Amino Acid Research Articles

Amino acid profile behavior during the fermentation of Criollo cocoa beans

The study investigated the behavior of seventeen amino acids during spontaneous (SF) and starter culture (SC) fermentation of Criollo cocoa beans from Copallín, Guadalupe and Tolopampa, Amazonas-Peru. For this purpose, liquid chromatography (UHPLC) was used to quantify amino acids. Multivariate analysis was used to differentiate the phases of the fermentation process. The percentage of essential amino acids during SC fermentation (63.4%) was higher than SF (61.8%); it was observed that the starter culture accelerated their presence and increased their concentration during the fermentation process. The multivariate analysis identified a first stage (day 0 to day 2), characterized by a low content of amino acids that increased due to protein hydrolysis. The study showed that adding the starter culture (Saccharomyces cerevisiae) to the fermentation mass increased the concentration of essential amino acids (63.0%) compared to the spontaneous process (61.8%). Moreover, this addition reduced the fermentation time (3–4 days less), demonstrating that the fermentation process with a starter culture allows obtaining a better profile of amino acids precursors of flavor and aroma.

Dynamics of industrial hemp vegetative growth and metabolite accumulation in response to light intensity

Industrial hemp (Cannabis sativa L.) is a high-yielding cash crop with strong stress resistance, and its fibrous and metabolic profile holds significant development potential. In this study, we measured various agronomic traits, amino acids, saccharides, and their derivatives under four different light intensities (30 μmol·m−2·s−1, 80 μmol·m−2·s−1, 130 μmol·m−2·s−1, 180 μmol·m−2·s−1). Results showed that plant height decreased with increasing light intensity and reached a minimum at 180 μmol·m−2·s−1, which was 27 % less than the control treatment. Further, stem diameter increased by 52 %, leaf fresh weight by 130 %, stem fresh weight by 116 %, and root fresh weight by 264 % were at their maximum at 130 μmol·m−2·s−1. Plant dry weight of leaf (113 %), stem (214 %), and root (427 %) were also found to be maximum at 130 μmol·m−2·s−1. Metabolic results revealed that the free amino acid content was 10 %, 61 %, and 60 % higher under 30 μmol·m−2·s−1 compared to 80, 130, and 180 μmol·m−2·s−1. Additionally, soluble protein contents were 42 %, 81 %, and 77 % higher under 30 μmol·m−2·s−1 compared to 80, 130, and 180 μmol·m−2·s−1. In contrast, the soluble sugar contents were 42 %, 61 %, and 75 % higher than in the other three treatments. In conclusion, industrial hemp morphological characteristics and metabolite profiles exhibit distinct responses to different light intensities. To achieve desired outcomes, it is imperative to carefully choose an optimized light intensity specifically tailored to align with the defined objectives. These findings provide a foundation for a deeper understanding of the characteristics of industrial hemp, optimizing cultivation techniques, and investigating the impact of light intensity on industrial hemp vegetative period growth, amino acids, and saccharides.

Electrostatics introduce a trade-off between mesophilic stability and adaptation in halophilic proteins.

Extremophile organisms have adapted to extreme physicochemical conditions. Halophilic organisms, in particular, survive at very high salt concentrations. To achieve this, they have engineered the surface of their proteins to increase the number of short, polar and acidic amino acids, while decreasing large, hydrophobic and basic residues. While these adaptations initially decrease protein stability in the absence of salt, they grant halophilic proteins remarkable stability in environments with extremely high salt concentrations, where non-adapted proteins unfold and aggregate. The molecular mechanisms by which halophilic proteins achieve this, however, are not yet clear. Here, we test the hypothesis that the halophilic amino acid composition destabilizes the surface of the protein, but in exchange improves the stability in the presence of salts. To do that, we have measured the folding thermodynamics of various protein variants with different degrees of halophilicity in the absence and presence of different salts, and at different pH values to tune the ionization state of the acidic amino acids. Our results show that halophilic amino acids decrease the stability of halophilic proteins under mesophilic conditions, but in exchange improve salt-induced stabilization and solubility. We also find that, in contrast to traditional assumptions, contributions arising from hydrophobic effect and preferential ion exclusion are more relevant for haloadaptation than electrostatics. Overall, our findings suggest a trade-off between folding thermodynamics and halophilic adaptation to optimize proteins for hypersaline environments.

Role of dietary nutrients and metabolism in colorectal cancer.

Colorectal cancer (CRC) is one of the most common malignancies and the leading causes of cancer related deaths worldwide. The development of CRC is driven by a combination of genetic and environmental factors. There is growing evidence that changes in dietary nutrition may modulate the CRC risk, and protective effects on the risk of developing CRC have been advocated for specific nutrients such as glucose, amino acids, lipid, vitamins, micronutrients and prebiotics. Metabolic crosstalk between tumor cells, tumor microenvironment components and intestinal flora further promote proliferation, invasion and metastasis of CRC cells and leads to treatment resistance. This review summarizes the research progress on CRC prevention, pathogenesis, and treatment by dietary supplementation or deficiency of glucose, amino acids, lipids, vitamins, micronutri-ents, and prebiotics, respectively. The roles played by different nutrients and dietary crosstalk in the tumor microenvironment and metabolism are discussed, and nutritional modulation is inspired to be beneficial in the prevention and treatment of CRC.

Optimal growth conditions to enhance Chlorella vulgaris biomass production in indoor phyto tank and quality assessment of feed and culture Stock

Commercial microalgae cultivation is a dynamic field with ongoing efforts to improve efficiency, reduce costs, and explore new applications. We conducted a study to examine how different light exposure periods affect Chlorella vulgaris's growth. We employed a Phyto tank batch system of approximately 3.5 liters with LED light control, controlled airflow, and sterilized bags, maintained at 22.0 ± 2.0°C indoors. Various methods, including spectrophotometry, and cell counter were employed to monitor Chlorella vulgaris growth under different light exposure cycles. Additionally, quality analysis as feed source was employed by proximate, amino acid, beta-glucan, and microbial content analysis. The results revealed significant variations in C. vulgaris biomass production based on light exposure duration. Notably, the 16:8-hour light-dark photoperiod exhibited the highest biomass concentration, reaching 6.48 × 107 ± 0.50 cells/mL with an optical density (OD) of 1.165 absorbance at 682 nm. The 12:12-hour light-dark photoperiod produced the second-highest biomass concentration, with 2.305 × 106 ± 0.60 cells/mL at an OD of 0.489. Proximate analysis of dry algae powder revealed low lipid content (0.48%), high protein content (37.61%), variable ash concentration (average 10.75%), and a significant carbohydrate fraction (51.16%) during extended daylight and shorter dark periods. Amino acid analysis identified nine essential amino acids, with glutamic acid being the most abundant (17.7%) and methionine the least (0.4%). Furthermore, quality analysis and microbiological assays demonstrated that the C. vulgaris biomass is well-suited for fish and livestock use as a feed source and possibility as human nutraceuticals. These findings can be considered more environmentally friendly and ethically sound due to the absence of genetic modification.

Development and Challenges of Cyclic Peptides for Immunomodulation.

Cyclic peptides are polypeptide chains formed by cyclic sequences of amide bonds between protein-derived or non-protein-derived amino acids. Compared to linear peptides, cyclic peptides offer several unique advantages, such as increased stability, stronger affinity, improved selectivity, and reduced toxicity. Cyclic peptide has been proved to have a promising application prospect in the medical field. In addition, this paper mainly describes that cyclic peptides play an important role in anti-cancer, anti-inflammatory, anti-virus, treatment of multiple sclerosis and membranous nephropathy through immunomodulation. In order to know more useful information about cyclic peptides in clinical research and drug application, this paper also summarizes cyclic peptides currently in the clinical trial stage and cyclic peptide drugs approved for marketing in the recent five years. Cyclic peptides have many advantages and great potential in treating various diseases, but there are still many challenges to be solved in the development process of cyclic peptides.

Fractionation and characterisation of sialylated-mucin glycoprotein from edible birds' nest hydrolysates through anion exchange chromatography

Edible bird's nest (EBN) is made up of sialylated-mucin glycoprotein with various health benefits due to its high antioxidative activity. However, as a macromolecule with distinct charged sialic acid and amino acids, fractions with different charges would have varied physicochemical properties and antioxidant activity, which have not been studied. Therefore, this study aimed to fractionate and purify the enzymatic hydrolysed of cleaned EBN (EBNhc) and EBN by-product (EBNhbyp) through anion exchange chromatography (AEC), and determine their molecular weights, physicochemical properties, and antioxidative activities. Overall, 26 fractionates were collected from enzymatic hydrolysate by AEC, which were classified into 5 fractions. It was found that the positively charged fraction of EBNhc (CF1) and EBNhbyp (DF1) showed the significantly highest (p < 0.05) soluble protein contents (22.86 and 18.40 mg/g), total peptide contents (511.13 and 800.47 mg/g) and ferric reducing antioxidant power (17.44 and 6.96 mg/g) among the fractionates. In conclusion, a positively charged fraction (CF1 and DF 1) showed more desired physicochemical properties and antioxidative activities. This research suggests the potential of AEC fractionation as a technology to purify EBN and produce positively charged EBN fractionates with antioxidative potential that could be applied as food components to provide health benefits.

Integrated multiple-omics reveals the regulatory mechanism underlying the effects of homologous Bacillus tequilensis (GCB-3) on growth performance of grass carp (Ctenopharyngodon idellus)

The purpose of this study was to investigate the regulatory mechanism of Bacillus tequilensis (GCB-3) in improving the growth performance of grass carp by microbiome and targeted metabolomics analysis. Fish were fed with different amounts of GCB-3 including B0 (0 CFU/g), B6 (1×106 CFU/g), B7 (1×107 CFU/g), B8 (1×108 CFU/g), and B9 (1×109 CFU/g) for 8 weeks. The results indicate that with the increase of dietary GCB-3 concentration, grass carp showed a significant linear trend in growth performance, and appetite factors showed a significant linear and quadratic trend, the growth performance and appetite-promoting factors in the B8 group reach their maximum value in the B8 group. Microbiome analysis found that Proteobacteria and Actinobacteria were the most abundant phylum in the gut microbiota of grass carp. The functional potential predicted that GCB-3 affected the synthesis and degradation of carbohydrates, amino acids, fatty acids and lipids. It found that GCB-3 also affected the tricarboxylic acid (TCA) cycle VII (acetate-producers) pathway. The results of targeted metabolomics showed a significant linear and quadratic increase in intestinal acetate with GCB-3 concentration and a maximum in the B8 group. The results of the correlation analysis showed a significant positive correlation between Actinobacteria abundance and acetic acid content, and also between TCA cycle VII (acetate-producers) pathway abundance and Actinobacteria abundance and acetic acid content. In B8 group, grass carp showed a faster regulation speed of blood glucose levels after glucose injection, and the GLP-1 content remained stable. The activities of glycolytic enzymes, lipolytic enzymes, protein metabolism enzymes, and their related mRNA expression levels in grass carp showed a linear or quadratic increasing trend. However, the expression levels of gluconeogenic enzymes and fatty acid synthesis enzyme related mRNA showed an opposite trend. In summary, the addition of 1×108 CFU/g GCB-3 in grass carp feed can improve the abundance of dominant bacterial phyla in the intestine, especially Actinobacteria, increase the concentration of intestinal acetate, provide substrate for the TCA cycle VII (acetate-producers) pathway, thereby regulating the nutrient metabolism of grass carp and ultimately promoting growth.

Effects of essential amino acids supplementation in a low-protein diet on growth performance, intestinal health and microbiota of juvenile blotched snakehead (Channa maculata)

Developing a low-protein feed is important for the sustainable advancement of aquaculture. The aim of this study was to investigate the effects of essential amino acid (EAA) supplementation in a low-protein diet on the growth, intestinal health, and microbiota of the juvenile blotched snakehead, Channa maculata in an 8-week trial conducted in a recirculating aquaculture system. Three isoenergetic diets were formulated to include a control group (48.66 % crude protein (CP), HP), a low protein group (42.54 % CP, LP), and a low protein supplementation EAA group (44.44 % CP, LP-AA). The results showed that significantly lower weight gain (WG), specific growth rate (SGR), protein efficiency ratio (PER), and feed efficiency ratio (FER) were observed in fish that were fed LP than in the HP and LP-AA groups (P < 0.05). The HP and LP-AA groups exhibited a significant increase in intestinal villus length, villus width, and muscular thickness compared to the LP group (P < 0.05). Additionally, the HP and LP-AA groups demonstrated significantly higher levels of intestinal total antioxidant capacity (T-AOC), catalase (CAT), and superoxide dismutase (SOD) and lower levels of malondialdehyde (MDA) compared to the LP group (P < 0.05). The apoptosis rate of intestinal cells in the LP group was significantly higher than those in the LP and HP groups (P < 0.05). The mRNA expression levels of superoxide dismutase (sod), nuclear factor kappa B p65 subunit (nfκb-p65), heat shock protein 70 (hsp70), and inhibitor of NF-κBα (iκba) in the intestine were significantly higher in the LP group than those in the HP and LP-AA groups (P < 0.05). The 16s RNA analysis indicated that EAA supplementation significantly increased the growth of Desulfovibrio and altered the intestinal microflora. The relative abundances of Firmicutes and Cyanobacteria were positively correlated with antioxidant parameters (CAT and T-AOC), whereas Desulfobacterota was negatively correlated with sod and T-AOC. The genera Bacillus, Bacteroides, and Rothia were associated with the favorable maintenance of gut health. In conclusion, dietary supplementation with EAAs to achieve a balanced amino acid profile could potentially reduce the dietary protein levels from 48.66 % to 44.44 % without adversely affecting the growth and intestinal health of juvenile blotched snakeheads.

Medicinal Properties of Crotalaria burhia: A Review

Background: Natural products have a crucial role in the discovery and development of drug molecules. The secondary metabolites derived from the plant have potential therapeutic value and reveal many pharmacological effects. Crotalaria burhia Buchham, a Leguminaceae herb, is mostly present in arid parts of India, Pakistan, China, and Afghanistan. Shinio, Bhip, Bhata, Ghugato, and Ban Sutra are just a few of the names it goes by. It is used to cure gout, eczema, hydrophobia, and inflammation. Plant root porridge is used for typhoid, wounds and cuts, rheumatism, flatulence, earaches, nasal bleeding, and stone problems. The powder form of the plant is used for digestive disorders. The plant has potent pharmacological activity and is also carcinogenic and anticarcinogenic in nature. Various phytoconstituents are present, like tannins, amino acids, alkaloids, steroids, triterpenoids, mucilage, flavonoids, gum, polyphenols, anthraquinones, carbohydrates, glycosides, and saponins. Croburhine is a novel pyrrolizidine alkaloid obtained from it. This review focuses on the existing knowledge of the phytochemistry and pharmacological action of C. burhia, which will provide broad information for a proper assessment of the plant as a medicinal agent. Methods: The experimental analysis and data reported on the species of Crotalaria were reviewed from 2005 to 2022. Various databases were used as article sources, like Medline and Pubmed, and a literature review was done using the keywords Crotalaria burhia, habitat, species, pharmacological action, and traditional uses. Result: On the basis of our perception, we have stated that the species Crotalaria burhia is a golden herb that possesses potent pharmacological activities. It is widely used as a conventional drug for the cure of various diseases, but its active constituents need to be evaluated more for future drug development and pharmacological action. Conclusion: There were a total of 21 research articles about the phytochemical screening of herbs, which showed the presence of different active constituents like alkaloids, tannins, flavonoids, steroids, glycosides, mucilage, gum, polyphenols, carbohydrates, terpenoids, etc., which are responsible for different pharmacological activities like antioxidant, antimicrobial, anti-inflammatory, antinociceptive, antihypertensive, antitumor, and analgesic activity.

A europium MOF-based turn-off fluorescent sensor for Cr2O72−, l-tryptophan, Dimethylnidazole detection in water

Various pollutants in water are a serious threat to human health. It is very important to identify and detect harmful substances in water. In this paper, an unprecedented new coordination polymer of luminescent sensor, [[Eu(L)2] Cl·H2O] n, (Eu-MOF), with excellent fluorescence sensing abilities for anions, amino acid and antibiotics was developed. It was synthesized by N-heterocyclic carboxylic acid ligands 1, 3-bis (4-carboxylbenzyl) triazole chloride and lanthanide metal ion Eu3+ under solvothermal conditions. The fluorescence results indicate that Eu-MOF has excellent fluorescence sensing abilities for Cr2O72−, l-tryptophan, and Dimethylnidazole via fluorescence quenching. The limit of detection (LOD) is 8.98 × 10−6 mol·L−1, 4.94 × 10−5 mol·L−1 and 5.50 × 10−5 mol·L−1, respectively. Furthermore, the possible mechanisms of fluorescence quenching in the sensing process were systematically studied by UV–vis and DFT. It is revealed that the rational design and utilization of such one multifunctional Eu-MOF can offer guidance for fluorescence detection with wide applications.

Dietary supplementation of ornithine improved the growth, immune response and acute NH3 stress tolerance of Pacific white shrimp, Penaeus vannamei

Ornithine is an arginine family amino acid and is not incorporated into protein. This study evaluated its role in growth, immune response and acute NH3 stress tolerance of Penaeus vannamei. Six diets with graded levels (0.6, 0.9, 1.2, 1.5, 1.8, 2.1gkg-1) of L-ornithine were formulated, and fed the shrimp for 8 weeks. The growth of shrimp was significantly improved with the dietary ornithine levels increased from 0.6 to 1.5gkg-1. Feed efficiency, protein efficiency ratio and protein productive value showed a similar trend to the growth performance of shrimp. Whole body protein and muscle lipid of shrimp were significantly increased in shrimp fed 1.8gkg-1 ornithine compared to those fed 0.6gkg-1 ornithine. The contents of several amino acids were increased with the dietary ornithine increased to 1.5gkg-1. Elevated levels of ornithine increased the Nitric Oxide (NO) content and suppressed arginase activity in the hemolymph. The relative mRNA expression of superoxide dismutase, heat shock protein 70, dorsal and caspase 3 were increased with the dietary addition of ornithine, while the mRNA level of arginase was decreased with the increasing dietary ornithine levels. After 9h acute NH3 stress, mortality of shrimp reduced with the increasing dietary ornithine levels. The relative mRNA levels of superoxide dismutase, heat shock protein 70, dorsal and catalase were increased with the dietary addition of ornithine after the acute NH3 stress. The second-order regression analysis of weight gain and survival after acute stress revealed that the suitable ornithine content in the shrimp diet was 1.4-1.5gkg-1.

PHYLOGENETIC CONNECTIONS AND COMPARISONS OF THE AMINO ACID SEQUENCES OF SEVERAL PEPTIDE INDUCERS OF MICROBIAL ORIGIN- A FOUNDATION STUDY FOR DEVELOPMENT OF NOVEL BIOPESTICIDES

Peptides of microbial origin stand out as one of theprominent tools to elicit plant immunity. These peptides comprise one of thekey strategies of integrated pest management and are considered as candidatesto develop novel biopesticides. Manyresearch investigations have proved their potential in fending off plantpathogens and were described as sustainable plant protection strategies. Thepresent study was attempted to discover phylogenetic relationships and compareamino acid sequence alignments of various peptide elicitors of microbialorigin. Phylogenetic analysis of 33 microbial peptide elicitors resulted in twoclusters, one cluster contained 19 flagellin sequences, which is furtherdivided into one major (15 peptide sequences) and one (4 peptide sequences)minor subclusters. Further amino acid sequence alignments were carried outbased on the evolutionary relationships among the peptides. The amino acidsequence alignment of flagellin sequences using Clustal W did not present conservativeamino acid sequences except Serine (S), Alanine (A) and Aspartic acid (D).These conserved amino acids (SAxD) that are positioned in the protruding loopmay play a vital role in recognition by plant surface receptors. Alignment ofamino acid sequences of cold shock protein, xylanase, elongation factor andharpin from various sources did not present conservative amino acid sequencesexcept glycine. These investigations lay a theoretical foundation for exploringmany more microbial peptides for inducing plant resistance.

Quality improvement of soybean meal by simultaneous microbial fermentation and enzymolysis and untargeted metabolomic analysis of its metabolites

Twelve lactobacillus strains, four yeast strains, and eight proteases were screened, and their effects on oligosaccharide degradation, lactic acid production, and proteolytic capacity were investigated in soybean meal. The simultaneous microbial fermentation and enzymolysis (SMFE) conditions were optimized as feed/water ratio of 1:0.8 (w/v), yeast inoculum of 2.1×105 CFU/g, lactobacillus inoculum of 2.1×107 CFU/g, protease addition of 200 U/g, and fermentation at 37 °C for 48 h. The results showed that SMFE soybean meal exhibited superior performance over unprocessed soybean meal, showing increases in crude protein content from 49.8% to 56.3%, acid-soluble protein content from 3.7% to 10.4%, lactic acid production from 0 to 71.5 g/kg, and a decrease in oligosaccharide content from 6.4% to 0%. The comprehensive performance of SMFE soybean meal is higher than that from enzymolysis or fermentation alone Non-targeted metabolomic analysis revealed a diversity of metabolites in fermented soybean meal produced by different processes, and 8366 detected metabolites were mainly amino acids and their metabolites. Furthermore, among the six experimental groups, the metabolites of the SMFE group, and the yeast and lactic acid bacteria fermentation groups were the most similar. This study facilitates the assessment of overall quality of soybean meal and reveals the superiority of SMFE over traditional methods in enhancing nutritional profile of soybean meal.

Site-directed mutagenesis reveals the interplay between stability, structure, and enzymatic activity in RidA from Capra hircus.

Reactive intermediate deaminase A (RidA) is a highly conserved enzyme that catalyzes the hydrolysis of 2-imino acids to the corresponding 2-keto acids and ammonia. RidA thus prevents the accumulation of such potentially harmful compounds in the cell, as exemplified by its role in the degradation of 2-aminoacrylate, formed during the metabolism of cysteine and serine, catalyzing the conversion of its stable 2-iminopyruvate tautomer into pyruvate. Capra hircus (goat) RidA (ChRidA) was the first mammalian RidA to be isolated and described. It has the typical homotrimeric fold of the Rid superfamily, characterized by remarkably high thermal stability, with three active sites located at the interface between adjacent subunits. ChRidA exhibits a broad substrate specificity with a preference for 2-iminopyruvate and other 2-imino acids derived from amino acids with non-polar non-bulky side chains. Here we report a biophysical and biochemical characterization of eight ChRidA variants obtained by site-directed mutagenesis to gain insight into the role of specific residues in protein stability and catalytic activity. Each mutant was produced in Escherichia coli cells, purified and characterized in terms of quaternary structure, thermal stability and substrate specificity. The results are rationalized in the context of the high-resolution structures obtained by x-ray crystallography.

Effects of dietary carbohydrate levels on growth performance, feed utilization, liver histology and intestinal microflora of juvenile tiger puffer, Takifugu rubripes

An 8-week feeding trial was conducted to investigate the effects of dietary carbohydrate levels on growth performance, body composition, and plasma biochemical parameters, liver and gut histology, and intestinal microbiota of Takifugu rubripes. Five isonitrogenous and isolipidic experimental diets were formulated with graded levels of cassava starch: 0% (C0), 8% (C8), 16% (C16), 24% (C24), and 32% (C32). The results showed that weight gain rate, specific growth rate (SGR) and protein efficiency ratio significantly increased in C16 and C24 groups (P < 0.05), while the feed conversion ratio expressed an opposite trend. Feeding rate is decreased with the increasing of carbohydrate levels (P < 0.05). Liver lipid content was dramatically increased in C24 treatment (P < 0.05). The intestinal amylase activity and muscle glycogen content were higher than other groups (P < 0.05). Plasma glucose levels were increased in C24 and C32 groups (P < 0.05). In addition, the abundance of intestinal potentially pathogenic bacteria (Staphylococcus) decreased, and the intestinal probiotics (Lactobacillus) increased with the increasing of dietary carbohydrate level (P < 0.05). The intestinal bacteria, indicated by Sobs, Chao and Shannon indexes, were more diverse with the increasing of the carbohydrate level (P < 0.05), meanwhile the abundance of transport and metabolism of carbohydrate, lipid and amino acid were enhanced with the increasing of carbohydrate levels (P < 0.05). In conclusion, appropriate supplementation of carbohydrate in diet promoted growth performance and feed utilization of juvenile Takifugu rubripes, and increased the liver lipid and muscle glycogen content, as well as intestinal microbial diversity. Based on the quadratic regression between SGR and carbohydrate levels, the optimal dietary carbohydrate level of juvenile Takifugu rubripes is 21.6%.

In-silico approach to characterize the structure and function of a hypothetical protein of Monkeypox virus exploring Chordopox-A20R domain-containing protein activity.

Background: Monkeypox has emerged as a noteworthy worldwide issue due to its daily escalating case count. This illness presents diverse symptoms,including skin manifestations, which have the potential to spread through contact. The transmission of this infectious agent is intricate and readily transfers between individuals.Methods: The hypothetical protein MPXV-SI-2022V502225_00135 strain of monkeypox underwent structural and functional analysis using NCBI-CD Search, Pfam, and InterProScan. Quality assessment utilized PROCHECK, QMEAN, Verify3D, and ERRAT, followed by protein-ligand docking, visualization, and a 100-nanosecond simulation on Schrodinger Maestro.Results: Different physicochemical properties were estimated, indicating a stable molecular weight (49147.14) and theoretical pI (5.62) with functional annotation tools predicting the target protein to contain the domain of Chordopox_A20R domain. In secondary structure analysis, the helix coil was found to be predominant. The three-dimensional (3D) structure of the protein was obtained using a template protein (PDB ID:6zyc.1), which became more stable after YASARA energy minimization and was validated by quality assessment tools like PROCHECK, QMEAN, Verify3D, and ERRAT. Protein-ligand docking was conducted using PyRx 9.0 software to examine the binding and interactions between a ligand and a hypothetical protein, focusing on various amino acids. The model structure, active site, and binding site were visualized using the CASTp server, FTsite, and PyMOL. A 100 nanosecond simulation was performed with ligand CID_16124688 to evaluate the efficiency of this protein.Conclusion: The analysis revealed significant binding interactions and enhanced stability, aiding in drug or vaccine design for effective antiviral treatment and patient management.

In-vitro Antioxidant, In-vitro and In-silico Ovarian Anticancer Activity (Ovarian Cancer Cells-PA1) and Phytochemical Analysis of Cissus quadrangularis L. Ethanolic Extract.

Cissus quadrangularis is a valuable natural source of traditional medicines. An in vitro investigation was performed to determine whether the ethanolic extract from the whole portions of C. quadrangularis had anticancer and free radical scavenging activities against ovarian cancer cells-PA1. C. quadrangularis is a herb collected from rural areas in Andhra Pradesh, India. C. quadrangularis was air-dried and crushed, and the powder and ethanol (0.5 kg) were used in a Soxhlet device for continuous extraction. Phytochemical analysis of the extracts was performed using a standard procedure. The antioxidant activity of the ethanolic extract of C. quadrangularis was evaluated using DPPH. An in vitro anticancer study used an ethanolic extract against the PA1 cell line. Apoptosis of ovarian cancer cells was studied using DAPI and carboxy-H2DCFDA staining. From LC-MS analysis, quercetin-3-O-alpha-Lrhamnopyranoside and erucic acid were docked with the threonine tyrosine kinase (TTK) enzyme using auto docking. The ethanolic extract of C. quadrangularis demonstrated significant dose-dependent antioxidant activity compared to ascorbic acid. The ethanolic extract of C. quadrangularis was found to have high anticancer activity against ovarian cancer cell lines (PA1), with an IC50 value of 482.057 ± 113.857 μg/ml. DAPI and carboxy-H2DCFDA staining confirmed that C. quadrangularis ethanolic extract induced apoptosis in ovarian cancer cells (p < .001). Molecular docking studies helped identify the binding affinities between the protein and ligand complexes, such as Quercetin-3-O-alpha-Lrhamnopyranoside binding sites of target proteins 5N7V (MET602, GLN672) and erucic acid 5N7V (GLY354). Quercetin-3-O-alpha-L-rhamnopyranoside was reported to bind with 5N7V by hydrogen bonding at MET602 and GLN672 amino acids with 2.02, 2.99 Å bonding length distance and binding affinity of -7.9 kcal/mol. Erucic acid was reported to bind with 5N7V by hydrogen bonding at GLY354 amino acid with 3.18, 2.93 Å bonding length (Å) distance and binding affinity of -4.3 kcal/mol. The current analysis showed that the ethanolic extracts of C. quadrangularis L. exhibited antioxidant and anticancer properties against ovarian PA1 cells. The experimental results confirmed that C. quadrangularis L. is a promising, safe chemotherapeutic plant for ovarian cancer PA1 cells. The docking results demonstrated that Quercetin-3-O-alpha-L-rhamnopyranoside strongly binds threonine tyrosine kinase at the MET602 and GLN672 positions. This study showed that the C. quadrangularis ethanolic extract has Quercetin-3-O-alpha-L-rhamnopyranoside, which can be used as an anticancer agent.

Bioactive peptides released by lactic acid bacteria fermented pistachio beverages

Fruit and vegetable are the raw materials for many popular fermented beverages with potential health benefits. In the present study, a novel pistachio beverage fermented with selected strains of lactic acid bacteria belonging to the species Leuconostoc pseudomesenteroides and Companilactobacillus paralimentarius was produced, and the extent of proteolysis was evaluated. Physico-chemical and microbiological analyses, together with peptidomics and proteomics, were carried out on the beverage after 24 h of fermentation, using both the non-inoculated and chemically acidified beverage as controls. Amino acid sequence of hundreds of peptides mainly released from 2S albumin, 11S and 7S globulin were characterized. The number and frequency of identified peptides were higher in beverage started with Leuconostoc pseudomesenteroides, followed by Companilactobacillus paralimentarius and acidic beverage. These results suggest that, in addition to endogenous proteases active at acidic pH, the proteolytic system of LAB directly participated to peptide degradation to some extent. According to the BIOPEP database, a group of 31 peptides were potentially bioactive, and primarily associated with antioxidative properties, ACE and DPP-IV inhibition. The beverage started with Leuconostoc pseudomesenteroides showed the highest amount and number of bioactive peptides. This study lays the foundations for the design of novel pistachio-fermented beverage with potential health properties.

Investigating the Precise Identification of Citrullination Sites with High-Performance Score Metrics using a Powerful Computation Predicting Tool.

To elucidate the detailed mechanisms of citrullination at the molecular level and design drugs applicable to major human diseases, predicting protein citrullination sites (PCSs) is essential. Using experimental approaches to predict PCSs is time-consuming and costly. However, there is a limited scope of the current PCS predictors. In particular, most predictors are commonly used for PCS prediction and have limited performance scores. This work aims to provide an improved sophisticated predictor of citrullination sites using a benchmark dataset in a machine learning platform. This study presents a reliable citrullination site predictor based on a benchmark dataset containing a 1:1 ratio of positive and negative samples. We classified citrullination sites using the Composition of the K-Spaced Amino Acid Pairs (CKSAAP) and Support Vector Machine (SVM). We developed PCS predictors using integrated machine-learning methods that produced the highest average scores. Using 10-fold cross-validation on test datasets, the True Positive Rate (TPR) was 98.34%, the True Negative Rate (TNR) was 99.44%, the accuracy was 98.89%, the Mathew Correlation Coefficient (MCC) was 98.21%, the Area Under the ROC Curve (AUC) was 0.999, and the partial Area Under the ROC Curve (pAUC) was 0.1968. According to overall performance, our developed predictor has a significantly higher implementation in comparison with the current tools on the same benchmark dataset. Moreover, it showed better performance metrics on both test and training datasets. Our developed predictor is promising and can be implemented as a complementary technique for identifying fast and precise citrullination sites.