Amino Acids Glutamate Research Articles

The causal effect of serum amino acids on the risk of prostate cancer: a two-sample mendelian randomization study

Prostate cancer (PCa) is the second most common malignancy affecting men globally. Recent advances in metabolomics have highlighted significant alterations in specific amino acid (AA) metabolism linked to PCa, indicating their potential utility in diagnosis and therapy. However, no direct causal association between serum AA levels and PCa risk has been established. A total of 35 patients with PCa and 30 individuals with benign prostatic hyperplasia (BPH) were recruited for this study. Targeted metabolomic analysis was performed using ultra-high-performance liquid chromatography-tandem mass spectrometry on serum samples. Two-sample Mendelian randomization (MR) was applied to explore potential causal links between serum AA levels and PCa risk, including mediator effects using dual-phase MR and assessing reverse causality through reverse MR. Results Targeted metabolomic profiling identified six amino acids—glutamate (Glu), Ser, histidine (His), arginine (Arg), aspartic acid (Asp), and glycine (Gly)—that showed significant area under the ROC curve in differentiating between BPH and PCa cases. Notably, Glu demonstrated an inverse association with PCa risk, distinct from the other AAs identified. However, definitive evidence supporting a causal relationship between low Glu levels and increased PCa risk was not observed. Our results suggest a protective role of Glu against PCa development, which may have implications for disease prognosis. Increasing dietary Glu intake may present a potential preventive or therapeutic approach for PCa.

Altered amino acid levels in young hypopituitarism: impact of NAFLD and insulin resistance

Elevated concentrations of amino acids (AAs) are commonly observed in patients with nonalcoholic fatty liver disease (NAFLD). Individuals with hypopituitarism (HP) are at a heightened risk of developing NAFLD due to factors such as visceral obesity, increased insulin resistance (IR), and disturbances in lipid metabolism. However, the changes in AAs concentrations associated with HP remain poorly understood. Therefore, our study aimed to investigate whether individuals with HP, who were not receiving growth hormone replacement therapy (GHRT), exhibited altered AAs compared to controls (CTs), and whether these AAs were associated with IR, the presence of NAFLD, and the Metabolic Syndrome (MetS) score. The AAs profiles of 133 young males with HP (age: 24.5 ± 5.9; 57 with NAFLD and 76 without NAFLD) and 90 age and BMI-matched CTs were analyzed using untargeted metabolomics. The results revealed that most AAs were found to be elevated in subjects with HPs compared to CTs. Glutamate, glutamine, norleucine, and branched-chain amino acids (BCAAs) (leucine and valine) were correlated with the homeostasis model assessment of insulin resistance (HOMA-IR), with glutamate and norleucine showing independent linkage. Glutamate and proline levels were specifically associated with MetS score, while alanine and proline linked to NAFLD. Given that elevated glutamate and BCAAs levels have higher prevalence of NAFLD, we hypothesized that the changes in AAs observed in HPs may be attributed to the impact of NAFLD and IR.

Potential PDE4A inhibition-mediated neuroprotective effects of psoralidin.

This study aimed to investigate the effect of psoralidin, a natural phenolic coumarin compound, on MK-801-induced neurotoxicity that may cause Alzheimer's disease and to determine the phosphodiesterase(PDE)-related molecular mechanism of action. In this study, neurotoxicity was performed using the MK-801 in the HT-22 cell line. The effects of compounds on the proliferation of HT-22 cells were determined by Real-Time Cell Analysis (RTCA). After measuring the total protein concentration, the PDE4A protein level was determined using the Western blot method. Psoralidin (100, 200, 400 µM) has been shown to have a neuroprotective effect against MK-801-induced neurotoxicity, as indicated by Real-Time Cell Analysis. In HT-22 cells, the half maximal effective concentration (EC50) value of psoralidin was calculated to be 230.4 µM, IC50 value of MK-801 was calculated to be 62.4 µM at 24 hours. It has been determined that psoralidin (200, 400 µM) inhibits PDE4A by using the Western blot method. This research uncovers that psoralidin has neuroprotective effects in MK801-associated accumulation of the excitatory amino acid glutamate neurodegeneration and Alzheimer's disease.

Evaluation of Xanthone and Cinnamoylbenzene as Anticancer Agents for Breast Cancer Cell Lines through In Vitro and In Silico Assays

Breast cancer is a severe global disease for women as the number of deaths increases annually. Therefore, attempts to find new anticancer agents are critical and inevitable. In this work, we report the investigation on the anticancer activity of xanthone and cinnamoylbenzene compounds against two breast cancer cell lines, i.e., T47D and MCF-7, through experimental in vitro and theoretical in silico assays. Xanthone and cinnamoylbenzene exhibit anticancer activity with a half-maximal inhibitory concentration (IC50) of 136.7–194.3 and 235.8–262.4 µg/mL against T47D and MCF-7 cancer cells, respectively. Cinnamoylbenzene generates less cytotoxicity to normal Vero cells with a selectivity index of 1.095–2.102. The molecular docking studies agree with the experimental data in which cinnamoylbenzene is more active against T47D with an IC50 of 136.7 µg/mL due to Topoisomerase II inhibition through π-π stacked interactions with Adenine12 and Guanine13 nitrogen bases. Meanwhile, xanthone is more active against MCF-7 with an IC50 of 235.8 µg/mL due to EGFR inhibition through van der Waals interaction and hydrogen bond with Glutamic acid767 and Methionine769 amino acid residues, respectively. Additionally, the pharmacokinetic parameters of xanthone and cinnamoylbenzene are predicted through absorption, distribution, metabolism, excretion, and toxicity (ADMET) analysis, and they show better suitability than doxorubicin as the commercial anticancer drug.

Screening of the Candidate Metabolite to Evaluate the Mycelium Physiological Maturation of Lyophyllum decastes Based on Metabolome and Transcriptome Analysis.

Lyophyllum decastes is a commercially cultivated rare edible mushroom with high dietary and medicinal value. The mycelium physiological maturation was an important factor to the yield and quality of mushrooms obtained. However, it was impossible to obtain discriminative characteristics represented the maturity level of the mycelia from morphological features. In this article, we screened a candidate metabolite for evaluating the mycelium physiological maturation by metabolomic and transcriptomic analysis during mycelial vegetative growth stages of L. decastes. The results showed that mycelial vegetative growth 55 d had reached to physiological maturation according to the yield and quality of L. decastes, which the single bottle yield (307 g/bottle), the contents of flavonoids (2.94 mg/g dry weight), and 5 flavor amino acids (glutamate 177.90 μg/g, aspartate 95.74 μg/g, phenylalanine 31.68 μg/g, tyrosine 13.79 μg/g, and alanine 10.99 μg/g) were the highest. In addition, the metabolomic and transcriptomic analysis results showed that biosynthesis of secondary metabolites and amino acid had significant different during mycelial vegetative growth stages, and glutamine may be a candidate indicator to evaluate the mycelial physiological maturation. These findings contribute to improve our knowledge of mycelium physiological maturation and it will be used to increase mushroom yield and quality.

Substrate preference of Tetrasphaera in dual PAOs synergistic EBPR system and its phosphorus removal performance

In this study, an alternated anaerobic/aerobic sequencing batch reactor (SBR) was employed to investigate the effect of the substrate conversion from casein hydrolysate (Cas aa) to a mixture of amino acids (glutamate, aspartate, glycine and proline) on the efficacy of nitrogen and phosphorus removal in an enhanced biological phosphorus removal (EBPR) system characterized by dual polyphosphate-accumulating organisms (PAOs), mainly focusing on the substrate preference of fermentative PAOs. The results indicated that a stable removal efficiency was found to be above 90 % as the substrate was converted to the mixture of amino acids. Meanwhile, the removal efficiency of ammonium eventually increased to over 95 % in stage III, despite an increase of ammonia nitrogen by hydrolysis with the rising proportion of mixed amino acids in influent. Moreover, shortening the anaerobic duration did not significantly affect the phosphorus removal performance, even enhancing anaerobic phosphorus release capacity. Nevertheless, the ammonia nitrogen removal was influenced. 16 s rRNA high-throughput sequencing results demonstrated that the detected fermentative PAO like Tetrasphaera in the EBPR system exhibited a clear preference for utilizing mixed amino acids and could cooperate with traditional PAO like Accumulibacter for phosphorus removal. In addition, the fermentation of amino acids mixture by Tetrasphaera was more pronounced and its abundance increased under the condition of a shorter anaerobic duration, leading to a relatively stable and high abundance of Accumulibacter in the presence of sufficient substrates from Tetrasphaera. However, considering the nitrogen removal performance and steady operation of the EBPR system, a longer anaerobic duration was more reasonable.

Interaction between energy level and starch: fat ratio on intestinal energy metabolism of layer pullets

During the growing period, the gastrointestinal tract of layer pullets is not yet well developed and may be susceptible to dietary energy level. The energy level and composition might impact the intestinal energy metabolism of layer pullets. To test this hypothesis, a total of 480 “Jing Tint 6” layer pullets were used in an 8-week study and allocated to 4 groups, each consisting of 8 replicates, with 15 birds per replicate. Pullets were treated with low or high starch: fat ratios (LS, 10:1; HS, 20:1) in a 2 × 2 factorial arrangement with regular energy (RE, 11.85 and 11.68 MJ/kg for birds from 6 to 10 weeks of age and 11 to 14 weeks of age, respectively) or low energy (LE, 0.55 MJ/kg lower than RE) levels. A significant interaction (P < 0.05) showed that HS increased glandular stomach weight and the jejunal villus length to crypt depth ratio (VCR) in LE diets, but decreased these parameters in RE diets. Dietary energy reduction impaired energy metabolism in the ileum (P < 0.05) mainly via decreasing the gene expression of enzymes involved in the tricarboxylic acid (TCA) cycle (α-ketoglutarate dehydrogenase complex [α-KGDH]; isocitrate dehydrogenase (NAD (+)) [IDH] catalytic; citrate synthase [CS]) and adenosine triphosphate (ATP) synthesis, reducing contents of phosphoenolpyruvate (PEP) and adenylate energy charges (AEC) and down-regulating the adenosine monophosphate-activated protein kinase (AMPK) pathway. HS stimulated AMPKα phosphorylation, increased protein abundance of peroxisome proliferator activated-receptor gamma coactivator 1α (PGC1α) and improved contents of amino acids (aspartate, glutamate, glutamine, alanine and threonine) and malate in the ileum regardless of energy levels (P < 0.05). By an interaction (P < 0.05), the transition from LS to HS diets up-regulated ileal gene expression of AMPKα1 and decreased content of adenosine monophosphate (AMP), accompanied by higher AEC but only in birds fed with LE diets. Collectively, these results suggest that low energy feeding may not be enough for maintaining intestinal energy homeostasis in layer pullets and emphasizes the importance of a relatively high starch: fat ratio in restoring energy metabolism in the ileum.

Candidatus Thiothrix phosphatis SCUT-1: A novel polyphosphate-accumulating organism abundant in the enhanced biological phosphorus removal system

A novel coccus Thiothrix-related polyphosphate-accumulating organism (PAO) was enriched in an acetate-fed enhanced biological phosphorus removal (EBPR) system. High EBPR performance was achieved for an extended period (>100 days). A high-quality draft genome (completeness 97.2 %, contamination 3.26 %) was retrieved, representing a novel Thiothrix species (with similarity<93.2 % to known Thiothrix species), and was denoted as 'Candidatus Thiothrix phosphatis SCUT-1'. Its acetate uptake rate (6.20 mmol C/g VSS/h) surpassed most Ca. Accumulibacter and known glycogen-accumulating organisms (GAOs), conferring their predominance in the acetate-fed system. Metatranscriptomic analysis suggested that Ca. Thiothrix phosphatis SCUT-1 employed both low- and high-affinity pathways for acetate activation, and both the conventional (PhaABC) pathway and the fatty acid β-oxidation pathway for PHA synthesis; additionally, a much more efficient FAD-dependent malate: quinone oxidoreductase (MQO) were encoded and employed than the traditional malate dehydrogenase (MDH) to oxidize malate to oxaloacetate in the TCA and glyoxylate cycle, collectively contributing to a higher acetate utilization and processing rate of this microorganism. Batch tests further demonstrated the versatile ability of this PAO in using VFA (acetate, propionate, and butyrate), lactate, amino acids (aspartate and glutamate), and glucose as carbon sources for EBPR, showing a partially overlapped but unique ecological niche of this microorganism comparing to Ca. Accumulibacter and known GAOs. A metabolic model was built for Ca. Thiothrix phosphatis SCUT-1 using the above-mentioned carbon sources for EBPR. Overall, this study represents the first comprehensive characterization of the physiology and metabolic characteristics of representative coccus Thiothrix-related PAOs, which are expected to provide new insights into PAO microbiology in EBPR systems.

Targeting harmful effects of non-excitatory amino acids as an alternative therapeutic strategy to reduce ischemic damage.

The involvement of the excitatory amino acids glutamate and aspartate in cerebral ischemia and excitotoxicity is well-documented. Nevertheless, the role of non-excitatory amino acids in brain damage following a stroke or brain trauma remains largely understudied. The release of amino acids by necrotic cells in the ischemic core may contribute to the expansion of the penumbra. Our findings indicated that the reversible loss of field excitatory postsynaptic potentials caused by transient hypoxia became irreversible when exposed to a mixture of just four non-excitatory amino acids (L-alanine, glycine, L-glutamine, and L-serine) at their plasma concentrations. These amino acids induce swelling in the somas of neurons and astrocytes during hypoxia, along with permanent dendritic damage mediated by N-methyl-D-aspartate receptors. Blocking N-methyl-D-aspartate receptors prevented neuronal damage in the presence of these amino acids during hypoxia. It is likely that astroglial swelling caused by the accumulation of these amino acids via the alanine-serine-cysteine transporter 2 exchanger and system N transporters activates volume-regulated anion channels, leading to the release of excitotoxins and subsequent neuronal damage through N-methyl-D-aspartate receptor activation. Thus, previously unrecognized mechanisms involving non-excitatory amino acids may contribute to the progression and expansion of brain injury in neurological emergencies such as stroke and traumatic brain injury. Understanding these pathways could highlight new therapeutic targets to mitigate brain injury.

The disruption of NEAT1-miR-125b-5p-SLC1A5 cascade defines the oncogenicity and differential immune profile in head and neck squamous cell carcinoma

Metabolic reprogramming sustains malignant head and neck squamous cell carcinoma (HNSCC) to overcome stressful microenvironments, and increased glutamine uptake is a common metabolic hallmark in cancers. Since metabolic reprogramming has been recognized as a new therapeutic target for tumor cells, understanding the regulatory axis of glutamine uptake in HNSCC and its potential downstream effects in its pathogenesis of HNSCC would be incredibly beneficial. Bioinformatic analysis of the Cancer Genome Atlas (TCGA)-HNSCC dataset and RNAseq analysis performed on HNSCC indicated that SLC1A5 was the most dysregulated transporter among the seven homologous glutamate or neutral amino acid transporters in the SLC1A family. To further clarify the role of SLC1A5 in HNSCC, we knocked down SLC1A5 expression. This knockdown decelerated cell growth, induced G0/G1 arrest, diminished tumorigenicity, and increased cleavage caspase3, LC3B, and intracellular Fe2+. Inhibitors against apoptosis, autophagy, or ferroptosis rescued the cell viability repressed by SLC1A5 knockdown. SLC1A5 knockdown also suppressed glutamine uptake, enhanced oxidative stress, and increased sensitivity to cisplatin. CRISPR/dCas9-mediated SLC1A5 induction conferred cisplatin resistance and reduced apoptosis, autophagy, and ferroptosis. Reporter assays and western blot data demonstrated that miR-125b-5p targets and attenuates SLC1A5, while the si-NEAT1 increases miR-125b-5p expression. Analysis of the TCGA-HNSCC databases showed concordant upregulation of NEAT1 and downregulation of miR-125b-5p, along with SLC1A5 upregulation in tumors. Analysis of transcriptomic data revealed that tumors harboring higher SLC1A5 expression had significantly lower immune scores in CD8+, monocytes, and dendritic cells, and higher scores in M0 and M1 macrophages. Disruptions in immune modulation, metabolism, and oxidative stress components were associated with SLC1A5 aberrations in HNSCC. This study concludes that the NEAT1/miR-125b-5p/SLC1A5 cascade modulates diverse activities in oncogenicity, treatment efficacy, and immune cell profiles in head and neck/oral carcinoma.

Theoretical study of a putative adsorption mechanism of arginine and glutamate on goldfish 5.24 and zebrafish Z06: Statistical physics modeling, thermodynamic study, and docking simulation

An adsorption phenomenon putatively involved in the sense of smell of arginine and glutamate amino acids in the goldfish olfactory receptor 5.24 and the zebrafish olfactory receptor Z06 was applied in this study to deeply interpret and characterize the olfaction process at a molecular level. As a result, the experimental concentration-olfactory response curves were adjusted using the mono-layer model with one energy (MLM1E). On the one hand, the physicochemical parameters involved in the chosen model were utilized to microscopically investigate the four olfactory systems. Indeed, the two amino acids were adsorbed via a non-parallel configuration, and the process of adsorption was multi-molecular. The molar adsorption energies, which ranged from 22.09 to 35.97 kJ/mol, could be utilized to define the energetic interaction types between the investigated amino acids and the two tested fish olfactory receptors. The size and adsorption energy heterogeneity of 5.24 and Z06 binding pockets were measured by estimating the pocket site size distributions or PSDs (stereographic characterization) and the adsorption energy distributions or AEDs (energetic characterization). On the other hand, these physicochemical parameters could also be applied to macroscopically investigate the four olfactory systems. Hence, three thermodynamic functions including the adsorption entropy Sa, the internal energy Eint, and the Gibbs free energy G were computed in order to investigate the disorder, the exothermicity, and the spontaneity of the studied systems, respectively. The docking modeling analysis of arginine and glutamate amino acids with goldfish 5.24 and zebrafish Z06 revealed simulated binding energies that closely matched the average adsorption energies (peaks of AEDs). These affinities corresponded to the adsorption energy bands known as olfactory bands or spectrums of vibration modes.

Guangxi sour bamboo shoots: A study on microbial diversity and flavor characteristics across regions

Sour bamboo shoots, traditional salt-free fermented vegetables widely consumed in Guangxi province, are valued for their regionally varied flavor profiles and microbial diversity. This study employed HPLC, HS-SPME–GC–MS, and Illumina MiSeq sequencing to explore both the flavor profiles and the underlying microbial communities of 55 sour bamboo shoots from diverse regions in Guangxi. Our findings reveal the Tropic of Cancer as a pivotal boundary influencing quality divergence. It is noteworthy that three organic acids (succinic acid, acetic acid, and oxalic acid), nine amino acids (alanine, glutamate, glycine, valine, methionine, leucine, isoleucine, phenylalanine, and proline), and three volatile compounds (2-methoxy-4-methylphenol, p-cresol, and acetic acid) were found to be present in higher concentrations in samples from the northern region, suggesting the influence of a distinct terroir effect. Spearman correlation analysis preliminarily linked Lactobacillus, Enhydrobacter, and Lactiplantibacillus to the biosynthesis of key metabolites like p-cresol and 2-methoxy-4-methylphenol, highlighting their significant role in flavor production, although the exact mechanisms require further investigation. This study contributes to our understanding of the regional characteristics of sour bamboo shoots and provides a foundation for future research on the influence of microbiota on flavor. The findings may inform the development of traditional fermented products with a consistent flavor profile across regions.

NMR Fingerprinting of Conventional and Genetically Modified Soybean Plants with AtAREB1 Transcription Factors.

Drought stress impacts soybean yields and physiological processes. However, the insertion of the activated form of the AtAREB1 gene in the soybean cultivar BR16, which is sensitive to water deficit, improved the drought response of the genetically modified plants. Thus, in this study, we used 1H NMR in solution and solid-state NMR to investigate the response of genetically modified soybean overexpressing AtAREB1 under water deficiency conditions. We achieved that drought-tolerant soybean yields high content of amino acids isoleucine, leucine, threonine, valine, proline, glutamate, aspartate, asparagine, tyrosine, and phenylalanine after 12 days of drought stress conditions, as compared to drought-sensitive soybean under the same conditions. Specific target compounds, including sugars, organic acids, and phenolic compounds, were identified as involved in controlling sensitive soybean during the vegetative stage. Solid-state NMR was used to study the impact of drought stress on starch and cellulose contents in different soybean genotypes. The findings provide insights into the metabolic adjustments of soybean overexpressing AREB transcription factors in adapting to dry climates. This study presents NMR techniques for investigating the metabolome of transgenic soybean plants in response to the water deficit. The approach allowed for the identification of physiological and morphological changes in drought-resistant and drought-tolerant soybean tissues. The findings indicate that drought stress significantly alters micro- and macromolecular metabolism in soybean plants. Differential responses were observed among roots and leaves as well as drought-tolerant and drought-sensitive cultivars, highlighting the complex interplay between overexpressed transcription factors and drought stress in soybean plants.

Metabolomic and Proteomic Analysis of ApoE4-Carrying H4 Neuroglioma Cells in Alzheimer's Disease Using OrbiSIMS and LC-MS/MS.

Growing clinical evidence reveals that systematic molecular alterations in the brain occur 20 years before the onset of AD pathological features. Apolipoprotein E4 (ApoE4) is one of the most significant genetic risk factors for Alzheimer's disease (AD), which is not only associated with the AD pathological features such as amyloid-β deposition, phosphorylation of tau proteins, and neuroinflammation but is also involved in metabolism, neuron growth, and synaptic plasticity. Multiomics, such as metabolomics and proteomics, are applied widely in identifying key disease-related molecular alterations and disease-progression-related changes. Despite recent advances in the development of analytical technologies, screening the entire profile of metabolites remains challenging due to the numerous classes of compounds with diverse chemical properties that require different extraction processes for mass spectrometry. In this study, we utilized Orbitrap Secondary Ion Mass Spectrometry (OrbiSIMS) as a chemical filtering screening tool to examine molecular alterations in ApoE4-carried neuroglioma cells compared to wild-type H4 cells. The findings were compared using liquid chromatography (LC)-MS/MS targeted metabolomics analysis for the confirmation of specific metabolite classes. Detected alterations in peptide fragments by OrbiSIMS provided preliminary indications of protein changes. These were extensively analyzed through proteomics to explore ApoE4's impact on proteins. Our metabolomics approach, combining OrbiSIMS and LC-MS/MS, revealed disruptions in lipid metabolism, including glycerophospholipids and sphingolipids, as well as amino acid metabolism, encompassing alanine, aspartate, and glutamate metabolism; aminoacyl-tRNA biosynthesis; glutamine metabolism; and taurine and hypotaurine metabolism. Further LC-MS/MS proteomics studies confirmed the dysfunction in amino acid and tRNA aminoacylation metabolic processes, and highlighted RNA splicing alterations influenced by ApoE4.

Interactions Involving Glycine and Other Amino Acid Neurotransmitters: Focus on Transporter-Mediated Regulation of Release and Glycine-Glutamate Crosstalk.

Glycine plays a pivotal role in the Central Nervous System (CNS), being a major inhibitory neurotransmitter as well as a co-agonist of Glutamate at excitatory NMDA receptors. Interactions involving Glycine and other neurotransmitters are the subject of different studies. Functional interactions among neurotransmitters include the modulation of release through release-regulating receptors but also through transporter-mediated mechanisms. Many transporter-mediated interactions involve the amino acid transmitters Glycine, Glutamate, and GABA. Different studies published during the last two decades investigated a number of transporter-mediated interactions in depth involving amino acid transmitters at the nerve terminal level in different CNS areas, providing details of mechanisms involved and suggesting pathophysiological significances. Here, this evidence is reviewed also considering additional recent information available in the literature, with a special (but not exclusive) focus on glycinergic neurotransmission and Glycine-Glutamate interactions. Some possible pharmacological implications, although partly speculative, are also discussed. Dysregulations in glycinergic and glutamatergic transmission are involved in relevant CNS pathologies. Pharmacological interventions on glycinergic targets (including receptors and transporters) are under study to develop novel therapies against serious CNS pathological states including pain, schizophrenia, epilepsy, and neurodegenerative diseases. Although with limitations, it is hoped to possibly contribute to a better understanding of the complex interactions between glycine-mediated neurotransmission and other major amino acid transmitters, also in view of the current interest in potential drugs acting on "glycinergic" targets.

A pharmacokinetic-pharmacodynamic analysis of l-glutamine for the treatment of sickle cell disease: Implications for understanding the mechanism of action and evaluating response to therapy.

The mechanisms of action of l-glutamine for the treatment of sickle cell disease (SCD) are not well understood and there are no validated clinical biomarkers to assess response. We conducted a three-week, dose-ascending trial of glutamine and measured the pharmacokinetic (PK) exposure parameters, peak concentration (Cmax) and area under the curve (AUC). We used a panel of biomarkers to investigate the pharmacodynamics (PD) of glutamine and studied PK-PD relationships. There was no plasma accumulation of glutamine, glutamate, arginine or other amino acids over time, but modestly improved arginine bioavailability was observed. In standard analysis by dose levels over time, there were no measurable effects on blood counts, viscosity, ektacytometry or reactive oxygen species (ROS). In PK-PD analysis, however, higher glutamine exposure (Cmax or AUC) was associated with increased whole blood viscosity and cellular dehydration, yet also with higher haemoglobin concentration, increased haematocrit-to-viscosity ratio, decreased reticulocyte ROS, improved RBC deformability and decreased point of sickling. This novel PK-PD analysis identified biomarkers reflecting the positive and negative effects of glutamine, helping to elucidate its mechanisms of action in SCD. PK-optimized dosing to achieve glutamine exposure (AUC or Cmax) that is associated with salutary biological effects should be studied to support its therapeutic use.

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.

Soil Microbial Functions Linked Fragrant Rice 2-Acetyl-1-Pyrroline with Soil Active Carbon Pool: Evidence from Soil Metagenomic Sequencing of Tillage Practices

Improved tillage management in fragrant rice cropping systems can enhance soil organic carbon (SOC) and the content of 2-Acetyl-1-Pyrroline (2-AP), a crucial volatile compound contributing to the aroma of fragrant rice. Despite this, the interplay between 2-AP content in fragrant rice and SOC metabolism, alongside the influences exerted by soil microbial functions, remains poorly understood. This study introduces a comprehensive 6-year field experiment which aims to correlate SOC with rice grain 2-AP content by analyzing soil microbial KEGG functions, such as carbon and amino acid metabolism, using metagenomic sequencing. The experiment assessed three tillage practices, conventional tillage (CT), reduced tillage (RT), and no tillage (NT), with soil samples collected on three dates in 2022. The results indicated that NT significantly (p &lt; 0.05) enhanced SOC content and modified carbon metabolism by upregulating the Calvin cycle (K01601) and reducing hemicellulose degradation (K01710). Additionally, NT notably increased the soil levels of alkaline amino acids, such as histidine and ornithine, which were 165.17% and 1218.42% higher, respectively, than those in CT, possibly linked to an increase in soil pH. Furthermore, the 2-AP content in fragrant rice under NT was significantly higher by 52.02% and 13.90% compared to under RT and CT, respectively. NT also upregulated K00250 (alanine, aspartate, and glutamate metabolism) and K00290 (valine, leucine, and isoleucine biosynthesis), leading to significantly higher levels of 2-AP biosynthesis-related amino acids proline and glutamate in fragrant rice grain. This study links SOC and 2-AP biosynthesis via soil microbial functions, presenting a novel strategy for improving the quality of fragrant rice through soil management practices.

Amino Acids Isolation from α-keratin of Javanese Goat (Capra hircus) Hair and Garut Sheep (Ovis aries) Hair Waste Using Acid Hydrolysis Method as BCAA Supplement

Javanese goat and Garut sheep hair contain α-keratin, a protein that can be broken by hydrolysis to produce simpler amino acids. Feather waste generates millions of tons of α-keratin biomass originating from animal slaughterhouses, thereby raising health concerns. The utilization of acid hydrolysis is considered to be more cost-effective compared to enzymatic hydrolysis, and it provides a broader range of amino acid cleavage sites compared to enzymes, which exhibit specific cleavage. This study aimed to isolate amino acids from Javanese goat and Garut sheep hair through acid hydrolysis. The methods included hair sample preparation, acid hydrolysis used 6 M HCL at 110°C, reflux isolation, amino acid separation based on isoelectric pH 4.9 –5.4, functional groups analysis using FTIR, and analysis of amino acid content by HPLC methods. The results showed that the yield produced after isolation on Javanese goat hair samples was 0.92% and Garut sheep hair 0.32%, respectively. The FTIR spectrum showed amino acid functional groups in both samples, including carboxyl (COOH), amine (C-N primer), (C-S disulfide), and amide I (-CONH2). Successful breakdown of α-keratin proteins into simpler amino acids was achieved for Javanese goat and Garut sheep hair. Amino acid analysis of Javanese goat hair isolates revealed the presence of aspartic acid, threonine, serine, glutamate, proline, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, lysine, and arginine amino acids, respectively. The highest content was isoleucine at 0.60% w/w. In conclusion, the isolated amino acids from Javanese goat hair can be used as a halal supplement that serves as nutrition in the body.

Being Stung Once or Twice by Bees (Apis mellifera L.) Slightly Disturbed the Serum Metabolome of SD Rats to a Similar Extent.

In most cases, the number of honeybee stings received by the body is generally small, but honeybee stings can still cause serious allergic reactions. This study fully simulated bee stings under natural conditions and used 1H Nuclear Magnetic Resonance (1H NMR) to analyze the changes in the serum metabolome of Sprague-Dawley (SD) rats stung once or twice by honeybees to verify the impact of this mild sting on the body and its underlying mechanism. The differentially abundant metabolites between the blank control rats and the rats stung by honeybees included four amino acids (aspartate, glutamate, glutamine, and valine) and four organic acids (ascorbic acid, lactate, malate, and pyruvate). There was no separation between the sting groups, indicating that the impact of stinging once or twice on the serum metabolome was similar. Using the Principal Component Discriminant Analysis ( PCA-DA) and Variable Importance in Projection (VIP) methods, glucose, lactate, and pyruvate were identified to help distinguish between sting groups and non-sting groups. Metabolic pathway analysis revealed that four metabolic pathways, namely, the tricarboxylic acid cycle, pyruvate metabolism, glutamate metabolism, and alanine, aspartate, and glutamate metabolism, were significantly affected by bee stings. The above results can provide a theoretical basis for future epidemiological studies of bee stings and medical treatment of patients stung by honeybees.