Liquid Chromatography-mass Spectrophotometry Research Articles

Postharvest blown-arc plasma treatment did not reduce Fusarium graminearum or its mycotoxins in field-infested wheat grain

The increasing prevalence of Fusarium graminearum, a major causative pathogen of Fusarium head blight in cereal crops, leads to contamination of grain with mycotoxins, which necessitates the development of effective management strategies. This study investigated whether postharvest Blown-Arc plasma treatment could reduce F. graminearum and associated mycotoxins in field-infested grain. Postharvest Blown-Arc plasma treatment for 60 or 180 s was trialled to reduce Fusarium graminearum and deoxynivalenol (DON) mycotoxins in field-infested wheat grain. Changes in fungal components of the grain microbiome following the treatment were assessed using metabarcoding analysis of the nuclear ribosomal internal transcribed spacer gene region, and liquid chromatography mass spectrophotometry was used to measure changes in concentrations of the mycotoxins deoxynivalenol (DON), DON-3-glucoside (DON3-G) and 3-acetyl-DON (3-ADON). Fusarium spp. were the most common taxa in the grain over two seasons, with the relative abundance being higher in 2020 (72%) compared to 2021 (45%). Postharvest plasma treatment of field-infested grain for 60 or 180 s did not significantly lower Fusarium abundance. Alpha and beta diversity of the natural mycobiota in the grain were also unaffected. The concentrations of mycotoxins were not significantly different after the treatments, except that in the 2021 trial, the 180 s treatment caused a significant increase in DON concentration. We concluded that the postharvest plasma treatment used in this study did not reduce Fusarium contamination or the concentration of mycotoxins, DON, DON3-G and 3-ADON in field-infested grain. The results highlight the importance of using field-infested grain in experiments, as opposed to grain inoculated under laboratory conditions, which provides only superficial infection. This approach is crucial for accurately understanding the dynamics of treatment efficacy.

Metformin-induced oxidative stress inhibits LNCaP prostate cancer cell survival.

Preclinical and clinical studies over the past several decades have indicated the potential value of metformin, a widely utilized treatment for Type 2 diabetes, in prostate cancer therapy. Notably, these studies demonstrated metformin's pleiotropic effects on several molecular and metabolic pathways, such as androgen signaling, cell cycle, and cellular bioenergetics. In this study we investigated the role of metformin in regulating intracellular redox status and cell survival in LNCaP prostate cancer cells. The cytotoxic effects of metformin with or without the presence of SBI0206965 (AMPK inhibitor) on LNCaP cells were determined using MTT and trypan blue exclusion assays. Seahorse XP extracellular analysis, Liquid Chromatography/ Mass Spectrophotometry (LC/MS), and 2,7- and Dichlorofluoresin diacetate (DCFDA) assay were used to assess the effects of metformin on cellular bioenergetics, redox status, and redox-related metabolites. mRNA expression and protein concentration of redox-related enzymes were measured using Real Time-qPCR and ELISA assay, respectively. Independently of AMP-activated protein kinase, metformin exhibited a dose- and time-dependent inhibition of LNCaP cell survival, a response mitigated by glutathione or N-acetylcysteine (ROS scavengers) treatment. Notably, these findings were concomitant with a decline in ATP levels and the inhibition of oxidative phosphorylation. The results further indicated metformin's induction of reactive oxygen species, which significantly decreased glutathione levels and the ratio of reduced to oxidized glutathione, as well as the transsulfuration metabolite, cystathionine. Consistent with an induction of oxidative stress condition, metformin increased mRNA levels of the master redox transcription factor Nrf-2 (nuclear factor erythroid-derived 2-like), as well as transsulfuration enzymes cystathionine beta-synthase and cystathionase and GSH synthesis enzymes γ-glutamylcysteine synthetase and glutathione synthetase. Our findings highlight multiple mechanisms by which metformin-induced formation of reactive oxygen species may contribute to its efficacy in prostate cancer treatment, including promotion of oxidative stress, Nrf2 activation, and modulation of redox-related pathways, leading to its anti-survival action.

Assessing malathion residue impact on poultry health, human safety, and production performance

Over the past decade, food safety has become a major concern due to the intensive use of pesticides. Pesticide contamination has been observed in poultry products when seeds are coated with pesticides or when stored products are exposed to pesticides in warehouses. In this experiment, the residue levels of malathion transferred from corn grain to the different parts of the chicken product, its transfer factors (TFs) and the human dietary risk for consumers were evaluated. Growth performance and carcass parameters of the chicken samples were also determined after different doses of malathion exposure. Malathion residues from different parts of chicken meat (breast, thigh, wing, liver and skin) were extracted by the QuEChERS method and analyzed by liquid chromatography-mass spectrophotometry (LC-MS/MS). A deterministic approach was used to calculate the acute and chronic risk assessment. Body weight, feed conversion ratio and feed intake decreased with increasing malathion dose. In addition to reduced feed intake, cold carcass and liver weights of the chicks were also decreased. The highest residues were found in the skin of the chicken followed by the breast, thigh, wing and liver. The TFs of malathion varied between 0.00 and 0.05 according to the different doses applied (4 mg/kg, 8 mg/kg, 16 mg/kg, 32 mg/kg). The chronic exposure assessment (HQ) showed that consumers of all ages and genders consumed 0.008–0.604% of the acceptable daily intake (0.3 mg/kg body weight (bw)/day) of malathion from chicken products. The acute intake assessment (aHQ) of consumers ranged from 0.00015 to 0.0135% of the acute reference dose (0.3 mg/kg bw). In conclusion the results suggest that the risk associated with the malathion residues in chicken meat was found to be low but the residue levels in meat should not be ignored.

Identification and Toxicity Profiling of Column Fractions of Ethanol Leaf Extract of Ziziphus mauritiana

Different plants may contain varying amounts of phytochemicals and also different degrees of toxicity. This study aimed to evaluate the phytochemicals present in the most active column fraction of ethanol leaf extract of the Ziziphus mauritiana plant and their toxic effects using brine shrimp lethality assay and animal model. Crude ethanol extract was obtained by maceration while fraction was achieved using a column chromatography experiment. Toxicity was evaluated with brine shrimp lethality assay and albino rat’s models while characterization was achieved with liquid chromatography-mass spectrophotometry. Ziziphus mauritiana leaves revealed 42 fractions pooled into seven fractions. Fraction three (3) was the most toxic with the brine shrimp lethality assay of (31.48ug/ml) and its toxicological evaluation revealed an adverse effect on the hematological parameter, biochemical indices, and histo-architecture of the liver and kidney of the experimental model studied. LCMS analysis of the most toxic fraction revealed the presence of Antirrhinoside, Lucidumol A, Apigenin 7-glucuronide-4'-(6''-malonylglucoside), Dioscoreside C, Camellioside D, and others which have been reported for various pharmacological effects including adverse effects, The mode of toxicity may be synergistic, individual, or antagonistic which may explain the moderate toxicity observed in animal model. Administration of these fractions may lead to toxicity despite their potential.

Association between altered tryptophan metabolism, plasma aryl hydrocarbon receptor agonists, and inflammatory Chagas disease.

Chagas disease causes a cardiac illness characterized by immunoinflammatory reactions leading to myocardial fibrosis and remodeling. The development of Chronic Chagas Cardiomyopathy (CCC) in some patients while others remain asymptomatic is not fully understood, but dysregulated inflammatory responses are implicated. The Aryl hydrocarbon receptor (AhR) plays a crucial role in regulating inflammation. Certain tryptophan (Trp) metabolites have been identified as AhR ligands with regulatory functions. We investigated AhR expression, agonist response, ligand production, and AhR-dependent responses, such as IDO activation and regulatory T (Treg) cells induction, in two T. cruzi-infected mouse strains (B6 and Balb/c) showing different polymorphisms in AhR. Furthermore, we assessed the metabolic profile of Trp catabolites and AhR agonistic activity levels in plasma samples from patients with chronic Chagas disease (CCD) and healthy donors (HD) using a luciferase reporter assay and liquid chromatography-mass spectrophotometry (LC-MS) analysis. T. cruzi-infected B6 mice showed impaired AhR-dependent responses compared to Balb/c mice, including reduced IDO activity, kynurenine levels, Treg cell induction, CYP1A1 up-regulation, and AhR expression following agonist activation. Additionally, B6 mice exhibited no detectable AhR agonist activity in plasma and displayed lower CYP1A1 up-regulation and AhR expression upon agonist activation. Similarly, CCC patients had decreased AhR agonistic activity in plasma compared to HD patients and exhibited dysregulation in Trp metabolic pathways, resulting in altered plasma metabolite profiles. Notably, patients with severe CCC specifically showed increased N-acetylserotonin levels in their plasma. The methods and findings presented here contribute to a better understanding of CCC development mechanisms and may identify potential specific biomarkers for T. cruzi infection and the severity of associated heart disease. These insights could be valuable in designing new therapeutic strategies. Ultimately, this research aims to establish the AhR agonistic activity and Trp metabolic profile in plasma as an innovative, non-invasive predictor of prognosis for chronic Chagas disease.

Predicting Clinically Significant Prostate Cancer Using Urine Metabolomics via Liquid Chromatography Mass Spectrometry.

Biomarkers predicting clinically significant prostate cancer (sPC) before biopsy are currently lacking. This study aimed to develop a non-invasive urine test to predict sPC in at-risk men using urinary metabolomic profiles. Urine samples from 934 at-risk subjects and 268 treatment-naïve PC patients were subjected to liquid chromatography/mass spectrophotometry (LC-MS)-based metabolomics profiling using both C18 and hydrophilic interaction liquid chromatography (HILIC) column analyses. Four models were constructed (training cohort [n=647]) and validated (validation cohort [n=344]) for different purposes. Model I differentiates PC from benign cases. Models II, III, and a Gleason score model (model GS) predict sPC that is defined as National Comprehensive Cancer Network (NCCN)-categorized favorable-intermediate risk group or higher (Model II), unfavorable-intermediate risk group or higher (Model III), and GS ≥7 PC (model GS), respectively. The metabolomic panels and predicting models were constructed using logistic regression and Akaike information criterion. The best metabolomic panels from the HILIC column include 25, 27, 28 and 26 metabolites in Models I, II, III, and GS, respectively, with area under the curve (AUC) values ranging between 0.82 and 0.91 in the training cohort and between 0.77 and 0.86 in the validation cohort. The combination of the metabolomic panels and five baseline clinical factors that include serum prostate-specific antigen, age, family history of PC, previously negative biopsy, and abnormal digital rectal examination results significantly increased AUCs (range 0.88-0.91). At 90% sensitivity (validation cohort), 33%, 34%, 41%, and 36% of unnecessary biopsies were avoided in Models I, II, III, and GS, respectively. The above results were successfully validated using LC-MS with the C18 column. Urinary metabolomic profiles with baseline clinical factors may accurately predict sPC in men with elevated risk before biopsy.

Defect-engineered Ni-Fe-LDH with porous flower-like architecture enables boosted advanced oxidation processes: Experimental and theoretical studies

In this paper, oxygen vacancies (VO) enriched flower-like Ni-Fe layered double hydroxide (VO-Ni-Fe-LDH) was prepared for activating peroxymonosulfate (PMS) to remove Tetracycline hydrochloride (TC). Sufficient experimental exploration and density functional theory (DFT) calculations clearly demonstrated the 3D morphology prevented the aggregation of VO-Ni-Fe-LDH and VO alter the electronic state of the surface. Thus, VO-Ni-Fe-LDH/PMS not only showed a 4 times enhancement in the reaction rate constant in comparison to Ni-Fe-LDH/PMS, but also exhibited a stable and efficient performance in continuous flow experiment up to 4.5 h. Quenching test and EPR experiments proved that degradation of TC was dominated by singlet oxygen (1O2), rather than sulfate radicals or hydroxyl radicals found by traditional LDH catalysts. The Chemical intermediates in TC oxidation process were identified through liquid chromatography-mass spectrophotometry, and degradation paths were provided for system optimization. In application, the VO-Ni-Fe-LDH/PMS system was able to withstand the effects of the water matrix, various pH values and remove the contaminants from real hospital wastewater with efficiency. This work emphasized the significance of rational architectural design combined with defect engineering in catalyst development, which holds the potential for advanced catalysis.

Mammalian cysteine-rich secretory proteins interact with plasma membrane Ca2+ exporter PMCA4b.

Mammalian cysteine-rich secretory proteins (CRISPs) are predominantly expressed in the male reproductive tract. Knockout mice lacking two or more CRISPs show defects in sperm transport, sperm-egg interaction and Ca2+ homeostasis. CRISPs play redundant and specific roles via their binding partners. To understand this, a comprehensive analysis of CRISP interactome needs to be undertaken. This study aimed to analyse CRISP4 binding partners on the plasma membrane of rat caudal spermatozoa. Total proteins from rat caudal spermatozoa were subjected to immunoprecipitation using anti-CRISP4 antibody followed by liquid chromatography-mass spectrophotometry analysis. Plasma membrane localised proteins were shortlisted, and a key target was validated by co-immunoprecipitation and co-localisation. Co-transfection followed by co-immunoprecipitation was carried out for studying the interaction of full-length as well as deletion mutants of CRISPs with human plasma membrane calcium ATPase, isoform b (hPMCA4b). Calcium assays were performed using Fura-2-AM. The cholesterol binding ability of different CRISPs was evaluated in silico. The membrane-specific interactome of rat CRISP4 (rCRISP4) from caudal spermatozoa revealed PMCA4b as a novel binding partner, and their interaction was validated in rat spermatozoa. Human CRISP1 (hCRISP1) and hCRISP3 also interacted with PMCA4b via the N-terminal domain. Interestingly, hCRISP1 and rCRISP4 delayed PMCA4b-mediated calcium extrusion but hCRISP3 did not. In silico analysis demonstrated that hCRISP1 and rCRISP4 have higher binding affinity towards cholesterol than hCRISP3. The secretion profile of different CRISPs also showed that the ratio of secreted to cell-associated proteins was highest for hCRISP3. Our study identifies PMCA4b as a target of multiple mammalian CRISPs and unravels a new role of CRISPs in regulating calcium homeostasis. Differences in the interaction of different CRISPs with cholesterol may regulate their enrichment in the lipid rafts and redistribution in the membrane post-capacitation, thereby affecting their interaction with PMCA4b.

Biochemical profiling of ‘toko’ (Livistona jenkinsiana griff.): An endangered underutilized fruit of north east India

Livistona jenkinsiana Griff., an endangered multi-purpose palm, is mostly confined to north-eastern India, cultivated and conserved for its edible fruit and other parts by tribal farmers of the state of Arunachal Pradesh. However, the nutritional composition of fruit is not known. Therefore, proximate estimation and metabolite profiling of fruits of this palm were carried out using spectrophotometry, gas chromatography-tandem mass spectrometry (GC-MS/MS), and liquid chromatography-mass spectrophotometry (LC-MS/MS). The major fatty acids were palmitic acid and oleic acid (approximately 45% and 33%, respectively). The purplish peel was due to malvidin (2032.6 μg/g), an anthocyanin. In the fruit pulp, the predominant sugar was fructose (158.7 μg/g); organic acid was citric acid (3856.2 μg/g); phenol was gentisic acid (660.8 μg/g); and flavonoid was rutin (115.8 μg/g). The bioactive compounds present in the fruit confer several health benefits, making L. jenkinsiana a potentially important species in terms of both nutrition and financial returns.

Proximate Analysis, Phytochemical Screening and High Resolution Liquid Chromatography Mass Spectrophotometry Analysis of Essential Oils of Hedychium Coronarium J. Koenig, a Medicinal and Aromatic Plant

Medicinal plants have been used in all cultures as a source of medicine since ages. When parts of plants such as rhizomes, leaves or barks and other natural materials are used as drugs to treat illnesses, they are called crude drugs and the study of crude drugs is called pharmacognosy. Proximate analysis in plants gives valuable information and help to assess the quality of a crude drug. Beneficial effects of crude drugs are believed to be attributed to plant phytochemicals. Hedychium coronarium J. Koenig, a medicinal and aromatic plant of high value is an endangered and red-listed plant. The rhizome of this plant is used for the treatment of various diseases. Crude extracts prepared from the rhizomes show antibacterial and antifungal properties. Essential oils obtained from Hedychium coronarium are found to be rich in terpenes and used for bactericidal, fungicidal, medicinal and cosmetic applications. The present study assesses the quality of crude drugs prepared from this highly medicinal plant. Proximate analysis and phytochemical screening of rhizomes of Hedychium coronarium, which is frequently consumed as food and as medicine were carried out. Ash content was found to be low) when compared to the moisture content. The water extractive value was more than alcohol extractive. Preliminary tests carried out on phytochemicals revealed the presence of terpenoids and oils. High Resolution Liquid Chromatography Mass Spectrophotometry was carried to ascertain the different components of essential oils and revealed the presence of eucalyptol (1,8-cineole), caryophyllene oxide, camphor, linoleic acid, ricinin, phloroglucinol, 6-gingerol, carvone and arjungenin.

Comparative metabolomics analysis shows key metabolites as potential biomarkers for selection of beef fat colour

Context Fat colour is one of the most important economic traits in the marketing of beef. There are many factors that affect fat colour, such as breed, age, diet and gender. Fat colour is observed in different ranges of colours, including white, yellow and brown. The main issue with improving fat colour is that consumer preferences of fat colour vary across the globe. Therefore, investigating the metabolic mechanisms of fat colour may provide new biomarkers for phenotyping, so as to develop effective selection strategies to achieve the locally desired fat colour. Aims This study aimed to perform a comparative metabolic analysis between white and yellow fat from crossbred cattle so as to identify potential biomarkers for the selection of fat colour and to better understand the metabolism of white and yellow fat depots. Methods Carcass samples of subcutaneous fat were collected from crossbred cattle (Simmental × Mongolian cattle) and scored for fat colour. Liquid chromatography–mass spectrophotometry analysis of extracted metabolites from the subcutaneous fat of six animals with white fat and six animals with yellow fat was performed. Key results The comparison between metabolites of white and yellow fat colour samples indicated that there were five categories of 235 significant metabolites, which included hydrocarbons, lipids and lipid-like molecules, organic acids and their derivatives, organic oxygen compounds and organoheterocyclic compounds. The principal-component analysis illustrated that yellow and white fat samples can be classified in groups; however, the metabolites of white fat samples showed greater variation than those in the yellow fat. In the white fat, there were 163 metabolites that had a higher relative abundance than in yellow fat and 72 that had a lower relative abundance than in yellow fat. 3-Hydroxyoctanoic acid, anethofuran, 9,10-DiHODE, furanoeremophilane, pregeijerene, N-glycolylneuraminic acid, and glycocholic acid were identified as the metabolites that differed the most in abundance between the white and yellow fat samples. Conclusions This study has provided insights into the metabolic differences between white and yellow fat depots and identified key metabolites associated with beef fat colour. Implications This study has provided potential biomarkers that may be useful for selection of beef fat colour in live animals.

Cardiolipin released by microglia can act on neighboring glial cells to facilitate the uptake of amyloid-β (1–42)

Cardiolipin is a mitochondrial phospholipid that is also detected in serum inferring its extracellular release; however, this process has not been directly demonstrated for any of the brain cell types. Nevertheless, extracellular cardiolipin has been shown to modulate several neuroimmune functions of microglia and astrocytes, including upregulation of their endocytic activity. Low cardiolipin levels are associated with brain aging, and may thus hinder uptake of amyloid-β (Αβ) in Alzheimer's disease. We hypothesized that glial cells are one of the sources of extracellular cardiolipin in the brain parenchyma where this phospholipid interacts with neighboring cells to upregulate the endocytosis of Αβ. Liquid chromatography-mass spectrophotometry identified 31 different species of cardiolipin released from murine BV-2 microglial cells and revealed this process was accelerated by exposure to Aβ42. Extracellular cardiolipin upregulated internalization of fluorescently-labeled Aβ42 by primary murine astrocytes, human U118 MG astrocytic cells, and murine BV-2 microglia. Increased endocytic activity in the presence of extracellular cardiolipin was also demonstrated by studying uptake of Aβ42 and pHrodo™ Bioparticles™ by human induced pluripotent stem cells (iPSCs)-derived microglia, as well as iPSC-derived human brain organoids containing microglia, astrocytes, oligodendrocytes and neurons. Our observations indicate that Aβ42 augments the release of cardiolipin from microglia into the extracellular space, where it can act on microglia and astrocytes to enhance their endocytosis of Aβ42. Our observations suggest that the reduced glial uptake of Aβ due to the decreased levels of cardiolipin could be at least partially responsible for the extracellular accumulation of Aβ in aging and Alzheimer's disease.

Phytoconstituent estimation and LC-MS studies of field grown Withania somnifera (L.) Dunal root extract in Jharkhand and Bihar

Plants in either raw form or their root extracts are utilized as complementary and alternative medicine in various disorders. The present study was undertaken for phytoconstituents estimation and to evaluate chemical compositions of methanolic extract of Withania somnifera (L.) Dunal (Ashwagandha) roots collected from BAU, Ranchi, Jharkhand and Sabour, Bhagalpur, Bihar. Phytochemicals through crude methanolic extract were analyzed in Ashwagandha roots. Alcoholic extract was prepared using Soxhlet extraction apparatus. On phytochemical evaluation, total phenolic, flavonoid, total tannin and alkaloids were estimated. Presence of bioactive components in extract was determined by Liquid Chromatography-Mass Spectrophotometry (ESI positive ion) technique. Qualitative screening of extract ensured the presence of alkaloids, phenols, tannins and flavonoids. Quantitatively these phytochemicals showed TPC (66.06 mg/gm), TFC (650.0 mg/gm), TAC (102 mg/gm) and TTC (5.89 mg/gm). However, the phytochemicals and yield did not accumulate in various fractions on polarity. This result was supported by LC-MS analysis which showed 17 definite peaks in their chromatogram but only 8 peaks offer a suitable approximation. Extract revealed forty five bioactive compounds where most of them were phenolic compounds, aryl propionic acid and cinnamide derivatives. Quantitative analysis of present phytochemical showed maximum quantity of TFC followed by TAC, TPC and TTC. On the other hand, presence of various bioactive compounds in root extract showed that plant is having very high medicinal value.

Detection and risk associated with organochlorine, organophosphorus, pyrethroid and carbamate pesticide residues in chicken muscle and organ meats in Jordan

The aims of this study were to: i) analyze the prevalence and concentration of pesticides belonging to 4 major classes, namely organochlorines, organophosphates, carbamates and pyrethroids in chicken meat, liver and kidney; ii) evaluate the impact of boiling, frying and freezing on pesticide levels in chicken meat, and iii) estimate the dietary exposure to pesticide residues in chicken meat and conduct a risk assessment. Liquid chromatography-mass spectrophotometry (LC – MS/MS) was used to determine the concentrations of pesticide residues in samples. A total of 300 domestic (local) and imported samples were collected from the marketplace in Jordan. About 2.3%, 48.3%, 83%, and 46.3% of the samples exceeded MRLs for the organochlorine, organophosphorus, carbamate and pyrethroid pesticide groups, respectively. About 90% of the local and imported muscle samples and almost all the local and imported liver samples contained one or more pesticide above the MRL. Among all the samples tested, two pesticides, namely deltamethrin and fenoxycarb, occurred in high concentrations. Domestic samples were observed to have higher concentrations of pesticides compared to their imported counterparts. The pesticides concentrations decreased by about 50, 75 and 15% after boiling, frying and freezing, respectively. Risk assessment has shown that there is no potential risk from consuming chicken meat, however, local chicken meat is of more concern than is imported product. This study will help food control authorities develop appropriate regulations regarding the use of pesticides in Jordan.

Combined transcriptome and metabolome analysis of the resistance mechanism of quinoa seedlings to Spodoptera exigua.

Quinoa has attracted considerable attention owing to its unique nutritional, economic, and medicinal values. The damage intensity of Spodoptera exigua at the seedling stage of quinoa fluctuates with the crop’s biological cycle and the environmental changes throughout the growing season. In this study, we used independently selected quinoa seedling resistant and susceptible cultivars to investigate the difference between insect resistance and insect susceptibility of quinoa at the seedling stage. Samples were collected when Spodoptera exigua 45 days after planting the seedlings, and broad targeted metabolomics studies were conducted using liquid chromatography-mass spectrophotometry combined with transcriptomic co-analysis. The metabolomic and genomic analyses of the insect-resistant and insect-susceptible quinoa groups revealed a total of 159 differential metabolites and were functionally annotated to 2334 differential genes involved in 128 pathways using the Kyoto Encyclopedia of Genes and Genomes analysis. In total, 14 metabolites and 22 genes were identified as key factors for the differential accumulation of insect-resistant metabolites in quinoa seedlings. Among them, gene-LOC110694254, gene-LOC110682669, and gene-LOC110732988 were positively correlated with choline. The expression of gene-LOC110729518 and gene-LOC110723164, which were notably higher in the resistant cultivars than in the susceptible cultivars, and the accumulations of the corresponding metabolites were also significantly higher in insect-resistant cultivars. These results elucidate the regulatory mechanism between insect resistance genes and metabolite accumulation in quinoa seedlings, and can provide a basis for the breeding and identification of new insect-resistant quinoa cultivars as well as for screening potential regulatory metabolites of quinoa insect-resistant target genes.

Investigation on Chloramphenicol Residues in Poultry Meat in Bangladesh Using a Validated Liquid Chromatography-Tandem Mass Spectrometry

Chloramphenicol (CAP) is a broad-spectrum antibiotic widely applied in veterinary practices. Continual use of CAP in livestock production may lead to antibiotic resistance and health-related hazards. In this study, a rapid and highly sensitive analytical method based on ultra-performance liquid chromatographytandem mass spectrometry (UPLC–MS/MS) was developed and validated to detect and quantify there is due of CAP in poultry meat regarding the safety of humans and animals. Total 80 samples of poultry meat were collected from different poultry farms, and poultry meat sellers in three upazilas namely- Sadar, Mirzapur and Ghatail of Tangail district in the Dhaka division. The test analysis was performed using matrix-matched liquid chromatography-mass spectrophotometry. The chromatographic separation of the CA Presidue was carried out at 40°C temperature on a reverse-phase C18 column using a binary gradient pump, and quantification was performed by LC-MS/MS in electrospray mode. Mobile phase constituents were solvent (a) deionized water, and (b) acetonitrile. The flow rate was 0.35 mL/min and the total run time was 5 min. The method was validated in terms of selectivity, linearity, recovery, and precision following the 2021/808/EC guidelines and acceptance criteria were met in all the cases. The relative standard deviation (RSD) for precision was <11%. The linearity of the calibration curves was excellent (R2 >0.999) at concentrations of 0.25, 0.50, 0.75, 1.0, 2.0, and 5.0 μg/kg for matrix-matched CAP standard, and the range of linearity of this method was 0.0-5.0 μg/kg with R2 value greater than 0.99. The decision limit (CC-a), and detection capability (CCβ) were 0.29 μg/ kg, and 0.31 μg/kg respectively, and the recovery percentages ranged from 94 to 100 %. In this study, the levels of CAP residue in tested poultry meat samples were found below the detection limit. The overall parameters of the proposed method met the validation criteria, and the method proved to be suitable for CAP residues determination in poultry meat samples. Thus, this method could be a precise and highly desirable analytical procedure for rapid and routine analysis of CA Presidues in poultry meat, and obtained tested results in this study could be an authentic data to ensure the chloramphenicol free safety poultry meat for human consumption.

Activated Serum Increases In Vitro Cellular Proliferation and Growth Factor Expression of Musculoskeletal Cells.

The purpose of this study was to investigate proteomic alteration that occurs to whole blood when converted to activated serum (AS) using an autologous thrombin system. This study further sought to evaluate the functional in vitro effect of AS on tenocytes, chondrocytes, subacromial bursal cells, and osteoblasts. The peptide/protein composition of AS was analyzed by liquid chromatography–mass spectrophotometry (LC-MS). The cell lines were treated with AS, and cellular proliferation was quantified 48 h after treatment. Platelet-derived growth factor (PDGF), insulin-like growth factor 1 (IGF-1), vascular endothelial growth factor (VEGF), interleukin-1 beta (IL-1β), and interleukin-1 receptor antagonist (IL-1Ra) were quantified utilizing enzyme-linked immunosorbent assays (ELISAs). LC-MS identified 357 proteins across the AS and whole blood. Fifty-four of the proteins identified had significant differences between the relative protein abundance of the AS samples compared to whole blood. Treatment with AS in all cell lines significantly increased proliferation compared to control cells at 48 h. Increased PDGF, VEGF, and IGF-1 in all cell lines exposed to AS compared to the control (p < 0.05) were observed. These findings suggest that treatment with AS increases in vitro cellular proliferation and the release of growth factors that may play a role in tissue repair.

Identification of Metabolite Compounds and Biological Activity of Diplazium esculentum

Diplazium esculentum is one of the medicinal plants used by Dayak tribes in Central Kalimantan to cure acne. The potential of D. esculentum can be proved through information on the active compounds of the extract obtained in decoction and infusa methods. Thus, a liquid chromatography-mass spectrophotometry (LC-MS)-based method is developed to identify the active compounds of D. esculentum extract in either decoction method or infusa method. The chemical compound potential of D. esculentum extract was then analysed using bioinformatics approach based upon the database of PASS online server. Results showed that the D. esculentum extract contained 81 chemical compounds in decoction method and 68 compounds in infusa method, in which the dominant compound was flavonoid. Moreover, PASS online web server analysis found 7 flavonoid compound groups potential as anti-acne containing antisebor, AR expression inhibitor and CYP1A1 inhibitor. This information could be very useful for designing a clinical test on plant natural compound potential for traditional drug development

Metagenomic-Metabolomic Mining of Kinema, a Naturally Fermented Soybean Food of the Eastern Himalayas.

Kinema is a popular sticky fermented soybean food of the Eastern Himalayan regions of North East India, east Nepal, and south Bhutan. We hypothesized that some dominant bacteria in kinema may contribute to the formation of targeted and non-targeted metabolites for health benefits; hence, we studied the microbiome–metabolite mining of kinema. A total of 1,394,094,912 bp with an average of 464,698,304 ± 120,720,392 bp was generated from kinema metagenome, which resulted in the identification of 47 phyla, 331 families, 709 genera, and 1,560 species. Bacteria (97.78%) were the most abundant domain with the remaining domains of viruses, eukaryote, and archaea. Firmicutes (93.36%) was the most abundant phylum with 280 species of Bacillus, among which Bacillus subtilis was the most dominant species in kinema followed by B. glycinifermentans, B. cereus, B. licheniformis, B. thermoamylovorans, B. coagulans, B. circulans, B. paralicheniformis, and Brevibacillus borstelensis. Predictive metabolic pathways revealed the abundance of genes associated with metabolism (60.66%), resulting in 216 sub-pathways. A total of 361 metabolites were identified by metabolomic analysis (liquid chromatography-mass spectrophotometry, LC-MS). The presence of metabolites, such as chrysin, swainsonine, and 3-hydroxy-L-kynurenine (anticancer activity) and benzimidazole (antimicrobial, anticancer, and anti-HIV activities), and compounds with immunomodulatory effects in kinema supports its therapeutic potential. The correlation between the abundant species of Bacillus and primary and secondary metabolites was constructed with a bivariate result. This study proves that Bacillus spp. contribute to the formation of many targeted and untargeted metabolites in kinema for health-promoting benefits.

Quantification of mycotoxin residues in domestic and imported chicken muscle, liver and kidney in Jordan

This study aimed to detect mycotoxins (aflatoxin B1, ochratoxin A, zearalenone, fumonisin B1 and deoxynivalenol) residues in imported frozen/local fresh chicken muscle and local fresh liver/kidney samples in Jordan. A total of 300 samples composed of 50 imported frozen and 150 locally produced fresh chicken muscle, 50 fresh liver and 50 kidney samples were analyzed. These were examined for mycotoxins using liquid chromatography-mass spectrophotometry (LC–MS/MS). The overall concentrations of mycotoxin residues in the tested samples ranged from 0.03 to 2920.00 μg/kg (mean 0.23–324.24 μg/kg). Zearalenone and aflatoxin B1 exceeding the maximum residue level (MRL) were found in 14 and 2% of the imported chicken muscle samples, respectively. Aflatoxin B1, zearalenone and deoxynivalenol above the MRL were found in 0.7%, 10.7% and 8.7% of local chicken muscle samples, respectively. Zearalenone (>MRL) was the only toxin detected in the local liver samples whereas zearalenone, fumonisin B1 and deoxynivalenol (>MRL) were present in local kidney samples. The results revealed a significant difference in the prevalence of deoxynivalenol between local and imported chicken muscle samples ( P < 0.05), while fumonisin B1 was significantly higher in local chicken kidney samples than local chicken muscle or liver samples. Results also showed that zearalenone was significantly higher in liver (30.0%) than in kidney samples (8.0%). These data may be useful to food control authorities in Jordan to implement mycotoxins residue monitoring and controls in all chicken meat products. • Zearalenone (4%) and aflatoxin B1 (2%) (>MRL) were found in imported chicken. • Aflatoxin B1 (>MRL) was found at low level (0.7%) in local chicken samples. • Zearalenone (10.7%) and deoxynivalenol (8.7%) (>MRL) were found in local chicken. • Zearalenone (>MRL) was detected in both liver (15%) and kidney (8%) samples. • Fumonisin B1 (8%) and deoxynivalenol (4%) >MRL were also found in kidney samples.