  
Ileal Digestibility ◽  
Bacterial Protease ◽  
Wide Range ◽  
Functional Patterns ◽  
The Mean

SummaryA meta-analysis of the effect of a mono-component bacterial protease (RONOZYME® ProAct) on the apparent ileal digestibility of amino acids in poultry and swine diets was conducted to examine functional patterns, mean effects and variability of response. A total of 25 independently-conducted experiments were included comprising a total of 804 datapoints. The mean response to protease was +3.74% (SE 1.1%, P < 0.001) and this ranged from +5.6% for Thr (SE 1.2%, P < 0.001) to +2.7% for Glu (SE 1.2%, P < 0.05). For the most economically critical amino acids (Lys, Cys, Met and Thr) the mean response was 4.5%. The effect of protease was independent of geography, animal species and diet composition (P > 0.05). However, the inherent digestibility of amino acids in the control diet as a single explanatory term explained around 47% of the variance (P < 0.001) in effect. When the inherent digestibility of amino acids in the control diet was less than 70% protease addition improved amino acid digestibility in 90% of cases with a mean improvement of around 10%. When the inherent digestibility of amino acids in the control diet was more than 90% there was a protease-mediated improvement in digestibility in only 60% of cases with a mean improvement of around 2%. It can be concluded that the inherent digestibility of amino acids in the diet without protease supplementation is the primary explanatory term for the efficacy of this exogenous protease, demonstrating that it is highly effective in improving the digestibility of amino acids across a wide range of feed ingredients. Benchmarking diets or feed ingredients as to their relative nutritional value would enhance the ability of nutritionists to determine the likely return on investment on use of bacterial proteases in their operation.

Standardized ileal amino acid digestibility in wheat, barley, pea and flaxseed for broiler chickens

2011 ◽  
Vol 91 (1) ◽  
pp. 103-111 ◽  
Author(s):  
A. Bandegan ◽  
A. Golian ◽  
E. Kiarie ◽  
R. L. Payne ◽  
G. H. Crow ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Broiler Chickens ◽  
Sole Source ◽  
Broiler Chicks ◽  
Test Diet ◽  
Nutrient Excretion ◽  
Ileal Digestibility ◽  
Coefficients Of Variation ◽  
The Mean

Bandegan A., Golian, A., Kiarie E., Payne R. L., Crow G. H., Guenter W. and Nyachoti C. M. 2011. Standardized ileal amino acid digestibility in wheat, barley, pea and flaxseed for broiler chickens. Can. J. Anim. Sci. 91: 103–111. Experiments were conducted to estimate ileal amino acids (AA) digestibility in six wheat (exp. 1; 288 birds), seven barley (exp. 2; 252 birds), five pea (exp. 3; 240 birds) and five flaxseed (exp. 4; 216 birds) samples in male Ross broiler chicks. Wheat, barley and pea were the sole source of dietary AA in exps. 1, 2 and 3, whereas, for exp. 4, flaxseed and wheat (50:50 wt/wt) or wheat provided dietary AA. Chromic oxide was the indigestible marker. Birds were fed chick starter for the first 14 d; on day 15 each test diet was randomly assigned to six cages, each with six (exps. 2 and 4) or eight (exps. 1 and 3) birds. On day 21, birds were killed and digesta were sampled to determine the apparent ileal AA digestibility (AID). The standardized ileal digestibility (SID) values were calculated using ileal endogenous AA losses previously determined in our laboratory from birds fed N-free diet. The mean SID values for indispensable AA ranged from 83.7% (Lys) to 93.8% (Phe), 80.4% (Arg) to 90.9% (Phe), 86.0% (Met+Cys) to 94.2% (Arg) and 47.4% (Thr) to 66.7% (Met) for wheat, barley, peas and flaxseed samples, respectively. Overall, SID AA estimates for flaxseed had higher (> 13%) coefficients of variation (CV) compared with other ingredients (CV<6%). These SID AA data will help nutritionists to formulate broiler diets that more closely match the birds' requirements and minimize nutrient excretion.

scholarly journals Real ileal digestibility of sunflower protein and amino acids in humans

2020 ◽  
Vol 79 (OCE2) ◽  
Author(s):  
Romain Tessier ◽  
Juliane Calvez ◽  
Nadezda Khodorova ◽  
Alain Quinsac ◽  
Romain Kapel ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Healthy Volunteers ◽  
Protein Isolate ◽  
Nitrogen Flow ◽  
Dual Isotope ◽  
Ileal Digestibility ◽  
Isotope Method ◽  
Indispensable Amino Acids ◽  
Sunflower Protein

AbstractIntroduction:Sunflower protein is not used in human nutrition despite a relatively good amino acid composition. However, the bioavailability of sunflower isolate has never been measured in Humans. The goal of this work was to determine ileal digestibility of protein and amino acids from a sunflower isolate in healthy volunteers and to challenge newly developed dual isotope method.Materials and methods:Eight healthy volunteers were equipped with a naso-ileal tube. They received during four hours twelve doses of biscuits containing, in total, 25 g of 15N sunflower protein isolate together with 400 mg of a mixture of free 13C amino acids incorporated in chocolates. Polyethylene glycol was perfused as non-absorbable marker and ileal contents were collected during 8 hours after ingestion of the first meal. Real ileal digestibility was measured by assessing nitrogen and carbon content as well as 15N and 13C enrichments by EA-IRMS. Amino acid digestibility was determined by measuring 15N and 13C enrichments by GC-C-IRMS and quantity of amino acids by UPLC. Blood was collected for 8 h to determine 15N and 13C enrichments by GC-C-IRMS.Results:The ileal nitrogen flow was 2.7 ± 0.5 mL/min (mean ± SD). In average, 53.1 ± 12.0 mmol of exogenous nitrogen was recovered during the eight hours of experiment, resulting in an ileal digestibility of sunflower isolate was 85.6 ± 2.6 % of nitrogen ingested. 13C amino acids were also recovered at the ileal level. The mixture of free 13C revealed an ileal digestibility of 94.9 ± 0.9 %. Ileal indispensable amino acids digestibility and DIAAS are in progress.Discussion:Ileal digestibility of sunflower isolate incorporated in toasted biscuits was lower than the value found or a raw isolate in a rat model (94.5%). The study revealed that 5 % of free amino acids were not absorbed in the ileum. Amino acid digestibility will complete the study to evaluate the DIAAS of sunflower isolate and to compare values obtained with the standard method and with the dual isotope method.

scholarly journals Apparent and standardised ileal digestibility coefficients of amino acids in wheat, soybean meal and rapeseed meal for broilers

2021 ◽  
Vol 30 (2) ◽  
Author(s):  
Sini Perttilä ◽  
Taina Jalava ◽  
Marketta Rinne ◽  
Gabriel Da Silva Viana ◽  
Jarmo Valaja
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Soybean Meal ◽  
Broiler Chicken ◽  
Rapeseed Meal ◽  
Ileal Digestibility ◽  
Endogenous Amino Acids ◽  
The Mean ◽  
The Difference ◽  
Protein Free Diet

The apparent (AID) and(SID) ileal amino acid digestibilities in wheat, soybean meal, and rapeseed meal were determined with Ross 308 broiler chicken (n = 64) using the slaughter technique with chromium mordanted straw as an indigestible marker. The recovery of endogenous amino acids at the distal ileum was determined with a protein-free diet and it was used to calculate the SID digestibilities of the studied feed ingredients. The mean amino acid AID and SID were higher in soybean meal and wheat than in rapeseed meal (p<0.05). The mean amino acid flow measured from the ileum was the highest in rapeseed meal, intermediate in soybean meal, and the lowest in wheat (p<0.05). The main amino acids in the basal endogenous secretion were aspartic and glutamic acid and the lowest ones present were methionine and histidine (p<0.05). The difference between amino acid AID and SID values varied among ingredients and was higher in wheat than in soybean meal and rapeseed meal (p<0.05).

scholarly journals Confrontation of the “Dual Tracer” Indirect Method With Direct Ileal Sampling for Indispensable Amino Acid Digestibility of Sunflower Isolate in Humans

2021 ◽  
Vol 5 (Supplement_2) ◽  
pp. 884-884
Author(s):  
Romain Tessier ◽  
Juliane Calvez ◽  
Nadezda Khodorova ◽  
Alain Quinsac ◽  
Romain Kapel ◽  
...  
Keyword(s):  
Amino Acid ◽  
Indirect Method ◽  
Enrichment Ratio ◽  
Tracer Method ◽  
National Agency ◽  
Isoleucine Leucine ◽  
Ileal Digestibility ◽  
Less Invasive ◽  
Indispensable Amino Acid ◽  
Dual Tracer

Abstract Objectives The direct assessment of ileal samples of amino acid (AA) digestibility is invasive in humans. A less invasive but indirect method, namely « dual tracer » was recently developed. It relies on the plasma isotopic enrichment ratio of two labeled protein, a reference protein or alternately AAs labeled with 13C and the test protein labeled with 15N. This recent method has not yet been challenged against direct measurement of ileal digestibility. Methods Seven healthy volunteers were intubated with naso-ileal tube. Every 30min for 4h, they ingested sunflower biscuits containing a total of 25g of 15N intrinsically labeled sunflower protein isolate. They also ingested 60g chocolate containing a total of 400mg of a mix of 13C algal individual AAs. Ileal contents were collected continuously for 8h following the first meal and plasma was sampled every 30min for 4h and hourly between 4 and 8h. 15N and 13C indispensable amino acid (IAA) ileal digestibility were determined by measuring 15N and 13C enrichment in AAs by GC-C-IRMS and AA content by UHPLC in ileal effluent. Plasma and meal 15N and 13C IAA enrichment were measured by GC-C-IRMS. Isotopic 15N/13C ratio were determined using area under the curve value for each isotope. Results Using direct ileal sampling, average IAA ileal digestibility was: (i) 88.5 ± 5.0% for sunflower isolate (15N) with values ranged from 85.8 ± 5.1% for threonine to 91.1 ± 5.8% for methionine, and (ii) 97.6 ± 1.7% for free AAs (13C) with values ranged from 95.9 ± 2.3% for lysine to 98.8 ± 0.8% for phenylalanine. With the “dual tracer” method, digestibility of isoleucine, leucine, threonine and valine was significantly lower than with ileal determination (from 7.9% for threonine to 24.3% for leucine), Methionine and phenylalanine values were aberrant (over 100%) For lysine, the difference between the two methods was not statistically different (4.7%, p = 0.49). Conclusions With our methodological conditions, the “dual tracer” method provides physiological values for most IAA except methionine and phenylalanine. However, values were low compared to ileal digestibility (about 10%) and interindividual variability was high. This less invasive method is promising but requires methodological improvements. Funding Sources French Research National Agency (ANR), financial support of SOFIPROTEOL under the FASO Project PRODIAL.

scholarly journals The Complementarity of Amino Acids in Cooked Pulse/Cereal Blends and Effects on DIAAS

Plants ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 1999
Author(s):  
Fei Han ◽  
Paul James Moughan ◽  
Juntao Li ◽  
Natascha Stroebinger ◽  
Shaojie Pang
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Mung Bean ◽  
Protein Quality ◽  
Latin Square ◽  
Growing Pigs ◽  
Adzuki Bean ◽  
Ileal Digestibility ◽  
Indispensable Amino Acids ◽  
The Mean

The aim was to study the complementary effect between cereals and pulses on protein quality. The values for the digestible indispensable amino acid score (DIAAS) in cooked cereals and pulses, given alone, and blends of cooked cereals and pulses, were determined. True ileal digestibility (TID) values of amino acids for adult humans were obtained. It is difficult to determine ileal amino acid digestibility in humans directly, and for this reason, the growing pig is often used to obtain such values, as a preferred animal model. Seven growing pigs fitted with a T-cannula at the terminal ileum were allotted to a 7 × 6 incomplete Latin square with seven semi-synthetic diets (cooked mung bean, adzuki bean, millet, adlay, mung bean + millet, adzuki bean + adlay, and an N-free diet) and six 7-day periods. The mean TID values for crude protein differed significantly (p < 0.05), with millet having the highest digestibility (89.4%) and the adzuki bean/adlay mixture having the lowest (79.5%). For lysine, adzuki bean had the highest TID (90%) and millet had the lowest (70%). For the mean of all the amino acids, there was a significant (p < 0.05) effect of diet, with the TID ranging from 72.4% for the adzuki bean/adlay mixture to 89.9% for the adzuki beans. For the older child, adolescent, and adult, the DIAAS (%) was 93 for mung beans, 78 for adzuki beans, 22 for millet, 16 for adlay, and 66 for mung beans + millet, and 51 for adzuki beans + adlay. For mung beans, valine was first-limiting, and the SAA for adzuki beans, while lysine was first-limiting for the other foods. Chinese traditional diets, containing both cereals and pulses, are complementary for most, but not all of the indispensable amino acids.

scholarly journals Comparison of Dual Isotope and Ileal Sampling Methods for the Determination of the Amino Acid Digestibility of Pea Protein and Casein in Adult Humans

2021 ◽  
Vol 5 (Supplement_2) ◽  
pp. 873-873
Author(s):  
Florence Guillin ◽  
Claire Gaudichon ◽  
Laetitia Guerin-Deremaux ◽  
Catherine Lefranc-Millot ◽  
Gheorghe Airinei ◽  
...  
Keyword(s):  
Amino Acid ◽  
Sampling Method ◽  
Protein Quality ◽  
Plasma Samples ◽  
Tracer Method ◽  
Pea Protein ◽  
Dual Isotope ◽  
Reference Protein ◽  
Dual Tracer

Abstract Objectives The measurement of amino acid (AA) digestibility of protein through direct ileal sampling is highly invasive and inappropriate for vulnerable populations, such as children or elderly. The new dual tracer method relies on comparing meal and plasma isotopic ratios of 1/a test protein 2/a reference protein (or AA mix) of known digestibility, each one being labelled with a different isotope. The aim of this study was to compare this new indirect dual tracer method to standard ileal method, for the determination of AA digestibility of pea protein and milk casein. Methods Fifteen healthy adult volunteers completed the study and were equipped with a naso-ileal tube. They were given 9 portions of mashed potatoes containing either pea protein or casein isolates that were intrinsically labelled with 15N and 2H. A 13C algal free AA mix was added in the meals as the reference for dual tracer method. Plasma samples were collected regularly from before the first ingestion to 8-h later, while ileal digesta were collected continuously. For ileal sampling method, the AA digestibility (RIDAA) was determined using the recovery a non-absorbable marker (PEG-4000) perfused in the ileum, and the measurement of 15N enrichment of the digesta. For the dual tracer method, the amount of AA absorbed (AbAA) was calculated by the ratio of 2H/13C enrichments in plasma and in meals. The isotopic enrichments were evaluated in digesta, plasma samples and meals by GC-C-IRMS. The AA content was measured in digesta and meals by U-HPLC. Results Mean AbAA and RIDAA of pea protein were 102.2 ± 3.1% and 94.3 ± 1.5%, respectively. Mean AbAA and RIDAA of casein were 91.9 ± 2.0% and 97.1 ± 0.8%, respectively. The dual tracer method overestimated by 10% and 5% the AA digestibility of pea protein and casein, respectively, and the variability was high. The mean ileal AA digestibility of the 13C free AA mix was high (98.1 ± 1.1%), which validated our choice to use it as the reference ‘protein’ in the dual tracer method. Conclusions Several AA digestibilities obtained with dual tracer method were in the same range as the digestibilities from ileal sampling method. The variability was high and the effect of the protein source was inconsistent. After further research and validation, the dual tracer method could lead to notable advances in the determination of protein quality in humans. Funding Sources Roquette.

scholarly journals Methodological considerations in measuring human calcium absorption: relevance to study the effects of inulin-type fructans

2005 ◽  
Vol 93 (S1) ◽  
pp. S105-S110 ◽  
Author(s):  
Ian J. Griffin ◽  
Steven A. Abrams
Keyword(s):  
Calcium Absorption ◽  
Plasma Samples ◽  
Mass Balances ◽  
Tracer Method ◽  
Dual Isotope ◽  
Isotope Tracer ◽  
Advantages And Disadvantages ◽  
Isotopic Methods ◽  
Methodological Considerations ◽  
Isotope Tracer Method

During the last 50 years, a variety of methods have been developed to estimate Ca absorption in man. Mass balances were initially used, but these were unable to accurately measure fractional Ca absorption because they cannot distinguish unabsorbed dietary Ca from endogenous faecal Ca excretion (excretion of previously absorbed Ca back into the gut). A number of isotopic methods have been developed that can measure true fractional Ca absorption, employing radioisotopes, stable isotopes, or both. Different methods involve collection of urine, faecal or plasma samples. Of the currently available methods, the dual isotope tracer method with a timed urine collection is probably the most precise and reliable. It is also relatively straightforward to carry out and avoids the need for a faecal collection. The purpose of the present paper is to discuss the general advantages and disadvantages of the different methods of Ca absorption. In addition, the limitations the different methods have in examining the possible effects of non-digestible oligosaccharides on Ca absorption will be discussed.

scholarly journals Protein-quality evaluation of complementary foods in Indian children

2019 ◽  
Vol 109 (5) ◽  
pp. 1319-1327 ◽  
Author(s):  
Nirupama Shivakumar ◽  
Sindhu Kashyap ◽  
Satvik Kishore ◽  
Tinku Thomas ◽  
Aneesia Varkey ◽  
...  
Keyword(s):  
Amino Acid ◽  
Mung Bean ◽  
Finger Millet ◽  
Protein Quality ◽  
Complementary Feeding ◽  
Food Protein ◽  
Complementary Foods ◽  
Dual Isotope ◽  
Isotope Tracer ◽  
Nationally Representative

ABSTRACT Background The types of food in complementary feeding of infants and young children are important for growth and development. Food protein quality, as measured by the Digestible Indispensable Amino Acid Score (DIAAS), requires the determination of true ileal digestibility of indispensable amino acids (IAAs) in children. Objectives First, the aim of this study was to measure the true ileal IAA digestibility of 4 (rice, finger millet, mung bean, and hen egg) commonly consumed complementary foods in children aged <2 y using the dual-isotope tracer method. Second, we calculated the DIAAS of complementary feeding diets and their relation to stunting in a representative Indian rural population. Design Rice, finger millet, and mung bean were intrinsically labeled with deuterium oxide (2H2O), whereas egg was labeled through oral dosing of hens with a uniformly 2H-labeled amino acid mixture. True ileal IAA digestibility was determined by the dual-isotope tracer technique. The DIAAS of complementary food protein was calculated in children aged 1–3 y from a nationally representative survey to evaluate its relation with stunting. Results True ileal IAA digestibility was lowest in mung bean (65.2% ± 7.1%), followed by finger millet (68.4 %± 5.3%) and rice (78.5% ± 3.5%), and was highest for egg (87.4% ± 4.0%). There was a significant inverse correlation of complementary food DIAAS with stunting in survey data (r = −0.66, P = 0.044). The addition of egg or milk to nationally representative complementary diets theoretically improved the DIAAS from 80 to 100. Conclusions The true ileal IAA digestibility of 4 foods commonly consumed in complementary diets showed that the DIAAS was associated with stunting and reinforces the importance of including animal source food (ASF) in diets to improve growth. This trial was registered at http://ctri.nic.in/clinicaltrials/login.php as CTRI/2017/02/007921.

Sign in / Sign up

Export Citation Format

Share Document