Aspartate Research Articles

Two newly established and mutually related subfamilies GH13_48 and GH13_49 of the α-amylase family GH13

Currently, the main α-amylase family GH13 has been divided into 47 subfamilies in CAZy, with new subfamilies regularly emerging. The present in silico study was performed to highlight the groups, represented by the maltogenic amylase from Thermotoga neapolitana and the α-amylase from Haloarcula japonica, which are worth of creating their own new GH13 subfamilies. This enlarges functional annotation and thus allows more precise prediction of the function of putative proteins. Interestingly, those two share certain sequence features, e.g. the highly conserved cysteine in the second conserved sequence region (CSR-II) directly preceding the catalytic nucleophile, or the well-preserved GQ character of the end of CSR-VII. On the other hand, the two groups bear also specific and highly conserved positions that distinguish them not only from each other but also from representatives of remaining GH13 subfamilies established so far. For the T. neapolitana maltogenic amylase group, it is the stretch of residues at the end of CSR-V highly conserved as L-[DN]. The H. japonica α-amylase group can be characterized by a highly conserved [WY]-[GA] sequence at the end of CSR-II. Other specific sequence features include an almost fully conserved aspartic acid located directly preceding the general acid/base in CSR-III or well-preserved glutamic acid in CSR-IV. The assumption that these two groups represent two mutually related, but simultaneously independent GH13 subfamilies has been supported by phylogenetic analysis as well as by comparison of tertiary structures. The main α-amylase family GH13 has thus been expanded by two novel subfamilies GH13_48 and GH13_49.Key points• In silico analysis of two groups of family GH13 members with characterized representatives• Identification of certain common, but also some specific sequence features in seven CSRs• Creation of two novel subfamilies—GH13_48 and GH13_49 within the CAZy database

Genetically predicted 1091 blood metabolites and 309 metabolite ratios in relation to risk of type 2 diabetes: a Mendelian randomization study.

Metabolic dysregulation represents a defining characteristic of Type 2 diabetes (T2DM). Nevertheless, there remains an absence of substantial evidence establishing a direct causal link between circulating blood metabolites and the promotion or prevention of T2DM. In addressing this gap, we employed Mendelian randomization (MR) analysis to investigate the potential causal association between 1,091 blood metabolites, 309 metabolite ratios, and the occurrence of T2DM. Data encompassing single-nucleotide polymorphisms (SNPs) for 1,091 blood metabolites and 309 metabolite ratios were extracted from a Canadian Genome-wide association study (GWAS) involving 8,299 participants. To evaluate the causal link between these metabolites and Type 2 diabetes (T2DM), multiple methods including Inverse Variance Weighted (IVW), Weighted Median, MR Egger, Weighted Mode, and Simple Mode were employed. p-values underwent correction utilizing False Discovery Rates (FDR). Sensitivity analyses incorporated Cochran's Q test, MR-Egger intercept test, MR-PRESSO, Steiger test, leave-one-out analysis, and single SNP analysis. The causal effects were visualized via Circos plot, forest plot, and scatter plot. Furthermore, for noteworthy, an independent T2DM GWAS dataset (GCST006867) was utilized for replication analysis. Metabolic pathway analysis of closely correlated metabolites was conducted using MetaboAnalyst 5.0. The IVW analysis method utilized in this study revealed 88 blood metabolites and 37 metabolite ratios demonstrating a significant causal relationship with T2DM (p < 0.05). Notably, strong causal associations with T2DM were observed for specific metabolites: 1-linoleoyl-GPE (18:2) (IVW: OR:0.930, 95% CI: 0.899-0.962, p = 2.16 × 10-5), 1,2-dilinoleoyl-GPE (18:2/18:2) (IVW: OR:0.942, 95% CI: 0.917-0.968, p = 1.64 × 10-5), Mannose (IVW: OR:1.133, 95% CI: 1.072-1.197, p = 1.02 × 10-5), X-21829 (IVW: OR:1.036, 95% CI: 1.036-1.122, p = 9.44 × 10-5), and Phosphate to mannose ratio (IVW: OR:0.870, 95% CI: 0.818-0.926, p = 1.29 × 10-5, FDR = 0.008). Additionally, metabolic pathway analysis highlighted six significant pathways associated with T2DM development: Valine, leucine and isoleucine biosynthesis, Phenylalanine metabolism, Glycerophospholipid metabolism, Alpha-Linolenic acid metabolism, Sphingolipid metabolism, and Alanine, aspartate, and glutamate metabolism. This study identifies both protective and risk-associated metabolites that play a causal role in the development of T2DM. By integrating genomics and metabolomics, it presents novel insights into the pathogenesis of T2DM. These findings hold potential implications for early screening, preventive measures, and treatment strategies for T2DM.

Ultra-high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry-characterized extract of Aerides odorata Lour alleviates paracetamol-induced hepatotoxicity in animal model evidenced by biochemical, molecular, and computational studies.

Many kinds of orchids have significant health benefits although adequate research on their biological functions is yet to be carried out. This study investigated the paracetamol-induced liver damage-protecting effect of epiphytic Aerides odorata methanol extract (AODE). The protective effects of AODE were studied by analyzing its effect on liver function parameters, messenger RNA (mRNA) expression, and tissue histopathological architecture. The results were confirmed by ligand-receptor interaction of molecular docking and multitarget interaction of network pharmacological analyses. AODE significantly (p < 0.05) minimized the dose-dependent increase in acid phosphatase, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, γ-glutamyl transferase, lactate dehydrogenase, and total bilirubin compared to the reference drug silymarin. Malondialdehyde level decreased, and the antioxidant genes catalase (CAT), superoxide dismutase (SOD), β-actin, paraoxonase-1 (PON1), and phosphofructokinase-1 (PFK-1) were upregulated in AODE-treated paracetamol-intoxicated rats. A total of 376 compounds comprising phenols and flavonoids were identified using ultra-high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry (UPLC-qTOF-MS). The online toxicity assessment using SwissADME and admetSAR exhibited drug-like, nontoxic, and potential pharmacological properties. Additionally, in silico analysis showed that isoacteoside, one of the identified compounds, exhibited the best docking score (-11.42) with the liver protein human pituitary adenylate cyclase-1 (Protein Data Bank ID: 3N94). Furthermore, network pharmacology analysis identified the top 10 hub genes, namely AKT1 (protein kinase B), CTNNB1 (catenin beta-1), SRC (proto-oncogene c-Src), TNF (tumor necrosis factor), EGFR (epidermal growth factor receptor), HSP90AA1 (heat shock protein 90α), MAPK3 (mitogen-activated protein kinase 3), STAT3 (signal transducer and activator of transcription 3), CASP3 (caspase protein), and ESR1 (estrogen receptor 1), which are responsible for hepatoprotective activity. The findings demonstrate that AODE could be a novel hepatoprotective target in drug-induced liver damage with a further single compound-based animal study.

Preclinical safety assessment of modified gamma globin lentiviral vector-mediated autologous hematopoietic stem cell gene therapy for hemoglobinopathies.

Previously, we reported the development of a human Aγ-globin gene lentivirus (LV), GbG, which expresses high levels of HbF to correct the sickle cell anemia (SCA) phenotype in the Berkeley SCA mouse model, and then modified the γ-globin gene by substituting glycine at codon 16 with aspartic acid in the Aγ-globin gene to generate GbGM LV. In the present study, we evaluated the long-term safety of human Aγ-globin gene carrying GbGM LV in wild-type mice after primary and secondary transplants of GbGM-modified hematopoietic stem cells (HSC) over 18 months. The safety of the GbGM bone marrow transplant was assessed by monitoring the effects on body weight, hematology, histopathology, malignancy formation, and survival. Mice transplanted with Mock-transduced and spleen focus forming virus (SFFV) γ-retroviral vector (RV)-transduced HSC served as negative and positive controls, respectively. The mean donor-cell engraftment was comparable across Mock, GbGM LV, and SFFV RV groups. There were no significant differences in body weight, clinical signs, immunophenotype, or histopathology in the GbGM-treated mice compared to controls. Four SFFV RV-treated mice, but none of the GbGM-treated mice, developed donor-derived, vector-positive lymphomas as demonstrated by flow cytometry analysis and in situ hybridization. These results highlight the safety of the administration of GbGM LV-modified HSC with long-term follow-up after primary and secondary transplants in mice. This data supported the initiation of phase 1/2 first-in-human SCA clinical trial in the United States.

Effects of biochanin A on lactational performance, nitrogen metabolism, and blood metabolites in dairy cows

Optimizing nitrogen utilization efficiency and mitigating nitrogen losses in cows plays a pivotal role in fostering economic sustainability within contemporary agricultural systems. Biochanin A (BCA), a natural component in red clover, has the potential to improve nitrogen metabolism in dairy cows. The primary objective of this study was to probe the impact of biochanin A supplementation on lactational performance, nitrogen metabolism, and blood metabolites in dairy cows. A complete randomized block design experiment was conducted over 28 d, involving 36 multiparous Holstein cows (comparable milk yield = 37.1 ± 2.90 kg, BW = 642 ± 70.0 kg, days in milk = 92 ± 8.0 d, and parity = 2.4 ± 0.50), which were allocated to three treatment groups: the Control group (with 0 g/d BCA), the Low group (with 10 g/d per cow BCA), and the High group (with 40 g/d per cow BCA). Biochanin A supplementation improved the lactational performance of cows by increasing milk yield by 6.3% (P = 0.007) and feed efficiency by 12.7% (P = 0.009). Total intestinal apparent digestibility was unaffected by BCA supplementation (P > 0.05), but microbial nitrogen was increased by 30.0% (P = 0.002) for promoting nitrogen utilization efficiency by 20.7% (P = 0.004). Milk competent yields (protein, lactose, and non-fat milk solid) were increased with increasing BCA supplementation (P < 0.05). Urea nitrogen levels in plasma and milk were both decreased by BCA supplementation (P < 0.05). Blood routine parameters and plasma biochemical parameters both received no effect by BCA supplementation (P > 0.05). BCA did not affect body health of dairy cows. Additionally, none of the plasma endocrine hormones were affected (P > 0.05). A total of 95 significantly different metabolites were screened from the plasma metabolites of cows in the BCA-added and non-added groups. After performing an enrichment analysis of the metabolic pathways associated with the different metabolites, six specific pathways were identified: bile acid biosynthesis, aspartate metabolism, pyrimidine metabolism, arginine and proline metabolism, the urea cycle, and ammonia recycling. The inclusion of BCA is suggested to enhance milk yield and modulate nitrogen metabolism by influencing relevant metabolites within the metabolic pathways.

Evaluation of Serum Amino Acid and Carnitine Profile in Dogs with Transmissible Venereal Tumor

The presented study aimed to reveal the changes in serum amino acid and carnitine profiles in dogs with transmissible venereal tumor (TVT). The study material comprised 40 female dogs ranging in age from 3 to 5 years. The dogs were divided into two groups based on genital organ examinations. Group 1 (n=20) consisted of healthy dogs, while Group 2 (n=20) consisted of TVT-positive dogs. The definite diagnosis of transmissible venereal tumor was made based on the detection of intracytoplasmic vacuoles and numerous mitotic figures in the collected vaginal smear samples. Blood samples were taken from dogs in both study groups and serum was obtained. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used for the determination of carnitine and amino acid profiles. The obtained data were compared using independent samples t-test. The serum amino acid profiles of Lysine, Aspartic Acid, Tyrosine, Asparagine, Alanine, Arginine, Citrulline, Glutamic Acid, Glycine, Methylglutaryl, Phenylalanine, and Ornithine were found to be lower in the TVT group (P

Nutritional and techno-functional properties of the brown seaweed Sargassum filipendula

With the increasing need to promote healthy and sustainable diets, seaweeds emerge as an environmentally friendly food source, offering a promising alternative for food production. The aim of this study was to characterize the brown seaweed Sargassum filipendula from the coast of São Paulo, Brazil, regarding its nutritional and techno-functional properties using two dehydration methods, oven drying and lyophilized. A commercial dried sample was used as a control. Analyses of proximate composition, mineral determination, amino acid determination, antioxidant capacity, pH, color, scanning electron microscopy, X-ray diffraction, thermal properties, Fourier-transform infrared spectroscopy, and techno-functional properties were performed. Seaweed flours showed significant differences in physicochemical composition, with dietary fiber content of seaweed flours exceeding 70 %. Glutamic and aspartic acids were the most abundant amino acids, with contents of 88.56 and 56.88 mg/g of protein in Sargassum oven drying. Both for antioxidant potential and bioactive compounds, Sargassum lyophilized flours showed the highest levels of compounds. Sargassum lyophilized exhibited lighter color compared to Sargassum oven drying and Sargassum commercial. Emulsion formation, foam formation capacity and stability were higher in Sargassum lyophilized, as well as water and oil absorption. The results suggest that seaweeds can be used to formulate a wide variety of food products, such as sausages, bread, cakes, soups, and sauces.

Synthesis of a novel polymer and design of carboxylate-terminated hyperbranched PEI-incorporated PVDF membranes for efficient oil-in-water emulsion separation

Polyvinylidene fluoride (PVDF) membranes utilizing ultrafiltration (UF) and microfiltration (MF) configurations have widely been used in many separation processes, including oil/water emulsions. Recently, there have been tremendous research interests in designing PVDF membranes on their structural-property relations as well as high separation performances. Herein we report the synthesis of a novel carboxylate-terminated hyperbranched polyethylenimine (CHPEI) using a facile three-step one-pot strategy to enhance the characteristics of PVDF membranes for oil/water emulsion separation. The first step involved the production of one amide and one carboxylate functional group per maleic anhydride molecule on the outer surface of polyethylenimine (PEI). The second step involved producing two additional carboxylate groups via amine-based Michael addition of alkenes using aspartic acid dimethyl ester hydrochloride, followed by basic hydrolysis. CHPEI-n-PVDF membranes having various CHPEI contents are further fabricated by a phase-inversion process, obtaining as free-stable flat-sheet membranes. The synthesized polymer and membranes were thoroughly characterized by FTIR and 1H NMR, SEM, EDX, mapping, AFM, and contact angle analysis. The incorporation of CHPEI into PVDF positively enhanced the hydrophilicity, and the WCA was significantly reduced from 80.6° to 59.5°. The maximum permeation flux was obtained using CHPEI-3-PVDF membranes, which was 200 % more than that obtained using pristine PVDF. The CHPEI-2-PVDF membranes exhibited a rejection performance of >99 % for separating oil-in-water emulsions. The incorporation of CHPEI into PVDF significantly improved the antifouling characteristics of the membranes. After 5 h of operation in separating oil-in-water emulsions, the flux recoveries of CHPEI-2-PVDF and CHPEI-3-PVDF were found to be 92 % and 97 %, respectively. CHPEI has demonstrated a considerable positive influence in improving the PVDF membrane’s performance, making it an excellent candidate for oily wastewater treatment because of its high rejection performance and excellent flux recovery ratio.

Total Synthesis of a Peptide Diatom Sex Pheromone Bearing a Sulfated Aspartic Acid.

The peptide sex-inducing pheromone SIP+ (1) bearing an unusual sulfated aspartic acid residue induces sexual reproduction in diatom populations. Herein, we report the first total synthesis of SIP+ using both a sulfated building block approach and a solid-phase peptide synthesis (SPPS)-compatible late-stage sulfation strategy to assemble the natural product. The modular approaches provide concise routes to useful quantities of the natural product for future structure activity relationship studies examining the role of SIP+ in diatom biology.

Zinc Oxide Nanoparticles Attenuated Neurochemical and Histopathological Alterations Associated with Aluminium Chloride Intoxication in Rats.

The present investigation explored the potential neuroprotective role of zinc oxide nanoparticles (ZnONPs) on aluminum chloride (AlCl3)-mediated Alzheimer's disease (AD)-like symptoms. Rats were distributed into four treatment groups equally: control, ZnONPs (4mg/kg b.wt.), AlCl3 (100mg/kg b.wt.), and ZnONPs + AlCl3 groups. Rats were treated for 42 consecutive days. ZnONPs injection into AlCl3-treated rats suppressed the development of oxidative challenge in the cortical and hippocampal tissues, as demonstrated by the decreased neuronal pro-oxidants (malondialdehyde and nitric oxide), and the increased glutathione and catalase levels. Additionally, ZnONPs injection showed anti-inflammatory potency in response to AlCl3 by decreasing levels of tumor necrosis factor-α and interleukin-1β. Moreover, pretreatment with ZnONPs prevented neuronal cell loss by decreasing the level of pro-apoptotic caspase-3 and enhancing the anti-apoptotic B cell lymphoma 2. Furthermore, ZnONPs ameliorated the disturbed acetylcholinesterase activity, monoamines (norepinephrine, dopamine, and serotonin), excitatory (glutamic and aspartic acids), and inhibitory amino acids (GABA and glycine) in response to AlCl3 exposure. These findings indicate that ZnONPs may have the potential as an alternative therapy to minimize or prevent the neurological deficits in AD model by exhibiting antioxidative, anti-inflammation, anti-apoptosis, and neuromodulatory effects.

The Short-term Exposure Effect of Chlorpyrifos (20% EC) on Haematological, Biochemical and Histopathological Response of Striped Catfish Pangasianodon hypophthalmus

An experiment of acute toxicity was conducted to find out the lethal concentration 50 (LC50) of chloropyrifos (CPF) for Pangasianodon hypophthalmus. Our findings indicated that the 96-h LC50 value of CPF for P. hypophthalmus was 0.106 mg L-1. Effects of different concentrations of CPF (0.09, 0.1, 0.11, 0.12, and 0.13 mg L-1) for 96-h on hemato-biochemical, enzyme and histological effects were evaluated in this study. Acute exposure induced significant differences in enzymes acid phosphatase, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase and acetylcholine of blood serum of the treated group as compared to control group. However, significant decreases in serum total protein, globulin, albumin, and triglycerides levels while serum glucose enhanced with consequent increasing doses of CPF exposure. Also, CPF induced significant declines in RBC count, haemoglobin and hematocrit level whereas the exposure groups showed significant rises in WBC and blood indices levels as compared to control group. Furthermore, CPF exposure causes gill curling, epithelium lifting, necrosis, and lamellae degeneration. Hemosiderin, sinusoid dilations, necrosis, malanomacrophage, hepatocyte hypertrophy were observed in liver histology. Reduction of lumen tubule, hyperplasia, increased bowman space and degenerations were seen in kidney. Overall findings indicates that the acute CPF exposure to P. hypophthalmus resulted in significant adverse changes in biochemical, hematological, and histopathological response.

Chiral gypsum with high‐performance mechanical properties induced by self‐assembly of chiral amino acid on an amorphous mineral

AbstractFunctional chiral suprastructures are common in biology, including in biomineralization, and they are frequently found in many hardened structures of both marine and terrestrial invertebrates, and even in pathologic human otoconia of the inner ear. However, the biological processes by which they form remain unclear. Here, we show that chiral hierarchical suprastructures of calcium sulfate dihydrate (gypsum) can be induced by the chiral Aspartic acid (Asp). Left‐handed (clockwise) morphology of gypsum is induced by the d‐enantiomer of Asp, while right‐handed (counterclockwise) morphology is induced by the l‐enantiomer. A layer‐by‐layer, oriented inclination mineral growth model controlled by continuous self‐assembly of chiral Asp enantiomers on an amorphous calcium sulfate mineral surface of gypsum platelet layers is postulated to produce these chiral architectures. This hybrid amorphous‐crystallized chiral and hierarchical suprastructure of gypsum displays outstanding mechanical properties, including high‐performance strength and toughness. Furthermore, the induction of chiral gypsum suprastructures can be more generally extended from specific acidic amino acids to other (nonamino acid) molecules. These findings contribute to our understanding of the molecular mechanisms by which biomineral‐associated enantiomers exert structural control over chiral architectures commonly seen in biominerals and in biomimetically synthesized functional materials.

Nutritional components of 5 potatoes in Anqing City in 2021

To analyze the nutrient composition and nutritional value evaluation of 5 potatoes in Anqing City. According to the requirements of the Technical Manual for Food Composition Monitoring Projects, 5 types of potato samples were collected from Anqing City, Anhui Province. National standard detection method were used to determine the nutritional components such as water, ash, protein, fat, dietary fiber, sugar, minerals, vitamins, and amino acids in the samples. The index of nutritional quality(INQ) method was used to evaluate proteins, vitamins, and minerals, and the amino acid scoring standard mode(FAO/WHO mode) was used to evaluate the nutritional value of amino acids. Among the 5 types of potatoes, purple potato had the highest protein(2.3 g/100 g) and dietary fiber content(3.6 g/100 g). Sweet potato(red) had the highest carotene content(4003 μg/100 g), sweet potato(white) had the highest content of vitamin C(15.4 mg/100 g). Sugar in potatoes mainly existed in three forms: fructose, glucose, and sucrose; Purple potatoes had the highest levels of calcium(47 mg/100 g) and phosphorus(74 mg/100 g), respectively. Potatoes(white) had the highest content of potassium(401 mg/100 g), while sweet potatoes(red) had the highest content of magnesium(31 mg/100 g). Sodium(104.0 mg/100 g), iron(0.9 mg/100 g), copper(0.17 mg/100 g), and manganese(0.40 mg/100 g) had the highest content in sweet potatoes(white). The Na/K ratio range of the 5 potato varieties was 0.003-0.456, and the INQ of phosphorus, potassium, magnesium, copper, and manganese were greater than 1. The detection result of 5 potatoes all contain 18 amino acids, and aspartic acid was the highest. The amino acid score(AAS) was 0.29-1.35, and the ratio coefficient(RC) was 0.47-1.69. The 5 types of potatoes are rich in dietary fiber, vitamin C and minerals, and belong to the high potassium and low sodium type of food. Potatoes can meet the daily needs of the human body for phosphorus, potassium, magnesium copper, and manganese elements. Lysine is rich in content and can be used as a nutritional supplement for grains. The AAS score and RC are close to 1, and the AAS evaluation mode is closer to the human amino acid composition mode, which can meet the daily needs of the human body for this essential amino acid.

Exploring the taste presentation and receptor perception mechanism of salty peptides from Stropharia rugosoannulata based on molecular dynamics and thermodynamics simulation

The taste presentation and receptor perception mechanism of the salty peptide of Stropharia rugosoannulata were predicted and verified using peptide omics and molecular interaction techniques. The combination of aspartic acid (D) and glutamic acid (E), or peptide fragments composed of arginine (R), constitute the characteristic taste structural basis of salty peptides of S. rugosoannulata. The taste intensity of the salty peptide positively correlates with its concentration within a specific concentration range (0.25–1.0 mg/mL). The receptor more easily recognizes the first amino acid residue at the N-terminal of salty peptides and the aspartic acid residue in the peptides. GLU513, ASP707, and VAL508 are the critical amino acid residues for the receptor to recognize salty peptides. TRPV1 is specifically the receptor for recognizing salty peptides. Hydrogen bonds and electrostatic interactions are the main driving forces for the interactions between salty peptides and TRPV1 receptors. KSWDDFFTR has the most potent binding capacity with the receptor and has tremendous potential for application in sodium salt substitution. This study confirmed the taste receptor that specifically recognizes salty peptides, analyzed the receptor-peptide binding interaction, and provided a new idea for understanding the taste receptor perception of salty peptides.

Dracocephalum moldavica L. Extract Ameliorates Hypertension Through Modulating the Interaction Between miRNAs and Gut Microbiota in 2K1C Rats

Aim: To investigate the therapeutic effects of Dracocephalum moldavica L. extract (DME) against hypertension and elucidate the underlying molecular mechanism. Method: A two-kidney one-clip (2K1C) model was established in male Wistar rats, and DME or saline was administered via oral gavage for 4 weeks. Subsequently, the blood pressure was monitored. Fecal microRNAs (miRNAs), gut microbes, and host metabolites were evaluated through miRNA sequencing, 16S rRNA gene sequencing, and serum metabolomics analysis, respectively. Results: DME administration attenuated 2K1C-induced hypertension in rats. Additionally, DME alleviated the dysregulation of miRNA expression and gut microbiota dysbiosis in 2K1C rats. The correlation analysis confirmed the association of the differentially expressed miRNAs with gut microbes. Furthermore, non-targeted metabolomics analysis revealed different metabolic profiles in the serum of 2K1C rats compared to control rats. Interestingly, the changes in those serum metabolites were partially attenuated by DME. The metabolite enrichment analysis demonstrated that the differentially expressed metabolites were mainly involved in three metabolic pathways (Arachidonic acid metabolism, TCA cycle, and Alanine, aspartate and glutamate metabolism). Conclusion: DME can be suggested as an effective intervention in the prevention of hypertension through modulating the miRNAs, gut microbiota, and host metabolites. Our findings may give new insights regarding the treatment of hypertension.

The Increased aspartate levels in transgenic cotton (Gossypium hirsutum L.) lead to improved tolerance against whitefly (Bemisia tabaci, Gennadius).

The whitefly, a polyphagous insect pest feeding on nearly 1328 plant species, is a major threat to global cotton production and incurs up to 50% yield losses in cotton production in Pakistan. We investigated whether increased aspartate in phloem sap imparts whitefly toxicity and protects cotton plants from intense damage. The enzymatic step for aspartate production is carried through aspartate aminotransferase (AAT). In this study, we constitutively overexpressed the Oryza sativa cytoplasmic AAT (OsAAT2) under the CaMV35S promoter in Gossypium hirsutum cv. CIM-482. Real-time PCR analysis of the AAT transcripts revealed a 2.85- to 31.7-fold increase in mRNA levels between the different cotton lines. A substantial increase in the free-amino acid content of the major N-assimilation and transport amino acids (aspartate, glutamate, asparagine, and glutamine) was seen in the phloem sap of the transgenic cotton lines. The bioassay revealed that the two transgenic cotton lines with the highest free aspartate content in the phloem sap exhibited 97 and 94% mortality in the adult whitefly population and a 98 and 96% decline in subsequent nymph populations, respectively. There was also a significant change in the physiological behaviour of the transgenic cotton lines, with an increased net assimilation (A), gaseous exchange (Gs) and rate of transpiration (E). Improved morphological characteristics like plant height, total number of bolls and fiber yield were recorded in transgenic cotton lines. The AAT gene shows promise in mitigating whitefly infestations and enhancing the overall health and yield of cotton plants.

Dietary protein re-alimentation following restriction improves protein deposition via changing amino acid metabolism and transcriptional profiling of muscle tissue in growing beef bulls

This study aimed to develop a compensatory growth model using growing beef cattle by changing dietary protein and to investigate the underlying mechanisms of compensatory protein deposition in muscle tissue. Twelve Charolais bulls were randomly assigned to one of two groups with two periods: 1) a control group (CON) fed a 13% crude protein (CP) diet for 6 weeks; 2) a treatment group (REC) fed a 7% CP diet for 4 weeks (restriction period) and fed a 13% CP diet in the following 2 weeks (re-alimentation period). Growth performance, digestibility, nitrogen balance, targeted metabolomics of amino acids (AA) in plasma, and transcriptional profiling in muscle tissue were analyzed. Protein restriction decreased average daily gain (ADG; P < 0.05), while protein re-alimentation increased ADG relative to the CON (P < 0.05). Compared to the CON, REC reduced retained N (P < 0.05), and protein re-alimentation increased retained N and N utilization efficiency (P < 0.05), due to reduced urinary urea, hippuric acid, and creatinine excretions (P < 0.05). Ruminal NH3-N in the REC was lower than that in the CON in the protein re-alimentation period (P < 0.05). However, there was no difference in microbial protein and plasma urea nitrogen concentrations. Dietary protein restriction decreased plasma valine and aspartic acid concentrations relative to the CON (P < 0.05), and increased proline and 3-methyl-L-histidine concentrations (P < 0.05). After dietary protein re-alimentation, REC increased plasma citrulline concentrations (P < 0.05). The transcriptional profiling revealed that REC upregulated the AA transporter SLC3A1, and protein re-alimentation downregulated SLC7A8 in the muscle cell membrane. Within the muscle cell, upregulated cytosolic arginine sensor for mTORC1 subunit 2 (CASTOR2) inhibited protein synthesis by inhibiting the mammalian target of rapamycin complex 1 phosphorylation in the protein restriction period, while DNA-damage-inducible transcript 4 (DDIT4) activated the mTOR signaling pathway and promoted protein synthesis in the protein re-alimentation period. In summary, the targeted metabolomics and transcriptomics analyses demonstrated that protein re-alimentation following restriction can promote protein synthesis and reduce muscle breakdown by regulating AA metabolism and functional transcripts related to AA transporters and the mTOR signaling pathway.

The difference in gene expression network based on gender in the recovery of knee cruciate ligament injuries during exercise

Anterior cruciate ligament (ACL) injuries are common and often debilitating, requiring extensive rehabilitation. The recovery process involves complex physiological, biomechanical, and biochemical factors. Gender differences in the incidence and recovery from ACL injuries are well-documented, with recent advances in molecular biology suggesting that gene expression networks play a crucial role in tissue repair and regeneration. This study aims to compare gene expression networks between genders to provide strategies for better-managing sports injuries. Gene expression data were sourced from the Gene Expression Omnibus (GEO) database, with samples divided into four groups based on gender and age. Differentially expressed genes (DEGs) were identified using GEO2R, and biological interaction networks were constructed using the STRING (Search Tool for the Retrieval of Interacting Genes/Proteins). Network analysis was performed using Gephi, focusing on betweenness centrality and overall centrality. Two common genes, TRNA Aspartic Acid Methyltransferase 1 (TRDMT1) and NCF1, were identified in both sexes but exhibited different centrality measures. NCF1, associated with the production of superoxide anion and linked to chronic granulomatous disease, was expressed ten times higher in men than women. TRDMT1, responsible for RNA methylation, was twice as highly expressed in men. The study underscores the importance of gender-specific molecular mechanisms in ACL injury recovery and highlights the need for personalized treatment strategies.

Influence of Amino Acids on Calcium Oxalate Precipitation in Systems of Different Chemical Complexity

The mechanisms and conditions under which urinary stones, pathological biominerals in the kidneys and bladder, are formed have not yet been fully clarified. This study aims to understand the role of the system complexity and seven different amino acids (alanine, phenylalanine, glycine, serine, cysteine, histidine, and aspartic acid) in the spontaneous precipitation of calcium oxalate. To elucidate these effects, the conditions simulating hyperoxaluria (ci(Ca2+) = 7.5 mmol dm−3 and ci(C2O42−) = 6.0 mmol dm−3) were used for the first time. In this work, systematic research on calcium oxalate precipitation was performed in three systems of different chemical complexities: (a) only calcium and oxalate ions, (b) increased ionic strength, and (c) artificial urine at two initial pHs (pHi = 5.0 and 9.0). In all the investigated systems, the dominant precipitation of calcium oxalate monohydrate (COM) was observed, except in the artificial urine system at pHi = 9.0, in which a mixture of COM and calcium oxalate dihydrate (COD) was obtained. In all the investigated systems, a significant influence of the selected amino acids on the morphology and crystal growth of COM was observed, with more pronounced changes at pHi = 9.0. Overall, polar amino acids and nonpolar phenylalanine inhibit the growth of COM, which is a more pathogenic hydrate form. The artificial urine system proved to be more relevant for the observation of effects relevant to kidney stone formation in the human body.

Impact of a novel caseinolytic protease single mutation on Lactiplantibacillus pentosus growth performance

The caseinolytic protease (ClpP)-producing strain CHFM, isolated from Algerian goat milk, was identified as Lactiplantibacillus pentosus through 16 S rRNA sequence analysis. The optimal conditions for ClpP expression by Lpb. pentosus CHFM were determined in a modified MRS medium, where peptone was replaced with casein. The most favourable conditions were a pH of 8.0, a temperature between 25°C and 55°C, and an incubation period of 48 h. PCR screening and subsequent cloning and sequencing of the ClpP-encoding gene from the Lpb. pentosus CHFM genome revealed two conservative substitutions: threonine to serine at position 161, and aspartic acid to glutamic acid at position 182. Bioinformatic analysis indicated an instability index value of 40.51 for ClpP CHFM, suggesting stability, with an alpha-helix being the predominant secondary structure element, followed by a random coil. The aliphatic index was 97.60, indicating thermal stability. The GRAVY index was −0.23, suggesting enhanced interaction with water. The molecular weight was found to be 25.5 kDa with an isoelectric point (pI) of 4.78. The structural and docking analysis demonstrated that ClpP CHFM possesses the ability to hydrolyze αS1-casein.