Acids Leucine Research Articles

Extracellular Enzymatic Activities of Oceanic Pelagic Fungal Strains and the Influence of Temperature.

Although terrestrial and aquatic fungi are well-known decomposers of organic matter, the role of marine fungi remains largely unknown. Recent studies based on omics suggest that marine fungi potentially play a major role in elemental cycles. However, there is very limited information on the diversity of extracellular enzymatic activities performed by pelagic fungi in the ocean and how these might be affected by community composition and/or critical environmental parameters such as temperature. In order to obtain information on the potential metabolic activity of marine fungi, extracellular enzymatic activities (EEA) were investigated. Five marine fungal species belonging to the most abundant pelagic phyla (Ascomycota and Basidiomycota) were grown at 5 °C and 20 °C, and fluorogenic enzymatic assays were performed using six substrate analogues for the hydrolysis of carbohydrates (β-glucosidase, β-xylosidase, and N-acetyl-β-D-glucosaminidase), amino acids (leucine aminopeptidase), and of organic phosphorus (alkaline phosphatase) and sulfur compounds (sulfatase). Remarkably, all fungal strains were capable of hydrolyzing all the offered substrates. However, the hydrolysis rate (Vmax) and half-saturation constant (Km) varied among the fungal strains depending on the enzyme type. Temperature had a strong impact on the EEAs, resulting in Q10 values of up to 6.1 and was species and substrate dependent. The observed impact of temperature on fungal EEA suggests that warming of the global ocean might alter the contribution of pelagic fungi in marine biogeochemical cycles.

Coadministration of tianeptine alters behavioral parameters and levels of neurotrophins in a chronic model of Maple Syrup Urine disease.

Maple Syrup Urine Disease (MSUD) is caused by the deficiency in the activity of the branched-chain α-ketoacid dehydrogenase complex (BCKDC), resulting in the accumulation of the branched-chain amino acids (BCAA) leucine, isoleucine, and valine, and their respective branched-chain α-keto acids. Patients with MSUD are at high risk of developing chronic neuropsychiatric disorders; however, the pathophysiology of brain damage in these patients remains unclear. We hypothesize that MSUD can cause depressive symptoms in patients. To test our hypothesis, Wistar rats were submitted to the BCAA and tianeptine (antidepressant) administration for 21days, starting seven days postnatal. Depression-like symptoms were assessed by testing for anhedonia and forced swimming after treatments. After the last test, the brain structures were dissected for the evaluation of neutrophins. We demonstrate that chronic BCAA administration induced depressive-like behavior, increased BDNF levels, and decreased NGF levels, suggesting a relationship between BCAA toxicity and brain damage, as observed in patients with MSUD. However, the administration of tianeptine was effective in preventing behavioral changes and restoring neurotrophins levels.

Abstract P4-01-03: Multiomics data reveal novel biomarkers for CDK4/6 resistance

Abstract Background: Cyclin-dependent 4/6 kinase inhibitors (CDK4/6i) and endocrine therapy (ET) have improved progression-free survival (PFS) and overall survival in hormone-receptor-positive (HR+) metastatic breast cancer (MBC), but endocrine resistance is a major challenge. PROMISE [NCT0281902; n=63] is a multicenter study that enrolled women with HR+ HER2- MBC commencing palbociclib (Pb) with letrozole (1st line [1L]) or fulvestrant (2nd line [2L]), and was designed to perform a comprehensive “omic” assessment of prospectively collected biospecimens (pre-treatment (M1), at 2 months (M2), and at disease progression). The goal is to identify novel genomic variants and pathways associated with resistance to CDK4/6i and ET and PFS outcomes. Here we report the association between the proteomic, metabolomics, and lipidomics data generated from pre-Pb and 2-month serum samples and PFS. Methods: Untargeted mass spectrometry data was generated from Metabolon, assaying 1308 metabolites and 831 lipids. Additionally, 1436 proteins were assayed on the Olink platform. Cox proportional hazard models were used to evaluate the univariate hazard ratio (HR) for all features with respect to PFS. The analyses were performed on samples from 45 patients (N=33 1Lwith 9 progression events and 12 2L with 8 progression events), obtained from M1 and M2 timepoints on Pb + ET. Enrichment analysis p-values are calculated using Fisher’s exact test. Results: Proteomics: In the M1 timepoint, 93 and 43 proteins were associated with PFS in the 1L and 2L settings, respectively; inflammation genes were enriched among the 1L setting (p= 0.034); 33 proteins presented HRs ranging between 0.026 and 0.56. The FABP9 protein (HR of 1.98, 95% CI 1.02-3.83) was associated with worse PFS. Conversely, inflammation genes were not observed to be enriched in 2L. In the M2 timepoint, we observed 60 and 21 proteins significantly associated with PFS, but no biological function was enriched in 1L and 2L. Metabolites: In the M1 timepoint, metabolism of the sulfur-containing amino acids (methionine, cysteine, SAM and taurine) were enriched in the 1L setting (p= 0.035, HR range 0.15-0.33); and the branched-chain amino acids (leucine, isoleucine, and valine) were significantly associated with PFS in the 2L setting (p= 0.028, HR range 0.013-0.33). At the M2 timepoint, the amino acids were no longer enriched, but fatty acid metabolism was significantly enriched for both 1L and 2L (p= 0.048 and 0.067, respectively). Pathways involving lipids, amino acids, and xenobiotics were enriched in metabolites related to PFS (p <0.05) for both treatment lines at M1 and M2. Lipidomics: In the M1 timepoint, 10 and 19 lipids were associated with PFS for 1L and 2L, respectively. The most notable lipid associated with worse PFS in the 1L was an 18 carbon phosphatidylinositol, PI(18:1/18:2), (HR 7.34 (CI 1.27-42.50); 8 triglycerides were associated with improved PFS (HR range 0.39 and 0.55). In 2L, the 19 lipids associated with PFS included 12 phosphatidylcholines (enrichment p = 5.6X10-8). In the M2 timepoint, 15 and 8 lipids were significantly associated with PFS for 1L and 2L. An enrichment of phosphatidylinositols was observed in 1L (p= 1.2X10-5); none were observed in the 2L.Future Directions: Networks are being constructed using the proximity scores of the proteins, lipids, and metabolites associated with PFS in M1 and M2 for 1L and 2L. Network similarities and analyses will be conducted.Conclusion: Distinct multi-omic changes identified in serum samples obtained from PROMISE participants M1 and M2 on Pb correlate with disease progression in both 1L and 2L settings. Additionally, validation studies will determine the significance of these findings. Citation Format: Krishna R. Kalari, Kevin J. Thompson, Jason Sinnwell, Xiaojia Tang, Vera J. Suman, Jun He, Seul Kee Byeon, Akhilesh Pandey, Abe Eyman Casey, Peter T. Vedell, Ann M. Moyer, Alvaro Moreno-Aspitia, Donald W. Northfelt, Minetta C. Liu, Tufia C. Haddad, Saranya Chumsri, Prema Peethambaram, Kathryn J. Ruddy, Karthik V. Giridhar, Roberto A. Leon-Ferre, Richard M. Weinshilboum, Liewei Wang, Ciara C. O’ Sullivan, Matthew P. Goetz. Multiomics data reveal novel biomarkers for CDK4/6 resistance [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-01-03.

Using sheep selection to improve the nutritional value of lambmeat

Relevance. The article provides a justification for the use of sheep selection taking into account the intra-breed type of Prekos ewes in purebred breeding and crossing with producers of semi-fine wool breeds Romney-Marsh and Kuibyshevskaya to improve the biological usefulness of meat in offspring.Methods: scientific and economic experiment in the use of different selection options, taking into account the productive type of sheep of the Precos breed.Results. The article presents the results of a study on the influence of the selection of sheep on the composition of lambmeat, increasing the usefulness and quality of meat and, thus, the efficiency of its production. It was found that the content of water in the meat of crossbred animalswas lower compared to purebred Precos. There is a lower water content in the meat of rams obtained from sheep of the Precos meat-wool type, both in purebred and crossbred animals. In terms of the protein content in meat, there is an advantage in rams with purebred breeding and crossing compared to meat obtained from the wool-meat type of queens. In the hip bran of purebred animals and crossbreeds with the Kuibyshev breed, this superiority was 1.1% (Р > 0.95), and in the lumbar bran of Romney-Marsh crossbreeds, this difference was 1.6% (Р > 0.95). Mutton from the offspring of the sheep of meat-wool type is characterized by a significantly high content of the essential amino acids threanine, lysine, leucine, phenylanin and isoleucine in comparison with meat from the offspring of the wool-meat type of sheep. A significant superiority was also established in the content of interchangeable amino acids — oxyproline, serine, tyrosine, proline, glycine, aspartic and glutamic acid, cystine and analin.

Biopotential of collagen-containing hydrolysates obtained from fish raw materials for protein sports nutrition

Low-molecular peptides of collagen origin are of crucial importance for the body metabolism. This work aims to substantiate the use of the hydrolysates of collagen-containing fish raw materials (scale) for protein sports nutrition. Sublimated low-molecular weight peptide water-soluble and dried water-insoluble protein-mineral supplements from the sardine scale were obtained by enzymatic thermal hydrolysis. The biopotential of scale and auxiliary food additives (apple pomace, flaxseed cake, pine nuts) was investigated using standard methods. The protein content and amino acid composition of the peptide supplement and the mineral composition of the protein-mineral supplement were analysed; their high biopotential according to the above criteria was shown. The peptide supplement contains all essential amino acids having a biological value of 59.9%. The protein-mineral supplement contains no tryptophan but a high amount of calcium (22.2 g/100 g) and phosphorus (12.0 g/100 g). The high content of functional food ingredients (protein, pectin, cellulose, polyphenols, minerals, vitamin C) in auxiliary vegetable raw materials was shown. The shape of a sports nutrition product (chocolate glazed protein bar) was substantiated. The basic recipe of a 60 g protein bar having the required protein content, a given structure and high consumer properties was established. Having a protein content of 23.5%, the bar provides a 24.1% calorific value, which corresponds to the "protein-rich food" for sports nutrition according to GOST 34006-2016. The biological protein value in the bar increased relative to the peptide supplement by 12% and amounted to 71.38%. The developed protein sports nutrition bar is dietary balanced by the content of amino acids (isoleucine, leucine, valine, methionine and cystine, phenylalanine and tyrosine, tryptophan, lysine, threonine, alanine, arginine, histidine, glycine, carnosine, taurine, ornithine, citrulline), fibre, polyphenols, calcium and phosphorus and is recommended for athletes and people having an active lifestyle.

Influence of an increased content of pea and yellow lupin protein in the diet of pigs on meat quality

BackgroundThe aim of the study was to test the effect of replacing soya beans with pea and yellow lupin seeds in the diet of pigs on meat quality. The meat for the tests was obtained from 60 fattening crossbred pigs F1 (Polish Large White × Polish Landrace) × F1 (Pietrain × Duroc). The animals belonged to three feeding groups depending on the feed used with the total share of soybeans and its reduction. Water holding capacity, colour, and tenderness were measured and visual and tactile evaluation (colour, marbling and firmness) was performed for meat samples collected from the longissimus lumborum muscle. The chemical composition of the meat and the content of minerals were determined. The content of amino acids, fatty acids and cholesterol was determined.ResultsThere was no significant differences among the assessed physicochemical characteristics of the meat. The obtained meat was of good quality, regardless of the proportion of proteins from legumes in the diet of pigs. The results of the subjective evaluation of meat, its colour, and the content of muscle pigments were uniform in all food groups. Similarly, a uniform, high protein content was found in all tested groups (C—24.98%; E1—24.82%; E2—25.09%) and the content of macro- and micronutrients in the tested meat was equivalent. The profile of fatty acids was not significantly affected by dietary treatment. Palmitoleic acid content was significantly higher (P < 0.05) in the E2 group compared to the E1 group (3.279% compared to 2.844%). The content of amino acids in meat samples was influenced by dietary treatment. Almost all the monitored essential amino acids (threonine, valine, leucine, phenylalanine and lysine) and some of nonessential amino acids’ proportion was increased in the experimental groups (E1, E2).ConclusionReplacing soya bean protein in the pigs’ diet with legume protein (peas and yellow lupin) did not adversely affect meat quality. This applies to both the physicochemical characteristics, the basic composition and the fatty acid profile. The meat of fattening pigs fed with the highest proportion of pea and lupin in the ration was characterized by more favourable proportion of essential amino acids.

Metabolic responses of plasma to extreme environments in overwintering Tibetan frogs Nanorana parkeri: a metabolome integrated analysis

Many animals lower their metabolic rate in response to low temperatures and scarcity of food in the winter in phenomena called hibernation or overwintering. Living at high altitude on the Tibetan Plateau where winters are very cold, the frog Nanorana parkeri, survives in one of the most hostile environments on Earth but, to date, relatively little is known about the biochemical and physiological adjustments for overwintering by this species. The present study profiled changes in plasma metabolites of N. parkeri between winter and summer using UHPLC-QE-MS non-target metabolomics in order to explore metabolic adaptations that support winter survival. The analysis showed that, in total, 11 metabolites accumulated and 95 were reduced in overwintering frogs compared with summer-active animals. Metabolites that increased included some that may have antioxidant functions (canthaxanthin, galactinol), act as a metabolic inhibitor (mono-ethylhexylphthalate), or accumulate as a product of anaerobic metabolism (lactate). Most other metabolites in plasma showed reduced levels in winter and were generally involved in energy metabolism including 11 amino acids (proline, isoleucine, leucine, valine, phenylalanine, tyrosine, arginine, tryptophan, methionine, threonine and histidine) and 4 carbohydrates (glucose, citrate, succinate, and malate). Pathway analysis indicated that aminoacyl-tRNA biosynthesis, phenylalanine, tyrosine and tryptophan biosynthesis, and nitrogen metabolism were potentially the most prominently altered pathways in overwintering frogs. Changes to these pathways are likely due to fasting and global metabolic depression in overwintering frogs. Concentrations of glucose and urea, commonly used as cryoprotectants by amphibians that winter on land, were significantly reduced during underwater hibernation in N. parkeri. In conclusion, winter survival of the high-altitude frog, N. parkeri was accompanied by substantial changes in metabolomic profiles and this study provides valuable information towards understanding the special adaptive mechanisms of N. parkeri to winter stresses.

Phenotypic and Genetic Characterization of Temperature-Induced Mutagenesis and Mortality in Cupriavidus metallidurans.

Cupriavidus metallidurans strains display a decreased viability when incubated in rich medium at a temperature of 37°C compared to their normal growth temperature of 30°C, a phenomenon coined “temperature-induced mortality and mutagenesis” (TIMM). To scrutinize this aberrant phenotype further, the contributions of specific inducers and protective agents were determined. Different growth media, including lysogeny broth (LB) and Schatz, and components, including casamino acids, in particular amino acids (proline, cysteine, glycine, glutamine, leucine, histidine and phenylalanine) and ammonium, were found to induce TIMM at 37°C. Sorbitol was found to counteract TIMM. Furthermore, although TIMM is well conserved within the C. metallidurans species, multiple and strain-specific TIMM inducers exist. Twenty-nine percent of the TIMM survivors inherited resistance to TIMM. Whole-genome sequencing of two resistant derivatives revealed an important role of an uncharacterized oxidoreductase, indicating putative metabolic poisoning when grown in high-concentration nitrogen-containing media at 37°C.

Abstract 2561: Tumorigenic and metabolomic impacts of adipose-derived monocyte chemotactic protein-1 in a model of obesity-enhanced, male breast cancer

Abstract Breast cancer is a rare but aggressive disease in men. Approximately, 90% of all breast cancer in men are invasive carcinomas, 25% of which have distant metastasis at the time of clinical presentation. Obesity is a risk factor for breast cancer. Monocyte chemotactic protein-1 (MCP-1) is an adipose-derived cytokine whose concentrations are elevated by obesity. This study tested the hypothesis that adipose-derived MCP-1 contributes to male breast cancer. In a 2x2 design, male MMTV-PyMT mice with or without adipose specific Mcp-1 knockdown [designated as Mcp-1-/- or wild-type (WT)] were fed the standard AIN93G diet or an obesogenic, high-fat diet (HFD) for 25 weeks. The AIN93G diet and HFD contained 16% and 45% of energy from soybean oil, respectively. Mcp-1-/- mice had lower adipose Mcp-1 expression than WT mice. Adipose Mcp-1 deficiency reduced plasma concentrations of MCP-1 in mice fed the HFD. Mice fed the HFD had a greater tumor progression rate than mice fed the AIN93G diet (4679% vs 2430%), regardless of genotype. Mcp-1-/- mice, compared to WT mice, had a longer tumor latency (25.2 weeks vs 18.0 weeks) and lower tumor incidence (19% vs 56%), tumor progression rate (2317% vs 4792%), and tumor weights (0.23 g vs 0.64 g). Metabolomic analysis of plasma identified 87 metabolites, 56 of which differed among the four groups, including 24 that differed between the diets and 22 that differed between the genotypes. Sparse partial least squares-discriminant analysis identified 10 major metabolites that were altered the most among the four groups. These included amino acids (alanine, isoleucine, leucine, phenylalanine, threonine, and valine) and carbohydrate metabolites (fumaric acid, glucuronic acid, hexuronic acid, and malic acid). The integrated pathway analysis of the 87 identified metabolites showed that protein biosynthesis and carbohydrate metabolism pathways are the most disturbed in MMTV-PyMT mice. In conclusion, adipose-derived MCP-1 contributes to mammary tumorigenesis and alters metabolism in male MMTV-PyMT mice. Citation Format: Lin Yan, Sneha Sundaram, Bret M. Rust, Matthew J. Picklo, Michael R. Bukowski. Tumorigenic and metabolomic impacts of adipose-derived monocyte chemotactic protein-1 in a model of obesity-enhanced, male breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2561.

White Paper - Plant Based Meat - Texturizing Plant Proteins

FAO forecasts that the global meat production needs to increase by 50% to provide quality protein for the ten billion people living on planet earth by 2050. Meat fits in a healthy diet and is a good source of essential proteins, B-vitamins and several minerals (i.e. selenium and iron). A growing world population will require in 2050 about twice more food, produced on twice less arable land. Meat production is seen as less sustainable with negatives impacts on animal welfare. There is a search for alternative proteins to replace part of the animal proteins that are currently used in abundance in the diet of most developed and affluent countries. Good digestible proteins may be a limiting factor in the future. Protein is an important part of the human diet, and the Essential Amino Acids (EAA) (histidine, isoleucine, leucine, lysine, methionine (+cystine), phenylalanine (+tyrosine), threonine, tryptophan and valine), which cannot be synthesized de novo, need to be supplied through the diet. The FAO recommends that amino acids be treated as individual nutrients and that adjustments for digestibility be made when designing dietary guidelines or evaluating actual diets (Food and Agriculture Organization of the United Nations (FAO), 2013). Animal protein contains more EAA, and is of slightly higher human intestinal digestibility, than most plant-based proteins. To meet the 2050 challenge of feeding a rapidly growing human population, it is essential to carefully consider how to produce enough protein. To help consumers reduce their meat consumption, several strategies have been developed: One strategy is encouragement of ‘meatless days’ or smaller portion sizes. As a second strategy, meat in a meal can be replaced by the consumption of vegetables, beans, pulses, and/or nuts. In processed meat products, it can be replaced by using plant-based meat extenders as well [1]. Meat extenders are plant-based ingredients that act as fillers in processed meats. By doing so, the actual meat content in the product is lowered, leading to reduced meat intake. A third strategy is based on the development of structured vegetable protein products. Examples of traditionally structured products are tofu and tempeh, which are produced and consumed in East Asia for centuries. The consumer acceptance of these products is, however, lower in Western countries. Therefore, another category of structured products has emerged, referred to as meat analogues. Meat analogues are products that can replace meat in its functionality, being similar in product properties/sensory attributes, and that can also be prepared by consumers as if they were meat. The resemblance of these products to meat, in terms of texture, taste, appearance, and smell, is important for consumers that currently mostly choose meat [2]. At the moment a large number of textured vegetable proteins have already entered the market as meat analogues and substitutes. Textured structures based on soy proteins were one of the first to enter the market. At the moment there are textured vegetable proteins based on (mixtures of) pea, wheat, carrot, sunflower, onion etc. The main issue with vegetable based alternative proteins are the texture and taste of the final products. Producing steak like structures is possible with textured proteins. Most off flavours can be masked by spices and taste ingredients. In this white paper the current state of the art of the technical production processes of plant- based proteins, with special focus on the texturizing processes, will be described. This will include the established techniques that have been operated on commercial scale, and novel techniques that are less mature.

Potential of potatoes as a source of nutritional value : Amino acid composition of protein in stubbies of potatoes of the Ural region, Russia

The evaluation of the selection material of the Ural selection was carried out for 17 amino acids that are part of the potato protein: lysine, histidine, arginine, aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, isoleucine, leucine, tyrosine, phenylalanine, cystine and meteonin. Analysis of 30 promising cultivars showed that the content of some amino acids in potato tubers, such as arginine, glutamic acid, proline, glycine and valine significantly affected by the genotype and environment. On an average, about 1/3 of the total amino acid composition of potato tubers was made up of essential amino acids, of which most of all were arginine (upto 0.56%), lysine (upto 0.16%) and leucine (upto 0.15%). These maximum indicators were noted in the Mishka variety. The main content of nonessential amino acids was aspartic (0.25 to 0.65%) and glutamic (0.21 to 0.45%) acids. The maximum value in terms of the total composition of amino acids (2.70%) was noted in the Mishka variety. varieties with high indicators were noted in Gornyak (2.64%), Legenda (2.53%), Terra (2.45%) and, hybrids 10-22-23 (2.58%) and 10-11-39 (2.48%). Correlation links were direct, average ones were found between the protein content and amino acids lysine (r = 0,371), arginine (r = 0,347), histidine (r = 0,345) and meteonin (r = 0,341). In other cases the correlation was direct and weak (r = 0,015-0.298). A direct and moderately positive correlation was noted between the content of reducing sugars and amino acids glycine (r = 0,411), arginine (r = 0,388), leucine (r = 0,366), histidine (r = 0,348) and lysine (r = 0,333). There was a direct high correlation dependence on the content of vitamin C and the amino acid cystine in potato tubers (r = 0,706).

Use of chicken feather meal fermented with Bacillus subtilis in diets to increase the digestive enzymes activity and nutrient digestibility of silver pompano Trachinotus blochii (Lacepede, 1801).

Background: Feather has the potential to be used as a fish feed ingredient because it has high protein content (80-85%), and is rich in amino acids arginine, leucine, isoleucine and valine. However, the protein consists mainly of keratin, which is classified as fiber that is difficult to digest. Therefore, to improve digestibility, the keratin protein is degraded using microbial Bacillus subtilis. This study aimed to determine the digestibility of fermented feather meal (FFM) in silver pompano diets and to observe the histological structure of their intestines after digestion. Methods: The method used was a one factor experiment with five treatments and three replications each, which were: diet without FFM (P0), diet containing 10% FFM (P1), 20% (P2), 30% (P3) and 40% (P4). The diets were given to juvenile silver pompano (with average body weight of 8.56 ± 0.18 g) and stocked with 10 fish in a 100 L capacity container. The experimental diets were given three times daily at approximately 8.00 AM, 12.00 PM and 5.00 PM to apparent satiation for 60 days. Results: The results showed that the use of FFM increased the activity of digestive enzymes (protease and lipase), but reduced the amylase activity of silver pompano, which was significantly different between treatments (P <0.05). Meanwhile, the diet containing 20% FFM produced the highest feed and protein, which are 37.05% and 67.24%, respectively. This was significantly different from other treatments (P <0.05), and was effectively absorbed by fish intestines. Conclusion: The addition of chicken feather meal fermented with Bacillus subtilis could increase the activity of digestive enzymes and nutrient digestibility of silver pompano.

Synthesis of BODIPY-Amino Acids and the Potential Applications as Specific Dyes for the Cytoplasm of Langerhans β-Cells

In this work, we explored the synthesis of 4,4-difluoro-4-bora-3a,4a-diazas-indacene (BODIPYs) bound to five different amino acids (BODIPY-FAA) (glycine, alanine, leucine, phenylalanine, and tyrosine) (amino group is kept protected with fluorophore Fmoc) and evaluated these conjugates in terms of (i) their photophysical properties and (ii) their potential application as cell staining agents of suspension and adherent cells at healthy and stress conditions. In general, all synthesized BODIPY-FAA (3a-3e) were found to emit fluorescence in the blue and green regions of the spectrum (depending on the solvent conditions). However, BODIPY-FTyr(trt) (3e) showed the best molar extinction coefficient (ε = 28,198 M-1 cm-1) and quantum yield (Φ = 0.17). Biologically speaking, all synthesized conjugates demonstrated a selective affinity for the cytoplasm of Langerhans β-cells employed as a model, being the BODIPY-FLeu conjugate the one displaying the highest observed intensity. As such, our results reveal the BODIPY-FAA as a novel attractive tool for the specific staining of the cell cytoplasm, demonstrating not only a dual fluorescence emission but also a sensing capability to recognize different cell states.

Amino acid composition of wheat proteins from different growing areas after long-term storage

The results of the research on the assessment of the biological value of wheat after storage by the composition and ratio of amino acids are presented. It is shown that after 6.5 years of storage in elevator silos, the protein content in wheat met the requirements of the standard for class 3 wheat. The sum of all amino acids was from 11.5 to 13.1 g / 100 g, including essential amino acids — 4.0-4.5g/100g and non-essential — 7.8-8.7 g / 100 g. The proportion of essential amino acids was 33.7-35.3% of the total amino acid content. The calculation of the amino acid score demonstrated that the main amino acid limiting the nutritional value of wheat after storage is lysine with an amino acid score of 45-53%. The essential amino acids threonine, valine, isoleucine, and leucine are also classified as deficient, but with a higher value of the amino acid score, close to the complete protein.

Understanding the Effect of Resistant Training and Amino Acid Supplement on Weight Perception and Body Image

The purpose of this study is to demonstrate if the combination of resistance training (RT) and an intake of amino acid supplement will have a positive impact on weight preoccupation, self-classified weight and body satisfaction. A total of 33 usable data sets were obtained from the survey among 40 women who participated in this 10-week RT program, and they were given either 5 grams of placebo or amino acid (leucine). RT and the amino acid supplement played an important role in body satisfaction overall. Determining the most effective combination of approaches to tailor recommendations for adults for optimal quality of life is crucial.

Molecular Simulation of the Separation of Some Amino Acid Enantiomers by β-Cyclodextrin in Gas-Phase.

The complexes formed by β-cyclodextrin and some amino acids (alanine, valine, leucine, and isoleucine) in vacuo are studied by molecular mechanics and dynamics simulations. These methods have been improved with respect to our previous studies with amino acids, regarding the determination of molecular structures or initial enantiomer dispositions in the molecular dynamics trajectories. The greatest contribution to the interaction energy is from the van der Waals term, although the discrimination between enantiomers is due mainly to the electrostatic contribution. The lowest energy structures of the complexes obtained from molecular mechanics are inclusion complexes in which the carboxylic end of amino acids is pointing toward the narrow (D-) or wide rim (L-) of β-cyclodextrin. The position probability density provided by molecular dynamics also confirms inclusion complex formation, because the guests spend most time inside the cavity of β-cyclodextrin along its axis, with the carboxylic end pointing toward the narrow rim. The L-amino acids are the first eluted enantiomers in all cases and chiral discrimination increases with the size of guests, except leucine, which has the lowest capacity to discriminate. During the simulation, Ala and Val remain in weakly enantioselective regions, while Leu and Ile stay in zones with great chiral selectivity.

Metabolomics and its application in the mechanism analysis on diabetic bone metabolic abnormality.

This study is aimed at analysing the endogenous metabolites profiling of patients with diabetic osteoporosis, so as to provide the reference for pathogenesis research of diabetic osteoporosis. The 1H-NMR metabolomics technology, combined with pattern recognition analysis and SIMCA-P 12.0 statistical analysis, were employed to identify the metabolites differences between diabetic patients with disordered bone metabolism (research group) and healthy volunteers (normal group) in this study. Compared with normal group, the results show that in research group, the levels of O-acetyl glycoprotein, proline, 1-methyl histidine, tricarboxylic acid cycle (TCA cycle) product (citric acid and α-ketoglutaric acid) decline, while the levels of branched chain amino acids (leucine, isoleucine, valine), glucose, choline, creatine, inositol, glutamine, aspartic acid, alanine, glycine, and citrulline increase. There are disordered metabolic pathways and imbalanced bone synthetic materials and regulatory substances in diabetic patients with bone metabolic abnormality. These metabolic abnormalities could be the specific indicators in early diagnosis of diabetic osteoporosis.

Effects of the lactation period, breed and feed on amino acids profile of mare’s milk

The effects of the lactation period, breed, and feed on amino acids profile of mare’s milk were investigated. The feed contained two major essential amino acids (EAAs) leucine (7.31 – 10.3 g.kg-1) and arginine (6.37 – 9.59 g.kg-1); it also included minor EAAs methionine (2.11 – 3.05 g.kg-1) and histidine (2.48 – 3.60 g.kg-1). Glu+Gln, Asp+Asn, and proline, major nonessential amino acids (NEAAs), constituted approximately 60% of total NEAAs (TNEAAs). The ratio of total EAAs to NEAAs ranged from 1:1.2 to 1:1.4. Amino acids (AA) content throughout all milk samples varied due to mare’s different conditions and lactation days. Except for the 1P milk sample, total AA content in the 2 – 8Ps specimens caused by differences in breed oscillated from the 2nd to 28th day of lactation within the following limits: 21.9 – 54.6 g.kg-1, 33.6 – 70.7 g.kg-1, 38.1 – 71.2 g.kg-1, 29.46 – 74.2 g.kg-1, 52.2 – 87.1 g.kg-1, 37.9 – 70.3 g.kg-1 and 26.4 – 64.5 g.kg-1, respectively. In relation to TEAAs in milk, the highest EAAs levels were reached in arginine, leucine and lysine ranging between 2.41 – 4.35 g.kg-1, 3.36 – 5.59 g.kg-1 and 2.72 – 4.80 g.kg-1, respectively, while the lowest AAs amounts were indicated in histidine and methionine, 0.91 – 1.58 g.kg-1 and 1.23 – 2.04 g.kg-1 respectively. Total NEAAs content was slightly higher than that of EAAs; the TNEAAs to TEAAs ratio was 1:0.9 proximately. Glu+Gln, Asp+Asn and proline were determined as major NEAAs of milk ranging between, 6.77 – 11.0 g.kg-1, 3.21 – 5.60 g.kg-1 and 1.25 – 2.18 g.kg-1, respectively; levels of NEAAs such as cysteine and glycine oscillated between 0.89 – 1.52 g.kg-1 and 0.64 – 1.15 g.kg-1, respectively. The average TAAs contents caused by breed differences were 62.8 g.kg-1, 42.8 g.kg-1, 44.7 g.kg-1 and 44.8 g.kg-1, respectively, on the 2nd, 5th, 10th, and 28th lactation days.

1437-P: A Newly Developed Diabetes Risk Index, Based on Lipoprotein Subfractions and Branched-Chain Amino Acids, Is Associated with Incident Type 2 Diabetes Mellitus in the PREVEND Cohort

Objective: Evaluate the ability of the Diabetes Risk Index (DRI) to predict incident type 2 diabetes mellitus (T2DM) in a large cohort. Methods: The DRI was developed by combining the 6 lipoprotein subfractions in the Lipoprotein Insulin Resistance Index (LP-IR) and the branched chain amino acids valine and leucine, all of which have been shown previously to be associated with future T2DM. DRI scores were calculated in a total of 6134 nondiabetic men and women in the Prevention of Renal and Vascular End-Stage Disease (PREVEND) Study. Cox proportional hazards regression was used to evaluate the association of DRI scores with incident T2DM. Results: During a median follow-up of 8.5 years, 306 new T2DM cases were ascertained. In analyses adjusted for age and sex, there was a significant association between DRI scores and incident T2DM with the hazard ratio (HR) for the highest versus lowest tertile being 7.49 (95% confidence interval: 5.01-11.19), P &amp;lt;0.001. After additional adjustment for BMI, family history of T2DM, alcohol consumption, diastolic blood pressure, total cholesterol, triglycerides, HDL cholesterol and HOMA-IR, the HR was attenuated but remained significant (HR 1.67 [1.01-2.76], P = 0.04). Similar results were obtained when DRI was analyzed as HR per 1 SD increase (HR 1.37 [1.14-1.65], P = 0.001). The Kaplan-Meier plot demonstrated that patients in the highest tertile of DRI scores presented at higher risk (p-value for log-rank test &amp;lt;0.001). Conclusions: Higher DRI scores are associated with an increased risk of T2DM. The association is independent of clinical risk factors for T2DM including HOMA-IR, BMI and conventional lipids. Disclosure J. Flores-Guerrero: None. M.A. Connelly: Employee; Self; LabCorp. I.Y. Shalaurova: None. E.G. Gruppen: None. J.D. Otvos: Employee; Self; LabCorp. R.P.F. Dullaart: None. S.J.I. Bakker: None.

169-OR: Differential Effects of Amount, Intensity, and Mode of Exercise Training on the Diabetes Risk Index

Objective: Examine the effects of amount, intensity, and mode of exercise training on type 2 diabetes (T2D) risk. Methods: A total of 531 men and women with dyslipidemia [STRRIDE I (n=222), STRRIDE AT/RT (n=142) or prediabetes [STRRIDE-PD (n=167)] were randomized to control group or one of 10 exercise interventions, ranging from doses of 8-22 kcal/kg/week; intensities of 50-75% V?O2peak; and duration of 6-9 months. Two groups included resistance training and one included dietary intervention (weight loss goal of 7%). Fasting plasma samples were obtained at baseline and 16-24 h after last exercise bout. The Diabetes Risk Index (DRI) and its components—Lipoprotein Insulin Resistance Index (LP-IR) and branched chain amino acids (BCAA) valine and leucine—were determined by NMR spectroscopy. DRI scores range from 1-100, the latter representing those at greatest T2D risk. Paired t-tests determined significance within groups (p&amp;lt;0.05). Study-specific ANCOVA determined differences among groups. Results: The inactive control group did not significantly change DRI score. After training, four of ten exercise groups significantly improved DRI scores, ranging from -2.8 ± 8.2 to -8.3 ± 10.4. In STRRIDE AT/RT, only the aerobic-plus-resistance group significantly improved DRI. In STRRIDE-PD, the moderate-intensity-plus-diet group had a greater DRI change than the other exercise-only moderate intensity groups. Reductions in DRI appeared to be driven largely by decreases in insulin resistance (LP-IR) but not dysmetabolism (BCAA), except for the moderate-intensity-plus-diet group, which exhibited a reduction in both. Conclusions: Multiple exercise interventions led to improvements in DRI scores. Results provide compelling evidence that adding resistance to aerobic training elicits a synergistic effect on insulin resistance and T2D risk. In individuals with prediabetes, incorporating dietary intervention with aerobic training results in the most robust improvement in T2D risk. Disclosure L.M. Ross: None. C.A. Slentz: None. I.Y. Shalaurova: None. J.D. Otvos: Employee; Self; LabCorp. M.A. Connelly: Employee; Self; LabCorp. C.W. Bales: None. J.A. Houmard: None. W.E. Kraus: None. Funding National Institutes of Health (R01HL057354, R01DK081559, T32HL007101)