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The COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has resulted in hundreds of millions of infections and millions of deaths globally. Although vaccination campaigns are mitigating the pandemic, emerging viral variants continue to pose challenges. The spike (S) protein of SARS-CoV-2 plays a critical role in viral entry by binding to the angiotensin-converting enzyme 2 (ACE2) receptor, making both proteins essential targets for therapeutic and vaccine development. The glycosylation of these proteins influences their structure and function. This underscores the need for detailed site-specific glycoproteomic analysis. In this study, we characterized the N- or O-glycosylation profiles of the recombinant receptor-binding domain (RBD) of spike protein and ACE2 proteins expressed from Expi293F cells, as well as the S2 subunit of spike protein expressed in plant (N. benthamiana) cells. Using a high-resolution Orbitrap Eclipse Tribrid mass spectrometer equipped with the Ultimate 3000 RSLCnano and I-GPA (Integrated GlycoProteome Analyzer) developed in a previous study, 148 N- and 28 O-glycopeptides from RBD, 71 N-glycopeptides from the S2 subunit, and 139 N-glycopeptides from ACE2 were characterized. In addition, we report post-translational modifications (PTMs) of glycan, including mannose-6-phosphate (M6P) and GlcNAc-1-phosphate-6-O-mannose in N-glycan of RBD and ACE2, and O-acetylation in O-glycan of RBD, identified for the first time in these recombinant proteins. The relative abundance distribution according to glycosites and glycan types were analyzed by quantified site-specific N- and O (only from RBD)-glycopeptides from RBD, S2, and ACE2 using I-GPA. Asn331 for RBD, Asn1098 for S2, and Asn103 for ACE2 were majorly N-glycosylated, and dominant glycan-type was complex from RBD and ACE2 and high-mannose from S2. These findings will provide valuable insights into the glycosylation patterns that influence protein function and immunogenicity and offer new perspectives for the development of vaccines and antibody-based therapies against COVID-19. [ABSTRACT FROM AUTHOR]

We have previously shown that 2-thiouridine (S2U), either as a single nucleoside or as an element of RNA chain, is effectively desulfurized under applied in vitro oxidative conditions. The chemically induced desulfuration of S2U resulted in two products: 4-pyrimidinone nucleoside (H2U) and uridine (U). Recently, we investigated whether the desulfuration of S2U is a natural process that also occurs in the cells exposed to oxidative stress or whether it only occurs in the test tube during chemical reactions with oxidants at high concentrations. Using different types of eukaryotic cells, such as baker's yeast, human cancer cells, or modified HEK293 cells with an impaired antioxidant system, we confirmed that 5-substituted 2-thiouridines are oxidatively desulfurized in the wobble position of the anticodon of some tRNAs. The quantitative LC-MS/MS-MRMhr analysis of the nucleoside mixtures obtained from the hydrolyzed tRNA revealed the presence of the desulfuration products of mcm5S2U: mcm5H2U and mcm5U modifications. We also observed some amounts of immature cm5S2U, cm5H2U and cm5U products, which may have indicated a disruption of the enzymatic modification pathway at the C5 position of 2-thiouridine. The observed process, which was triggered by oxidative stress in the living cells, could impair the function of 2-thiouridine-containing tRNAs and alter the translation of genetic information. [ABSTRACT FROM AUTHOR]

A QuEChERS (quick, easy, cheap, effective, rugged, and safe)-based multi-mycotoxin method was developed, analyzing 24 (17 free and 7 modified) Alternaria and Fusarium toxins in cereals via ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). A modified QuEChERS approach was optimized for sample preparation. Quantification was conducted using a combination of stable isotope dilution analysis (SIDA) for nine toxins and matrix-matched calibration for ten toxins. Quantification via a structurally similar internal standard was conducted for four analytes. Alternariol-9-sulfate (AOH-9-S) was measured qualitatively. Limits of detection (LODs) were between 0.004 µg/kg for enniatin A1 (ENN A1) and 3.16 µg/kg for nivalenol (NIV), while the limits of quantification were between 0.013 and 11.8 µg/kg, respectively. The method was successfully applied to analyze 136 cereals and cereal-based foods, including 28 cereal-based infant food products. The analyzed samples were frequently contaminated with Alternaria toxins, proving their ubiquitous occurrence. Interestingly, in many of those samples, some modified Alternaria toxins occurred, mainly alternariol-3-sulfate (AOH-3-S) and alternariol monomethyl ether-3-sulfate (AME-3-S), thus highlighting the importance of including modified mycotoxins in the routine analysis as they may significantly add to the total exposure of their parent toxins. Over 95% of the analyzed samples were contaminated with at least one toxin. Despite the general contamination, no maximum or indicative levels were exceeded. [ABSTRACT FROM AUTHOR]

The composition of the protein corona covering any nanoparticle (NP) when it enters a biological fluid determines the parameters of the NP's interaction with the body. To "control" these parameters, it is important to know the composition of the protein corona, the determination of which is a complex task associated with the two-layer organization of the corona (hard and soft coronas). In a previous publication, we reported obtaining lipid-coated NPs with a full protein corona, isolating them, and proving the presence of the corona on the surface of the NPs. This work reports on the preparation, isolation, and purification of lipid-coated NPs bearing a hard corona. The protein corona composition was determined by using the LC–MS/MS method. Thirty-seven serum proteins were identified with a high degree of reliability. The hard corona contained various apolipoproteins, including apolipoprotein E, which can potentially affect the penetration of NPs into the cell. [ABSTRACT FROM AUTHOR]

Wildlife faces an increasing threat from extreme climatic events, such as heatwaves, which can have a severe impact on various species, including crucial pollinators like bumblebees. Bumblebees are cold‐adapted and heterothermic, possessing the ability to regulate their internal temperature. The impact of heat stress seems species specific in bumblebees. While most species are impacted, some bumblebee species manage to survive, potentially by employing physiological mechanisms, including the modulation of their protein profile (e.g. Heat Shock Proteins). However, there is limited understanding of how their protein profiles are associated with heat exposure. In this study, we examined the global variation in the protein profile of males from two bumblebee species sampled in the wild: the heat‐tolerant Bombus terrestris and the heat‐sensitive Bombus magnus. After subjecting them to heat stupor at 40°C in controlled condition, it was observed that nearly all B. terrestris survived the stress, while over 50% of B. magnus individuals succumbed to the heat exposure. Through off‐gel bottom‐up proteomics and LC–MS/MS analysis of the hemolymph proteome, we identified 164 proteins in both species with a large part of differentially expressed proteins after heat exposure. Additionally, quantitative analysis of fat bodies revealed that the relative mass was stable in B. terrestris, while it was significantly lower in B. magnus exposed to heat stress. Our data suggest that compared with B. magnus, B. terrestris displays a higher adaptability of its hemolymph proteome in response to heat stress. This adaptability could be a key factor contributing to the high physiological resistance of B. terrestris and its ability to adapt to new, stressful environments expected due to climate change. Understanding these mechanisms of protein regulation in bumblebees could provide valuable insights into their resilience and vulnerability facing environmental stresses. [ABSTRACT FROM AUTHOR]

Arisarum vulgare O. Targ. Tozz. (Araceae), locally known as "Elbgouga," holds significant traditional importance in Algeria for the treatment of various human ailments, including pain, infections, inflammation, digestive disorders, cancer, skin problems, eczema, wounds, and burns. The aim of this study was to explore for the first time the phytochemical profile, antioxidant, antibacterial, and inhibitory effects on acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes, DNA protection, and capacity to promote wound healing. Preliminary phytochemical experiments were conducted to evaluate the major classes of bioactive compounds, in addition to the total phenol and flavonoid amounts in AVEE. An LC‐MS/MS analysis was conducted to clarify the phytochemical composition of this particular botanical species. The antioxidant capacity was assessed using DPPH· and ABTS·+ radical scavenging tests. The agar diffusion approach was used to ascertain the antibacterial efficacy against four bacteria (Staphylococcus aureus, Enterococcus faecium, Escherichia coli, and Salmonella typhimurium). Cholinesterase inhibition was evaluated using a colorimetric method that relies on Ellman's reaction. The protective effects of AVEE on pBR322 plasmid DNA damaged by H2O2 and UV treatment were assessed by their DNA‐breaking forms. The in vivo acute dermal toxicity and wound healing potential of the AVEE ointment (1–5% AVEO) were also investigated, and histological analyses were carried out on biological samples. Five protein targets were the subject of an in silico molecular docking investigation (TNFα, TGFBR1, IL‐1ß, GSK‐3ß, and NOS) and ADMET studies of the main components of the extracts were performed. The screening of phytochemicals revealed a significant concentration of phenolic compounds (176.00 ± 1.08 mg of gallic acid equivalents/g of dry extract), mainly flavonoids (81.21 ± 1.24 mg of quercetin equivalents/g of dry extract). Three major compounds were identified by LC‐MS/MS analysis, belonging mainly to the class of flavonoids, with rutin as the most abundant compound (11.85 mg/g extract), followed by hesperidin (4.125 mg/g extract). A conspicuous presence of isoquercitrin (1.327 mg/g extract) was also found. Small amounts of flavonoids and phenolic acids were also detected (catechin, chlorogenic acid, syringic acid, salicylic acid, kaempferol, and luteolin). DPPH assay showed higher antioxidant capacity compared to the ABTS·+ assay (142.60 ± 5.52 µg/mL and 236.10 ± 0.22 µg/mL, respectively). AVEE was effective against all selected bacterial strains; however, the highest zone of inhibition (36.00 ± 0.1 mm) was noted against E. faecium. The extract of the plant significantly inhibited both AChE and BChE (89.98 ± 18.76 µg/mL inhibitory activity against AChE and 98.28 ± 44.68 µg/mL inhibitory activity against BChE, respectively). The ethanolic extract of A. vulgare and quercetin exhibited more significant DNA protection action in form I, with percentages of 90.41 and 94.23%, respectively, compared to form II, where the percentages were 27.91 and 51.92%, respectively. The AVEO formulation may be safely administered by topical application. A statistically significant wound contraction was discovered in the group treated with 5% AVEO compared to the groups that were not treated or treated with petroleum jelly. Furthermore, there was no notable disparity detected between the group treated with 5% AVEO and the group treated with the reference drug. The 5% AVEO‐treated group showed, after the research was complete, the most significant percentage of wound contraction (96.90 ± 0.42%). Moreover, the topical application of the formulation improved histological parameters. In silico study showed that rutin, hesperidin, and isoquercitrin had a high affinity to the five main targets that might contribute to the wound healing potential of A. vulgare ethanolic extract. The impressive biological capabilities of A. vulgare indicate that the plant has the ability to be a valuable source of bioactive chemicals with various medical applications. [ABSTRACT FROM AUTHOR]

Salt is a potent abiotic stress that arrests plant growth by impairing their physio-biochemical and molecular processes. However, it is unknown how the ABA signaling system and vacuolar-type Na+/H+ antiporter proteins induce stress tolerance in wheat (Triticum aestivum L.) seedlings. The present study aimed to identify salt-responsive proteins and signaling pathways involved in the resistance of wheat to salt stress. We explored the proteome profile, 20 amino acids, 14 carbohydrates, 8 major phytohormones, ion content, and salt tolerance genes in wheat (Triticum aestivum L., cv.) under 200 mM NaCl with control plants for six days. The results showed that amino acids such as alanine, serine, proline, glutamine, and aspartic acid were highly expressed under salt stress compared with control plants, suggesting that amino acids are the main players in salinity tolerance. The ABA signaling system was activated in response to salinity stress through the modulation of protein phosphatase 2C (PP2C) and ABA-responsive element binding factor (ABF), resulting in an ABA-mediated downstream response. Additionally, the vacuolar-type Na+/H+ antiporter was identified as a key protein in salt stress tolerance via compartmentalizing Na+ in the vacuole. Furthermore, a significant increase in the abundance of the 14-3-3 protein was noticed in salt-fed plants, suggesting that this protein plays an important role in Na+ compartmentalization. Moreover, up-regulation of ascorbate peroxidase (APX), glutathione-S-transferase (GST), and thioredoxin-scavenged reactive oxygen species resulted in improved plant growth under salt stress. These data will help to identify salt-responsive proteins that can be used in future breeding programs to develop salt-tolerant varieties. [ABSTRACT FROM AUTHOR]

Sacha inchi (Plukenetia volubilis) is a valuable oilseed crop with a high content of polyunsaturated fatty acids (PUFAs). However, there is a lack of in-depth understanding of the lipidomics in Sacha inchi seeds (SIDs). Saturated fatty acids occupied more than half of the proportion (59.31%) in early development, while PUFAs accounted for 78.92% at maturation. The main triacylglycerols were TAG(18:3/18:3/18:3), TAG(18:2/18:2/18:3), and TAG(16:0/18:2/18:2). The corresponding species (18:3/18:3, 18:2/18:2, and 16:0/18:2) were also the main ingredients in diacylglycerol and phosphatidic acid, indicating high PUFA composition in the sn-1 and sn-2 positions of TAG. Only LPC(18:3), LPC(18:2), and LPC(16:0) were identified in SIDs, implying that those PUFAs on the sn-2 positions of the PC(18:3/-), PC(18:2/-), and PC(16:0/-) categories were released into the acyl-CoA pool for the Kennedy pathway. Conversely, the PC(18:1/-) and PC(18:0/-) categories might be responsible for the generation of PC-derived DAG and TAG. The lipidomics data will contribute to understanding the TAG assembly in developing SIDs, especially for PUFAs. [ABSTRACT FROM AUTHOR]

Limited or absent activity of the enzyme α-galactosidase A (α-Gal A), due to mutation in the related gene on the X chromosome, leads to the development of a rare hereditary and genetic disease known as Fabry disease (FD). This pathology involves a progressive accumulation in various organs of the substrates of the enzyme e.g., globotriaosylceramide (Gb3) and its deacylated form, globotriaosylsphingosine (Lyso-Gb3), suggesting these molecules as biomarkers of Fabry disease. The present paper describes the development of an analytical strategy for the identification and quantification of Gb3 and Lyso-Gb3, in serum and blood samples by using liquid chromatography (LC) coupled to mass spectrometry in multiple reaction monitoring (MRM/MS) ion mode. The best experimental conditions were obtained by extracting the glycolipids with chloroform/methanol/H2O (2/1/0.3) and by separating them on a C4 column with a linear gradient (A: H2O with 2 mM ammonium formate. B: methanol with 1 mM ammonium formate, both acidified with 0.2% formic acid). The best transitions (a combination of precursor and fragment ions—m/z) were 786.8 m/z > 268.3 m/z for Lyso-GB3, 1137.3 m/z > 264.3 m/z for Gb3, 1039.3 m/z > 264.4 m/z for N-heptadecanoyl-ceramide trihexoside, and 843.5 m/z > 264.3 m/z for N-glycinated lyso-ceramide trihexoside, the latter being used as an internal standard. The developed method provided a reliable, fast, and effective procedure for direct measurements of GB3 and Lyso-GB3 in serum and blood for diagnosis of Fabry disease, suggesting this method as a complementary assay to the current enzymatic test. Therefore, this approach could open new insights into the clinical diagnostics of lysosomal storage disorders. [ABSTRACT FROM AUTHOR]

Sulfoxaflor is a systemic sulfoximine insecticide, used to control sucking pests in different agricultural crops including wheat. However, the information regarding its residual fate in wheat agroecosystem under tropical environment and the associated dietary risk are still obscure. Therefore, a supervised field experiment was conducted in wheat comprising the application of sulfoxaflor 12% (w/v) SC at 30 and 60 g a.i. ha−1 in two consecutive seasons. An optimised method for estimation of sulfoxaflor residues using modified QuEChERS technique was developed and validated. The extracting solvent ethyl acetate + cyclohexane (9 + 1 v/v) along with the clean-up combinations viz. 75 mg PSA+25 mg GCB+150 mg magnesium sulphate for wheat plant and straw and 75 mg PSA+75 mg C18 + 150 mg magnesium sulphate for grain produced satisfactory recovery, minimum matrix interference and optimum peak shape. The average recovery, repeatability and within-laboratory reproducibility of sulfoxaflor in wheat substrates ranged between 90.66% and 103.55%, 2.67–5.17% and 3.05–8.15%, respectively, and the limit of quantification was estimated 0.01 mg kg−1. Sulfoxaflor was stable up to 60 days in wheat matrices under storage (−20°C). The dissipation of sulfoxaflor in wheat plants followed first-order kinetics and the overall shorter persistence in Season I and II ranged between 1.05–1.14 and 2.44–2.47 days (T1 – T2), respectively. The deviation in the persistence of the compound between two seasons due to variation in climatic parameters was observed to be statistically significant. The estimated pre-harvest interval of sulfoxaflor ranged between 4.95–5.34 (Season I) and 11.37–11.55 days (Season II). Besides, the terminal residues of sulfoxaflor were below quantification limit in the harvested wheat products, and the estimated long-term dietary risk of sulfoxaflor was observed to be negligible. Therefore, it was expected that the application of this insecticide in wheat cultivation should pose no residual toxicity and the harvested wheat products can be considered safe for human consumption. [ABSTRACT FROM AUTHOR]

Straw from no-till cropping systems, in addition to increasing the soil organic matter content, may also impede the movement of applied herbicides into the soil and, thus, alter the behavior and fate of these compounds in the environment. Rain or irrigation before or after an herbicide treatment can either help or hinder its movement through the straw, influencing weed control. Our objective was to develop a system for herbicide application and rain simulation, enabling the evaluation of the movement of various herbicides either in dry or wet straw under different rainfall volumes (25, 50, 75, and 100 mm). The amount of the applied herbicides that moved through the straw were collected and measured using a liquid chromatograph with a tandem mass spectrometry system (LC-MS/MS). Measurements obtained with the developed system showed a high herbicide treatment uniformity across all replications. The movement of the active ingredients through the straw showed variability that was a function of the applied herbicide, ranging from 17% to 99%. In wet straw, the collected herbicide remained constant from 50 to 100 mm of simulated rainfall. For the wet straw, the decreasing percentages of the herbicide movement through straw to the soil were sulfentrazone (99%), atrazine and diuron (91% each), hexazinone (84%), fomesafen (80.4%), indaziflam (79%), glyphosate (63%), haloxyfop-p-methyl (45%), and S-metolachlor (27%). On the dry straw, the decreasing percentages of the herbicide movement were fomesafen (88%), sulfentrazone (74%), atrazine (69.4%), hexazinone (69%), diuron (68.4%), glyphosate (48%), indaziflam (34.4%), S-metolachlor (22%), and haloxyfop-p-methyl (18%). Overall, herbicide movement was higher in wet straw (with a previous 25 mm simulated rainfall layer) than in dry straw. Some herbicides, like haloxyfop-p-methyl and indaziflam, exhibited over 50% higher movement in wet straw than dry straw after 100 mm of simulated rain. The developed system can be adapted for various uses, serving as a valuable tool to evaluate the behavior of hazardous substances in different agricultural and environmental scenarios.

A method was developed for the determination of 26 drugs of abuse from different classes, including illicit drugs in quantitative dried blood spots (qDBSs), with the aim to provide a convenient method for drug testing by using only 10 μL of capillary blood. A satisfactory limit of quantification (LOQ) of 2.5 ng/mL for 9 of the compounds and 5 ng/mL for 17 of the compounds and a limit of detection (LOD) of 0.75 ng/mL for 9 of the compounds and 1.5 ng/mL for 17 of the compounds were achieved for all analytes. Reversed-phase liquid chromatography was applied on a C18 column coupled to MS, providing selective detections with both +ESI and -ESI modes. Extraction from the qDBS was performed using AcN-MeOH, 1:1 (v/v), with recovery ranging from 84.6% to 106%, while no significant effect of the hematocrit was observed. The studied drugs of abuse were found to be stable over five days under three different storage conditions (at ambient temperature 21 °C, at −20 °C, and at 35 °C), thus offering a highly attractive approach for drug screening by minimally invasive sampling for individuals that could find application in forensic toxicology analysis. [ABSTRACT FROM AUTHOR]

Straw from no-till cropping systems, in addition to increasing the soil organic matter content, may also impede the movement of applied herbicides into the soil and, thus, alter the behavior and fate of these compounds in the environment. Rain or irrigation before or after an herbicide treatment can either help or hinder its movement through the straw, influencing weed control. Our objective was to develop a system for herbicide application and rain simulation, enabling the evaluation of the movement of various herbicides either in dry or wet straw under different rainfall volumes (25, 50, 75, and 100 mm). The amount of the applied herbicides that moved through the straw were collected and measured using a liquid chromatograph with a tandem mass spectrometry system (LC-MS/MS). Measurements obtained with the developed system showed a high herbicide treatment uniformity across all replications. The movement of the active ingredients through the straw showed variability that was a function of the applied herbicide, ranging from 17% to 99%. In wet straw, the collected herbicide remained constant from 50 to 100 mm of simulated rainfall. For the wet straw, the decreasing percentages of the herbicide movement through straw to the soil were sulfentrazone (99%), atrazine and diuron (91% each), hexazinone (84%), fomesafen (80.4%), indaziflam (79%), glyphosate (63%), haloxyfop-p-methyl (45%), and S-metolachlor (27%). On the dry straw, the decreasing percentages of the herbicide movement were fomesafen (88%), sulfentrazone (74%), atrazine (69.4%), hexazinone (69%), diuron (68.4%), glyphosate (48%), indaziflam (34.4%), S-metolachlor (22%), and haloxyfop-p-methyl (18%). Overall, herbicide movement was higher in wet straw (with a previous 25 mm simulated rainfall layer) than in dry straw. Some herbicides, like haloxyfop-p-methyl and indaziflam, exhibited over 50% higher movement in wet straw than dry straw after 100 mm of simulated rain. The developed system can be adapted for various uses, serving as a valuable tool to evaluate the behavior of hazardous substances in different agricultural and environmental scenarios. [ABSTRACT FROM AUTHOR]

Studying the mutagenesis mechanism is crucial for pulsed light use in the food processing industry. After being exposed to pulsed light, the original strain Y Lactobacillus Plantarum CICC6048 was transformed into the high acid-producing mutant G10. The differing levels of protein expression between the two strains were compared using the LC–MS/MS analysis. The bacterium displayed a distinct differential protein composition after pulsed light treatment, according to GO analysis. A KEGG analysis revealed that the pathways for cofactor biosynthesis, starch, sucrose metabolism, and phosphate transfer systems were considerably different in the proteins of high acid-producing strains (PTS). In the protein interaction network, A0A0R2G2S1 showed the highest level of enhanced connectivity among the differentially expressed proteins. These pathways improve the efficiency of crucial metabolism and lessen DNA repair. They may be a key mechanism for increasing the growth rate and acid production of Lactobacillus Plantarum by pulsed light. [ABSTRACT FROM AUTHOR]

Sacha inchi (Plukenetia volubilis) is a valuable oilseed crop with a high content of polyunsaturated fatty acids (PUFAs). However, there is a lack of in-depth understanding of the lipidomics in Sacha inchi seeds (SIDs). Saturated fatty acids occupied more than half of the proportion (59.31%) in early development, while PUFAs accounted for 78.92% at maturation. The main triacylglycerols were TAG(18:3/18:3/18:3), TAG(18:2/18:2/18:3), and TAG(16:0/18:2/18:2). The corresponding species (18:3/18:3, 18:2/18:2, and 16:0/18:2) were also the main ingredients in diacylglycerol and phosphatidic acid, indicating high PUFA composition in the sn-1 and sn-2 positions of TAG. Only LPC(18:3), LPC(18:2), and LPC(16:0) were identified in SIDs, implying that those PUFAs on the sn-2 positions of the PC(18:3/-), PC(18:2/-), and PC(16:0/-) categories were released into the acyl-CoA pool for the Kennedy pathway. Conversely, the PC(18:1/-) and PC(18:0/-) categories might be responsible for the generation of PC-derived DAG and TAG. The lipidomics data will contribute to understanding the TAG assembly in developing SIDs, especially for PUFAs.

Since antiquity, Centaurea species have been used in folk medicine to treat several diseases owing to their potential biological activities that distinguish this genus such as antioxidant, anticancer, and anti-inflammatory effect. The current study aimed to investigate the possible neuroprotective effects of the n-butanol extract of Centaurea maroccana (BECM) against cisplatin (CP) induced neurotoxicity in mice. BECM's potential neuroprotective properties were studied in vitro and in vivo models. Male Swiss albino mice were orally received BECM (200 mg/kg) for 10 days before a single intraperitoneal injection of cisplatin (8 mg/kg). Vitamin E (100 mg/kg) was given daily by gavage as a positive control. In vitro results revealed that BECM inhibited lipid peroxidation (LPO) levels and acetylcholinesterase (AChE) activity. In vivo findings showed that BECM pretreatment was able to regulate lactate dehydrogenase (LDH) levels and to improve CP-induced cholinergic dysfunction by inhibiting AChE activity in mice brains. Moreover, BECM attenuated CP-provoked oxidative stress by suppressing LPO levels, increasing total antioxidant capacity (TAC) and enhancing the activities of antioxidant enzymes (catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH), glutathione peroxidase (GPx) and glutathione S-transferase (GST)) in both brain cytosolic and mitochondrial fractions. The histological analysis exhibited neurotoprotective effect of BECM by protecting the cerebral cortex and reducing the histomorphological alterations resulted by cisplatin. Interestingly, our extract achieved neuroprotection comparable to vitamin E in most evaluated parameters. It appears that protective potency of BECM against CP-induced neurotoxicity could be related to its richness in polyphenols confirmed by liquid-chromatography tandem mass spectrometry analysis (LC-MS/MS). [ABSTRACT FROM AUTHOR]

Acne vulgaris is a chronic inflammatory disease with high incidence, diverse clinical manifestations, poor clinical efficacy, and easy recurrence. Recent studies have found that the occurrence of acne is related to metabolic factors such as insulin resistance; however, the specific mechanism of action remains unclear. This study aimed to identify significantly different metabolites and related metabolic pathways in the serum of acne vulgaris patients with or without insulin resistance. LC–MS/MS was used to analyze serum samples from patients about acne with insulin resistance (n = 51) and acne without insulin resistance (n = 69) to identify significant metabolites and metabolic pathways. In this study, 18 significant differential metabolites were screened for the first time. In the positive-ion mode, the upregulated substances were creatine, sarcosine, D-proline, uracil, Phe–Phe, L-pipecolic acid, and DL-phenylalanine; the downregulated substances were tridecanoic acid (tridecylic acid), L-lysine, cyclohexylamine, sphingomyelin (d18:1/18:0), gamma-L-Glu-epsilon-L-Lys, and 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine. In the negative-ion mode, the upregulated substance was cholesterol sulfate, and the downregulated substances were D(-)-beta-hydroxybutyric acid, myristic acid, D-galacturonic acid, and dihydrothymine. Cholesterol sulfate showed the most significant expression among all differential metabolites (VIP = 7.3411). Based on the KEGG database, necroptosis and ABC transporters were the most significantly enriched metabolic pathways in this experiment. The differential metabolites and pathways identified in this study may provide new possibilities for the clinical diagnosis and development of targeted drugs for acne patients with insulin resistance. [ABSTRACT FROM AUTHOR]

In the present study, two new Bacillus subtilis phages, BSTP4 and BSTP6, were isolated and studied further. Morphologically, BSTP4 and BSTP6 are podoviruses with complete genome of 19,145 (39.9% G + C content) and 19,367 bp (39.8% G + C content), respectively, which became among the smallest Bacillus phages. Three most prominent structural proteins were separated and identified as pre-neck appendage, major head, and head fiber proteins using LC–MS/MS. Both phages encode putative terminal proteins (TP) and contain short inverted terminal repeats (ITRs) which may be important for their replication. In addition, non-coding RNA (pRNA) and parS sites were identified which may be required for DNA packaging and their maintenance inside the host, respectively. Furthermore, the phage genome sequences show significant similarity to B. subtilis group species genome sequences. Finally, phylogenomic and phylogenetic analyses suggest that BSTP4 and BSTP6 may form a new species in the genus Salasvirus, subfamily Picovirinae of family Salasmaviridae. Considering the small numbers of ICTV-accepted B. subtilis phages and the importance of the host in the food industry and biotechnology, the current study helps to improve our understanding of the diversity of B. subtilis phages and shed light on the phage–host relationships. [ABSTRACT FROM AUTHOR]

Abstract Background Poplar fungal infections are difficult to control and result in severe economic loss. As a viable alternative to chemical pesticides, biocontrol is an effective safe method for disease control. Results Inhibitory activity of Bacillus velezensis 33RB and Aspergillus niger 46SF was evaluated against numerous phytopathogens. The bacterial strain displayed the highest inhibitory activity toward Colletotrichum gloeosporioides BJ02 and Fusarium oxysporum 20RF (61.2 and 49.4%, respectively). Also, the maximum inhibitory activity of A. niger 46SF was exhibited (75.51 and 70.83%) against C. gloeosporioides BJ02 and F. oxysporum 20RF, respectively. The minimum volume (6.25 ml) of sterilized cultural filtrate of bacterial and fungal strains significantly inhibited the growth of C. gloeosporioides BJ02 by 73.3 and 83.3%, respectively, and F. oxysporum 20RF reached 40.4 and 78.8%, respectively. B. velezensis 33RB and A. niger 46SF displayed the highest inhibition toward C. gloeosporioides BJ02 and F. oxysporum 20RF at neutral pH and pH 5, respectively. Moreover, the highest inhibitory activity of B. velezensis 33RB and A. niger 46SF was achieved at 37 °C and 28 °C, respectively. Pathogenicity tests on sterilized detached leaves indicated that these isolates could potentially affect anthracnose and fusarium wilt diseases. Several secondary bioactive metabolites that assured the biocontrol efficacy of tested microbes were detected by liquid chromatography–mass spectrometry (LC–MS). The most detectable compounds included organic acids such as fumaric, DL-malic, citric, isobutyric, and glutamic acids. Also, numerous fatty acids such as lauric, linoleic, oleic, stearic, and myristic acids with diverse biological functions, including antimicrobial properties, were determined. Conclusions Bacillus velezensis 33RB and A. niger 46SF were potential alternatives to chemical pesticides as biological control agents for the phytopathogens C. gloeosporioides BJ02 and F. oxysporum with environmentally friendly and sustainable properties.

This work highlights the methodology for the development of diffusive gradients in thin films (o-DGT) through its adaptation for 12 natural and synthetic hormones belonging to three different families (estrogens, progestins, and androgens). A reliable strategy must be applied during o-DGT lab adaptation to avoid issues related to the analysis (i.e., presence of matrix effects in grab or passive samples) but also to the o-DGT configuration (i.e., undesirable sorption or desorption, lack of performance with insufficient elution or unreliable diffusion coefficient). To avoid analytical issues due to the presence of salts in grab samples, CaCl2 exposure solutions must be used on a lab-scale development to monitor the hormone concentration. The selected o-DGT was composed of an Oasis® HLB binding gel and a diffusive gel in agarose because they provided better performance than polyacrylamide gels (i.e., higher elution factors and more repeatable diffusion coefficients). The elution factors of the binding gel were then from 0.79 ± 0.13 to 1.04 ± 0.13 (RSD < 15%) and the diffusion coefficients at 25 °C were from 4.07 ± 0.24 to 5.49 ± 0.28 × 10–6 cm2 s−1 (RSD < 9%). A laboratory exposure to a synthetic solution was performed to check the consistency with the DGT quantification model validating the calibration parameters for all hormones (except 17α-ethinylestradiol with a bias of 40%). Therefore, the o-DGT configuration is suitable for sampling hormones in the natural environment with LOQDGT ranging from 0.3 to 6.6 ng L−1. [ABSTRACT FROM AUTHOR]

Chenopodium murale (Syn. Chenopodiastrum murale) (amaranthaceae) is used in the rural Egypt to treat oral ulcers in newborn children. The current study aimed to discover new natural products suitable for treating candidiasis disease with minimal side effects. Characterization of bioactive compounds by LC-QTOF-HR-MS/MS from Chenopodium murale fresh leaves' juice (CMJ) was carried out in order to elucidate their potential anti-fungal and immunomodulatory effects in oral candidiasis in immunosuppressed rats. An oral ulcer candidiasis model was created in three stages: (i) immunosuppression by drinking dexamethasone (0.5 mg/L) for two weeks; (ii) Candida albicans infection (3.00 × 106 viable cell/mL) for one week; and (iii) treatment with CMJ (0.5 and 1.0 g/kg orally) or nystatin (1,000,000 U/L orally) for one week. Two doses of CMJ exhibited antifungal effects, for example, through a significant reduction in CFU/Petri (236.67 ± 37.86 and 4.33 ± 0.58 CFU/Petri), compared to the Candida control (5.86 × 104 ± 1.21 CFU/Petri), p ≤ 0.001. In addition, CMJ significantly induced neutrophil production (32.92% ± 1.29 and 35.68% ± 1.77) compared to the Candida control level of 26.50% ± 2.44. An immunomodulatory effect of CMJ at two doses appeared, with a considerable elevation in INF-γ (103.88 and 115.91%), IL-2 (143.50, 182.33%), and IL-17 (83.97 and 141.95% Pg/mL) compared with the Candida group. LC-MS/MS analysis operated in negative mode was used for tentative identification of secondary (SM) metabolites based on their retention times and fragment ions. A total of 42 phytoconstituents were tentatively identified. Finally, CMJ exhibited a potent antifungal effect. CMJ fought Candida through four strategies: (i) promotion of classical phagocytosis of neutrophils; (ii) activation of T cells that activate IFN-γ, IL-2, and IL-17; (iii) increasing the production of cytotoxic NO and H2O2 that can kill Candida; and (iv) activation of SOD, which converts superoxide to antimicrobial materials. These activities could be due to its active constituents, which are documented as anti-fungal, or due to its richness in flavonoids, especially the active compounds of kaempferol glycosides and aglycone, which have been documented as antifungal. After repetition on another type of small experimental animal, their offspring, and an experimental large animal, this study may lead to clinical trials. [ABSTRACT FROM AUTHOR]

Milk protein concentrate (MPC) is considered an ideal substitute of cow milk because of its similar protein and nutrition. In this study, MPC was hydrolyzed to peptides by alcalase and neutrase, and the properties of hydrolysate were evaluated. When MPC was hydrolyzed at the ratio of alcalase and neutrase of 1:1 and enzyme to substrate ratio of 6000 U/g MPC at 50°C, pH 8.5 for 3 h, the proportion of peptides with molecular weights <1 kDa was 85.31%, and the antigenicity reduction rates of casein and β‐lactoglobulin were 33.76% and 22.38%, respectively. Moreover, LC–MS/MS peptide identification revealed that the alcalase and neutrase disrupted a total of 65 epitopes of casein and 21 epitopes of whey protein, which further elucidated the mechanism of complex enzyme hydrolysis to reduce milk protein allergenicity. [ABSTRACT FROM AUTHOR]

This study describes the analysis of five quinolizidine alkaloids (QA), i.e. 13-OH-lupanine, lupanine, lupinine, angustifoline and sparteine in 30 samples of lupine flours, lupine seeds and derived products collected 2019–2021 on the German retail market. QA occur as secondary metabolites in plants of lupine species. Certain QA are of toxicological relevance. The analytical determination performed by LC-MS/MS revealed some samples with high QA concentrations (up to 21,000 mg/kg), particularly in bitter lupine seeds. As those concentrations would result in substantial exceedances of the maximum tolerable intake values proposed by health authorities, they must be considered as a health concern. [ABSTRACT FROM AUTHOR]

Zivzik pomegranate (Punica granatum) has recently sparked considerable interest due to its nutritional and antioxidant properties. To evaluate the antioxidant capacities of P. granatum juice, ethanol (EEZP), and water (WEZP) extracts from peel and seed, the antioxidant methods of 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid radical (ABTS•+) scavenging, 1,1-diphenyl-2-picrylhydrazyl free radical (DPPH•) scavenging, Fe3+-2,4,6-tris(2-pyridyl)-S-triazine (TPTZ) reducing, Fe3+ reducing, and Cu2+ reducing methods were used. The antioxidant capacities of samples were compared with the most commonly used synthetic antioxidants, i.e., BHA, BHT, α-tocopherol, and Trolox. In terms of setting an example, the IC50 values of EEZP for ABTS•+ and DPPH• scavenging activities were found to be lower than standards, at 5.9 and 16.1 μg/mL, respectively. The phenolic and flavonoid contents in EEZP peel were 59.7 mg GAE/g and 88.0 mg QE/g, respectively. Inhibition of α-glycosidase, α-amylase, acetylcholinesterase, and human carbonic anhydrase II (hCA II) enzymes was also investigated. EEZP demonstrated IC50 values of 7.3 μg/mL against α-glycosidase, 317.7 μg/mL against α-amylase, 19.7 μg/mL against acetylcholinesterase (AChE), and 106.3 μg/mL against CA II enzymes. A total of 53 phenolic compounds were scanned, and 30 compounds were determined using LC-MS/MS. E. coli and S. aureus bacteria were resistant to all four antibiotics used as standards in hospitals. [ABSTRACT FROM AUTHOR]

The determination of the selected antihypertensive drugs in human plasma samples with the novel solvent front position extraction (SFPE) technique is presented. The SFPE procedure combined with LC-MS/MS analysis was used for the first time to prepare a clinical sample containing the drugs mentioned above from different therapeutic groups. The effectiveness of our approach was compared with the precipitation method. The latter technique is usually used to prepare biological samples in routine laboratories. During the experiments, the substances of interest and the internal standard were separated from other matrix components using a prototype horizontal chamber for thin-layer chromatography/high-performance thin-layer chromatography (TLC/HPTLC) with a moving pipette powered by a 3D mechanism, which distributed the solvent on the adsorbent layer. Detection of the six antihypertensive drugs was performed by liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS) in multiple reaction monitoring (MRM) mode. Results obtained by SFPE were very satisfactory (linearity R2 ≥ 0.981; %RSD ≤ 6%; LOD and LOQ were in the range of 0.06–9.78 ng/mL and 0.17–29.64 ng/mL, respectively). The recovery was in the range of 79.88–120.36%. Intra-day and inter-day precision had a percentage coefficient of variation (CV) in the range of 1.10–9.74%. The procedure is simple and highly effective. It includes the automation of TLC chromatogram development, which significantly reduced the number of manual operations performed, the time of sample preparation and solvent consumption. [ABSTRACT FROM AUTHOR]

The demand for skin tissue allografts to treat burns and other types of injuries increases each year to the extent that categories of donors formerly deemed "unsuitable", such as victims of suicide by polytrauma or poisoning, are now considered. Patients who died by ingestion of/exposure to toxic substances can be accepted as tissue donors after assessment of graft safety to rule out any risks of transferring toxic substances to the recipient. A cadaveric skin donation was obtained from a 57-year-old woman who died from intoxication after ingesting colchicine tablets (0.2 mg/kg). To determine the safety of cadaveric skin allografts, high-performance liquid chromatography-mass spectrometry (LC–MS/MS) was used to identify and quantify colchicine in procured skin. Results revealed that colchicine concentrations were lower than the instrument limit of detection (LOD) of 0.5 ng/mg both in epidermis and dermis. Cell viability assessed through the MTT ([3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]) test was within standard limits. Thanks to accurate tests performed, which are routinely applied also in clinical diagnostics and forensic toxicology, it was possible to ascertain the safety and suitability of skin tissue for donation. [ABSTRACT FROM AUTHOR]

Simple Summary: In our work, we described and proposed a novel LC-MS/MS based approach for early lung cancer screening based on the VOC marker. Through this approach, two key volatile organic compounds (VOCs) used in the early screening and diagnosis of lung cancer were determined as 3-hydroxy-2-butanone and 2-pentanone. 3-hydroxy-2-butanone in the human body was found for the first time to be derived from the sugar metabolism of oral bacteria and we found a correlation between the bacteria that metabolize 3-hydroxy-2-butanone and lung cancer. (1) Background: lung cancer is the world's deadliest cancer, but early diagnosis helps to improve the cure rate and thus reduce the mortality rate. Annual low-dose computed tomography (LD-CT) screening is an efficient lung cancer-screening program for a high-risk population. However, LD-CT has often been characterized by a higher degree of false-positive results. To meet these challenges, a volatolomic approach, in particular, the breath volatile organic compounds (VOCs) fingerprint analysis, has recently received increased attention for its application in early lung cancer screening thanks to its convenience, non-invasiveness, and being well tolerated by patients. (2) Methods: a LC-MS/MS-based volatolomics analysis was carried out according to P/N 5046800 standard based breath analysis of VOC as novel cancer biomarkers for distinguishing early-stage lung cancer from the healthy control group. The discriminatory accuracy of identified VOCs was assessed using subject work characterization and a random forest risk prediction model. (3) Results: the proposed technique has good performance compared with existing approaches, the differences between the exhaled VOCs of the early lung cancer patients before operation, three to seven days after the operation, as well as four to six weeks after operation under fasting and 1 h after the meal were compared with the healthy controls. The results showed that only 1 h after a meal, the concentration of seven VOCs, including 3-hydroxy-2-butanone (TG-4), glycolaldehyde (TG-7), 2-pentanone (TG-8), acrolein (TG-11), nonaldehyde (TG-19), decanal (TG-20), and crotonaldehyde (TG-22), differ significantly between lung cancer patients and control, with the invasive adenocarcinoma of the lung (IAC) having the most significant difference. (4) Conclusions: this novel, non-invasive approach can improve the detection rate of early lung cancer, and LC-MS/MS-based breath analysis could be a promising method for clinical application. [ABSTRACT FROM AUTHOR]

Due to the growing global incidence of allergy to nuts and peanuts, the need for better protection of consumers sensitive to those products is constantly increasing. The best strategy to defend them against adverse immunological reactions still remains the total removal of those products from their diet. However, nuts and peanuts traces can also be hidden in other food products, especially processed ones, such as bakery products, because of cross-contamination occurring during production. Precautionary labelling is often adopted by producers to warn allergic consumers, usually without any evaluation of the actual risk, which would require a careful quantification of nuts/peanuts traces. In this paper, the development of a multi-target method based on liquid chromatography-tandem high resolution mass spectrometry (LC-MS, MS/MS), able to detect traces of five nuts species (almonds, hazelnuts, walnuts, cashews and pistachios) and of peanuts in an in-house incurred bakery product (cookie) through a single analysis is described. Specifically, allergenic proteins of the six ingredients were used as the analytical targets, and the LC-MS responses of selected peptides resulting from their tryptic digestion, after extraction from the bakery product matrix, were exploited for quantification, following a bottom-up approach typical of proteomics. As a result, nuts/peanuts could be detected/quantified down to mg·kg−1 levels in the model cookie, thus opening interesting perspectives for the quantification of hidden nuts/peanuts in bakery products and, consequently, for a more rational use of precautionary labelling. [ABSTRACT FROM AUTHOR]

Veterinary drugs are widely used to treat livestock diseases, but the emergence of problems related to drug resistance prompted the government of the Republic of Korea to enact laws regulating the use of veterinary drugs in livestock. Monitoring the contents of residual drugs in livestock products at regular intervals is necessary to assess whether the regulations have resulted in the effective management of food product safety and prevent and manage unexpected problems related to this issue. A 2019–2023 livestock product post-marketing monitoring program launched by the Ministry of Food and Drug Safety (MFDS) of the Republic of Korea inspected 1135 livestock products. Sixty-two different veterinary drugs were detected among 167 monitored, with a detection rate of 28.2% (320/1135 samples; chicken [125], pork [114], milk [35], beef [30], and egg [16]). Diclazuril was detected most frequently, followed by flumequine, clopidol, and enrofloxacin. Over the past five years, two samples that did not conform to the regulations were identified to contain 20 and 215 ng/g, diclazuril and toltrazuril sulfone, respectively. The estimation of exposure and risk assessment revealed that the hazard index of diclazuril was below 75%. However, in case of toltrazuril sulfone, it showed a hazard index exceeding 138% in the scenario of the 95th percentile high consumption of chicken. The results of this study may provide a useful reference in initiating more targeted work in the future, such as the further fine-tuning of food safety monitoring programs and developing food safety standards. In future studies, the MFDS shall continue to monitor the statuses of residual veterinary drugs in commercial livestock products. • We investigated 151 veterinary drug residues in 1135 livestock samples from the Republic of Korea. • Among the analyzed samples, 28.2% (n = 320) contained veterinary drug residues. • Two violations of the maximum residual limits set by the Republic of Korea were observed. • Various residues of veterinary drugs were often detected in combination. • The current levels of exposure to diclazuril and toltrazuril sulfone did not present a significant risk. [ABSTRACT FROM AUTHOR]

Per- and polyfluoroalkyl substances, known as PFAS, are emerging contaminants detected in surface water, groundwater, drinking water, air, and food. Due to its chemical stability and high hydrophobicity, these compounds are used as surfactants, surface protectors, flame retardants, textile impregnation, fire-fighting foam, and pesticides and may be a risk for human health in the long-term exposure. Currently, a little research has been carried out for these compounds in Brazil. In this study, the concentration and distribution of 23 PFAS were analyzed in groundwater samples collected from water wells in urbanized areas and in surface water samples from Porto Alegre, one of the biggest cities in Brazil. The total concentrations of PFAS (ΣPFAS) in a groundwater sample varied between 22 and 718 ng L−1. Eleven PFAS species were detected in groundwater, including PFOA, PFHxA, PFPeA, PFBA, PFDA, PFNA, and PFOS. Most of PFAS were quantified in samples located in the aquifer formed by medium to coarse sands intercalated with silty-sandy lenses and low organic matter content that could facilitate the percolation of these compounds. The total concentrations of PFAS (ΣPFAS) in a surface water sample varied between 11 and 17 ng L−1. Ten species were founded in surface water being PFOA, PFOS, and PFHpA the most dominant. Considering all the samples, PFOA was the predominant species in the study. In addition, tributary water bodies had higher concentrations of PFAS than the main water body (Guaiba Lake) probably due dilution process. [ABSTRACT FROM AUTHOR]

Previous reports revealed that peel extracts of Ficus carica (fig) have a wide range of pharmacological and biological activities. The current study aimed to determine the phytochemical components of the ethanol extracts of Peggy Red fig (PRF) and Green fig (GF) peels by liquid chromatography–tandem mass spectrometry (LC–MS/MS) analysis, along with its antioxidant properties and neuroprotective effect in Caenorhabditis elegans. LC–MS/MS analysis confirmed 50 compounds in the extract, which revealed the presence of phenols, flavonoids, and anthocyanins, and exhibited in vitro antioxidant activity. PRF and GF peel had 163.25 (mg gallic acid equivalent [mg GAE]) g−1, 125.32 (mg GAE) g−1 of total phenolic content, 62.52 (mg rutin equivalent [mg RE]) g−1, and 43.36 (mg RE) g−1 flavonoids content, respectively. In all antioxidant assays, the extract of PRF peel showed higher antioxidant activity than the GF peel, and the extract of PRF peel could effectively reduce the aggregation of amyloid-beta (Aβ), decrease the paralysis of the body, and increase the antioxidant enzyme activities to reduce the toxicity of Aβ1–42 in Alzheimer's disease (AD) transgenic C. elegans CL4176. Therefore, PRF peel extract may have potential applications as a new source for drug development against AD. [ABSTRACT FROM AUTHOR]

The main objective of the present study is to estimate, through differential analysis, various biological activities of total phenolics content in alcoholic extracts of three date palm varieties sensitive or resistant to Fusarium oxysporum. sp Albidinis. Here, stilbene products with antioxidant and bioactive capacities were evidenced in resistant variety Taabdount (TAAR). Furthermore, the methanolic fraction of the TAAR-resistant date palm variety contains a significant product, determined by LC-MS/MS and 1H, 13C NMR, belonging to the family of hydroxystilbenes, which exhibits antioxidant capacities, inhibits the mushroom tyrosinase activity, and activates and exerts a protective effect on hypochlorite-induced damage in 20S proteasome of human dermal fibroblast aged cells. Altogether, the present results indicate that hydroxystilbene present in resistant Phoenix dactylifera L. should be studied to understand the way that the stilbene could exert anti-aging ability. [ABSTRACT FROM AUTHOR]

Grain filling is a pivotal grain weight determinant factor and has a great impact on rice yield potential. High grain weight was attributed to a high grain-filling rate. Proteomic analysis during the early ripening stage of rice grains from two cultivars that possess contrasting phenotypes in grain-filling rates was performed to identify proteins associated with high grain-filling rates. Two hundred and nineteen differentially expressed proteins (DEPs) were identified from the grain of two selected cultivars. We observed six more abundant enzymes involved in starch and sucrose metabolism during rice grain filling in the high grain-filling rate cultivar X42. Gene ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that most of the identified DEPs during grain filling were implicated in physiological and biochemical processes. These processes include carbohydrate metabolism, energy metabolism, secondary metabolites metabolism, and amino acids metabolism. Besides, most of those identified DEPs are differentially expressed in the two rice cultivars during grain filling. These findings indicate the essential roles of DEPs in rice grain development. During the grain-filling stage, starch, sucrose, fatty acids, and amino acids biosynthesis were more active and their metabolisms were also significantly enriched. Protein–protein interactions indicate the implication of proteins in several biological processes, for instance, carbohydrate metabolism, fatty acid metabolism, energy metabolism, and secondary metabolites biosynthesis. Data from our study provided valuable information about carbohydrate and amino acids biosynthesis, transport, and their metabolism in two genotypically grain filling. [ABSTRACT FROM AUTHOR]

Psychedelic mushrooms belonging to basidiomycota have gained prominence in research due to the range of hallucinogenic compounds. To combat the challenge of antimicrobial resistance, exploring alternative antimicrobial peptides has become crucial. In this study, we present the proteomic analysis of psychedelic mushroom. Psilocybe cubensis was identified by molecular characterisation using the ITS1 and ITS4 regions. Subsequently, LC-MS/MS and gene ontology analyses were used to characterise the proteome of P. cubensis . The proteomic analysis unveiled several antimicrobial peptides. The results revealed favourable binding scores, suggesting the potential efficacy of these peptides against Staphylococcus aureus . Hence the inhibition of bacterial growth, supporting the antimicrobial properties of the identified peptides. In our findings, the individual peptides from P. cubensis against S. aureus suggest a promising avenue for the discovery of novel antimicrobial peptides.

Chinese hamster ovary (CHO) cells remain the most widely used host cell line for biotherapeutics production. Despite their widespread use, understanding endoplasmic reticulum (ER) stress conditions in recombinant protein production remains limited, often creating bottlenecks preventing improved production titers and product quality. Ubiquitination not only targets substrates (e.g., misfolded proteins) for proteasome degradation but also has important regulatory control functions including cell cycle regulation, translation, apoptosis, autophagy, etc. and hence is likely to be central to understanding and controlling the productivity of recombinant biotherapeutics. This study aimed to uncover differentially expressed ubiquitinated proteins following artificial induction of ER-stress in recombinant CHO cells. CHO cells were treated with the stress inducer tunicamycin and the proteasome inhibitor MG132, followed by LC-MS/MS proteomic analysis. We identified >4000 ubiquitinated peptides from CHO-DP12 cells under ER stress conditions and proteasome inhibition. Moreover, data analysis showed altered abundance levels of >900 ubiquitinated proteins under the combination of ER stress and proteasome inhibition compared to untreated controls. Gene Ontology (GO) analysis of these ubiquitinated proteins resulted in a significant enrichment of key pathways involving the proteasome, protein processing in the ER, N-glycan biosynthesis, and ubiquitin-mediated proteolysis. ER stress response proteins such as GRP78, HSP90B1, ATF6, HERPUD1, and PDIA4 were found to be highly ubiquitinated and exhibited a significant increase in abundance following induction of ER-stress conditions. This study broadens our comprehension of the roles played by protein ubiquitination in CHO cell stress responses, potentially revealing targets for tailored cell line engineering aimed at enhancing stress tolerance and production efficiency., (© 2024 The Author(s). Biotechnology Journal published by Wiley‐VCH GmbH.)

This study developed a suitable analytical method for kasugamycin residues in Achyranthes japonica using LC/MS/MS equipped with an amide column for polar substances. Extraction and cleanup processes were done at pH 4.5–5. Purification efficiency was assessed and confirmed step by step by selecting silica, hydrophilic-lipophilic balance (HLB), strong cation exchange (SCX), and double (HLB and SCX) cleanup SPE cartridges. The results indicated that silica SPE cartridge exhibited overloading tendency, while HLB SPE cartridge had low cleaning efficiency. Among SPE cartridges used, double cleanup and SCX were found sufficient with respective matrix effects of –15% and +14%, respectively. The LOD and LOQ were 0.008 ng and 0.04 mg/kg, respectively. The correlation coefficient (R2) was higher than 0.99, recovery rate ranges were 86.3–97.2%, and the RSD was below 8.8%. All methods are consistent with the Codex guidelines criteria. This study developed an appropriate LC/MS/MS analytical method for kasugamycin residue analysis in A. japonica with optimized, efficient extraction and purification conditions using a single SCX SPE cartridge, which is simple and time-efficient. In addition, the HLB and SCX SPE cartridges of the double cleanup methods were identified as primary methods that can be applied for the cleanup of other medicinal herbs.

Within the global monitoring plan (GMP) established by article 16 of the Stockholm Convention on Persistent Organic Pollutants, perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA), and perfluorohexane sulfonic acid (PFHxS) are recommended for analysis in core matrices to assess occurrence and changes geographically and with time. In 101 samples consisting of 86 national pools and 15 pools from States in Brazil obtained between 2008 and 2019, PFHxS was detected in 17% of the national pools and none in Brazil. PFOA and PFOS had a detection frequency of 100% and 92%, respectively. Other perfluoroalkane substances (PFAS) had either low detection frequencies and median values of zero (carboxylic acids C4–C11; except PFOA) or could not be quantified in any sample (sulfonic acids, C4–C10, and long-chain carboxylic acids, C12–C14). Correlation between PFOA and PFOS was moderately (r = 0.58). Whereas median values were almost identical (18.9 pg/g f.w. for PFOS; 18.6 pg/g f.w. for PFOA), PFOS showed larger ranges (< 6.2 pg/g f.w.−212 pg/g f.w.) than PFOA (< 6.2 pg/g f.w.−63.4 pg/g f.w.). It was shown that wealthier countries had higher PFOA concentrations than poorer countries. No difference in concentrations was found for samples collected in countries having or not having ratified the Stockholm Convention amendments to list PFOS or PFOA. The goal to achieve 50% decrease in concentrations within ten years was met by Antigua and Barbuda, Kenya, and Nigeria for PFOS and by Antigua and Barbuda for PFOA. In a few cases, increases were observed; one country for PFOS, four countries for PFOA. [ABSTRACT FROM AUTHOR]

Diabetes mellitus is a major endocrine dysfunction that severely threatens the global population. Litsea cubeba (Lour.) Pers. (Lauraceae), commonly known as "mountain pepper", has been utilized in many chronic disorders in different countries. However, the antidiabetic activity of this plant has not been explored to date. The present study aimed to explore the antidiabetic and antihyperlipidemic activities of the methanolic extract of fruits of L. cubeba (LCMF) from the Darjeeling Himalaya, India, through in vivo experimentations and network pharmacological analysis. The HFD-STZ induced diabetic model rats were treated with LCMF for 28 days and were examined for blood glucose, lipid profile, liver and kidney function parameters, hepatic antioxidant status and histopathology of liver, kidney and pancreas. The LCMF treatment effectively lowered (about 25%) high blood glucose levels and improved serum lipid profiles, liver glycogen content, and normalized antioxidant status. A significant increment in endogenous insulin (6.83 mU/L) and C-peptide levels (0.86 ng/mL), along with the regeneration of beta islets, were observed. LC-MS/MS and in silico network-pharmacological analysis showed the presence of some important metabolites, viz. thymol, scopoline, isosafrole, safrole, arecoline, etc. that interact with the target proteins (viz. IL6, ADRB1 etc.), transcription factors (viz. NFKB1, CYP1A1 etc.) and enzymes (viz. α-glucosidase, α-amylase etc.) associated with diabetes and hyperlipidemia. The study for the first time proved that the traditional edible fruits of L. cubeba have potent antidiabetic and antihyperlipidemic properties. The subsequent detailed characterization and mode of action studies would lead to discovering new antidiabetic and antihyperlipidemic drugs for human use. • Litsea cubeba fruits exerted significant hypoglycemic activities in vivo. • Promising antihyperlipidemic activities were also observed. • Regeneration of beta-islet cells of the pancreas was observed. • Endogenous insulin and C-peptide levels were found to be increased. • Network pharmacology revealed interactions of metabolites with target proteins. [Display omitted] [ABSTRACT FROM AUTHOR]

Mesobuthus tamulus (Indian Red Scorpion) sting is a severe but neglected health issue in India. The accomplishment of in-patient scorpion sting management is highly dependent on the safety, efficacy, and homogeneity of scorpion antivenom preparation. Therefore, in this study, the above qualities of commercial anti-scorpion antivenoms manufactured in India were assessed by in vitro laboratory analyses. Biophysical characterization of venom by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, size exclusion chromatography, and proteomics analysis demonstrated that anti-scorpion antivenoms (ASAs) mostly contain F(ab') 2 molecules with a trace amount of undigested immunoglobulin (Ig) G. The physicochemical characterization, electron microscopy, and dynamic light scattering studies revealed that ASAs were prepared according to the guidelines of World Health Organization (WHO), and were devoid of aggregate content and virus particles. ASAs did not show IgE contamination and bacterial endotoxin but demonstrated moderate complement activation properties, which may have adverse effects in treated patients. Spectrofluorometric and atomic force microscopy analyses showed poor binding of venom with commercial ASAs. The percent of antibodies raised against the venom toxins in commercial ASAs was determined at the range of 5.3–6.3%, which is a reason for their poor efficacy. This study advocates the importance of in vitro laboratory analyses for assessing commercial antivenom's quality and safety parameters before their pre-clinical research and clinical use to treat Indian red scorpion sting. [Display omitted] • Commercial anti-scorpion antivenoms (ASAs) manufactured in India contain F(ab') 2 with a trace quantity of immunoglobulin G. • Study showed that the ASAs were prepared according to the guidelines of the World Health Organization. • The ASAs were devoid of aggregate content and virus particles. • Biophysical analyses demonstrated the poor binding of venom with commercial ASAs. • Commercial ASAs contain less proportion of scorpion venom toxins-specific antibodies. [ABSTRACT FROM AUTHOR]

Manure is a major source of soil and plant contamination with veterinary drugs residues. The aim of this study was to evaluate the uptake of 14 veterinary pharmaceuticals by parsley from soil fertilized with manure. Pharmaceutical content was determined in roots and leaves. Liquid chromatography coupled with tandem mass spectrometry was used for targeted analysis. Screening analysis was performed to identify transformation products in the parsley tissues. A solid-liquid extraction procedure was developed combined with solid-phase extraction, providing recoveries of 61.9–97.1% for leaves and 51.7–95.6% for roots. Four analytes were detected in parsley: enrofloxacin, tylosin, sulfamethoxazole, and doxycycline. Enrofloxacin was detected at the highest concentrations (13.4–26.3 ng g−1). Doxycycline accumulated mainly in the roots, tylosin in the leaves, and sulfamethoxazole was found in both tissues. 14 transformation products were identified and their distribution were determined. This study provides important data on the uptake and transformation of pharmaceuticals in plant tissues. [ABSTRACT FROM AUTHOR]

The Indian monocled cobra (Naja kaouthia) is one of the most prevalent venomous snakes in northeast India (NEI) and is the cause of many fatalities. The composition of NEI N. kaouthia venom (NkV) was deciphered using two different proteomic approaches: (i) 1D SDS-PAGE coupled to label-free quantification of protein bands using stringent identification criteria and (ii) reversed-phase high-performance liquid chromatography (RP-HPLC) followed by quantification based on area under the RP-HPLC peaks. The proteomic data from both strategies were compared. Proteomic analyses from both workflows identified 32 proteins (toxins) distributed over 10–14 snake venom protein families in NEI NkV. The relative abundances of the venom proteins determined from the analytical workflows coincided with the densitometry band intensities of the NEI NkV. Phospholipase A 2 (13.1–16.0%) and three-finger toxins (58.5–64.2%) represented the most abundant enzymatic and non-enzymatic proteins in NEI NkV, respectively. Immuno-cross-reactivity studies by enzyme-linked immunoassay and immunoblot analyses pointed to the poor efficacy of commercial PAVs in recognizing the low molecular mass (<15 kDa) toxins of NEI NkV. Spectrofluorometric titration determined the presence of NEI NkV-specific antibodies in commercial PAV, at a level that was higher than that previously reported for eastern India NkV-specific antibodies in commercial antivenom. • Each method of venom de-complexation for protein identification and quantification has its pros and cons • The venom proteome composition of N. kaouthia from North-East India was deciphered using two proteomic workflows and compared • The study demonstrated that both analytical workflows can be considered reliable approaches for analyzing snake venom proteome • The 3FTx and PLA 2 represented the most abundant non-enzymatic and enzymatic proteins in the tested venom • Immunoprofiling demonstrated poor efficacy of commercial PAVs against the low molecular weight (<15 kDa) NEI NKVtoxins [ABSTRACT FROM AUTHOR]

A liquid chromatography tandem mass spectrometry (LC–MS/MS) multi-mycotoxin method was developed for the analysis of the Alternaria toxins alternariol (AOH), alternariol monomethyl ether (AME), tentoxin (TEN), altertoxin I (ATX I), altertoxin II (ATX II), alterperylenol (ALTP), and altenuene (ALT), as well as the modified toxins AOH-3-glucoside (AOH-3-G), AOH-9-glucoside (AOH-9-G), AME-3-glucoside (AME-3-G), AOH-3-sulfate (AOH-3-S), and AME-3-sulfate (AME-3-S) in barley and malt. The toxin tenuazonic acid (TeA) was analyzed separately as it could not be included into the multi-mycotoxin method. Quantitation was conducted by using a combination of stable isotope dilution analysis (SIDA) for AOH, AME, and TeA, and matrix-matched calibration for all other toxins. Limits of detection were between 0.05 µg/kg (AME) and 2.45 µg/kg (ALT), whereas limits of quantitation ranged from 0.16 µg/kg (AME) to 8.75 µg/kg (ALT). Recoveries between 96 and 107% were obtained for the analytes when SIDA was applied, while recoveries between 84 and 112% were found for analytes quantified by matrix-matched calibration. The method was applied for the analysis of 50 barley samples and their respective malts from the harvest years 2016–2020 for their mycotoxin content, showing the overall potential of toxin formation during the malting process. The toxins ALTP and ATX I were mainly found in the malt samples, but not in barley. [ABSTRACT FROM AUTHOR]

Intoxication by organophosphorus (OP) poisons, like nerve agents and pesticides, is characterized by the life-threatening inhibition of acetylcholinesterase (AChE) caused by covalent reaction with the serine residue of the active site of the enzyme (phosphylation). Similar reactions occur with butyrylcholinesterase (BChE) and serum albumin present in blood as dissolved proteins. For forensic purposes, products (adducts) with the latter proteins are highly valuable long-lived biomarkers of exposure to OP agents that are accessible by diverse mass spectrometric procedures. In addition, the evidence of poison incorporation might also succeed by the detection of remaining traces of the agent itself, but more likely its hydrolysis and/or enzymatic degradation products. These relatively short-lived molecules are distributed in blood and tissue, and excreted via urine. This review presents the mass spectrometry-based methods targeting the different groups of biomarkers in biological samples, which are already internationally accepted by the Organisation for the Prohibition of Chemical Weapons (OPCW), introduces novel approaches in the field of biomedical verification, and outlines the strict quality criteria that must be fulfilled for unambiguous forensic analysis.

Considering medicinal plants as an inexhaustible source of active ingredients that may be easily isolated using simple and inexpensive techniques, phytotherapy is becoming increasingly popular. Various experimental approaches and analytical methods have been used to demonstrate that the genus Calendula (Asteraceae) has a particular richness in active ingredients, especially phenolic compounds, which justifies the growing interest in scientific studies on this genus' species. From a chemical and biological viewpoint, Calendula aegyptiaca is a little-studied plant. For the first time, high-performance liquid chromatography combined with negative electrospray ionization mass spectrometry (HPLC-HESI-MS) was used to analyze methanolic extracts of Calendula aegyptiaca (C. aegyptiaca) fruits. Thirty-five molecules were identified. Flavonoids (47.87%), phenolic acids (5.18%), and saponins (6.47%) formed the majority of these chemicals. Rutin, caffeic acid hexoside, and Soyasaponin βg' were the most abundant molecules in the fruit methanolic extract, accounting for 17.49% of total flavonoids, 2.32 % of total phenolic acids, and 0.95% of total saponins, respectively. The antioxidant activity of the fruit extracts of C. aegyptiaca was investigated using FRAP, TAC, and DPPH as well as flavonoids and total phenols content. Because the phenolic components were more extractable using polar solvents, the antioxidant activity of the methanolic extract was found to be higher than that of the dichloromethane and hexane extracts. The IC50 value for DPPH of methanolic extract was found to be 0.041 mg·mL−1. Our findings showed that C. aegyptiaca is an important source of physiologically active compounds. [ABSTRACT FROM AUTHOR]

Objectives: The objectives of the research are to extract chemicals from Algerian fir needles, to determine the total phenolics and flavonoids contents in ethylacetate fraction (EAF) and to assess the biological activities of the extract. Methods: Several in-vitro biochemical assays were used to measure antioxidant and oxygen radical-scavenging abilities, and antibacterial activity was evaluated. Components of the extract were identified by mass spectrometry and comparison to reference compounds. Results: Results showed that EAF presented a powerful antioxidant activity in all assays and exhibited a potent inhibitory effect against all bacterial strains used. Fractionation and mass spectrometry identified a high amount of astragalin, hyperoside and quercitrin in the EAF from needles. Conclusion: The best of our knowledge, we report here for the first time, the evaluation of biological activities and the determination of phytochemical profile of Algerian fir leaves extract underlying the importance of this species as a source of health-promoting.

Dereplication of crude plant extracts through liquid chromatography-mass spectrometry is a powerful technique for the discovery of novel natural products. Unfortunately, this technique is often plagued by a low level of confidence in natural product identification. This is mainly due to the lack of extensive chromatographic and mass spectrometric optimizations that result in improper and incomplete MS/MS fragmentation data. This study proposes a solution to this problem by the optimization of chromatographic separation and mass spectrometry parameters. We report herein a direct and high-throughput strategy for natural product dereplication in five Salvia species using high-resolution ESI-QTOF-MS/MS data. In the present study, we were able to identify a total of forty-seven natural products in crude extracts of five Salvia species using MS/MS fragmentation data. In addition to dereplication of Salvia species, quantitative profiling of twenty-one bioactive constituents of the genus was also performed on an ion trap mass spectrometer. For the quantitation study, method development focused on chromatographic optimizations to achieve maximum sensitivity. The developed dereplication and quantitation strategy can be extended to develop comprehensive metabolic profiles of other plant genera and species and thus can prove useful in the field of drug discovery from plants.