Acids Lysine Research Articles

Abstract 1973: Patients with operable esophageal cancer and improved responses to combined chemoradiotherapy and immunotherapy display distinct microbiome profiles enriched in multiple Bacteroides species

Abstract Background: Preclinical and clinical data indicate that neoadjuvant chemoradiotherapy (CRT) may prime an anti-tumor immunological response in esophageal cancer driven by intratumoral CD8+ T cells and PD-L1 expression. LAG-3 is also highly expressed in esophagogastric cancers. The microbiome, a novel and potentially modifiable, biomarker of IO response, has not yet been examined in the neoadjuvant setting in esophageal cancer and is the goal of our study. Methods: Fecal samples were collected from patients with stage II/III esophageal or gastroesophageal junction carcinoma eligible for curative resection treated with the standard of care regimen of carboplatin paclitaxel (50mg/m2), radiation 50.4 Gy in 28 fractions and an Ivor-Lewis esophagectomy 6-10 weeks after last CRT and immunotherapy (IO) dose. Patients on arm A (n=11) received 2 cycles of induction with nivolumab plus 3 additional cycles on week 1, 3 and 5 of CRT. Patients on arm B (n=8) received nivolumab plus relatlimab on the same schedule (Clinical trial: NCT03044613). We examined longitudinal fecal samples from n=19 patients across both arms (n=90 samples) using 16S rRNA amplicon sequencing. Patients were classified based on pathological response: complete response (CR) and grades 1, 2, and 3 (G1, G2, G3) with increasing residual tumor visible in the resected specimen. Sequencing data was trimmed and filtered for contaminants, followed by high-resolution taxonomic assignment and normalization of reads across all samples. Analysis was performed using multiple metrics for alpha diversity and beta-diversity, with principal coordinates analysis/PERMANOVA, and pathway analysis using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt). Results: Patients with improved response in the neoadjuvant setting (CR/G1 vs G2/G3) grouped in distinct clusters using Bray-Curtis (p < 0.001). Patients with CR had higher alpha diversity, using both measures of richness and evenness, compared to patients with a G3 responses (p < 0.03). Specifically, family Bacteroidaceae and genus Bacteroides were enriched in patients with CR vs G3 (p < 0.02). At the species level, B. finegoldii, B. ovatus, and B. uniformis were enriched in patients with CR vs G3 (p < 0.02). In contrast, genus Klebsiella and Clostridium termitidis were enriched in patients with a poor response, G3 (p <0.001, both). Pathway analysis found two metabolic pathways enriched in patients with CR: secondary bile acid biosynthesis (p=0.005) and lysine biosynthesis (p=0.02). Conclusions: Patients with operable esophageal cancer and improved responses to combined CRT and IO had distinct microbiome profiles enriched in multiple Bacteroides species. Further analyses and validation efforts are underway to confirm metabolomic pathways. Citation Format: Fyza Y. Shaikh, James R. White, Ronan J. Kelly, Ali H. Zaidi, Jenna V. Canzoniero, Josephine L. Feliciano, Russell K. Hales, K Ranh Voong, Richard J. Battafarano, Blair A. Jobe, Stephen C. Yang, Stephen Broderick, Jinny Ha, Kellie N. Smith, Elizabeth Thompson, Eun J. Shin, Ali I. Amjad, Patrizia Guerrieri, Benny Weksler, Chen Hu, Valsamo Anagnostou, Vincent K. Lam, Cynthia L. Sears. Patients with operable esophageal cancer and improved responses to combined chemoradiotherapy and immunotherapy display distinct microbiome profiles enriched in multiple Bacteroides species [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1973.

Effects of different protein levels on growth performance and stress parameters in beef calves under heat stress

This study investigated the effects of dietary protein levels under various heat stress (HS) conditions on the growth performance and stress parameters in Korean native beef calves. Male calves (n = 40; initial BW = 202.2 ± 3.31 kg) were randomly assigned to climatic-controlled chambers with 3 × 3 factorial arrangements. Calves were assigned into three dietary protein levels (low protein; LP = 12.5%, medium protein; MP = 15%, and high protein; HP = 17.5%) and three HS levels [mild: temperature-humidity index (THI) = 74 to 76, moderate: THI = 81 to 83, and severe: THI = 89 to 91] with control (threshold: THI = 70 to 73 and dietary protein level 12.5%). The calves were subjected to ambient temperature (22 °C) for 7 days and subsequently to the temperature and humidity corresponding to the target THI level for 21 days. The data were analyzed using the repeated-measures analysis by the GLM procedure of SAS. As a result, average daily gain (ADG) was decreased (P < 0.05) under severe HS level compared to the mild and moderate HS stress levels. However, HP increased ADG (P < 0.05) than moderate levels (LP) and severe levels (LP and MP). Under different HS levels (mild, moderate, and severe), HR, RT, and blood cortisol were increased (P < 0.05) compared to a threshold level, but no differences were observed in the parameters among various protein levels. Varied HS levels decreased the levels of blood glucose, NEFA, and amino acids (AAs) (lysine and glutamic acid) compared to a threshold (P < 0.05). But, the HP group resulted in increased (P < 0.05) levels of blood glucose, NEFA, and AAs (lysine and glutamic acid) compared to LP and MP groups under severe HS stress. The expression level of the HSP70 gene in peripheral blood mononuclear cell (PBMC) and hair follicles was increased (P < 0.05) following an increase in moderate and severe HS levels. Also, HSP70 gene expression in the HP group was decreased (P < 0.05) compared with LP and MP groups under intense HS level. Overall, HS in Korean native beef calves exhibited negative effects on ADG, blood glucose, NEFA, and AA profile. However, 17.5% of dietary protein (HP) could compensate for the growth of heat-exposed Korean native beef calves through the regulation of homeostasis by protein and energy metabolism. Also, it was evident that adequate protein (HP) is used as a major nutrient for HSP70 synthesis in PBMC and hair follicles causing, a boost in the immune system of heat-exposed Korean native beef calves.

Amino Acid Composition of Whey and Cottage Cheese Under Various Rennet Enzymes

Rennet cheeses occupy an important place in the diets of the population of Ukraine. The technology of making cheeses depends on both the quality of raw materials and the quality of enzymes that are used to curdle milk. Therefore, the study of the effect of rennet enzymes obtained using advanced biotechnology on the transformation of milk protein and amino acids into a finished product is of scientific and practical importance. Considering the above, the purpose of this study was to investigate the amino acid composition of whey and cottage cheese for the use of various enzyme preparations. Three groups of samples (n=5) were formed to set up the experiment. Cow's milk for research was selected from clinically healthy cows during the milking period. In the control group of samples, rennet of microbial origin was used for milk clotting. In the first experimental group of samples, an enzyme preparation extracted from rennet of dairy calves was used according to the method of Yu.Ya. Svyrydenko. In the second experimental group, an enzyme preparation obtained by extraction of rennet enzymes according to the method of S.V. Merzlov was used. The content of amino acids in milk, whey, and cheeses was determined by capillary electrophoresis. Studying the milk used for the experiment, it was found that the content of amino acids (lysine, methionine + cystine, tryptophane, valine, leucine, isoleucine, phenylalanine + tyrosine, proline, serine, alanine, glycine, histidine, arginine, aspartic acid and glutamine) did not significantly differ from the typical indicators of milk obtained in central Ukraine. It was found that the use of rennet enzymes extracted according to the method of S.V. Merzlov is accompanied by a decrease in the content of amino acids in serum by an average of 15.9%

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 &gt; 0.95), and in the lumbar bran of Romney-Marsh crossbreeds, this difference was 1.6% (Р &gt; 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.

The Effect of Adding Protected Amino Acids in the Rumen (Methionine and Lysine and their Mixtures) on Some Physiological Characteristics of Male Arabi Lambs

This study was conducted at the animal field of the Agriculture College / University of Basra / Karma Ali site for the period from 7/10/2019 to 1/5/2020 to investigation the effect of adding protected amino acids (methionine and lysine and their mixtures) on some physiological trails of male Arabi lambs. Sixteen male lambs with an average weight of (27.95) kg and an age of 5-6 months were selected, and randomly divided into four group (4 animals per group). The results indicated that there were no significant differences in the concentration of hemoglobin, packed cell volume, the number of red and white blood cells, cholesterol, triglycerides and the activity of the liver enzyme (ALT and AST) between the different treatments, while there was a significant (p &lt;0.05) increase in the total protein and albumin concentration in the fourth treatment compared with the first treatment (control), while, there was a significant decrease (p &lt;0.05) in the urea concentration for the fourth, third and second treatments compared with the first treatment. The concentrations of glucose, thyroxine and growth hormone increased significantly (p &lt;0.05) in the fourth and second treatment compared with the first treatment. However, it can be concluded from the study that feeding the male lambs with 5g methionine or mixing it with lysine by 5g raises the concentration of glucose, growth hormones and thyroxine in the blood. Also, the levels of methionine and lysine used did not change the concentration of the studied biochemical parameters

The Morphological, Clinical and Radiological Outputs of the Preclinical Study After Treatment of the Osteochondral Lesions in the Porcine Knee Model Using Implantation of Scaffold Based on the of Calcium Phosphate Biocement

The symptomatic full-thickness cartilage lesions or cartilage degeneration leads to the destruction of the normal chondral architecture and bone structure in affected area, causes the osteoarthritis, and general damage to the health. Knee joints are most frequently affected by this condition. The permanent damage of the articular cartilage and subchondral bone has motivated many scientists and clinicians to explore new methods of regeneration of osteochondral defects, such as novel materials.We studied the potential of the biocement based on calcium phosphate consisting of a mixture of four amino acids (glycine, proline, hydroxyproline and lysine) in the regenerating process of the artificially created osteochondral defect on the porcine medial femoral condyle in the stifle joint. The mass ratio of the amino acids in biocement CAL was 4:2:2:1. The Ca/P ratio in cement was 1.67 which correspond with ratio in hydroxyapatite. We compared the results with spontaneous healing of an artificially created cyst with that of the healthy tissue. The animal group treated with biocement paste CAL presented completely filled osteochondral defects. The results were confirmed by histological and radiological assessments, which have shown regenerated chondral and bone tissue in the examined knee joints. Macroscopic evaluation showed that neocartilage was well integrated with the adjacent native cartilage in animal group with biocement CAL, compared with healing of the artificial cyst, where treated cartilage surfaces were visibly lower than the surrounding native cartilage surface and a border between native and restored tissue was apparent. The qualitative assessment of the implant histology specimens showed full regeneration of the hyaline cartilage and subchondral bone in animals with biocement CAL. The artificial cyst group showed remarkable fibrillation. The detailed MRI analysis of cross-section of osteochondral defect confirmed the complete cartilage and subchondral bone healing where the thickness of the regenerated cartilage was 1.5 mm. The MRI imaging of defects in the artificial cyst group showed incomplete healing, neo cartilage tissue reduced up to 50%.

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.

Effects of Escherichia coli Alkaline Phosphatase PhoA on the Mineralization of Dissolved Organic Phosphorus

Alkaline phosphatases, which play the key role in the mineralization of organic phosphorus, have been grouped into three distinct families, PhoA, PhoX, and PhoD. PhoA is still an important component of the Pho regulon for many microbes although its distribution is not as wide as that of PhoX and PhoD. However, several questions remain unclear about the effect of PhoA mineralization of dissolved organic phosphorus. In this study, the role of Escherichia coli alkaline phosphatase PhoA (hereinafter referred to as PhoA) in the mineralization of different organic phosphorus including phosphate monoesters, phosphate diesters, and phytic acids was investigated. The influence of the reaction time, organic phosphorus concentration, and L-amino acid on PhoA mineralization was examined. The results show that PhoA specifically hydrolyzes phosphate monoesters except for phytic acid and the optimal reaction time is around 12 h. The PhoA mineralization rate of glucose 6-phosphate disodium (G6P), 5′-adenosine monophosphate (AMP), and sodium glycerophosphate (BGP) significantly decreased by 38.01%, 55.31%, and 57.08%, respectively (p &lt; 0.01), while the concentration of organic phosphorus increased from 0.50 to 5.00 mg/L. Overall, L-amino acids inhibited PhoA mineralization in a concentration-independent manner. The inhibitory effect of neutral amino acids serine (L-Ser) and tyrosine (L-Tyr) was significantly higher than that of basic amino acids arginine (L-Arg), lysine (L-Lys), and histidine (L-His). All the five amino acids can inhibit PhoA mineralization of AMP, with the highest inhibition rate observed for L-Tyr (23.77%), the lowest—for L-Arg (1.54%). Compared with other L-amino acids, L-Tyr has the highest G6P and BGP mineralization inhibition rate, with the average inhibition rates of 12.89% and 11.65%, respectively. This study provides meaningful information to better understand PhoA mineralization.

The effects of tea plants-soybean intercropping on the secondary metabolites of tea plants by metabolomics analysis

BackgroundIntercropping, especially with legumes, as a productive and sustainable system, can promote plants growth and improves the soil quality than the sole crop, is an essential cultivation pattern in modern agricultural systems. However, the metabolic changes of secondary metabolites and the growth in tea plants during the processing of intercropping with soybean have not been fully analyzed.ResultsThe secondary metabolomic of the tea plants were significant influence with intercropping soybean during the different growth stages. Especially in the profuse flowering stage of intercropping soybean, the biosynthesis of amino acids was significantly impacted, and the flavonoid biosynthesis, the flavone and flavonol biosynthesis also were changed. And the expression of metabolites associated with amino acids metabolism, particularly glutamate, glutamine, lysine and arginine were up-regulated, while the expression of the sucrose and D-Glucose-6P were down-regulated. Furthermore, the chlorophyll photosynthetic parameters and the photosynthetic activity of tea plants were higher in the tea plants-soybean intercropping system.ConclusionsThese results strengthen our understanding of the metabolic mechanisms in tea plant’s secondary metabolites under the tea plants-soybean intercropping system and demonstrate that the intercropping system of leguminous crops is greatly potential to improve tea quality. These may provide the basis for reducing the application of nitrogen fertilizer and improve the ecosystem in tea plantations.

Detection of Isopeptide Bonds in Monoclonal Antibody Aggregates

PurposeA major difficulty in monoclonal antibody (mAb) therapeutic development is product aggregation. In this study, intermolecular isopeptide bonds in mAb aggregates were characterized for the first time. We aim to propose a mechanism of covalent aggregation in a model antibody using stressed studies at raised temperatures to aid in the understanding of mAb aggregation pathways.MethodsAggregate fractions were generated using raised temperature and were purified using size-exclusion chromatography (SEC). The fractions were tryptically digested and characterized using liquid chromatography hyphenated to tandem mass-spectrometry (LC–MS/MS).ResultsAn increased amount of clipping between aspartic acid and proline in a solvent accessible loop in the constant heavy 2 (CH2) domain of the mAb was observed under these conditions. Detailed peptide mapping revealed 14 isopeptide bonds between aspartic acid at that cleavage site and lysine residues on adjacent antibodies. Two additional isopeptide bonds were identified between the mAb HC N-terminal glutamic acid or a separate aspartic acid to lysine residues on adjacent antibodies.ConclusionsInter-protein isopeptide bonds between the side chains of acidic amino acids (aspartate and glutamate) and lysine were characterized for the first time in mAb aggregates. A chemical mechanism was presented whereby spontaneous isopeptide bond formation could be facilitated via either the aspartic acid side chain or C-terminus.

Amino acids in compound feed when rearing replacement young quails

In the technology of production of quail eggs it is important to prepare the replacement young quails for egg laying since the timing of the beginning and duration of the productive period, the level of egg production, and the quality of eggs depend on it. A prerequisite for obtaining high productivity of laying quails in the future is a complete feeding during the rearing period. One solution to this problem is the use of synthetic amino acids in poultry feeding, which increase the biological value of the feed protein to meet the poultry’s protein needs. The results of research on the use of compound feed with an increased content of amino acids (lysine and methionine with cystine) by 15 and 20 % in the nutritional content of replacement young quails of Omskaya breed of meat and egg productivity have been presented in the article. It has been found that the use of experimental compound feed contributed to the increase in the live weight of quails of the experimental groups by 3,94 and 2,97 %, the decrease in the average daily feed consumption by 11,21 and 7,56 %, and expenditures of compound feed for 1 kg of gain by 14,54 and 8,31 %. The higher content of amino acids has increased the yield of effective young quails in the experimental groups by 4,09 and 7,07 %. It has been found in the result of carrying out anatomical cutting of carcasses that feeding quails with experimental compound feed had a stimulating effect on the development of egg-forming organs. The calculation of the economic efficiency of rearing replacement young quails has shown that the use of compound feed with the increase in amino acids by 15 and 20 % in compound feed reduced the expenditures of their rearing by 8,04 and 5,28 % and the prime-cost of one replacement female quail by 12,86 and 11,63 %.

Separation and Identification of some amino acids in the bark, sapwood, and heartwood of Ceratonia siliqua L. trees growing in Mosul.

This study was performed to separate and identify a number of amino acids present in the bark, sapwood, and heartwood of Ceratonia siliqua L growing in Mosul, seven of the amino acids were identified in the bark, eight in the sapwood, and seven in the heartwood. In general, the number of identified acids in the carob understudy is eight, which are: Aspartic Acid, Phenylalanine Acid, Glumatic Acid, Lysine Acid, Glycine Acid, Tryptophan Acid, Lucien Acid, and Serine Acid. The highest quantities were in the sapwood and the lowest was in the bark. These results are consistent with the results of the study samples (bark, sapwood, and heartwood) of carob trees. The bark, sapwood, and heartwood are similar in terms of amino acids with the exception of serine, which did not appear in both bark and heartwood, and glutamic acid did not appear in sapwood, the presence of this large number of amino acids in carob wood may be due to the ability of carob trees to fix nitrogen in root nodes.

Abstract 325: Nascent proteome analysis of tumor cells and their microenvironment in cultured human tumor tissues

Abstract Solid tumors are often considered as abnormal organs composed of the cancerous cells and their surrounding tumor microenvironment (TME) containing fibroblasts, immune cells, blood and lymphatic vessels, and the extracellular matrix. The heterotypic interactions between this diversity of cell types within the TME are maintained through a wide variety of secreted proteins, resulting in a favorable milieu for the progression of the malignancy. The interactions between tumor cells and TME are complex and remain poorly understood. Here we investigated this by developing a unique nascent proteomic approach in tumor tissues.Precision cut cancer tissue slices (PCCTS) maintain tissue heterogeneity with different cell types and preserved TME. Cultivation of PCCTS provides an ex vivo model for tumor tissues. We developed an approach for PCCTS's nascent proteome analysis, using pulsed-SILAC (stable isotope labeling with amino acids in cell culture) labeling combined with click-chemistry to selectively isolate and quantify newly synthesized proteins in the TME upon applying a cellular perturbation. It is a powerful tool to selectively enrich secretory proteins from culture media even with presence of serum. Primary human ovarian tumors (phOVT) and patient derived xenografts (PDX) were used to produce the PCCTS with thickness of 150µm to 300µm. The different cell types and extracellular matrix of PCCTS make the depletion period of cells from the amino acids (methionine, lysine and arginine) prior the AHA-SILAC treatment difficult to define. The PCCTS need longer depletion periods than the 2D cell culture, the longer the depletion period the better depletion efficiency. Following, PCCTS were cultured in AHA-SILAC media and treated with cisplatin. PCCTS and culture media (containing secreted proteins) were harvested; newly synthesized proteins were enriched via click-chemistry and analyzed with mass spectrometry. The labeling time of 10 to 12h showed a good labeling efficiency of more than 60%, still further optimizations are needed. Different PCCTS showed various labeling efficiency indicating the patients heterogeneity. The nascent proteome analysis with cisplatin treatment demonstrated different protein regulations in patients suggesting different drug responses. STRING analysis can be applied to predict the protein-protein interactions. The immunohistochemical staining of the same PCCTS can be processed to further validate the results of the proteome analysis. In conclusion, we established a pulsed SILAC-AHA treatment approach for the PCCTS with the TME. This unique approach allows tracking the compositional and dynamic changes within the proteome and monitoring the direct proteome response at rapid time scale. It can be used to reveal a part of the proteome that has been poorly understood in the tumor tissues and contribute to studying cellular communication and finding new therapeutic targets. Citation Format: Meng Dong, Karim Aljakouch, Kathrin Böpple, Bernd Winkler, Julia Schüler, Frank Essmann, Hans-Georg Kopp, Jeroen Krijgsveld, Walter E. Aulitzky. Nascent proteome analysis of tumor cells and their microenvironment in cultured human tumor tissues [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 325.

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.

GARLAND CHRYSANTHEMUM IS A PROMISING NEW MEDICINAL PLANT

A brief overview of the relevance of using garland chrysanthemum (Glebionis coronaria (L.) Cass. ex Spach syn. Chrysanthemum coronarium (L.)) as a raw material in the preparation of medicaments is introduced in this article. High content of micro- and macroelements (K, Na, Ca, P, Zn, Fe, Cu, Mn, J) was found in garland chrysanthemum, as well as other biologically active compounds, of which anthraquinones (emodin, chrysophanol), flavonoids (quercetin, rutin), carotenoids and ascorbic acid (up to 56.0 mg%) can be considered the main ones, likewise vitamins B, PP and free amino acids (lysine, methionine, cysteine, histidine, arginine, threonine, serine, proline, glycine, valine, isoleucine, leucine, tyrosine, phenylalanine; nine of which are essential). These elements have a positive therapeutic effect on the cardiovascular system, the human gastrointestinal tract. The garland chrysanthemum is a very plastic and undemanding vegetable crop with medicinal properties. It is capable of growing on technogenic soils contaminated with cobalt, zinc and nickel, accumulating these elements to a lesser extent than traditional vegetable plants, which is very important in modern conditions. It is shown that in the conditions of the Moscow region it can be well grown by sowing seeds directly on field, continues to maintain the rhythms of growth and development, passes through all phenological phases and gives a stable high yield of green mass — up to 5.3 kg/m2, inflorescences — up to 1.4 kg/m2, seeds — up to 130.0 g/m2. Low laboratory and field germination of seeds up to 50 % has been established.

Changes in melon plant phytochemistry impair Aphis gossypii growth and weight under elevated CO2

Elevated CO2 (eCO2) modifies plant primary and secondary metabolism that subsequently impacts herbivore insect performance due to changes in its nutritional requirements. This laboratory study evaluated interactions between Aphis gossypii Glover (Hemiptera: Aphididae) and melon (Cucumis melo L., Cucurbitaceae), previously acclimated two or six weeks to different CO2 levels, eCO2 (700 ppm) or ambient CO2 (400 ppm). Under eCO2, melon plants decreased nitrogen foliar concentration and increased carbon to nitrogen ratio, independently of acclimation period, significantly reducing the content of some amino acids (alanine, asparagine, glycine, isoleucine, lysine, serine, threonine, and valine) and increasing the carbohydrate (sucrose) content in melon leaves. The dilution in some essential amino acids for aphid nutrition could have aggravated the reduction in A. gossypii population growth reared on melon previously acclimated two weeks to eCO2, as well as the loss of aphid body mass from two successive generations of A. gossypii reared under eCO2 on plants previously acclimated two or six weeks to eCO2. The response to eCO2 of phloem feeders, such as aphids, is actually variable, but this study highlights a negative response of A. gossypii to this climate change driver. Potential implications on control of this pest in a global change scenario are discussed.

Determination and Evaluation of Se-Rich High-Quality Rice Produced by Compound Nutrient Solution

Rice is one of the most important food crops in the world. Its biggest flaw is the low content of protein and essential amino acids, which severely limits its nutritional value. In order to produce high-quality rice with rich Se, we sprayed different concentrations of compound nutrient solution (containing Se (selenium), amino acid compound, zinc and boron) on the rice at different growth stages; and then determined the main nutrient content of their polished rice. The results showed that spraying low concentration compound nutrient solution (Each liter contained 20 mg of Se, 333 mg of complex amino acids, 33 mg of zinc and 33 mg of boron) to rice in the heading stage produced rice with the highest total starch and fat content and lower amylose content. Spraying high concentration compound nutrient solution to rice during the filling stage produced rice with the highest Se content. Multiple sprays of compound nutrient solution produced rice with low protein and low starch. Spraying low concentration compound nutrient solution on rice in milky stage significantly increased the content of protein, total starch, fat, all essential amino acids (Lysine increased by more than 57%), amylopectin and Se in rice; significantly reduced amylose content; significantly improved the nutritional value and taste quality of rice. The conclusion is that spraying low concentration compound nutrient solution on rice in the milky stage can produce rice with the highest content of protein and essential amino acids, higher content of total starch, fat and Se, and the lowest amylose content; significantly improve the nutritional value and taste quality of rice. The technical solution can comprehensively and effectively improve the nutritional value and flavor quality of rice, and has great development and application value.

Сложность патогенеза коагулопатии при COVID-19

Введение. Коронавирус SARS-CoV-2 является новым вирусом, обладающим способностью осуществлять трансмиссию воздушно-капельным путем, вызывая тяжелое течение атипичной пневмонии, нередко сочетающейся с коагулопатиями. Роль структурных белков коронавируса в их патогенезе неизвестна. Цель исследования: с помощью биоинформационного анализа выявить в структурных белках коронавируса SARS-CoV-2 последовательности, гомологичные белкам системы гемостаза, и рассмотреть возможные сценарии их участия в патогенезе коагулопатий при COVID-19, а также объяснить существование вирусостатического эффекта гепарина. Материалы и методы. Для компьютерного анализа были использованы доступные в Интернете базы данных первичных структур белков коронавирусов и их рецепторов, а также поверхностных белков других вирусов, белков системы гемостаза и иммунной системы. Сравнивали аминокислотный состав белков и распределение оснόвных аминокислот (аргинина и лизина) в их первичных последовательностях. С целью выявления пептидного (иммуноэпитопного) родства структурных белков коронавирусов с белками системы гемостаза человека был выполнен поиск гомологичных последовательностей в их белках. Результаты. В структурных белках коронавируса SARS-CoV-2 выявлено множество последовательностей, гомологичных белкам системы гемостаза и иммунной системы. В отличие от коронавирусов SARS-CoV и MERS-CoV, S1-субъединица S-белка коронавируса SARS-CoV-2 имеет положительную полярность. Заключение. Множество последовательностей в структурных белках коронавируса SARS-CoV-2, гомологичных белкам системы гемостаза, потенциально способны вы- зывать различные сценарии патогенеза коагулопатий. Положительная полярность S1-субъединицы S-белка коронавируса SARS-CoV-2 позволяет объяснить неспецифическое взаимодействие ее с гепарином и его вирусостатический (неантикоагулянтный) эффект. Background. The coronavirus SARS-CoV-2 is a new virus capable of human-human transmission and inducing a severe atypical pneumonia often associated with coagulopathy. A role of SARS-CoV-2 structural proteins in coagulopathy pathogenesis is unknown. Objectives: to use a bioinformation analysis to identify SARS-CoV-2 sequences in the structural proteins that are homologous to hemostasis system proteins, regard their possible participation in coagulopathy pathogenesis and explain the antiviral effect of heparin. Materials / Methods. For computer analysis, Internet databases were used of the primary structures of coronavirus proteins and their receptors, as well as surface proteins of other viruses, proteins of hemostasis and immune systems. The amino acid composition of proteins and the distribution of basic amino acids (arginine and lysine) in their primary sequences were compared. For detection of peptide (immunoepitopic) relationship of coronaviruses structural proteins with human hemostasis proteins, a search for homologous sequences in their proteins was performed. Results. Many sequences have been identified in structural proteins of SARS-CoV-2 coronavirus that are homologous to the proteins of hemostasis and immune systems. In contrast with SARS-CoV and MERS-CoV coronaviruses, the S1-subunit of SARS-CoV-2 coronavirus S-protein has a positive polarity. Conclusions. Many sequences in SARS-CoV-2 structural proteins that homologous to hemostasis system proteins are potentially responsible for coagulopathy pathogenesis. The positive polarity of the S1-subunit of SARS-CoV-2 S-protein explains its nonspecific interaction with heparin and its virostatic (non-anticoagulant) effect.

Characterization and Stress Response of the JmjC Domain-Containing Histone Demethylase Gene Family in the Allotetraploid Cotton Species Gossypium hirsutum.

Histone modification is an important epigenetic modification that controls gene transcriptional regulation in eukaryotes. Histone methylation is accomplished by histone methyltransferase and can occur on two amino acid residues, arginine and lysine. JumonjiC (JmjC) domain-containing histone demethylase regulates gene transcription and chromatin structure by changing the methylation state of the lysine residue site and plays an important role in plant growth and development. In this study, we carried out genome-wide identification and comprehensive analysis of JmjC genes in the allotetraploid cotton species Gossypium hirsutum. In total, 50 JmjC genes were identified and in G. hirsutum, and 25 JmjC genes were identified in its two diploid progenitors, G. arboreum and G. raimondii, respectively. Phylogenetic analysis divided these JmjC genes into five subfamilies. A collinearity analysis of the two subgenomes of G. hirsutum and the genomes of G. arboreum and G. raimondii uncovered a one-to-one relationship between homologous genes of the JmjC gene family. Most homologs in the JmjC gene family between A and D subgenomes of G. hirsutum have similar exon-intron structures, which indicated that JmjC family genes were conserved after the polyploidization. All G. hirsutum JmjC genes were found to have a typical JmjC domain, and some genes also possess other special domains important for their function. Analysis of promoter regions revealed that cis-acting elements, such as those related to hormone and abiotic stress response, were enriched in G. hirsutum JmjC genes. According to a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis, most G. hirsutum JmjC genes had high abundance expression at developmental stages of fibers, suggesting that they might participate in cotton fiber development. In addition, some G. hirsutum JmjC genes were found to have different degrees of response to cold or osmotic stress, thus indicating their potential role in these types of abiotic stress response. Our results provide useful information for understanding the evolutionary history and biological function of JmjC genes in cotton.

PREBIOTICS AND NUTRITIONAL CORRECTION IN INTEGRATED THERAPY OF PATIENTS WITH PULMONARY TUBERCULOSIS AND COMORBID PROTEIN METABOLIC DISORDERS

The purpose of the work is to study the effectiveness of prebiotics and nutritional correction as components of the integrated therapy of patients with pulmonary tuberculosis and protein metabolic disorders. 67 patients with pulmonary tuberculosis were examined and then divided into two groups: I group included 35 patients, who received standard antimycobacterial therapy in the intensive phase of the treatment in combination with prebiotic (lactulose) in a dose of 20 ml 3 times a day plus nutriological correction of metabolic disorders; the II group included 32 patients, who received standard antimycobacterial therapy. The control group consisted of 30 healthy individuals of comparable sex and age. In order to study the protein metabolism state, the content of certain substitutable amino acids (ornithine, aspartic acid, serine, glutamic acid, proline, glycine, alanine, cysteine, tyrosine, glutamine) and essential amino acids (lysine, histidine, arginine, threonine, valine, methionine, isoleucine, phenylalanine, leucine) and their total amount (mg per 100 ml of blood serum) were assessed. Results and discussion. It has been found that all patients in groups I and II, which were under our supervision before the treatment, had symptoms of intoxication and respiratory syndrome, the tuberculosis process in the lungs occupied more than three segments and single decaying and rupturing cavities prevailed. The patients with massive bacterial excretion dominated in both groups. Moreover, before the treatment, protein metabolic disorders were found out in both groups. They manifested by the imbalance of individual amino acids and led to a decrease in the total content of essential amino acids in 1,2 – 1,3 times, total content of substitutable amino acids in 1, 2 times that led to a decrease in total content of substitutable amino acids in patients of both groups in 1,2 – 1,3 in comparison with the control group. A comparative analysis of the effectiveness of prebiotics (lactulose) and nutritional correction of metabolic disorders in combination with antimycobacterial therapy in the TB patientsl with protein metabolic disorders showed that probiotics and dietary modifications contribute to improving protein metabolism.&#x0D; This has been evidenced by a significantly high (p &lt;0,05) level of the total content of essential amino acids in blood serum, total content of substitutable amino acids and total content of amino acids (reliably reaching the level of indicators in healthy individuals) compared with the relevant indicators in the group of the patients receiving only antimycobacterial therapy. Probiotics and nutritional correction promote the rapid dynamics of the disappearance of respiratory syndromes and intoxication, promote cavity healing, and reduce the duration of treatment. Conclusions. The results of our research obtained convincingly suggest that prior to the beginning of treatment; all patients with pulmonary tuberculosis have protein metabolic disorders that require selecting a way to correct these disorders in order to increase the effectiveness of TB treatment. Antimycobacterial therapy during two months contributes to the positive dynamics of protein metabolism correction, but complete normalization of the amino acid composition in blood serum does not occur. Applying prebiotics (lactulose) and nutriological correction of metabolic disorders in combination with antimycobacterial therapy promotes the normalization of protein metabolism and increases the effectiveness of the TB treatment.