Hybrid-genome sequence analysis of Enterobacter cloacae FACU and morphological characterization: insights into a highly arsenic-resistant strain.

Many organisms have adapted to survive in environments with high levels of arsenic (As), a naturally occurring metalloid with various oxidation states and a common element in human activities. These organisms employ diverse mechanisms to resist the harmful effects of arsenic compounds. Ten arsenic-resistant bacteria were isolated from contaminated wastewater in this study. The most efficient bacterial isolate able to resist 15,000 ppm Na2HAsO4·7H2O was identified using the 16S rRNA gene and whole genome analysis as Enterobacter cloacae FACU. The arsenic E. cloacae FACU biosorption capability was analyzed. To further unravel the genetic determinants of As stress resistance, the whole genome sequence of E. cloacae FACU was performed. The FACU complete genome sequence consists of one chromosome (5.7 Mb) and two plasmids, pENCL 1 and pENCL 2 (755,058 and 1155666 bp, respectively). 7152 CDSs were identified in the E. cloacae FACU genome. The genome consists of 130 genes for tRNA and 21 for rRNAs. The average G + C content was found to be 54%. Sequencing analysis annotated 58 genes related to resistance to many heavy metals, including 16 genes involved in arsenic efflux transporter and arsenic reduction (five arsRDABC genes) and 42 genes related to lead, zinc, mercury, nickel, silver, copper, cadmium and chromium in FACU. Scanning electron microscopy (SEM) confirmed the difference between the morphological responses of the As-treated FACU compared to the control strain. The study highlights the genes involved in the mechanism of As stress resistance, metabolic pathways, and potential activity of E. cloacae FACU at the genetic level.

Pseudochrobactrum asaccharolyticum mitigates arsenic induced oxidative stress of maize plant by enhancing water status and antioxidant defense system.

BACKGROUND: Oxidative stress mediated by reactive oxygen species (ROS) is a common denominator in arsenic toxicity. Arsenic stress in soil affects the water absorption, decrease stomatal conductance, reduction in osmotic, and leaf water potential, which restrict water uptake and osmotic stress in plants. Arsenic-induced osmotic stress triggers the overproduction of ROS, which causes a number of germination, physiological, biochemical, and antioxidant alterations. Antioxidants with potential to reduce ROS levels ameliorate the arsenic-induced lesions. Plant growth promoting rhizobacteria (PGPR) increase the total soluble sugars and proline, which scavenging OH radicals thereby prevent the oxidative damages cause by ROS. The main objective of this study was to evaluate the potential role of Arsenic resistant PGPR in growth of maize by mitigating arsenic stress. METHODOLOGY: Arsenic tolerant PGPR strain MD3 (Pseudochrobactrum asaccharolyticum) was used to dismiss the 'As' induced oxidative stress in maize grown at concentrations of 50 and 100 mg/kg. Previously isolated arsenic tolerant bacterial strain MD3 "Pseudochrobactrum asaccharolyticum was used for this experiment. Further, growth promoting potential of MD3 was done by germination and physio-biochemical analysis of maize seeds. Experimental units were arranged in Completely Randomized Design (CRD). A total of 6 sets of treatments viz., control, arsenic treated (50 & 100 mg/kg), bacterial inoculated (MD3), and arsenic stress plus bacterial inoculated with three replicates were used for Petri plates and pot experiments. After treating with this MD3 strain, seeds of corn were grown in pots filled with or without 50 mg/kg and 100 mg/kg sodium arsenate. RESULTS: The plants under arsenic stress (100 mg/kg) decreased the osmotic potential (0.8 MPa) as compared to control indicated the osmotic stress, which caused the reduction in growth, physiological parameters, proline accumulation, alteration in antioxidant enzymes (Superoxide dismutase-SOD, catalase-CAT, peroxidase-POD), increased MDA content, and H2O2 in maize plants. As-tolerant Pseudochrobactrum asaccharolyticum improved the plant growth by reducing the oxidation stress and antioxidant enzymes by proline accumulation. PCA analysis revealed that all six treatments scattered differently across the PC1 and PC2, having 85.51% and 9.72% data variance, respectively. This indicating the efficiency of As-tolerant strains. The heatmap supported the As-tolerant strains were positively correlated with growth parameters and physiological activities of the maize plants. CONCLUSION: This study concluded that Pseudochrobactrum asaccharolyticum reduced the 'As' toxicity in maize plant through the augmentation of the antioxidant defense system. Thus, MD3 (Pseudochrobactrum asaccharolyticum) strain can be considered as bio-fertilizer.

Pepper immunity against Ralstonia solanacearum is positively regulated by CaWRKY3 through modulation of different WRKY transcription factors.

BACKGROUND: Several WRKY transcription factors (TFs), including CaWRKY6, CaWRKY22, CaWRKY27, and CaWRKY40 are known to govern the resistance of pepper (Capsicum annuum L.) plants to Ralstonia solanacearum infestation (RSI) and other abiotic stresses. However, the molecular mechanisms underlying these processes remain elusive. METHODS: This study functionally described CaWRKY3 for its role in pepper immunity against RSI. The roles of phytohormones in mediating the expression levels of CaWRKY3 were investigated by subjecting pepper plants to 1 mM salicylic acid (SA), 100 µM methyl jasmonate (MeJA), and 100 µM ethylene (ETH) at 4-leaf stage. A virus-induced gene silencing (VIGS) approach based on the Tobacco Rattle Virus (TRV) was used to silence CaWRKY3 in pepper, and transiently over-expressed to infer its role against RSI. RESULTS: Phytohormones and RSI increased CaWRKY3 transcription. The transcriptions of defense-associated marker genes, including CaNPR1, CaPR1, CaDEF1, and CaHIR1 were decreased in VIGS experiment, which made pepper less resistant to RSI. Significant hypersensitive (HR)-like cell death, H2O2 buildup, and transcriptional up-regulation of immunological marker genes were noticed in pepper when CaWRKY3 was transiently overexpressed. Transcriptional activity of CaWRKY3 was increased with overexpression of CaWRKY6, CaWRKY22, CaWRKY27, and CaWRKY40, and vice versa. In contrast, Pseudomonas syringae pv tomato DC3000 (Pst DC3000) was easily repelled by the innate immune system of transgenic Arabidopsis thaliana that overexpressed CaWRKY3. The transcriptions of defense-related marker genes like AtPR1, AtPR2, and AtNPR1 were increased in CaWRKY3-overexpressing transgenic A. thaliana plants. CONCLUSION: It is concluded that CaWRKY3 favorably regulates phytohormone-mediated synergistic signaling, which controls cell death in plant and immunity of pepper plant against bacterial infections.

Investigating the therapeutic potential of aqueous extraction of curry plant (Murraya koenigi) leaves supplementation for the regulation of blood glucose level in type 2 diabetes mellitus in female human subjects.

Type 2 diabetes mellitus is characterized by hyperglycemia and insulin resistance. It is spreading around the globe like a pandemic. Major factors behind the development of diabetes can be genetics, environmental factors, dietary choices and obesity. Many medicinal plants have anti-diabetic potential. This study has investigated the anti-diabetic effect of curry leaves extract. This study also investigated the chemical characterization of curry leaves. Phytochemicals including saponins, tannins, alkaloids, flavonoids, phenols and glycosides were also investigated. Encapsulated 5mg per kg of the body weight and 10mg per kg of the body weight were given to treatment groups I and II. Random blood sugar, fasting blood sugar and HbA1c of 45 diabetic female adults were measured on the 0-day and 45th days. All results were analyzed using the two-sample t-test in IBM SPSS Statistics 20. Curry leaves contained moisture (24.1±1.78)%, ash (17.82±2.13)%, nitrogen free extract (36.12±3.52)%, crude protein (8.32±0.83)%, crude fiber (6.98±2.31)% and crude fat (6.87±0.21)%. Mineral analysis showed that magnesium and calcium were major minerals present in curry leaves. Curry leaves extract contained saponins 2.71±0.23, flavonoids 7.84±0.42, tannins 0.91±0.09, glycosides 0.17±0.01, phenols 3.89±0.12, alkaloids 2.01±0.87. These phytochemicals were expressed in mg/100 g of the sample. Curry leaf extract showed a significant (p<0.05) reduction in fasting blood sugar, random blood sugar and glycated hemoglobin in both treatment groups.

Investigating the nutraceutical potential of apple peel extract supplementation for regulating the glucose metabolism in hyperlipidemic Female human subjects.

Hyperglycemia is a condition often observed in diabetics, dyslipidemia and obese. It is a major factor behind the development of diabetes and the reasons can be genetics, environmental factors, dietary choices and obesity. Many medicinal plants have anti-diabetic potential. This study investigated the anti-hyperglycemic effect of apple peel extract. This study also investigated the chemical characterization of apple peel. Phytochemicals including total phenolics and flavonoids were determined. Encapsulated 350mg/day was given to treatment groups. Random blood sugar, fasting blood sugar and HbA1c of 45 diabetic female adults was measured on the 0-day and 45th day. Results showed that apple peel contained moisture (14.71±3.57)%, ash (17.82±2.13)%, nitrogen free extract (32.12±3.52)%, crude protein (6.89±0.83)%, crude fiber (19.17±0.21)% and crude fat (9.91±2.31)%. Findings showed that apple peel contains magnesium (6.61±1.088), calcium (8.17±0.32), zinc (14.08±1.21) and potassium (67.21±1.86). These findings were shown in mg in kg. Apple peel extract contained total phenolic content (TPC) of 8.14±1.07 and total phenolic content (TFC) of 4.89±1.81. Apple peel extract showed a significant reduction in all blood parameters of hyperglycemia. All results were significant at p<0.05.

Determination of chemical composition and investigation of potential of triphala powder in hypercholesterolemia in men in controlled randomized trial.

Hypercholesterolemia is a condition with elevated cholesterol and lipid profile. It is the leading reason behind myocardial infarction and coronary heart disease. It is observed in young people as well due to a sedentary lifestyle. Triphala powder has a hypolipidemic and anti-hypercholesterolemia effect. This study was designed to investigate the effect of triphala powder against hypercholesterolemia. This study also examined Triphala powder's chemical composition. Total phenolic and flavonoid content were examined. Encapsulated 400 mg and 600 mg Triphala powder were given to treatment groups I and II. Lipid profile parameters were measured and compared at 0 weeks and 10th weeks in all groups. All results were analyzed using ANOVA in IBM SPSS Statistics 20. Results of proximate analyses have shown that Okra pod powder contains moisture 12.27%, ash 11.25%, nitrogen-free extract 45.93%, crude protein 13.37%, crude fat 2.95% and crude fiber 14.23%. Mineral analysis showed that iron and manganese are major minerals in triphala powder. Triphala powder showed a significant reduction in lipid profile parameters in hypercholesterolemia. All results are taken significantly at p<0.05.

Evaluation of chemical characterization and antihyperlipidemic potential of orange peel powder (Citrus sinensis) in male human subjects.

Citrus sinensis is an important member of the genus Citrus which contains phenolic compounds and bioflavonoids which have antihyperlipidemic and antiatherogenic effects. It also has the potential to reduce oxidative stress. To investigate the antihyperlipidemic effect of orange peel powder was encapsulated and analyzed in hyperlipidemic patients. Results showed that it contains moisture (12.2%), ash content (7.9%), crude fat (0.78%), crude protein (12.37%) and crude fiber (13.2%). Total phenolic content and total flavonoid content were observed as 163.17 mg and 17.23mg in quercetin equivalent per gram a dry weight basis. Furthermore, the Orange peel powder was given in the form of medicinal capsules to hyperlipidemia male subjects. The experimental groups (G1 and G2) were given orange peel powder in capsules 400mg/d to the G1 group and 800mg/d to the G2 group for the time of 45 days. The serum lipid profile of patients was measured before and after the experimental trial. The result showed that G1 and G2 showed a decrease in plasma lipid parameters and increased high-density lipoprotein content in blood substantially as compared to G0. Thus, it was concluded from the results that orange peel powder depicts a significant impact on treating hyperlipidemia.

Omega-3 Polyunsaturated Fatty Acids Provoke Apoptosis in Hepatocellular Carcinoma through Knocking Down the STAT3 Activated Signaling Pathway: In Vivo and In Vitro Study.

Hepatocellular carcinoma (HCC) is a common type of liver cancer and is a leading cause of death worldwide. Signal transducer and activator of transcription 3 (STAT3) is involved in HCC progression, migration, and suppression of apoptosis. This study investigates the apoptotic effect of the dietary antioxidant (n-3 PUFAs) on HepG2 cells and analyzes the underlying molecular mechanisms of this effect both in vivo and in vitro. In vivo study: Seventy-five adult male albino rats were divided into three groups (n = 25): Group I (control): 0.9% normal saline, intraperitoneal. Group II: N-Nitrosodiethylamine (200 mg/kg b.wt) intraperitoneal, followed by phenobarbital 0.05% in drinking water. Group III: as group II followed by n-3 PUFAs intubation (400 mg/kg/day). In vivo study: liver specimens for biochemical, histopathological, and immunohistochemical examination. In vitro study: MTT assay, cell morphology, PCR, Western blot, and immunohistochemical analysis. n-3 PUFAs significantly improved the histopathologic features of HCC and decreased the expression of anti-apoptotic proteins. Further, HepG2 cells proliferation was suppressed through inhibition of the STAT3 signaling pathway, cyclin D1, and Bcl-2 activity. Here we report that n-3 PUFAs may be an ideal cancer chemo-preventive candidate by targeting STAT3 signaling, which is involved in cell proliferation and apoptosis.

In Vitro Study of the Phytochemical Composition and Antioxidant, Immunostimulant, and Hemolytic Activities of Nigella sativa (Ranunculaceae) and Lepidium sativum Seeds.

The Moroccan flora abounds and is an important reserve of medicinal plants. Nigella sativa and Lepidium sativum are plants that are widely used in traditional medicine for their multiple therapeutic properties. The current study aims to highlight the biological activities that can justify and valorize the use of these plants. Flavonoids, total phenols, condensed tannins, and sugars were determined. The biological activities tested were antioxidant by determining the IC50 (defined as the concentration of an antioxidant required to decrease the initial concentration by 50%; inversely related to the antioxidant capacity), hemagglutination, and hemolytic activities. Phytochemical quantification of the seed extracts indicated that the total phenol content was largely similar for both plants and in the order of 10 mg GAE (Gallic acid equivalent)/g. On the other hand, L. sativum seeds registered a higher content of flavonoids (3.09 ± 0.04 mg QE (quercetin equivalent)/g) as compared to Nigella saliva (0.258 ± 0.058). Concerning condensed tannins, N. saliva seeds present a higher amount with a value of 7.2 ± 0.025 mg/g as compared to L. sativum (1.4 ± 0.22 mg/g). Concerning the total sugar content, L. sativum shows a higher content (67.86 ± 0.87 mg/g) as compared to N. sativa (58.17 ± 0.42 mg/g); it is also richer in mucilage with a content of 240 mg as compared to 8.2 mg for N. saliva. Examination of the antioxidant activity using a DPPH (2.2-diphenyl 1-pycrilhydrazyl) test revealed that the EButOH (n-butanol extract) and EAE (ethyl acetate extract) extracts were the most active, with IC50 values of 48.7 and 50.65 µg/mL for the N. sativa extracts and 15.7 and 52.64 µg/mL for the L. sativum extracts, respectively. The results of the hemagglutination activity of the different extracts of the two plants prepared in the PBS (phosphate-buffered saline) medium showed significant agglutination for the L. sativum extract (1/50) compared to the N. sativa extract (1/20). An evaluation of the hemolytic effect of the crude extract of the studied seeds on erythrocytes isolated from rat blood incubated in PBS buffer compared to the total hemolysis induced by distilled water showed a hemolysis rate of 54% for Nigella sativa and 34% for L. sativum. In conclusion, the two plants studied in the current work exhibited high antioxidant potential, which could explain their beneficial properties.

Interleukin-18 up-regulates amino acid transporters and facilitates amino acid-induced mTORC1 activation in natural killer cells.

Upon inflammation, natural killer (NK) cells undergo metabolic changes to support their high energy demand for effector function and proliferation. The metabolic changes are usually accompanied by an increase in the expression of nutrient transporters, leading to increased nutrient uptake. Among various cytokines inducing NK cell proliferation, the mechanisms underlying the effect of interleukin (IL)-18 in promoting NK cell proliferation are not completely understood. Here, we demonstrate that IL-18 is a potent cytokine that can enhance the expression of the nutrient transporter CD98/LAT1 for amino acids independently of the mTORC1 pathway and thereby induce a dramatic metabolic change associated with increased proliferation of NK cells. Notably, treatment of IL-18-stimulated NK cells with leucine activates the metabolic sensor mTORC1, indicating that the high expression of amino acid transporters induces amino acid-driven mTORC1 activation. Inhibition of the amino acid transporter CD98/LAT1 abrogated the leucine-driven mTORC1 activation and reduced NK cell effector function. Taken together, our study identified a novel role of IL-18 in up-regulating nutrient transporters on NK cells and thereby inducing metabolic changes, including the mTORC1 activation by amino acids.

Anabasis setifera leaf extract from arid habitat: A treasure trove of bioactive phytochemicals with potent antimicrobial, anticancer, and antioxidant properties.

The main objective of this study was to evaluate the biological activities of Anabasis setifera extract, including its antimicrobial, anticancer, and antioxidant properties. In the current study, Anabasis setifera leaves extract was evaluated for antimicrobial, anticancer, antioxidant activities and phytochemical analyses. Ethyl acetate extract of Anabasis setifera (EA-AS) exhibited promising antimicrobial activity toward Escherichia coli, Staphylococcus aureus, Salmonella typhimurium, Bacillus subtilis, Candida albicans, Aspergillus brasiliensis, Aspergillus fumigatus with MICs 62.5, 125, 62.5, 31.25, 62.5, 125 and 125 µg/mL respectively. Moreover, EA-AS showed anticancer activity at safe concentrations, where IC50 were 36.4 and 44 µg/mL toward Hep-G2 and MCF-7 cancerous cell lines. EA-AS was found to contain 55 significant compounds identified through gas chromatography mass spectrophotometry (GCMS). The most abundant compounds were 1,4-dimethoxy-6,7,8,9-tetrahydro-5-benzocycloheptenone (26.04%), hexa-2,4-diyn-1-ylbenzene (8.40%), dihydrobenzo[b]fluoranthene (6.10%), ethanone, 1-[2,3-dihydro-2-(1-methylethenyl)-5-benzofuranyl (6.10%), and valerenol (4.08%). GC mass analysis confirmed the antioxidant properties of AS by detecting several compounds with antioxidant activity, including hexa-2,4-diyn-1-ylbenzene, nerolidol, spathulenol, -naphthalenem ethanol, decahydro-4-trimethyl-8-methylene, hexadecenoic acid, tremetone, desmethoxyencecalin, heptadecyn-1-ol, thunbergol, hexadecanol, dotriacontane, taylorione, ligulatin, retinoic acid, and falcarinol. The analysis of EA-AS reveals that it is a rich source of valuable phytochemicals: total Phenolic Content: a promising 4,264 µg/mL /, suggesting substantial biological and pharmacological potential. Total tannin content: 391.17 µg/mL, indicating potential applications in industries like nutraceuticals, pharmaceuticals, and cosmetics. Total flavonoid content exceptionally high at 5,163 µg/mL, while the total alkaloid content measured 1,036.26 µg/mL. Additionally, EA-AS demonstrated antioxidant activity with an EC50 of 30.6 µg/mL. In conclusion, the comprehensive analysis of the EA-AS reveals its immense potential as a rich source of valuable phytochemicals with diverse bioactivities, warranting further in-depth studies to unlock its full pharmaceutical and commercial prospects. Our results suggest substantial biological and pharmacological prospects for EA-AS as a promising antimicrobial, anticancer, and potent antioxidant.

Comprehensive multi-omics analysis and prognostic significance of fibroblast growth factor binding protein 1 (FGFBP1) in pancreatic adenocarcinoma.

BACKGROUND: Pancreatic adenocarcinoma (PAAD) is a highly aggressive cancer with poor prognosis and limited therapeutic options. Identifying molecular markers and understanding their role in PAAD pathogenesis is crucial for developing targeted therapies. This study integrates bioinformatics and molecular experiments to investigate the diagnostic, prognostic, and therapeutic significance of FGFBP1 in PAAD. METHODS: UALCAN, TNMplot, OncoDB, GEPIA2, HPA, GSCA, KM Plotter, TISIDB, TISCH2, CancerSEA, STRING, DAVID, cell culture, RT-qPCR analysis, western blot analysis, colony formation, cell proliferation, and wound healing assays. RESULTS: Expression analyses revealed a significantly elevated FGFBP1 levels in PAAD tissues compared to normal samples. Promoter methylation analysis indicated lower methylation levels in PAAD, inversely correlated with FGFBP1 expression, suggesting epigenetic regulation. Genetic alteration analysis showed that FGFBP1 is not significantly affected by single nucleotide variants, but copy number variations are present without impacting mRNA expression. Survival analysis using KM plotter demonstrated that high FGFBP1 expression is associated with poor overall and disease-free survival. A Cox regression-based prognostic model confirmed the negative impact of elevated FGFBP1 on patient outcomes. Correlation analysis with immune-related factors indicated that FGFBP1 may contribute to an immunosuppressive tumor microenvironment, affecting immune cell infiltration and function. Single-cell analysis highlighted FGFBP1 expression in malignant, endothelial, and fibroblast cells within the tumor microenvironment. Gene enrichment analysis revealed FGFBP1's involvement in various biological processes and pathways related to cancer progression. Experimental validation using RT-qPCR confirmed high FGFBP1 expression in PAAD cell lines. FGFBP1 knockdown in HEK293T cells significantly reduced cell proliferation, colony formation, and migration. CONCLUSION: These findings suggest that FGFBP1 plays a critical role in PAAD pathogenesis and could serve as a potential therapeutic target for improving patient outcomes.

Peripheral NK cell phenotypic alteration and dysfunctional state post hepatitis B subviral particles stimulation in CHB patients: evading immune surveillance.

Background: The relationship between chronic hepatitis B (CHB) infection and natural killer (NK) cell dysfunction is well-established, but the specific role of HBV viral antigens in driving NK cell impairment in patients with CHB remains unclear. This study investigates the modulatory effects of hepatitis B virus subviral particles (HBVsvp, a representative model for HBsAg) on the phenotypic regulation (activating and inhibitory receptors), cytokine production and cytotoxic potential of peripheral blood mononuclear cell-derived natural killer cells (PBMCs-derived NK cell), which contributes to NK cell dysfunction in CHB infection, potentially serving as an effective HBV immune evasion strategy by the virus. Methods: NK cells were isolated from peripheral blood of patients with CHB (n=5) and healthy individuals (n=5), stimulated with HBVsvp. Subsequent flow cytometric characterization involved assessing changes in activating (NKp46 and NKG2D) and inhibitory (CD94) receptors expression, quantifying TNF-α and IFN- γ cytokine secretion, and evaluating the cytotoxic response against HepG2.2.15 cells with subsequent HBVsvp quantification. Results: In CHB patients, in vitro exposure of PBMCs-derived NK cell with HBVsvp (represent HBsAg model) significantly reduced NK cell-activating receptors expression (P = 0.022), increased expression of CD94 + NK cells (p = 0.029), accompanied with a reduced TNF-α - IFN-γ cytokine levels, and impaired cytotoxic capacity (evidenced by increased cell proliferation and elevated HBVsvp levels in co-cultures with HepG2.2.15 cells in a time-dependent), relative to healthy donors. Conclusion: These findings suggest that HBVsvp may induce dysfunctional NK cell responses characterized by phenotypic imbalance with subsequent reduction in cytokine and cytotoxic levels, indicating HBVsvp immunosuppressive effect that compromises antiviral defense in CHB patients. These data enhance our understanding of NK cell interactions with HBsAg and highlight the potential for targeting CD94 inhibitory receptors to restore NK cell function as an immunotherapeutic approach. Further clinical research is needed to validate these observations and establish their utility as reliable biomarkers.

Phyto-assisted synthesis of zinc oxide nanoparticles using Bauhinia variegata buds extract and evaluation of their multi-faceted biological potentials.

Zinc oxide nanoparticles have wide range biological, biomedical and environmental applications. However, traditional nanofabrication of ZnONPs uses various toxic chemicals and organic solvents which limit their bio-applications. To overcome this hurdle, Bauhinia variegata derived buds extract was utilized to fabricate ZnONPs. The greenly generated ZnONPs were successfully prepared and extensively characterized using different analytical tools and the average crystalline size was calculated as 25.47 nm. Further, bioengineered ZnONPs were explored for multiple biological activities that revealed excellent therapeutic potentials. The antibacterial potential was determined using different bacterial strains. Pseudomonas aeruginosa (MIC: 137.5 µg/mL) was reported to be the most resistant variant while Bacillus subtilis (MIC: 34.38 µg/mL) was observed to be most susceptible bacterial strain. DPPH radical scavenging potential was measured to determine the antioxidant capacity of ZnONPs and the highest scavenging potential was observed as 82% at highest of 300 µg/mL. The fungicidal effect of green ZnONPs in comparison with Amphotericin B was assessed against five selected pathogenic fungal strains. The results revealed, Fusarium solani (MIC: 46.875 µg/mL) was least resistant and Aspergillus flavus (MIC: 187.5 µg/mL) was most resistant in fungicidal examination. Cytotoxicity potential of B.V@ZnONPs was analyzed against newly hatched nauplii of brine shrimps. The results for greenly produced ZnONPs was recorded as 39.78 µg/mL while 3.006 µg/mL was reported for positive control vincristine sulphate. The results confirmed the category of general cytotoxic for greenly synthesized nano sized B.V@ZnONPs.

New Pyrimidinone Bearing Aminomethylenes and Schiff Bases as Potent Antioxidant, Antibacterial, SARS-CoV-2, and COVID-19 Main Protease MPro Inhibitors: Design, Synthesis, Bioactivities, and Computational Studies.

New 2-thioxopyrimidinone derivatives (A1-A10) were synthesized in 87-96% yields via a simple three-component condensation reaction. These compounds were screened extensively through in vitro assays for antioxidant and antibacterial investigations. The DPPH assays resulted in the excellent potency of A6-A10 as antioxidants with IC50 values of 0.83 ± 0.125, 0.90 ± 0.77, 0.36 ± 0.063, 1.4 ± 0.07, and 1.18 ± 0.06 mg/mL, which were much better than 1.79 ± 0.045 mg/mL for the reference ascorbic acid. These compounds exhibited better antibacterial potency against Klebsiella with IC50 values of 2 ± 7, 1.32 ± 8.9, 1.19 ± 11, 1.1 ± 12, and 1.16 ± 11 mg/mL for A6-A10. High-throughput screenings (HTS) of these motifs were carried out including investigation of drug-like behaviors, physiochemical property evaluation, and structure-related studies involving DFT and metabolic transformation trends. The radical scavenging ability of the synthesized motifs was validated through molecular docking studies through ligand-protein binding against human inducible nitric oxide synthase (HINOS) PDB ID: 4NOS, and the results were promising. Furthermore, the antiviral capability of the compounds was examined by in silico studies using two viral proteins PDB ID: 6Y84 and PDB ID: 6LU7. Binding poses of ligands were discussed, and amino acids in the protein binding pockets were investigated, where the tested compounds showed much better binding affinities than the standard inhibitors, proving to be suitable leads for antiviral drug discovery. The stabilities of the molecular docked complexes in real systems were validated by molecular dynamics simulations.

Analyzing the mutational landscape of prostate cancer susceptibility genes through next-generation sequencing (NGS).

OBJECTIVES: Prostate cancer is characterized by diverse genetic mutations that influence disease progression and treatment response. This study was launched to explore the genetic basis of prostate cancer patients. METHODS: We employed Next Generation Sequencing (NGS) to analyze 14 cancer-susceptible genes in prostate cancer patients. RESULTS: Our study identified genetic mutations in BRCA1, BRCA2, TP53, and PMS2. In BRCA1 gene, we identified two pathogenic mutations, c.181T>G (p.Cys61Gly) and c.2457delC (p.Ala819fs), found in 10 patients, along with three benign mutations, c.5357T>G (p.Leu1786Arg), c.1111T>C (p.Leu371Pro), and c.1201C>G (p.Thr401Arg), present in 13, 11, and 15 patients, respectively. For the BRCA2 gene, one pathogenic mutation, c.6275_6276del (p.Val2092fs), was detected in 10 patients, and four benign mutations, c.5347A>T (p.Met1783Leu), c.5198A>G (p.Asp1733Gly), c.5158A>G (p.Thr1720Ala), and c.5117G>C (p.Gly1706Ala), were found in 17, 21, 34, and 12 patients, respectively. In the TP53 gene, we found two pathogenic mutations, c.1014_1015insT (p.Glu339Ter) and c.916C>T (p.Arg306Ter), in 10 and 11 patients, respectively, and two benign mutations, c.311T>C (p.Ser104Pro) and c.1129C>T (p.Arg377Cys), in 8 and 9 patients, respectively. Lastly, the PMS2 gene exhibited 16 benign mutations. Notably, the detected pathogenic mutations are rare in the broader Asian population according to the gnomAD database. Functional analyses using RT-qPCR and immunohistochemistry showed decreased expression of BRCA1, BRCA2, and TP53 in samples with pathogenic mutations, corroborating their impact on tumor suppressor function. Furthermore, drug sensitivity analysis revealed that BRCA1 and BRCA2 mutations are associated with increased sensitivity to a range of chemotherapeutic agents, supporting the concept of synthetic lethality. However, TP53 did not significantly impact drug sensitivity. CONCLUSION: This comprehensive analysis emphasizes the critical roles of BRCA1, BRCA2, TP53, and PMS2 in prostate cancer pathogenesis and highlights the importance of population-specific genetic screening.

MERS-CoV Infection and Its Impact on the Expression of TSLP Cytokine and IgG Antibodies: An In Vivo and In Vitro Study.

Purpose: Thymic stromal lymphopoietin (TSLP) is a proinflammatory cytokine produced by epithelial cells that is involved in the activation of allergic disorders. To date, no study has examined TSLP induction during Middle East respiratory syndrome coronavirus (MERS-CoV) infection. Herein, we aimed to study the effects of the recombinant spike protein of MERS-CoV on TSLP production. Additionally, the effects of recombinant human TSLP (rhTSLP) on B cell survival and antibody production were investigated. Patients and Methods: B cells were separated using the Human B Cell Enrichment Kit, and B cell survival was measured using the WST-1 Assay Kit. Enzyme-linked immunosorbent assay (ELISA) was used to measure TSLP levels in the sera of both MERS-CoV-infected (n=4; median age, 53 years) and healthy individuals (n=5; median age, 35 years). Results: We showed that the group of infected patients had significantly higher levels of TSLP than healthy controls (37.6 pg/mL vs 19.8 pg/mL, *p<0.05). The levels of TSLP in A549 cells were remarkably increased after 48 h of stimulation with recombinant full-length spike protein (rSP) (32.2 pg/mL, p=0.01). B cell survival was greatly enhanced by rhTSLP alone or in combination with rSP (0.02 vs 0.046, and 0.045; **p<0.01, respectively). Our data also showed a significant synergistic effect of rhTSLP and rSP on the augmented response of IgG antibodies against the spike protein of MERS-CoV compared with unstimulated cells (0.156 vs 0.22; *p<0.05). Conclusion: TSLP production is induced in vivo after MERS-CoV infection and in vitro after treatment with the rSP of MERS-CoV, which has a significant effect on the survival of B cells. Our data suggest that TSLP can be used as a strong mucosal adjuvant for vaccine development against MERS-CoV infection. However, further investigation is required to study the functional role of TSLP in MERS-CoV infection.

Synthesis and evaluation of pyridine-3-carboxamide analogs as effective agents against bacterial wilt in tomatoes.

This study focused on developing novel pyridine-3-carboxamide analogs to treat bacterial wilt in tomatoes caused by Ralstonia solanacearum. The analogs were synthesized through a multistep process and their structures confirmed using spectroscopy. Molecular docking studies identified the most potent analog from the series. A specific analog, compound 4a, was found to significantly enhance disease resistance in tomato plants infected with R. solanacearum. The structure-activity relationship analysis showed the positions and types of substituents on the aromatic rings of compounds 4a-i strongly influenced their biological activity. Compound 4a, with a chloro group at the para position on ring C and hydroxyl group at the ortho position on ring A, was exceptionally effective against R. solanacearum. When used to treat seeds, the analogs displayed remarkable efficacy, especially compound 4a which had specific activity against bacterial wilt pathogens. Compound 4a also promoted vegetative and reproductive growth of tomato plants, increasing seed germination and seedling vigor. In plants mechanically infected with bacteria, compound 4a substantially reduced the percentage of infection, pathogen quantity in young tissue, and disease progression. The analogs were highly potent due to their amide linkage. Molecular docking identified the best compounds with strong binding affinities. Overall, the strategic design and synthesis of these pyridine-3-carboxamide analogs offers an effective approach to targeting and controlling R. solanacearum and bacterial wilt in tomatoes.

Seed Priming with Glutamic-Acid-Functionalized Iron Nanoparticles Modulating Response of Vigna radiata (L.) R. Wilczek (Mung Bean) to Induce Osmotic Stress.

Rising soil salinity is a major concern for agricultural production worldwide, particularly in arid and semi-arid regions. To improve salt tolerance and the productivity of economic crop plants in the face of future climatic changes, plant-based solutions are required to feed the continuously increasing world population. In the present study, we aimed to ascertain the impact of Glutamic-acid-functionalized iron nanoparticles (Glu-FeNPs) on two varieties (NM-92 and AZRI-2006) of mung beans with different concentrations (0, 40 mM, 60 mM, and 80 mM) of osmotic stress. The result of the study showed that vegetative growth parameters such as root and shoot length, fresh and dry biomass, moisture contents, leaf area, and the number of pods per plant were significantly decreased with osmotic stress. Similarly, biochemicals such as protein, chlorophylls, and carotenes contents also significantly declined under induced osmotic stress. The application of Glu-FeNPs significantly (p ≤ 0.05) restored both the vegetative growth parameters and biochemical contents of plants under osmotic stress. The pre-sowing treatment of seeds with Glu-FeNPs significantly ameliorated the tolerance level of Vigna radiata to osmotic stress by optimizing the level of antioxidant enzymes and osmolytes such as superoxide dismutase (SOD), peroxidase (POD), and proline contents. Our finding indicates that Glu-FeNPs significantly restore the growth of plants under osmotic stress via enhancing photosynthetic activity and triggering the antioxidation system of both varieties.

Masitinib analogues with the N-methylpiperazine group replaced - A new hope for the development of anti-COVID-19 drugs.

Masitinib is an orally acceptable tyrosine kinase inhibitor that is currently investigated under clinical trials against cancer, asthma, Alzheimer's disease, multiple sclerosis and amyotrophic lateral sclerosis. A recent study confirmed the anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) activity of masitinib through inhibition of the main protease (Mpro) enzyme, an important pharmacological drug target to block the replication of the coronavirus. However, due to the adverse effects and lower potency of the drug, there are opportunities to design better analogues of masitinib. Herein, we substituted the N-methylpiperazine group of Masitinib with different chemical moieties and evaluated their drug-likeness and toxicities. The filtered analogues were subjected to molecular docking studies which revealed that the analogues with substituents methylamine in M10 (CID10409602), morpholine in M23 (CID59789397) and 4-methylmorpholine in M32 (CID143003625) have a stronger affinity to the drug receptor compared to masitinib. The molecular dynamics (MD) simulation analysis reveals that the identified analogues alter the mobility, structural compactness, accessibility to solvent molecules, and the number of hydrogen bonds in the native target enzyme. These structural alterations can help explain the inhibitory mechanisms of these analogues against the target enzyme. Thus, our studies provide avenues for the design of new masitinib analogues as the SARS-CoV-2 Mpro inhibitors.