Amino Acids Gln Research Articles

Specific inhibitory effects of exogenous d-Aspartate on the proliferation of intestinal epithelial cells

d-amino acids have been actively examined since improved analytical techniques revealed their presence in animal bodies. Although D-Asp was identified in mammals earlier than D-Ser, research on D-Asp has lagged behind that on D-Ser, mainly because the target protein of D-Asp remains unknown. To date, the only reported functions of D-Asp are its roles in reproduction and suggested neuromodulatory functions. Since d-amino acids are also present in food, it is important to clarify their effects on gastrointestinal epithelial cells, which are always contacted after ingestion. Therefore, the present study examined the effects of d-amino acids on gastrointestinal tract basal cells. The effects of 11 types of amino acids (Ala, Arg, Asn, Asp, Gln, Glu, Leu, Lys, Pro, Ser, and Val) on the proliferation of three types of gastrointestinal epithelial cells (HGC-27, IEC-6, and Caco-2) were assessed. Although the proliferation of HGC-27 and Caco-2 was not affected by any of the 11 types of L- and d-amino acids, D-Asp inhibited the proliferation of IEC-6, derived from small intestinal epithelial cells, in concentration- and exposure time-dependent manners. The present study also examined uptake transporters, metabolic enzymes, and insulin signaling pathways; however, the mechanisms underlying the inhibitory effects of D-Asp on the proliferation of IEC-6 were not elucidated. A more detailed understanding of these mechanisms may lead to the development of pharmaceuticals as main drugs or formulation materials. Further studies are warranted on the physiological effects of d-amino acids, including D-Asp.

New Thiazolidine-4-One Derivatives as SARS-CoV-2 Main Protease Inhibitors.

It has been more than four years since the first report of SARS-CoV-2, and humankind has experienced a pandemic with an unprecedented impact. Moreover, the new variants have made the situation even worse. Among viral enzymes, the SARS-CoV-2 main protease (Mpro) has been deemed a promising drug target vs. COVID-19. Indeed, Mpro is a pivotal enzyme for viral replication, and it is highly conserved within coronaviruses. It showed a high extent of conservation of the protease residues essential to the enzymatic activity, emphasizing its potential as a drug target to develop wide-spectrum antiviral agents effective not only vs. SARS-CoV-2 variants but also against other coronaviruses. Even though the FDA-approved drug nirmatrelvir, a Mpro inhibitor, has boosted the antiviral therapy for the treatment of COVID-19, the drug shows several drawbacks that hinder its clinical application. Herein, we report the synthesis of new thiazolidine-4-one derivatives endowed with inhibitory potencies in the micromolar range against SARS-CoV-2 Mpro. In silico studies shed light on the key structural requirements responsible for binding to highly conserved enzymatic residues, showing that the thiazolidinone core acts as a mimetic of the Gln amino acid of the natural substrate and the central role of the nitro-substituted aromatic portion in establishing π-π stacking interactions with the catalytic His-41 residue.

A novel hydrophobic peptide FGMp11: Insights into antimicrobial properties, hydrophobic sites on Staphylococcus aureus and its application in infecting pasteurized milk

Hydrophobic peptides have received considerable attention in food preservation. The elucidation of the hydrophobic points of hydrophobic peptides is helpful in revealing the antimicrobial mechanism, and screening strategies for novel antimicrobials such as molecular docking and deep learning. A novel antimicrobial peptide (AMP) FGMp11 was identified and screened from fermented goat milk with strong hydrophobic properties, β-turn structure, and the amino acid sequence of TPVVVPPFLQP. The hydrophobic properties of FGMP11 could significantly damage the integrity of the cell membrane, resulting in changes in cell morphology. Hydrophobic interactions of FGMp11 with Mur enzymes (MurC, MurD, MurE, MurF, and MurG) are ideal targets for antimicrobial therapy, as these targets are involved in peptidoglycan synthesis via hydrogen bond interactions. The electrostatic binding sites of FGMp11 and the key amino acids GLN, LEU, PRO, and VAL were mainly -OH, -NH2, and -C=O groups, reducing cell activity. FGMp11 was able to significantly inhibit the growth of S. aureus during the storage of pasteurized milk. Overall, these findings provide a scientific basis for the antimicrobial activity of FGMp11 against food-borne pathogens and its application in food preservation.

Fine mapping of TFL, a major gene regulating fruit length in snake gourd (Trichosanthes anguina L).

Fruit length is a crucial agronomic trait of snake gourd (Trichosanthes anguina L); however, genes associated with fruit length have not been characterised. In this study, F2 snake gourd populations were generated by crossing the inbred lines, S1 and S2 (fruit lengths: 110 and 20cm, respectively). Subsequently, bulk segregant analysis, sequencing, and fine-mapping were performed on the F2 population to identify target genes. Our findings suggest that the fruit length of snake gourd is regulated by a major-effect regulatory gene. Mining of genes regulating fruit length in snake gourd to provide a basis for subsequent selection and breeding of new varieties. Genotype-phenotype association analysis was performed on the segregating F2 population comprising 6,000 plants; the results indicate that the target gene is located on Chr4 (61,846,126-61,865,087bp, 18.9-kb interval), which only carries the annotated candidate gene, Tan0010544 (designated TFL). TFL belongs to the MADS-box family, one of the largest transcription factor families. Sequence analysis revealed a non-synonymous mutation of base C to G at position 202 in the coding sequence of TFL, resulting in the substitution of amino acid Gln to Glu at position 68 in the protein sequence. Subsequently, an InDel marker was developed to aid the marker-assisted selection of TFL. The TFL in the expression parents within the same period was analysed using quantitative real-time PCR; the TFL expression was significantly higher in short fruits than long fruits. Therefore, TFL can be a candidate gene for determining the fruit length in snake gourd. Collectively, these findings improve our understanding of the genetic components associated with fruit length in snake gourds, which could aid the development of enhanced breeding strategies for plant species.

Exploring metabolic effects of dipeptide feed media on CHO cell cultures by in silico model-guided flux analysis

There is a growing interest in perfusion or continuous processes to achieve higher productivity of biopharmaceuticals in mammalian cell culture, specifically Chinese hamster ovary (CHO) cells, towards advanced biomanufacturing. These intensified bioprocesses highly require concentrated feed media in order to counteract their dilution effects. However, designing such condensed media formulation poses several challenges, particularly regarding the stability and solubility of specific amino acids. To address the difficulty and complexity in relevant media development, the biopharmaceutical industry has recently suggested forming dipeptides by combining one from problematic amino acids with selected pairs to compensate for limitations. In this study, we combined one of the lead amino acids, L-tyrosine, which is known for its poor solubility in water due to its aromatic ring and hydroxyl group, with glycine as the partner, thus forming glycyl-L-tyrosine (GY) dipeptide. Subsequently, we investigated the utilization of GY dipeptide during fed-batch cultures of IgG-producing CHO cells, by changing its concentrations (0.125 × , 0.25 × , 0.5 × , 1.0 × , and 2.0 ×). Multivariate statistical analysis of culture profiles was then conducted to identify and correlate the most significant nutrients with the production, followed by in silico model-guided analysis to systematically evaluate their effects on the culture performance, and elucidate metabolic states and cellular behaviors. As such, it allowed us to explain how the cells can more efficiently utilize GY dipeptide with respect to the balance of cofactor regeneration and energy distribution for the required biomass and protein synthesis. For example, our analysis results uncovered specific amino acids (Asn and Gln) and the 0.5 × GY dipeptide in the feed medium synergistically alleviated the metabolic bottleneck, resulting in enhanced IgG titer and productivity. In the validation experiments, we tested and observed that lower levels of Asn and Gln led to decreased secretion of toxic metabolites, enhanced longevity, and elevated specific cell growth and titer.Key points• Explored the optimal Tyr dipeptide for the enhanced CHO cell culture performance• Systematically analyzed effects of dipeptide media by model-guided approach• Uncovered synergistic metabolic utilization of amino acids with dipeptideGraphical abstract

Human leukocyte antigen-DQA1*04:01 and rs2040406 variants are associated with elevated risk of childhood Burkitt lymphoma

Burkitt lymphoma (BL) is responsible for many childhood cancers in sub-Saharan Africa, where it is linked to recurrent or chronic infection by Epstein-Barr virus or Plasmodium falciparum. However, whether human leukocyte antigen (HLA) polymorphisms, which regulate immune response, are associated with BL has not been well investigated, which limits our understanding of BL etiology. Here we investigate this association among 4,645 children aged 0-15 years, 800 with BL, enrolled in Uganda, Tanzania, Kenya, and Malawi. HLA alleles are imputed with accuracy >90% for HLA class I and 85-89% for class II alleles. BL risk is elevated with HLA-DQA1*04:01 (adjusted odds ratio [OR] = 1.61, 95% confidence interval [CI] = 1.32-1.97, P = 3.71 × 10−6), with rs2040406(G) in HLA-DQA1 region (OR = 1.43, 95% CI = 1.26-1.63, P = 4.62 × 10−8), and with amino acid Gln at position 53 versus other variants in HLA-DQA1 (OR = 1.36, P = 2.06 × 10−6). The associations with HLA-DQA1*04:01 (OR = 1.29, P = 0.03) and rs2040406(G) (OR = 1.68, P = 0.019) persist in mutually adjusted models. The higher risk rs2040406(G) variant for BL is associated with decreased HLA-DQB1 expression in eQTLs in EBV transformed lymphocytes. Our results support the role of HLA variation in the etiology of BL and suggest that a promising area of research might be understanding the link between HLA variation and EBV control.

Penetrance of missense mutation of OR2AT4 and OR9G1/9 gene polymorphism increases risk factor in azoospermic patients

Objectives: In human, olfactory receptor (ORs) gene polymorphism regulate testicular development and physiological functions during spermatogenesis. However, the etiopathology of OR gene in male infertility is still restricted that required further studies. Basic mechanism involves the identification of peripheral signals based on specific chemoreceptors site that are expressed on the surface of mature spermatozoa inside the female reproductive tract. These receptors belong to G-protein couple receptors (GPCR) of superfamily and the roles of ORs gene linked to sperm chemotaxis through chemokinesis. In the present study, we have studied the role of Olfactory receptor family 2 subfamilies AT member 4 (OR2AT4) and Olfactory receptor family 9 subfamily G member 1/9 (OR9G1/9) genes polymorphism in clinically diagnosed azoospermic patients. Materials and Methods: Systematically, GTG banded karyotypes were developed using short term lymphocyte cultures, fluorescence in situ hybridization using SRY probe and PCR based microdeletion of Y-chromosome was characterized using STS marker (SY255). Whole exome sequencing (WES) based analysis was performed in azoospermic patients to identify the involvement of de-novo mutation in OR2AT4 and OR9G1/OR9G9 genes mutation followed by bioinformatics tools used for the analysis to identify changes in respective amino acids. Results: OR2AT4 showing missense mutation after substitution of nucleotide (G→A) followed by lack of change in amino acid (Gln→Gln), but in OR9G1/G9 gene showing changes of substitution of nucleotide thymidine into adenine (T→A), whereas valine is replaced by glutamine (Val→ Glu), similarly thymidine is substituted by cysteine (T→C), after decode phenylalanine is replaced by leucine (Phe→Leu). Further, 3D protein helical structure was developed with the help of bioinformatics on the basis of amino acids coded protein and compare with mutated amino acid or truncated protein residues with the help of iTASSER server. Conclusion: Present study demonstrate the role of OR2AT4 and OR9G1/9 gene polymorphism elucidates the functional role of protamine in ectopic receptors followed by increasing “risk” of developing male infertility that leads the clinical condition defined as non-obstructive azoospermia.

New imidazole-2-ones and their 2-thione analogues as anticancer agents and CAIX inhibitors: Synthesis, in silico ADME and molecular modeling studies

The present study involves the synthesis of a series of new imidazole-2-ones derivatives and their 2-thione analogs using conventional heating and the environmentally friendly benign technique, the microwave technique. Structure of the compounds was well elucidated by considering the data of both elemental and spectral analyses. The obtained data and theoretical values of the synthesized molecules correlated with the proposed molecular structure. Moreover, all the synthesized compounds were evaluated in vitro for antitumor activity against HCT-116 and HeP2 human cancer cell panels and assessed as selective carbonic anhydrase IX isozyme (CA9/CAIX) inhibitors, thereby providing useful preliminary evidence for drug development. In addition, computational techniques were used to investigate the molecular and electronic characteristics of the investigated organic compounds. The 4b compound exhibited the best quantum chemistry features, as the highest occupied molecular orbital, softness, energy gap, and dipole moment, indicating the highest biological activity. This was supported by the experimental findings. Moreover, the in silico evaluation of drug candidates was also investigated. Thereafter, the anticancer activity of the most reactive candidate was studied via molecular docking to determine the types of interactions between this molecule and CAIX. According to the docking experiments, the 4b molecule generates five hydrogen bond interactions with active amino acid residues, Gln 92, Gln 67, and Thr 200.

Molecular Characteristics of Chicken Infectious Anemia Virus in Central and Eastern China from 2020 to 2022.

To evaluate the recent evolution of CIAV in China, 43 flocks of chickens from the provinces of Henan, Jiangsu, Hubei, and Anhui were screened via polymerase chain reaction during 2020-2022. Of these, 27 flocks tested positive for CIAV nucleic acids, including 12 which were positive for other immunosuppression viruses. Additionally, 27 CIAV strains were isolated, and their whole genomes were sequenced. The AH2001 and JS2002 strains shared the highest identity at 99.56%, and the HB2102 and HB2101 strains shared the lowest identity at 95.34%. Based on the genome sequences of these strains and reference strains, a phylogenetic tree was constructed and divided into eight main branches. Most of the strains were grouped with the East Asian strains, whereas the HB2101 strain belonged to the Brazil and Argentina cluster. A recombination event was detected in multiple strains, in which AH2002 recombined from KJ728827/China/2014 (from Taiwan Province) and HN2203, and AH2202 recombined from KX811526/China/2017 (from Shandong Province) and HN2203. All the obtained strains had a highly pathogenic Gln amino acid site at position 394 of the VP1. Overall, our findings demonstrate the importance of CIAV monitoring and provide data that aid in understanding the evolution of CIAV.

Analysis of Urinary Amino Acids by High-Performance Liquid Chromatography with Fluorescence Detection Using 2,3-Naphthalenedicarboxaldehyde as Fluorescence Derivatization Reagent

Amino acids are involved in various chemical reactions in vivo, and changes in several amino acids in urine are related to certain disease states. Therefore, developing an efficient method to analyze the amino acids in urine is useful in the timely diagnosis of diseases. In this study, we developed a high-performance liquid chromatography (HPLC) fluorescence method for the quantitative analysis of urinary amino acids using the fluorescence derivatization reagent 2,3-naphthalenedicarboxaldehyde (NDA). NDA was selected because it does not require heating for the reaction and can react within a short time, rendering its use in clinical settings feasible. The reaction temperature, reaction time, and other derivatization conditions were optimized, and the reaction was found to be completed in 5 min at 25 °C. The separation of NDA–amino acids was investigated on an octadecylsilyl (ODS) column under gradient conditions. The mobile phase was a mixture of water–acetonitrile–trifluoroacetic acid. Eighteen NDA–amino acids (histidine (His), arginine (Arg), asparagine (Asn), glutamine (Gln), citrulline (Cit), serine (Ser), aspartic acid (Asp), threonine (Thr), glutamic acid (Glu), glycine (Gly), tyrosine (Tyr), alanine (Ala), tryptophan (Trp), valine (Val), phenylalanine (Phe), isoleucine (Ile), leucine (Leu), and 5-aminovaleric acid (internal standard)) were separated within 100 min under optimal conditions. The calibration curves showed good linearity in the range of 0.25–25 pmol per injection with correlation coefficients of >0.998. The limits of quantification for NDA–amino acids were 16.7–74.7 fmol. The developed analytical method was applied to a human urine sample and 16 amino acids (His, Arg, Asn, Gln, Cit, Ser, Thr, Glu, Gly, Tyr, Ala, Trp, Val, Phe, Ile, and Leu) were quantified. The urinary amino acid concentrations were 5–960 μM. Urinary amino acid analysis using this method is expected to be clinically applicable as a novel biomarker for diseases affecting the bladder, urinary tract, and kidneys.

Combined Effect of Feed and Housing System Affects Free Amino Acid Content of Egg Yolk and Albumen in Brown Layer Chickens.

In recent years, the market share for cage-free eggs has gradually increased. Because commercially available cage-free eggs are often produced not only by several housing systems but also with different feed crude protein (CP) levels, there are combined effects of feed and housing systems between cage-free and cage eggs. Therefore, using field data, this study aimed to determine the combined effects of feed and housing systems on egg traits and yolk and albumen amino acids in table eggs. Brown layers (n = 40) at the middle laying stage under two feed and housing systems (cage, CP 15.5% diet; barn, CP 17.0% diet) were used. One-way analysis of variance and Pearson's correlation analysis were used to evaluate 10 egg traits, 19 yolk amino acid traits, and 20 albumen amino acid traits. We observed significant effects of feed and housing on two egg traits (yolk weight and eggshell color redness), 16 yolk amino acids (Asp, Glu, Asn, Ser, Gln, His, Arg, Thr, Ala, Tyr, Met, Cys, Ile, Leu, Phe, and Lys), and 14 albumen amino acids (Asp, Asn, Ser, Gln, Gly, His, Arg, Thr, Ala, Val, Met, Cys, Ile, and Leu). This study revealed that eggs from the barn system (CP 17.0%) contained higher levels of free amino acids in 15 yolk and nine albumen amino acid traits. Phenotypic correlations among the 49 egg traits indicated similar correlation patterns in both systems, which implies that the balance of free amino acid content in yolk and albumen is similar in each system. Although some potential confounding factors may be present for comparing egg content between cage (CP 15.5%) and barn (CP 17.0%) systems, this study suggests that commercially available cage-free eggs may be different from cage eggs not only in external egg traits but also yolk and albumen amino acid traits.

Glutamate Metabolism Shapes Osteoclast Differentiation: New Potential Therapeutic Target to Block Osteoclastic Bone Resorption in Multiple Myeloma Patients

Multiple Myeloma (MM) cells are characterized by an impaired Glutamine (Gln) metabolism and Gln-addiction feature. As a result of this, MM patients have lower levels of Gln and higher levels of its main metabolite Glutamate (Glu) in the bone marrow (BM) microenvironment as compared with Smoldering MM (SMM) and Monoclonal Gammopathy of Uncertain Significance (MGUS) patients. Moreover, we have recently shown that MM-dependent Gln-exhausted microenvironment impairs osteoblast differentiation. Based on this evidence, we have hypothesized that alterations of Gln/Glu axis may have a significant impact on the activation of osteoclasts (OCLs) and in the development of osteolytic lesions. We purified BM CD14+ monocytes from a cohort of 30 patients with monoclonal gammopathies (5 MGUS, 12 SMM and 13 newly diagnosed MM (ND-MM)). Cells have been incubated in osteoclastogenic medium in the presence or absence of Glu. The samples were subsequently analyzed by real-time PCR, Western blot and TRAP staining after 3 or 14 days. Amino acid uptake and amino acid content have been assessed under the same conditions. Glu uptake inhibitors have been tested throughout osteoclastogenesis. Our data showed that the presence of Glu significantly increases the number of OCLs generated from MGUS/SMM CD14+ compared with cells differentiated in the absence of the amino acid (p<0.01). In these patients, Glu increases the expression of RANK as well as NF-κB and NFATc1 protein level pointing to a stimulatory effect on osteoclastogenic signaling pathway. On the contrary, CD14+ obtained from MM patients have a lower response to Glu probably as a consequence of the high Glu microenvironment imposed by MM cells. Consistently, MM CD14+ cells are prone to generate more OCLs also in the absence of Glu compared with CD14+ cells obtained from premalignant samples (median number of OCLs: MGUS/SMM 33, MM 123, p=0.02). The analysis of intracellular amino acids (Gln, Glu, and α-ketoglutarate (2-OG)) indicated that Glu content significantly increases after 8 days of differentiation while decreasing after 14 days pointing to a fast accumulation of Glu upon differentiation. The content of both Gln and 2-OG do not vary appreciably throughout the differentiation. These data suggest that OCLs differentiation is associated with an increased uptake of extracellular Glu. Consistently, the activity of the sodium-dependent Glu transporters EAATs, increases already after 3 days of incubation in differentiating medium without Glu, with a further increase in the presence of the amino acid. Expression analysis revealed that, among EAATs, EAAT1 (encoded by SLC1A3 gene) is responsible for the uptake of extracellular Glu upon differentiation both in the presence and in the absence of the amino acid. The expression of other Glu transporters do not change during differentiation. Selective inhibitors of EAAT1, UCPH101 and TFB-TBOA completely block the formation of multinucleated OCLs in the presence of Glu (p<0.01). In parallel to EAAT1 induction, Glu treatment increases the expression of Calcineurin, the serine/threonine phosphatase activated by increased intracellular calcium levels indicating the possible modulation of calcium signaling by Glu. In conclusion, our results indicate that high BM Glu level sustains OCLs formation by stimulating of NF-κB/NFATc1/calcineurin pathway. EAAT1 transporter is responsible for the uptake of extracellular Glu during osteoclastogenesis while its inhibition markedly blocks OCL formation. Therefore, targeting glutamate metabolism could be a therapeutic strategy to block OCL formation in MM patients.

In silico structural and interactional study of wheat proteins gliadin and glutenin with comparison to wheat toxin alpha-purothionin

Wheat is most important component of food throughout the world. Wheat is full of different proteins but most abundant and most studied proteins are gliadin and glutenin. Interaction of gliadin and glutenin is important in biological properties of the wheat flour. Wheat also produce a toxin known as purothionin, which is used by the plant to show defense against different types of pathogen attack. Current in silico study was done to investigate the interaction between gliadin and glutenin proteins against wheat toxin alpha-purothionin solely. Methodology that was used in the current study consists of RaptorX tool for protein modeling, PROCHECK for predicted 3D model verification. CASTp 3.0 online tool for active site prediction. Online tool Cluspro and offline tool ligplus+v2.1 for protein-protein docking and visualization of docked complex respectively. Results showed high interaction between gliadin and glutenin, which supported the results of previous studies. While interaction between gliadin and purothionin proteins was high as compared to interaction between glutenin and alpha-purothionin proteins. Different helices were noted in the 3D structure of gliadin and glutenin proteins, where most of the time Gln amino acid was involved in making the helices. Form the results it has been concluded that greater interaction between gliadin and alpha-purothionin was due to similar sequence of gliadin with viral protein so the interaction of toxin purothionin against pathogens enhance the interaction of gliadin with alpha-purothionin. Current in silico study will enhance the understanding about the interaction of important wheat proteins.

Functional Characterization of Serum Amyloid P Component (SAP) in Host Defense against Bacterial Infection in a Primary Vertebrate.

Serum amyloid P component (SAP), an ancient short pentraxin of the pentraxin family, plays an essential role in resistance to bacterial infection. In this study, the expression and functional characterization of SAP (OnSAP) in Nile tilapia (Oreochromis niloticus), a primary vertebrate, are investigated. The open reading frame of OnSAP is 645 bp of a nucleotide sequence encoding a polypeptide of 214 amino acids. As a calcium-binding protein, the structure and relative motif of OnSAP is highly similar to those of humans, containing amino acid residues Asn, Glu, Gln and Asp. In healthy fish, OnSAP mRNA is extensively distributed in all eleven tissues examined, with the highest level in spleen. The mRNA expression of OnSAP was significantly up-regulated after being challenged with gram-positive bacterium Streptococcus agalactiae and gram-negative bacterium Aeromonas hydrophila in vivo. In addition, recombinant OnSAP ((r)OnSAP) protein had capacities of binding S. agalactiae or A. hydrophila in the presence of Ca2+. Further, (r)OnSAP helped monocytes/macrophages to efficiently phagocytize bacteria. Moreover, the (r)OnSAP was able to enhance the complement-mediated lysis of the chicken red blood cells. Collectively, the evidence of SAP in tilapia, based on the results including its evolutionary conserved protein structure, bacterial binding and agglutination, opsonophagocytosis of macrophage and hemolysis enhancement, enriches a better understanding of the biological functions of the pentraxin family.

Nitrate fertilization may delay autumn leaf senescence, while amino acid treatments do not.

Fertilization with nitrogen (N)‐rich compounds leads to increased growth but may compromise phenology and winter survival of trees in boreal regions. During autumn, N is remobilized from senescing leaves and stored in other parts of the tree to be used in the next growing season. However, the mechanism behind the N fertilization effect on winter survival is not well understood, and it is unclear how N levels or forms modulate autumn senescence. We performed fertilization experiments and showed that treating Populus saplings with inorganic nitrogen resulted in a delay in senescence. In addition, by using precise delivery of solutes into the xylem stream of Populus trees in their natural environment, we found that delay of autumn senescence was dependent on the form of N administered: inorganic N (NO3−) delayed senescence, but amino acids (Arg, Glu, Gln, and Leu) did not. Metabolite profiling of leaves showed that the levels of tricarboxylic acids, arginine catabolites (ammonium, ornithine), glycine, glycine‐serine ratio and overall carbon‐to‐nitrogen (C/N) ratio were affected differently by the way of applying NO3 − and Arg treatments. In addition, the onset of senescence did not coincide with soluble sugar accumulation in control trees or in any of the treatments. We propose that different regulation of C and N status through direct molecular signaling of NO3 − and/or different allocation of N between tree parts depending on N forms could account for the contrasting effects of NO3 − and tested here amino acids (Arg, Glu, Gln, and Leu) on autumn senescence.

Effects of Different Dietary Carbohydrate Sources on the Meat Quality and Flavor Substances of Xiangxi Yellow Cattle.

This study investigated the dietary supplementation of starches with different carbohydrate sources on the proximate composition, meat quality, flavor substances, and volatile flavor substances in the meat of Chinese Xiangxi yellow cattle. A total of 21 Chinese Xiangxi yellow steers (20 ± 0.5 months, 310 kg ± 5.85 kg) were randomly divided into three groups (control, corn, and barley groups), with seven steers per group. The control steers received a conventional diet (coarse forage type: whole silage corn at the end of the dough stage as the main source), the corn group received a diet with corn as the main carbohydrate source, and the barley group received a diet with barley as the main carbohydrate source. The experiment lasted for 300 d. and the means of the final weights in the control, corn, and barley groups were 290 kg, 359 kg, and 345 kg. The diets were isonitrogenous. The corn and barley groups reduced the moisture (p = 0.04) and improved the intramuscular fat content of the meat (p = 0.002). They also improved meat color (a*) (p = 0.01) and reduced cooking loss (p = 0.08), shear force (p = 0.002), and water loss (p = 0.001). There was no significant difference in the 5'-nucleotide content (p &gt; 0.05), the equivalent umami concentration (EUC) (p = 0.88), and taste activity value (TAV) (p &gt; 0.05) among the three groups. The 5'-IMP (umami) content was the highest in the 5'-nucleotide and its TAV &gt; 1. The corn and barley groups improved the content of tasty amino acids (tAA, p &lt; 0.001). The corn group had a higher content of sweet amino acids (SAA, p &lt; 0.001) and total amino acids (TAA, p = 0.003). Corn and barley improved the levels of MUFA (p &lt; 0.001), PUFA (p = 0.002), n-3 PUFA (p = 0.005), and n-6 PUFA (p = 0.020). The levels of alcohols, hydrocarbons, and aldehydes in the corn group were higher than in the barley and control groups (p &lt; 0.001). The esters content in the corn group was higher than in the barley and control groups (p = 0.050). In conclusion, feeding corn or barley as a carbohydrate source can improve the nutrient content and taste. Feeding corn as a carbohydrate source can improve the content of free amino acids (Cys, Glu, Gly, Thr, Leu, Trp, Gln, Asn, and Asp), fatty acids (saturated fatty acid, monounsaturated fatty acid, polyunsaturated fatty acid, n-3PUFA, n-6PUFA, and total fatty acid), and volatile flavor substances (alcohols, aldehydes, acids, and hydrocarbons) to improve the flavor and meat quality.

Investigating pathogenic SNPs in androgen receptor with direct influence on polycystic ovary syndrome (PCOS) in women

BackgroundPolycystic ovary syndrome (PCOS) became one of the main reasons for infertility in women. It has an obvious effect on phenotype represented by hirsutism, increased body mass index, obesity, and acne, while biochemical tests show adverse hormonal imbalance with hyperandrogenism as testosterone levels increases. From molecular level point of view, pathogenic SNPs may change CAG repeats number along androgen receptor (AR) resulting in altered function of the gene causing different affinity to androgen hormones.MethodsRecruiting 150 patients diagnosed with PCOS for the study, genomic DNA was extracted and amplified using specifically designed exon 1 PCR primers employing gene walking technique. The resulting amplicons were sequenced and thoroughly analyzed for polymorphism and CAG repeats number.ResultsData obtained from recruiting 150 patients diagnosed with PCOS showed that sequences X:67545209–67545742; X:67545503–67545739 of exon 1 harbored 7 SNPs altered secondary structure of the resulting protein and forced toward the use of CAA as synonymous codon instead of the normal CAGs stretches. This led to produced alternative mRNA that eventually changed nonsense-mediated mRNA decay mechanism.ConclusionProbability of PCOS in women with polymorphic AR gene is higher than others, especially women with high number of CAG stretches. The new finding and highlight of this study is that alternative codon usage (CAAs) to produce the same amino acid (Gln) and compensate the reduced number of CAG repeats number may be attributed to epigenetic mechanism to mitigate the adverse effect of such change and maintain a normal function of AR gene. This finding was not previously reported in former studies.

Crosstalk between Selenium and Sulfur Is Associated with Changes in Primary Metabolism in Lettuce Plants Grown under Se and S Enrichment.

This study investigated the beneficial effects of selenium (Se) and sulfur (S) enrichment on the primary metabolism in butterhead lettuce. The plants were treated with three levels of Se via foliar application in the presence of two S levels in the nutrient solution under greenhouse conditions. The lettuce plants that were exposed to the lower selenate level (1.3 μM) in combination with the adequate and high S supplies (1 and 2 mM, respectively) accumulated 38.25 ± 0.38 µg Se g−1 DM and 47.98 ± 0.68 µg Se g−1 DM, respectively. However, a dramatic increase in the Se concentration (122.38 ± 5.07 µg Se g−1 DM, and 146.71 ± 5.43 µg Se g−1 DM, respectively) was observed in the lettuce heads that were exposed to the higher selenate foliar application (3.8 μM) in response to the varied sulfate concentrations (S1 and S2, respectively). Under higher Se and S supplies in the lettuce plants, the levels of organic acids, including malic acid and citric acid, decreased therein to 25.7 ± 0.5 and 3.9 ± 0.3 mg g−1 DM, respectively, whereas, in the plants that were subjected to adequate S and lower Se fertilization, the malic acid, and citric acid levels significantly increased to 47.3 ± 0.4 and 11.8 ± 0.4 mg g−1 DM, respectively. The two Se levels (1.3 and 3.8 μM) under the S1 conditions also showed higher concentrations of water-soluble sugars, including glucose and fructose (70.8.4 ± 1.1 and 115.0 ± 2.1 mg g−1 DM; and 109.4 ± 2.1 and 161.1 ± 1.0 mg g−1 DM, respectively), compared to the control. As with the glucose and fructose, the amino acids (Asn, Glu, and Gln) exhibited strikingly higher levels (48.7 ± 1.1 μmol g−1 DM) under higher S and Se conditions. The results presented in this report reveal that the “crosstalk” between Se and S exhibited a unique synergistic effect on the responses to the amino acids and the soluble sugar biosynthesis under Se and S enrichment. Additionally, the Se-and-S crosstalk could have an important implication on the final nutritional value and quality of lettuce plants.

Synthesis, DFT and In Silico Anti-COVID Evaluation of Novel Tetrazole Analogues

A new series of 3-aryl/heteroaryl-2-(1H-tetrazol-5-yl) acrylamides have been synthesized through catalyst-free, one-pot cascade reactions, utilizing click chemistry approach and evaluated for their anti-COVID activities against two proteins in silico. The structural properties of the synthesized molecules were evaluated based on DFT calculations. Total energy of the synthesized tetrazole compounds were obtained through computational analysis which indicate the high stability of the synthesized compounds. The Frontier Molecular Orbitals (FMO) and associated energies and molecular electrostatic potential (MEP) surfaces were generated for the compounds. Spectral analysis by DFT gave additional evidence to the structural properties of the synthesized molecules. All tetrazole analogues come under good ADMET data as they followed the standard value for ADMET parameters. Docking studies offered evidence of the molecules displaying excellent biological properties as an anti-Covid drug. Compound 4 g exhibited excellent anti-COVID-19 properties with four hydrogen binding interactions with amino acids GLN 2.486 Å, GLN 2.436 Å, THR 2.186 Å and HSD 2.468 Å with good full-fitness score (–1189.12) and DeltaG (–7.19). Similarly, compound 4d shown potent activity against anti-COVID-19 mutant protein (PDB: 3K7H) with three hydrogen binding interactions, i.e., SER 2.274 Å, GLU 1.758 Å and GLU 1.853 Å with full-fitness score of –786.60) and DeltaG (–6.85). The result of these studies revealed that the compounds have the potential to become lead molecules in the drug discovery process.

Antidiabetic Formulation Development Based on Natural Materials As α-Glucosidase Enzyme Inhibitor

Salam (Syzygium polyanthum), stevia (Stevia rebaudiana), tea (Camellia sinensis), and yakon (Smallanthus sonchifolius) leaves act as inhibitors of the α-glucosidase enzyme. The aims of this research were to (i) determine the best combination of polyherbals that could inhibit the α-glucosidase enzyme, (ii) determine the phytochemical content of the teraactive fraction, and (iii) predict potential bioactive compounds that were responsible for the inhibition of the α-glucosidase enzyme. The novelty of this research was its use of several types of herbs, combined with various compositions, to obtain the IC50 value. Also, this research predicted the activity of the compound through a molecular docking approach with Molegro Virtual Docker software. The extraction was carried out with the maceration method with 96% ethanol and then partitioned with ethyl acetate and water. Two fractions with high inhibition percentages were combined with a ratio of 1:1, 1:2, 2:1, 1:3, and 3:1, and they were tested against the α-glucosidase inhibitory enzyme. The best fraction was identified for chemical compound content by Liquid Chromatography Mass Spectroscopy (LC-MS/MS). The compounds were analyzed in silico by using Molegro Virtual Docker software, which obtained the best reranc score. The results showed that the ethanol extract of tea leaves had the highest activity, at 65.51%, and salam, stevia, and yakon leaves had inhibition percentages of 44.11%, 34.74%, and 28.85%, respectively. The combination of water fractions of tea and salam leaves had the best activity at a ratio of 3:1 with IC50 of 16.53 ppm, which was almost equivalent to acarbose as a positive control with IC50 of 14.62 ppm. The analysis of LC-MS/MS for the water fraction of tea contained nicotiflorin with m/z 594, a reranc score of -118.24, and amino acid residues Arg, Asp, Asn, Cys, Glu, His, Thr, and Tyr. The water fraction of salam leaves contained quercetagetin with m/z 319, the reranc score was –89.123, and the amino acid residues Asn, Asp, Gln, Lys, Phe, Pro, Thr, and Trp.