Multi-omics reveals the regulatory mechanism of nutrients accumulation in broccoli head under preharvest arginine application.

Scorpions are characterized by their venomous adaptations, including specialized stingers, and their ecological diversity. Some families, such as Buthidae, have medically significant species and their venoms possess a diverse array of chemicals. In Mexico, Centruroides suffusus and Centruroides vittatus coexist, with C. suffusus considered medically important due to its highly toxic venom. This study describes the metabolomic and microbiomic profiles of C. suffusus and C. vittatus. The metabolomic profiling (12 amino acids and 28 acylcarnitines) reveals significant differences between the two species, hinting at metabolic and ecological variations. Ornithine (ORN) and arginine (ARG) were the most abundant in C. vittatus, while tyrosine (TYR) was the most abundant amino acid molecule in C. suffusus. The microbiome analysis (by Next-Generation Sequencing of the 16S ribosomal gene) indicates similarities in gut bacteria composition between the two species (Phyla: Actinobacteria, Bacteroidetes, Cyanobacteria, Firmicutes, Proteobacteria and Tenericutes).

A new natural deep eutectic solvent (DES) based on arginine (ARG) and glutaric acid (GLU) was prepared. The eutectic point phase diagram showed that the best ratio for preparation of ARG/GLU-DES is one mole of ARG to three moles of GLU. Then, the new DES was characterized by eutectic points, FT-IR, 1H NMR, TGA/DTA, and densitometry techniques. Additionally, it was demonstrated to be an effective catalyst for the green synthesis of pyrano[2,3-d]-pyrimidines through a one-pot three-component condensation reaction of aromatic aldehydes, barbituric acid, and 4-hydroxycoumarin at 80 °C under solvent-free conditions with high yields and short reaction times.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-19681-5.

Abstract This study examined the effects of heat stress (HS) (41°C) and amino acid treatments, including citrulline (CIT) and arginine (ARG), on muscle-related gene expression in bovine myocytes. We hypothesized that CIT would increase ARG availability, which increases protein synthesis and myotube formation under HS conditions. Bovine satellite cells were isolated from a female fetus (23.1 kg) and plated at a density of 4 × 10⁴ cells in DMEM containing 10% FBS. The cells were incubated at 38°C with 5% CO₂ until they reached 80% confluency. The culture medium was replaced with differentiation medium (control media) consisting of 2% HS SILAC DMEM Flex Media (DMEM without ARG and CIT). The treatment groups were as follows: T1 (control media), T2 (control media with 120 µM ARG), and T3 (control media with 120 µM CIT). The cells were exposed to 38°C or 41°C, and harvested at 48 hours after temperature exposure. Real-time q-PCR was used to determine the gene responses related to HS and muscle growth for all treatment groups. The cells were stained to determine the differentiation fusion index and muscle fiber size. Data was analyzed using two-way ANOVA to evaluate the effects of treatments, temperature, and their interaction on gene expression, differentiation fusion index, and muscle fiber size. Tukey’s test was used for post-hoc comparisons. Results are presented as significant at P < 0.05. Analyses were conducted using GraphPad Prism 9.4.1 to assess treatment effects under normal and HS conditions. Gene expression of IGF-I was affected by treatment (P = 0.013) and its interaction with temperature (P = 0.003), with the highest expression in the CIT group under HS. Myf5 decreased with heat and ARG treatment (P < 0.001, P = 0.006). MyoD increased at 41°C (P = 0.015), with the highest levels in control. Myogenin was upregulated under HS, especially with CIT (P < 0.001, P = 0.004). Expression of MHC I was elevated with ARG at 41°C (P < 0.001). MHC ⅡA was affected by the interaction (P = 0.018), while MHC ⅡX expression was influenced by treatment (P = 0.01) and temperature (P= 0.015). Heat stress altered gene expression, with CIT enhancing IGF-1 and myogenin, while ARG increased MHC I. At 38°C, the treatments were statistically different (P=0.004) from one another with CIT (34.0%) being the highest. There were no differences between treatments in myotube length (P=0.820) or width (P=0.34) at 38°C. At 41°C, CIT and ARG treatments were both significantly different (P< 0.001) than control and both caused a greater myotube length (P=0.007). However, there was no difference in myotube width (P = 0.760). Therefore, CIT and ARG can increase fusion index and myotube length during times of HS while not affecting myotube width.

ABSTRACTAquilaria sinensis, a plant of the genus Aquilaria in the family Thymelaeaceae, can form resinous and fragrant agarwood in response to wounding or infection. The resin physically occludes xylem vessels and ray parenchyma, limiting pathogen mobility and resource loss. The quality of agarwood is often evaluated using highly oxygenated 2‐(2‐phenylethyl)chromones, such as agarotetrol, 2‐[2‐(4‐methoxy)phenylethyl]chromone, 4′‐methoxyagarotetrol and 6,7‐dimethoxy‐2‐(2‐phenylethyl)chromone. Among the derivatives of 2‐(2‐phenylethyl)chromone (PEC), the majority possess a hydroxyl or methoxy group at the C‐6 position of the chromone, with subsequent substitutions occurring at the C‐5, C‐7 and/or C‐8 positions. The hydroxylation at the C‐6 position is a critical step in the biosynthesis of these 2‐(2‐phenylethyl)chromones (PECs). In this study, we investigated the gene expression differences and accumulation patterns of PECs in two‐year‐old wild A. sinensis and Qi‐Nan A. sinensis, using transcriptomic and LC–MS/MS analysis. Based on these analyses, we identified 52 candidate genes encoding cytochrome P450 (CYPs). Further investigations using a yeast expression system and functional analysis in Nicotiana benthamiana and A. sinensis revealed that AsCYP82G1 could hydroxylate 2‐(2‐phenylethyl)chromone to form 6‐hydroxy‐2‐(2‐phenylethyl)chromone. Homology modelling and molecular docking demonstrated that the residues methionine (MET) and arginine (ARG) at positions 222 and 223 of the AsCYP82G1 protein are likely to play an important role in the catalysis of 2‐(2‐phenylethyl)chromone to 6‐hydroxy‐2‐(2‐phenylethyl)chromone. Finally, the reaction mechanism of AsCYP82G1 was studied by quantum mechanical calculation. Our research provides new insights into the molecular mechanism of PECs biosynthesis and lays the foundation for the breeding of high‐quality agarwood varieties and large‐scale production of PECs using synthetic biology.

Autophagy induced by itraconazole and ritonavir was found involved in the pathogenesis of C. albicans. This study was designed to explore the possible molecular mechanism of itraconazole and ritonavir in the treatment of Candida albicans infection through autophagy pathway. The overlapping targets of itraconazole and ritonavir, and those-related to C. albicans and autophagy were screened. Then the core targets were identified by protein-protein interaction (PPI) network analysis. Gene enrichment analysis of targets and the drug-target-pathway-disease network was constructed. The interactions between itraconazole, ritonavir and core targets were analyzed by molecular docking and molecular dynamics simulation. Finally, the core target-miRNA interaction network was constructed to predict candidate miRNAs. PPI network showed that PIK3R1, RELA, STAT3, HSP90AA1, TP53, JUN, GRB2, EGFR, ESR1 and TNF were potential core targets of autophagy therapy for C. albicans infection with itraconazole and ritonavir. Enrichment analysis showed that the two drugs may regulate the autophagy process through pathways including PI3K-AKT, IL-17, MAPK, Toll-like receptor, JAK-STAT and NF-κB. Molecular docking analysis indicated that itraconazole and ritonavir possess strong binding affinities with the cote target proteins, with binding free energies ranging from -5.6 to -9.5kcal/mol. Key interactions were identified at the active sites of the targets, suggesting stable ligand-receptor complex formation. Itraconazole docked to PIK3R1 through SER-78 and GLU-82 (-9.3kcal/mol), and ritonavir docked to PIK3R1 through ASN-85, GLU-1011 and arginine (ARG)-1088 (-7.7kcal/mol). Molecular dynamics simulation of itraconazole and ritonavir with representative target genes lasted for 100ns showed the structures of the formed complexes remained stable throughout. Finally, the candidate miRNAs including miR-486-5p, miR-411-5p.1 and miR-296-5p were identified. Network pharmacological analysis showed a multi-target and multi-pathway molecular mechanism of itraconazole and ritonavir in the treatment of C. albicans infection, and provided a theoretical basis for subsequent studies.

Introduction and Objective: Arginine (ARG) is an important amino acid in T2D as a potent insulin secretagogue and precursor for nitric oxide (NO). Citrulline (CIT), the substrate for de novo ARG synthesis, is mostly produced from glutamine (GLN). Here, we aimed to investigate their metabolism in T2D using a novel stable isotope tracer approach. Methods: We studied 42 individuals (21 with T2D, 21 controls). After an overnight fast, we collected blood samples following pulse administration of ARG, CIT, and GLN stable amino acid tracers. Plasma concentrations and isotopic enrichments were measured by LC-MS/MS and lean soft tissue (LST) by DXA. Compartmental analysis was performed to calculate their WBP. Data are mean (SD) or [95% CI] (t-test and ANCOVA by JASP). Results: The cohort was 60% female, mean age 64.4 (7.5) years, and BMI 33.0 (4.3) kg/m2 (all p>0.05). After adjusting for sex and age, T2D group had lower plasma concentrations of ARG (57.7 [48.9, 66.4] vs. 75.8 [66.9, 84.5] µM, p=0.005), CIT (23.4 [18.9, 27.9] vs. 34.4 [29.9, 38.9] µM, p=0.001), and GLN (463.5 [421.1, 506] vs. 561.7 [519, 604.3] µM, p=0.002). In T2D, WBP was lower for CIT (11.8 [10.1, 13.6] vs. 15.5 [13.8, 17.2] µmol/min, p=0.004) but higher for GLN (537 [503, 571] vs. 481.9 [448.3, 515.5] µmol/min, p=0.004) after controlling for age, sex and LST. T2D group also had lower CIT intracellular production (p=0.003), but higher GLN clearance (p=0.001) and intracellular pool size (p=0.007), and a trend towards higher ARG clearance (p=0.06). Conclusion: In T2D, significant dysregulation exists in ARG, CIT and GLN metabolism. We hypothesize that increased muscle GLN synthesis stimulates gluconeogenesis, increases GLN consumption, and contributes to T2D pathogenesis via blunted insulin secretion, sensitivity and incretin response by having less GLN available for CIT production that may affect ARG (and NO) metabolism. Interventions aiming to reduce GLN production but increase CIT availability may be useful in T2D. Disclosure M. Tosur: None. R. Wierzchowska-McNew: None. M.P. Engelen: None. N.E. Deutz: None. Funding National Institutes of Health (K23-DK129821)(MT)

Prolonged disturbances in citrulline metabolism following resistance exercise in COPD.

Abstract Two hundred and thirty-two PIC (PIC 1050, Genus, Hendersonville, TN) sows were used to investigate the effects of arginine supplementation in lactation diets on sow performance and litter growth rate. Sows were allotted by randomized complete block design to one of two dietary treatments formulated to different standard ileal digestible (SID) arginine levels. The control (CON) diet contained 0.9% SID arginine, while the experimental diet contained 1.07% SID arginine (ARG). Diets were formulated to meet or exceed all other NRC (2012) dietary recommendations. Sow body weight (BW), backfat (BF), and body condition score (BCS) were recorded upon entering the farrowing room and at weaning. Piglet litter weights were recorded after cross-fostering and at weaning to assess litter growth rate. Data were analyzed using generalized linear mixed models, with treatment as a fixed effect and group as a random effect. Models were fitted using R (v4.4.1), and results were considered significant at P ≤ 0.05 and a tendency at 0.50 < P ≤ 0.10. Sow and litter were considered the experimental unit. Sow lactation length and BW were not significantly different between treatments (P = 0.440 and 0.519, respectively). There was no significant difference in ADFI (P = 0.931). The supplementation of Arg did not change BCS at weaning (2.46, 2.47 (scale of 1-5); P = 0.919). Upon entering the farrowing room, the sow BF was 10.15 and 9.99 mm for the CON and Arg diets respectively. Change in BF after weaning was not significantly different between the CON (-0.946 mm) and the ARG (-1.31 mm) diets (P = 0.236). The number of pigs weaned per sow was not statistically different (P = 0.486). Starting litter weight was 19.1 kg for CON and 20.2 kg for ARG. Litter wean weight did not differ significantly between treatments (P = 0.695). Piglet average daily gain (ADG) differed significantly between treatments with a mean of 0.25 kg/d for CON and 0.24 kg/d for ARG (P = 0.029). Post-foster mortality was not changed with the addition of Arg with the CON post-foster mortality at 5.33% and the ARG mortality at 5.79% (P = 0.616). This resulted in the percent total livability of 78.5 (CON) and 75.3 (ARG) which was not significantly different (P = 0.258). In summary, arginine supplementation during lactation did not significantly influence sow performance or overall piglet growth and livability, except for a minor difference in piglet ADG.

Design of indomethacin novel small molecule hydrogels for concomitant release and permeability increases.

This study investigated the impact of an increased arginine (ARG) level in a boar diet on semen production, sperm quality, and seminal plasma proteome. Adult Nebraska Index Line boars were assigned to two groups, one receiving a control diet with 0.77% arginine (n = 4) and the other a high-arginine diet with 1.77% arginine (n = 5). Semen was collected twice a week over the whole experiment, including one week before, six weeks during, and six weeks after the supplementation. Parameters such as semen volume and concentration were assessed immediately after collection, alongside sperm motility and morphology. Centrifugation of raw semen samples yielded seminal plasma for a gel-based proteome analysis. The seminal plasma proteins were extracted, quantified, and separated via 2D gel electrophoresis, allowing protein identification through mass spectrometry. Data analysis involved two-way ANOVA for comparisons (p < 0.05). Results showed that arginine supplementation improved semen volume and total sperm counts, with averages of 21 ± 3 doses in the control group versus 24 ± 2 in the ARG group (p = 0.05). Although sperm motility and morphology remained unaffected (p > 0.05), dietary arginine upregulated ten proteins and downregulated two. In summary, increased dietary arginine did not significantly alter key parameters of semen output or sperm quality but significantly impacted seminal plasma proteome, warranting further research on sperm viability.

Background/Objectives: Tannic acid (TA) is a well-known natural phenolic acid composed of ten gallic acids linked to each other with ester bonding possessing excellent antioxidant properties in addition to antimicrobial and anticancer characteristics. Arginine (ARG) is a positively charged amino acid at physiological pH because of nitrogen-rich side chain. Method: Here, poly(tannic acid-co-arginine) (p(TA-co-ARG)) particles at three mole ratios, TA:ARG = 1:1, 1:2, and 1:3, were prepared via a Mannich condensation reaction between TA and ARG by utilizing formaldehyde as a linking agent. Results: The p(TA-co-ARG) particles in 300-1000 nm size range with smooth surfaces visualized via SEM analysis were attained. Abundant numbers of functional groups, -OH, -NH2, and -COOH stemming from TA and ARG constituent confirmed by FT-IR analysis. The isoelectric point (IEP) of the particles increased from pH 4.98 to pH 7.30 by increasing the ARG ratios in p(TA-co-ARG) particles. The antioxidant capacity of p(TA-co-ARG) particles via gallic acid (GA) and rosmarinic acid (RA) equivalents tests revealed that particles possess concentration-dependent antioxidant potency and increased by TA content. The α-glucosidase inhibition of p(TA-co-ARG) particles (2 mg/mL) 1:1 and 1:2 mole ratios revealed significant enzyme inhibition ability, e.g., 91.3 ± 3.1% and 77.6 ± 12.0%. Interestingly, p(TA-co-ARG) (1:3 ratio) possessed significant antibacterial effectiveness against Escherichia coli (ATCC 8739) and Staphylococcus aureus (ATCC 6538) bacteria. Furthermore, all p(TA-co-ARG) particles at 1000 mg/mL concentration showed >80% toxicity on L929 fibroblast cells and increased as ARG content of p(TA-co-ARG) particles is increased. Conclusions: p(TA-co-ARG) showed significant potential as natural biomaterials for biomedical use.

Salinity stress disrupts water uptake and nutrient absorption, causing reduced photosynthesis, stunted growth, and decreased crop yields in plants. The use of indole acetic acid (IAA), arginine (AN), and mango fruit waste biochar (MFWB) can be effective methods to overcome this problem. Indole acetic acid (IAA) is a natural auxin hormone that aids cell elongation and division, thereby increasing plant height and branching. L-arginine, an amino acid, is crucial for plant defense mechanisms, forming proline, polyamines, and nitric oxide, which regulate biological activities and prevent oxidative damage. Mango fruit waste biochar enhances soil fertility and water retention, thereby enhancing fruit development and yield. This study investigates the effects of combining IAA and AN as amendments to fenugreek, with and without MFWB. Four treatments (control, 2mM IAA, 250 mg/L AN, and 250 mg/L AN + 2mM IAA) study were conducted in four replications using a completely randomized design. Results demonstrate that the 250 mg/L AN + 2mM IAA with MFWB treatment led to a significant rise in fenugreek plant length (30.26%), plant fresh weight (36.37%), and plant dry weight (15.78%) over the control under salinity stress. There was a notable increase in chlorophyll a (5.13%), chlorophyll b (14.06%), total chlorophyll (7.79%), and shoot N, P, K from the control under salinity stress also showing the potential of 250 mg/L AN + 2mM IAA with MFWB. In conclusion, applying 250 mg/L AN + 2mM IAA with MFWB is a strategy for alleviating salinity stress in fenugreeks.

Overcoming solubility limitations of nutraceutical compositions based on poorly-soluble phosphoethanolamine using ionic liquids approach

Amino acid parameters provide important and relevant data for objective assessment of physiological and biochemical status of organism and patterns of formation of economically useful traits of animals. The objective of this work is to model phenotypic and genetic patterns of variability of main amino acid parameters of blood of pig hybrids within the framework of mixed equation model. Main 19 amino acid parameters of blood serum of 58 heads of three-breed hybrids (Large White × Landrace × Duroc) were obtained and studied: content (g/100 ml) of aspartic acid and its amide (ASP+ ASN) was 0.64; glutamic acid and its amide (GLU+GLN) ‒ 1.04; alanine (ALA) – 0.46; glycine (GLY) – 0.28; threonine (THR) – 0.34, serine (SER) – 0.39; cysteine (CYS) – 0.21; methionine (MET) –0.07; valine (VAL) – 0.52; isoleucine (ILE) – 0.28; leucine (LEU) –0.83; tyrosine (TYR) – 0.46; phenylalanine (PHE – 0.50); histidine (HIS) – 0.31; lysine (LYS) – 0.69; arginine (ARG) – 0.51; proline (PRO) – 0.41. To calculate the correlation values, covariance and variance of features, models using the REMLF90 programs were fulfilled. It was shown that all amino acids have moderate and strong both phenotypic and genetic correlations (r&gt; 0.5). All amino acids have a fairly close connection with the main biochemical indices. From the above, we can conclude that the equation of the mixed model allows us to obtain a deep understanding of the nature of the variability of the main amino acid indices of the blood, connected in a complex labile biochemical system.

Growth hormone (GH) secretion is pulsatile, entropic, and nycthemeral and is mainly controlled by the hypothalamus through two neurohormones, the stimulating growth hormone releasing hormone (GHRH) and the inhibiting somatostatin. Shortly after its discovery and synthesis, GHRH was intensely investigated diagnostically to define GH secretion. The nascent enthusiasm for using GHRH as a single diagnostic tool to investigate GH deficiency (GHD) dropped down quickly due to a flawed reproducibility. The subsequent combinatory use of molecules implicated in GH secretion through inhibition of the somatostatinergic tone, such as arginine (ARG), or the synthesis of receptor-orphan pharmaceutical compounds capable of stimulating pituitary somatotrophs to release GH, such as the GH secretagogues (GHSs), improved the reproducibility of GH response to GHRH alone, thus gaining access into the clinical practice by means of different diagnostic approaches. This review will focus on the history of the GHRH test, with main emphasis on GHRH plus ARG as a dynamic testing for the diagnosis of GHD. Our attention will extend crosswise from studies aimed at validating GHRH-based tests for the clinical practice, to address main pitfall conditions capable of affecting per se GH secretion, such as obesity, hypothalamic damage, and ageing. The history of GHRH test has been progressively dismantled due to the cease of its production for business reasons, opening a gap in the diagnostic workup of patients with GHD. In the urgency to seek further robust, safe, and validated diagnostic tests or tools, we hope to stimulate attention on a so important peptide for the health of our patients suffering from pituitary diseases.

BackgroundNutritional stress is a mechanism that allows tumor cells to evade the immune system. Arginine (ARG), an amino acid involved in immunomodulation, aids in regulating T-lymphocyte cell activity and the...

Nutraceuticals can affect transactions immunologically for health and endocrinologically for production and reproduction, specifically, referring to bioactive peptidomemics and saccharomemics in disease resistance and immunlogical balance. The nutraceuticals: Vit D2/D3, fructans and WSCs, PUFAs, alpha-lactoferrin and polysaccharides can affect SCI and as indicated by blood biomarkers. The functional amino acids (FAAs): histidine (HIS), arginine (ARG), lysine (LYS) and leucine (LEU) can affect lean body mass (LBM) accretion and milk production with bovine growth hormone (bGH)/bovine growth-releasing hormone (bGRH) and prolactin. The two prebiotic nutraceuticals referred to can be applied to “designer” oligomers from enriched seed proteins and polysaccharides to improve feed nutritive value (NV). High non-fibrous carbohydrate (NFC) and water-soluble carbohydrate (WSC) grasses can also provide higher-end energy forages. There are, thus, proteinogenic approaches that can be used for supplemental feeding. It is suggested that lower quality residual feedstocks can be converted to food-feed applications and can involve pretreating of fibrous carbohydrates (FC) and NFC, conversion to natural sugars and sweeteners, and “shuffling” copolymerization. Applications can be made to fishmeal for production and health. Slowed reaction enzymes (SRE) can be used with so-called osmolytic resins to study behaviour of pre-formed amino acids (PFAAs) in the rumen milieu to fit rumen protein solubility with use of inhibitors to both plant and microbial proteases, modulating as a result the MCP and “escape” protein flows to the lower GI tract. Clean tech can produce seed-derived proteins using “bulk” cell culturing (processed and unprocessed) and extracted/enriched yeast culturing. There is a need to verify the prebiotic binding receptors in the small intestines (SI) (e. g. gut-associated lymphoid tissues, GALT, and as speculated the SI’s jejunum).

Coamorphous systems of rebamipide: Selection of amino acid coformers based on protein-ligand docking, in vitro assessment and study of interactions by computational and multivariate analysis

The purpose of this study was to determine the effects of dietary supplementation of arginine (ARG) or guanidinoacetic acid (GAA), with or without phenylalanine (PHE), on the ascites susceptibility in the cold-stressed broilers fed canola meal (CM)-based diet. A total of 450 one-day-old male broiler chicks were randomly allocated to 30 floor pens with 6 replications for each of the 5 treatments. The dietary treatments were as follows: CM-based diet (control), CM-based diet+2.57g/kg ARG, CM-based diet+1.8g/kg GAA, CM-based diet+2.57g/kg ARG+1.5g/kg PHE and CM-based diet+1.8g/kg GAA+1.5g/kg PHE. The groups experienced cold stress induction starting at 32°C on day one, with gradual reductions to 15°C on days 21. The temperature was then held at 15°C for the remainder of the experiment. The supplements ARG+PHE and GAA+PHE resulted in improved feed conversion ratios (FCR) when compared to the control group. In comparison with the control group, supplementation of ARG and ARG+PHE decreased the ascites mortality by increasing the plasma nitric oxide level (NO), blood O2 partial pressure, blood O2 saturation (SO2), and decreasing the blood CO2 partial pressure (PCO2) and right ventricle to total ventricle (RV:TV) ratio. Supplementation of GAA and GAA+PHE also declined ascites mortality by reducing blood PCO2 while increasing blood SO2 and plasma NO levels. Although plasma corticosterone level and RV:TV ratio were similar between the GAA and control groups, adding GAA+PHE significantly reduced both compared to the control group. In summary, supplementing cold-stressed broiler chicken diets with 2.57g/kg ARG may alleviate hypertension. Additionally, 1.8g/kg GAA proves to be an effective substitute for dietary ARG in low-ARG diets, alleviating adverse effects from cold stress. Proper PHE formulation, at 1.5g/kg in this study, is crucial when using ARG and GAA supplements.