Search Results (958)

Amino acids (AAs), a type of nitrogen-based organic compounds in the atmosphere, are directly and indirectly related to climate change, and as their link to allergic diseases becomes more known, the need for quantitative analysis of ultrafine dust (PM_2.5) will become increasingly necessary. When sensing water-soluble AAs using a gas chromatograph combined with a tandem mass spectrometer (GC-MS/MS), derivatization should be considered to increase the volatility and sensitivity of target analytes. In this study, two methods were used to compare and evaluate 13 AA derivatives in PM_2.5 samples: N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide with 1% tert-butyldimethylchlorosilane (MTBSTFA w/1% t-BDMCS), which is preferred for silylation, and ethyl chloroformate (ECF) with methanol (MeOH) for chloroformate derivatization. The most appropriate reaction conditions for these two derivative methods, such as temperature and time, and the analytical conditions of GC-MS/MS for the qualitative and quantitative analysis of AAs were optimized. Furthermore, the calibration curve, detection limit, and recovery of both methods for validating the quantification were determined. The two derivative methods were applied to 23 actual PM_2.5 samples to detect and quantify target AAs. The statistical significances between pairwise measurements of individual AAs detected by both methods were evaluated. This study will help in selecting and utilizing appropriate derivative methods for the quantification of individual AAs in PM_2.5 samples. Full article

TasA gene, encoding a functional amyloid protein critical for biofilm formation and antimicrobial activity, was cloned from the endophytic strain Bacillus amyloliquefaciens BS-3, isolated from rubber tree roots. This study identified the shortest functional TasA variant (483 bp, 160 aa) reported to date, featuring unique amino acid substitutions in conserved domains. Bioinformatics analysis predicted a signal peptide (1–27 aa) and transmembrane domain (7–29 aa), which were truncated to optimize heterologous expression. Two prokaryotic vectors (pET28a and pCZN1) were constructed, with pCZN1-TasA expressed solubly in Escherichia coli Arctic Express at 15 °C, while pET28a-TasA formed inclusion bodies at 37 °C. Purified recombinant TasA exhibited potent antifungal activity, achieving 98.6% ± 1.09 inhibition against Colletotrichum acutatum, 64.77% ± 1.34 against Alternaria heveae. Notably, TasA completely suppressed spore germination in C. acutatum and Oidium heveae Steinmannat 60 μg/mL. Structural analysis via AlphaFold3 revealed that truncation enhanced protein stability. These findings highlight BS-3-derived TasA as a promising biocontrol agent, providing molecular insights for developing protein-based biopesticides against rubber tree pathogens. Full article

Tianshanhong (TSH), black tea products originating from the Ningde Tianshan Mountain, has gained significant recognition in the market. However, the chemical characteristics contributing to the flavor of TSH have not yet been reported. To systematically investigate the non-volatile and volatile compounds in TSH, four grades of TSH were evaluated using national standard sensory methods, revealing that overall quality improved with higher grades. Based on the detection of ultra-performance liquid chromatography–mass spectrometry (UPLC-MS), the content of ester-type catechins was relatively high and decreased with lower grades. A total of 19 amino acids (AAs) were clustered, among them, three amino acids, L-Theanine (L-Thea), Arg, and GABA, showed highly significant correlations with the refreshing taste of TSH. Notably, the content of Arg had the highest correlation with TSH grade, with a coefficient of 0.976 (p < 0.01). According to gas chromatography mass spectrometry (GC-MS) analysis, a total of 861 kinds of volatile compounds were detected, with 282 identified and aroma-active compounds across grades selected using the PLS model. Methyl salicylate and geraniol were particularly notable, showing strong correlations with TSH grades at 0.975 and 0.987 (p < 0.01), respectively. Our findings show that non-volatile and volatile compounds can rationally grade TSH and help understand its flavor quality. Full article

Pleurotus eryngii is an edible mushroom with previously characterized β-glucans. Its potential to ameliorate postprandial glycemia and regulate appetite at the postprandial state has been previously shown. However, its effect on protein digestion remains unexplored. We aimed to investigate the effect of baked P. eryngii with a known β-glucan content (4.5 g) on plasma free amino acids of patients with central obesity and metabolic abnormalities at a postprandial state. In this acute, randomized controlled cross-over study, thirteen healthy male volunteers consumed one meal that was prepared with P. eryngii and one control meal; each meal was separated by one month. Blood was collected, and plasma was isolated at different timepoints before and after the consumption. Gas chromatography–mass spectrometry was used to quantify 24 free amino acids in the plasma samples. The area under the curve with respect to increase (AUCi) was computed, and the AUCi for aromatic amino acids was found to be higher after the consumption of the control meal compared to the P. eryngii meal (p = 0.027 for phenylalanine, p = 0.008 for tyrosine, and p = 0.003 for tryptophan). The above novel findings suggest that the β-glucans present in P. eryngii mushrooms are potential modulators of AA release into the bloodstream. Full article

Isotopic analyses are useful tools with a wide range of applications, including environmental studies, archaeology and biomedicine. Founded in 2019 at the University of Campania “Luigi Vanvitelli”, the iCONa laboratory specialises in stable isotope mass spectrometry, with a particular focus on cultural heritage. The laboratory performs carbon, nitrogen and oxygen isotopic analyses, including the most recent advances in compound-specific stable isotope analysis of amino acids (CSIA-AAs). In addition to these analytical services, iCONa provides chemical and physical sample preparation for a variety of sample types. This paper focuses on our applications in the field of cultural heritage, exploring how the analysis of stable isotopes performed on archaeological remains can be used to reconstruct past subsistence strategies and human behaviours. We also discuss the challenges inherent in isotopic analysis and recent methodological advances in the field. Full article

Serotoninomics is an expanding field that focuses on the comprehensive study of the serotoninergic system, including serotonin’s biosynthesis, metabolism, and regulation, as well as related scientific methodologies 5-hydroxytryptamine (5-HT). This field explores serotonin’s complex roles in various physiological and pathological contexts. The essential amino acid tryptophan (Trp) is a precursor for several metabolic and catabolic pathways, with the kynurenine (KYN) pathway being particularly significant, representing about 95% of Trp metabolism. In contrast, only a small portion (1–2%) of dietary Trp enters the serotonin pathway. Anthranilic acid (AA), a metabolite in the KYN pathway, has emerged as a potential biomarker and therapeutic target for schizophrenia. Elevated serum AA levels in patients with schizophrenia have been associated with neurotoxic effects and disruptions in neurotransmission, suggesting AA’s critical role in the disorder’s pathophysiology. Furthermore, the 5-HT_2A receptor’s involvement is particularly noteworthy, especially in relation to schizophrenia’s positive symptoms. Recent findings indicate that 5-HT_2A receptor hyperactivity is linked to positive symptoms of schizophrenia, such as hallucinations and delusions. This study investigates serotoninomics’ implications for neuropsychiatric disorders, focusing on AA in schizophrenia and analysing recent research on serotonin signalling pathways and AA’s neurochemical effects. Understanding the roles of the 5-HT_2A receptor and AA in neuropsychiatric disorders could lead to the development of more precise and less invasive diagnostic tools, specific therapeutic strategies, and improved clinical outcomes. Ongoing research is essential to uncover these pathways’ exact mechanisms and therapeutic potential, thereby advancing personalised medicine and innovative treatments in neuropsychiatry. Full article

A novel barna-like virus was found to be associated with field-collected Afrina sporoboliae plant-parasitic nematodes. The positive-sense, single-stranded RNA genome of this virus, named Afrina barna-like virus (AfBLV), comprises 4020 nucleotides encoding four open reading frames (ORFs). ORF 1 encodes a protein product spanning a transmembrane, a peptidase, and VPg domains, whereas an overlapping ORF 2 encodes an RNA-dependent RNA polymerase (RdRP). ORF2 may be expressed via a −1 translational frameshift. In phylogenetic reconstructions, the RdRP of AfBLV was placed inside a separate clade of barna and barna-like viruses related to but distinct from the genera in the Solemoviridae and Alvernaviridae families, within the overall lineage of Sobelivirales. ORF 3 of AfBLV encodes a protein product of 206 amino acids (aa) long with homology to a putative protein encoded by a similarly positioned gene of an uncharacterized virus sequence identified previously as Barnaviridae sp. ORF 4 encodes a 161 aa protein with no significant similarities to sequences in the GenBank databases. AfBLV is the first barnavirus found in a nematode. Sequence comparisons of the AfBLV genome and genomes of other barna-like viruses suggested that a recombination event was involved in the evolution of AfBLV. Analyses of the phylogeny of RdRPs and genome organizations of barna-like and solemo-like viruses support the re-classification of Barnavirus and Dinornavirus genera as members of the Solemoviridae family. Full article

The metabolic hyperactivity of tumor cells demands a substantial amount of energy and molecules to build new cells and expand the tumor, diverting these resources from healthy cells. Amino acids (AAs) are the only totipotent and essential molecules for protein construction. Previous in vitro studies in human and murine cancer cells, along with in vivo studies in mice, have shown that an excess of essential amino acids (EAAs) exerts an inhibitory effect on tumor proliferation by promoting apoptosis and autophagy. In this study, both in vitro and in vivo, we evaluated whether a mixture based on EAA can influence the development of human colon cancer (HCT116). To this end, in vitro, we assessed the proliferation of HCT116 cells treated with a special mix of EAA. In vivo, immunosuppressed athymic nude mice, injected with HCT116 cells subcutaneously (s.c.) or intraperitoneally (i.p.), were given a modified EAAs-rich diet (EAARD) compared to the standard laboratory diet (StD). In vitro data showed that the EAA mix impairs cancer growth by inducing apoptosis and autophagy. In vivo, the results demonstrated that EAARD-fed mice developed s.c. tumors significantly smaller than those of StD-fed mice (total mass 3.24 vs. 6.09 g, respectively). Mice injected i.p. and fed with EAARD showed a smaller and more limited number of intra-peritoneal tumors than StD-fed mice (total mass 0.79 vs. 4.77 g, respectively). EAAs prevents the growth of HCT116 cells by inducing autophagy and apoptosis, increasing endoplasmic reticulum stress, and inhibiting inflammation and neo-vascularization. In addition, the EAARD-fed mice, maintained muscle mass and white and brown adipose tissues. A diet with an excess of EAAs affects the survival and proliferative capacity of human colon cancer cells, maintaining anabolic stimuli in muscular cells. Full article

Optimizing the metabolizable protein level in ruminant diets represents a promising strategy to increase nitrogen use efficiency and mitigate environmental pollution. This study explored the impacts of varying metabolizable protein (MP) levels on amino acid (AA) balance, nitrogen (N) utilization, and the ruminal microbiota in Hu lambs. Fifty-four female Hu lambs of 60 d old, with an average body weight (BW) of 18.7 ± 2.37 kg, were randomly allocated to three dietary MP groups: (1) low MP (LMP, 7.38% of DM), (2) moderate MP (MMP, 8.66% of DM), and (3) high MP (HMP, 9.93% of DM). Three lambs with similar BW within each group were housed together in a single pen, serving as one experimental replicate (n = 6). The feeding trial lasted for 60 days with 10 days for adaptation. The final BW of lambs in the MMP and HMP groups increased (p < 0.05) by 5.64% and 5.26%, respectively, compared to the LMP group. Additionally, lambs fed the MMP diet exhibited an 11.6% higher (p < 0.05) average daily gain than those in the LMP group. Increasing dietary MP levels enhanced (p < 0.05) N intake, urinary N, retained N, and percent N retained, but decreased apparent N digestibility (p < 0.05). Urinary uric acid, total purine derivatives, intestinally absorbable dietary protein, microbial crude protein, intestinally absorbable microbial crude protein, and actual MP supply all increased (p < 0.05) with higher MP values in the diet. The plasma concentrations of arginine, lysine, methionine, phenylalanine, threonine, aspartic acid, proline, total essential AAs, and total nonessential AAs were the lowest (p < 0.05) in the LMP group. In the rumen, elevated MP levels led to a significant increase (p < 0.05) in the ammonia N content. The relative abundances of Candidatus_Saccharimonas, Ruminococcus, and Oscillospira were the lowest (p < 0.05), whereas the relative abundances of Terrisporobacter and the Christensenellaceae_R-7_group were the highest (p < 0.05) in the MMP group. In conclusion, the moderate dietary metabolizable protein level could enhance growth performance, balance the plasma amino acid profiles, and increase nitrogen utilization efficiency in Hu lambs, while also altering the rumen bacterial community by increasing beneficial probiotics like the Christensenellaceae_R-7_group. Full article

The integration of HS-SPME-GC/MS and UPLC-MS/MS techniques enabled the profiling of volatile organic compounds (VOCs) and amino acids (AAs) in 18 crabapple flower cultivars, facilitating the development of a novel VOC–AA model. Among the 51 identified VOCs, benzyl alcohol, benzaldehyde, and ethyl benzoate were predominant, categorizing cultivars into fruit-almond, fruit-sweet, and mixed types. The amino acids, namely glutamic acid (Glu), asparagine (Asn), aspartic acid (Asp), serine (Ser), and alanine (Ala) constituted 83.6% of the total AAs identified. Notably, specific amino acids showed positive correlations with key VOCs, suggesting a metabolic regulatory mechanism. The Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) model, when combined with volatile organic compounds (VOCs) and amino acid profiles, enabled more effective aroma type classification, providing a robust foundation for further studies on aroma mechanisms and targeted breeding. Full article

Insects and their endosymbionts have a close mutualistic relationship. However, the precise nature of the bacterial endosymbiont-mediated interaction between host plants and whitefly Bemisia tabaci MEAM1 is still unclear. In the present study, six populations of Bemisia tabaci MEAM1 sharing the same genetic background were established by rearing insects for ten generations on different host plants, including poinsettia, cabbage, cotton, tomato, and tobacco, and an additional population was reared on cotton and treated with antibiotics. The physiological and nutritional traits of the insects were found to be dependent on the host plant on which they had been reared. Systematic analysis was conducted on the endosymbiont titers, the amino acid molecules and contents, as well as developmental and oviposition changes in the MEAM1 populations reared on each host plant tested. The results indicate that B. tabaci contained the primary symbiont Portiera and the secondary symbionts Hamiltonella and Rickettsia. In addition, the titer of endosymbiotic bacteria in females is higher than that in males. Among the MEAM1 populations reared on each host plant, the variation pattern of Portiera titer generally corresponded with changes in biological characteristics (body length, weight and fecundity) and AA contents. This suggests that changes in the amino acid contents and biological characteristics of different B. tabaci populations may be due to changes in the Portiera content and the differences in the nutrition of the host plants themselves. Our findings were further confirmed by the reduction in Portiera with antibiotic treatment. The amino acids, body size, body weight, and fecundity of B. tabaci were all reduced with the decrease in the Portiera titer after antibiotic treatment. In summary, our research revealed that host plants can affect the content of symbiotic bacteria, particularly Portiera, and subsequently affect the nutrition (i.e., the essential amino acids content) of host insects, thus changing their biological characteristics. Full article

Caffeic acid-O-methyltransferase (COMT) is a key enzyme in lignin synthesis and secondary metabolism in plants, and it participates in the regulation of plant growth and development as well as plants’ stress response. To further investigate the function of COMT in grapevine, a total of 124 COMT family genes were identified from three Vitis species in this study, namely Pinot noir (Vitis vinifera L.), Vitis amurensis, and Vitis riparia. The amino acid sequence encoded by these genes ranged from 55 to 1422 aa, and their molecular mass ranged from 6640.82 to 77,034.43 Da. Subcellular localization prediction inferred that they were mainly located in the plasma membrane and cytoplasm. The prediction of secondary structures showed that α-helix and irregular coiled-coil were primary structural elements. These genes were unevenly distributed across 10 different chromosomes, respectively. Phylogenetic tree analysis of the amino acid sequences of VvCOMT, VaCOMT, VrCOMT, and AtCOMT proteins showed that they were closely related and were divided into four subgroups. The motif distribution was similar among the cluster genes, and the gene sequence was notably conserved. The 124 members of the COMT gene family possessed a variable number of exons, ranging from 2 to 13. The promoter region of all of these COMTs genes contained multiple cis-acting elements related to hormones (e.g., ABA, IAA, MeJA, GA, and SA), growth and development (e.g., endosperm, circadian, meristem, light response), and various stress responses (e.g., drought, low temperature, wounding, anaerobic, defense, and stress). The intraspecies collinearity analysis suggested that there were one pair, three pairs, and six pairs of collinear genes in Va, Pinot noir, and Vr, respectively, and that tandem duplication contributed more to the expansion of these gene family members. In addition, interspecific collinearity revealed that the VvCOMTs had the strongest homology with the VaCOMTs, followed by the VrCOMTs, and the weakest homology with the AtCOMTs. The expression patterns of different tissues and organs at different developmental stages indicated that the VvCOMT genes had obvious tissue expression specificity. The majority of VvCOMT genes were only expressed at higher levels in certain tissues. Furthermore, we screened 13 VvCOMT genes to conduct qRT-PCR verification according to the transcriptome data of VvCOMTs under abiotic stresses (NaCl, PEG, and cold). The results confirmed that these genes were involved in the responses to NaCl, PEG, and cold stress. This study lays a foundation for the exploration of the function of the COMT genes, and is of great importance for the genetic improvement of abiotic stress resistance in grapes. Full article

Background: Symbiotic gut microbiota can enhance cancer therapy efficacy, while treatment-induced dysbiosis may reduce effectiveness or increase toxicity. Our preclinical study compared the anticancer effects and impact on fecal microbiota and metabolites of two water-soluble SN-38 derivatives (BN-MePPR and BN-MOA), with those observed after treatment with Irinotecan, and the FOLFOX regimen in NOD scid gamma mice bearing patient-derived colon adenocarcinoma xenografts (CRC PDX). Methods: Five individual experiments with Irinotecan and its derivatives and eight individual experiments with FOLFOX were conducted using eight CRC PDX models. Chemotherapeutics were administered intraperitoneally 4–5 times at 5-day intervals. Fecal samples were collected before and after treatment. Microbiota composition was analyzed by 16S rRNA gene (V3–V4 regions) sequencing. Mass spectrometry was used to quantify short-chain fatty acids (SCFAs) and amino acids (AAs). Results: All treatments significantly inhibited tumor growth versus controls. However, no significant changes were observed in gut microbiota α- and β-diversity between treated and untreated groups. Tumor progression in controls was associated with increased abundance of Marvinbryantia, Lactobacillus, Ruminococcus, and [Eubacterium] nodatum group. FOLFOX-treated mice showed increased Marvinbryantia, Bacteroides, and Candidatus Arthromitus, and decreased Akkermansia. No distinct taxa changes were found in the Irinotecan or derivative groups. SCFA levels remained unchanged across groups, while BN-MePPR, BN-MOA, and Irinotecan all increased AA concentrations. Conclusions: Contrary to earlier toxicological data, these findings indicate a relatively limited impact of the tested chemotherapeutics on the gut microbiome and metabolome, emphasizing the importance of research method selection in preclinical studies. Full article

Background/Objectives: Gestational diabetes mellitus (GDM) represents an increased metabolic risk in future life for both mother and child. We hypothesize free fatty acids (FFAs) and amino acids (AAs) disturbances in plasma during second trimester might be indicating high risk of persisting glucose intolerance (PGI). The aim of study was to determine plasma FFAs and AAs during pregnancy in women with normal pregnancy and GDM and also in post-GDM women with PGI after delivery and to find potential association of altered FFAs and AAs profile with adverse peripartal outcomes and PGI after GDM. Material and Methods: A total of 54 pregnant women were included in the study. Of those 34 participants had GDM. PGI was diagnosed by oGTT up to one year after delivery. Plasma FFAs were determined using GC-FID and plasma AAs levels were determined using CE-MS method. Results: Decreased levels of tetradecanoic acid and several AAs were found in GDM group during pregnancy. Oleic and docosahexaenoic acid correlated positively while almost all AAs negatively correlated with oGTT values in the pregnancy (all p < 0.05, Spearman). Logistic regression model (using AAs, FFAs and BMI) identified higher citrulline and glutamate levels and lower tetradecenoic acid and choline as the best predictors for postpartum PGI. Some differences in AA levels were detected in women with macrosomic babies. Conclusions: Data support a possible link between GDM development and PGI after delivery and selected metabolite levels. The predictive potential of plasma FFAs and AAs levels on a diabetes risk in future life requires further validation. Full article

The aim of this study was to explore the effects of different lactic acid bacteria additives (Lactiplantibacillus plantarum or Lentilactobacillus buchneri) on the fermentation quality, chemical composition, in vitro digestibility, bacterial community structure, and predictive function of S. perfoliatum silage feed. Fresh S. perfoliatum was wilted overnight, then its moisture content was adjusted between 65 and 70%. The experiment was performed in three groups as follows: (1) the control group (CK group), which lacked a Lactobacillus preparation; (2) the Lactiplantibacillus plantarum (L. plantarum) group (LP group), which was inoculated with L. plantarum at 5 × 10⁶ cfu/g FW; and (3) the Lentilactobacillus buchneri (L. buchneri) group (LB group), which was inoculated with L. buchneri at 5 × 10⁶ cfu/g FW. The results showed that L. plantarum significantly reduced pH and increased lactic acid (LA) content in S. perfoliatum silage compared with the control. L. buchneri, on the other hand, excelled in reducing ammonia nitrogen (NH₃-N) content and significantly increased acetic acid (AA) content. At 60 days of fermentation, the CP content was significantly higher (p < 0.05) in the LP and LB groups than in the CK group (19.29 vs. 15.53 and 15.87). At 60 days of fermentation, the ivCPD was significantly higher (p < 0.05) in the LB group than in the CK and LP groups (57.80 vs. 54.77 and 55.77). The 60-day silage process completely altered the bacterial community of S. perfoliatum silage. In the fresh samples, the dominant genera were Weissella_A and Pantoea_A. Weissella_A and Pantoea_A were gradually replaced by Lentilactobacillus and Lactiplantibacillus after S. perfoliatum ensiling. After 45 days of fermentation, L. buchneri became the dominant strain in CK, LP and LB groups. Inoculation with L. plantarum altered the succession of the bacterial community from 7 to 15 days of fermentation of S. perfoliatum. In contrast, inoculation with L. buchneri affected the succession of the bacterial community from 30 to 60 days of S. perfoliatum fermentation. In S. perfoliatum silage aged 7 to 60 days, the amino acid metabolic pathway in the LB group remained upregulated. The experimental results revealed that inoculation with L. buchneri had a stronger effect on S. perfoliatum silage than inoculation with L. plantarum. Thus, L. buchneri should be selected as an additive for S. perfoliatum silage fermentation in practical production. Full article