Evaluation of the Effect of Carvedilol on the Thioredoxin Pathway in H9C2 Rat Cardiomyocytes

The thioredoxin (Trx) system is an endogenous antioxidant system that affects cell function and survival through controlling cellular redox status. Trx and TrxR are the main enzymes in this system while thioredoxin interacting protein (TXNIP) is a negative regulator. This study’s goal was to better understand the Trx system’s involvement in the cardiovascular disease and modulate the pathway through drug treatment. Carvedilol is a non-selective β-blocker that also exhibits antioxidant properties, but the exact mechanism of the antioxidant effect is still unclear. H9c2 rat cardiomyocytes were used to examine the effect of carvedilol on the Trx system under normal conditions. Interestingly, carvedilol was able to decrease TXNIP not through its expression or proteasomal degradation but through increased TXNIP nuclear localization. Immunoprecipitation also showed an increase in TXNIP-PARP complexation in the nucleus and a decrease in Trx-TXNIP complexation in the cytosol. The results indicate that carvedilol may exhibit its antioxidant activity through altering TXNIP subcellular localization. TXNIP is known to be important in both physiologic and pathophysiologic conditions. Western blot data showed that TXNIP in the cytosol will increase with increasing glucose concentration. Oxidative stress inducers such as doxorubicin, hypoxiareoxygenation, and radiation were able to decrease cytosolic TXNIP. Doxorubicin is a commonly utilized anticancer drug that induces oxidative stress and therefore causes cardiac toxicity. A study was conducted to determine if carvedilol could protect against doxorubicin-induced cardiotoxicity through TXNIP modulation. Carvedilol and doxorubicin alone reduced cytosolic TXNIP. Doxorubicin increased mitochondrial translocation of TXNIP accompanied by the induction of apoptosis. However, carvedilol was not able to prevent TXNIP mitochondrial translocation, but it did protect against doxorubicin-induced apoptosis. The complex of Trx2 and the proapoptotic ASK1 in the mitochondria was increased with carvedilol pretreatment followed by doxorubicin exposure. The increase in the ASK1-Trx2 complex can reduce apoptosis through decreased ASK1 activation. This was confirmed through Western blot of cleaved PARP. The findings are consistent with reports of TXNIP’s response to mild oxidative stress conditions. In conclusion, this study shows for the first time that carvedilol impacts TXNIP localization and complexation and that the Trx pathway may be involved in carvedilol’s observed cardioprotective effect

Pharmacological Potential of Hippophae rhamnoides L. Nano-Emulsion for Management of Polycystic Ovarian Syndrome in Animals’ Model: In Vitro and In Vivo Studies

The most common female endocrinopathy, polycystic ovarian syndrome (PCOS), generally affects women of childbearing age. Hippophae rhamnoides L. has been traditionally used to improve menstrual cyclicity. Gas chromatography by flame ionization detection analysis showed that it contained various phytoconstituents such as omega-3 fatty acid, phytosterols, palmitic acid, oleic acid, and linoleic acid. H. rhamnoides L. (HR) nano-emulsion was also formulated. HR and its encapsulated nano-emulsion (HRNE) were evaluated for the treatment of PCOS. Thirty-five healthy female adult albino rats were acquired and divided into seven groups (n = 5). Letrozole (1 mg/kg) was used for 5 weeks to induce the disease. To confirm disease (PCOS) induction, the animals were weighed weekly and their vaginal smears were analyzed daily under a microscope. After PCOS induction, animals were treated with metformin, HR, and HRNE with two different doses (0.5/kg and 1 g/kg, p.o.) for 5 weeks. At the end of the treatment, animals were euthanized, and blood was collected for hormonal assessment, lipid profiling, and liver functioning test assessment. Both the ovaries were preserved for histopathology and liver for the purpose of assessment of antioxidant potential. The results revealed that HR and HRNE at both doses improved the hormonal imbalance; follicle-stimulating hormone, estrogen, and progesterone levels are increased, while luteinizing hormone surge and testosterone level are controlled. Insulin sensitivity is improved. Ovarian histopathology showed that normal ovarian echotexture is restored with corpus luteum and mature and developing follicles. HR and HRNE also improved the lipid profile and decreased lipid peroxidation (MDA) with improved antioxidant markers (SOD, CAT, and GSH). Results were statistically analyzed by one-way analysis of variance and were considered significant only if p < 0.05. In conclusion, it can be postulated that H. rhamnoides L. proved effective in the management of PCOS and its nano-emulsion effects were statistically more significant, which might be due to better bioavailability

Comparative genomics of food-derived probiotic Lactiplantibacillus plantarum K25 reveals its hidden potential, compactness, and efficiency

This study aimed to investigate the intricate genetic makeup of the Lactiplantibacillus plantarum K25 strain by conducting a comprehensive analysis of comparative genomics. The results of our study demonstrate that the genome exhibits a high-level efficiency and compactness, comprising a total of 3,199 genes that encode proteins and a GC content of 43.38%. The present study elucidates the evolutionary lineage of Lactiplantibacillus plantarum strains through an analysis of the degree of gene order conservation and synteny across a range of strains, thereby underscoring their closely interrelated evolutionary trajectories. The identification of various genetic components in the K25 strain, such as bacteriocin gene clusters and prophage regions, highlights its potential utility in diverse domains, such as biotechnology and medicine. The distinctive genetic elements possess the potential to unveil innovative therapeutic and biotechnological remedies in future. This study provides a comprehensive analysis of the L. plantarum K25 strain, revealing its remarkable genomic potential and presenting novel prospects for utilizing its unique genetic features in diverse scientific fields. The present study contributes to the existing literature on Lactiplantibacillus plantarum and sets the stage for prospective investigations and practical implementations that leverage the exceptional genetic characteristics of this adap organism

GC–MS profiling of Bacillus spp. metabolites with an in vitro biological activity assessment and computational analysis of their impact on epithelial glioblastoma cancer genes

Background: Bacterial metabolites play a crucial role in human health and have proven effective in treating various diseases. In this study, the 16S rRNA method and streaking were employed to isolate and molecularly identify a bacterial strain, with the goal of characterizing bioactive volatile metabolites extracted using nonpolar and polar solvents.Methods: Gas chromatography–mass spectrometry (GC–MS) analysis was conducted to identify 29 compounds in the bacterial metabolites, including key compounds associated with Bacillus spp. The main compounds identified included 2-propanone, 4,4-ethylenedioxy-1-pentylamine, 1,2-benzenedicarboxylic acid, 1,1-butoxy-1-isobutoxy-butane, and 3,3-ethoxycarbonyl-5-hydroxytetrahydropyran-2-one.Results: The literature indicates the diverse biological and pharmacological applications of these compounds. Different concentrations of the metabolites from Bacillus species were tested for biological activities, revealing significant inhibitory effects on anti-diabetic activity (84.66%), anti-inflammatory activity (99%), antioxidant activity (99.8%), and anti-hemolytic activity (90%). Disc diffusion method testing also demonstrated a noteworthy inhibitory effect against tested strains.Conclusion:In silico screening revealed that 1,2-benzenedicarboxylic acid exhibited anticancer activity and promising drug-designing properties against epithelial glioblastoma cancer genes. The study highlights the potential of Bacillus spp. as a valuable target for drug research, emphasizing the significance of bacterial metabolites in the production of biological antibacterial agents

The therapeutic potential of skin mucus from Asian swamp eel (Monopterus albus): In vivo evaluation and histological evidence

Objectives The Asian swamp eel (Monopterus albus), is commonly distributed in Asian countries. However, its therapeutic potential has not been thoroughly investigated yet. The current study aimed to evaluate the in-vivo therapeutic properties of the skin mucus of this fish. Methods The eel mucus was collected fleshly and topical gel with carbopol 934 was formulated to study the antibacterial activity on the infected skin of the rats. Sprague Dawley rats were used in the study and divided into 4 groups negative, positive, normal control, and treated groups. Results Intracutaneous injections of pathogenic bacteria (Streptococcus pyogenes, Staphylococcus aureus) and fungi (Microsporum gypseum, Candida albicans) were injected into the rats. The development of tinea capitis, impetigo, and cutaneous candidiasis in the animal model was confirmed based on clinical and histopathological observations. To treat the infected rats, a formulated gel of eel skin mucus was applied on the infected rat’s skins topically. The histological analysis confirms a complete recovery in the skin tissues similar to commercial antifungal and antibacterial agents used in the positive control groups. Conclusion The present novel eel skin mucus is an efficient therapeutic candidate in treating skin infections associated with pathogenic microbes

Analysis of codon usage bias of lumpy skin disease virus causing livestock infection

Lumpy skin disease virus (LSDV) causes lumpy skin disease (LSD) in livestock, which is a double-stranded DNA virus that belongs to the genus Capripoxvirus of the family Poxviridae. LSDV is an important poxvirus that has spread out far and wide to become distributed worldwide. It poses serious health risks to the host and causes considerable negative socioeconomic impact on farmers financially and on cattle by causing ruminant-related diseases. Previous studies explained the population structure of the LSDV within the evolutionary time scale and adaptive evolution. However, it is still unknown and remains enigmatic as to how synonymous codons are used by the LSDV. Here, we used 53 LSDV strains and applied the codon usage bias (CUB) analysis to them. Both the base content and the relative synonymous codon usage (RSCU) analysis revealed that the AT-ended codons were more frequently used in the genome of LSDV. Further low codon usage bias was calculated from the effective number of codons (ENC) value. The neutrality plot analysis suggested that the dominant factor of natural selection played a role in the structuring of CUB in LSDV. Additionally, the results from a comparative analysis suggested that the LSDV has adapted host-specific codon usage patterns to sustain successful replication and transmission chains within hosts (Bos taurus and Homo sapiens). Both natural selection and mutational pressure have an impact on the codon usage patterns of the protein-coding genes in LSDV. This study is important because it has characterized the codon usage pattern in the LSDV genomes and has provided the necessary data for a basic evolutionary study on them

Utilization of aqueous broccoli florets extract for green synthesis and characterization of silver nanoparticles, with potential biological applications

The process of creating nanoparticles using chemicals is not eco-friendly. However, a more environmentally conscious approach known as green chemistry, which involves using vegetable-mediated nanoparticle production, combines nanotechnology with biotechnology. In this study, the researchers aimed to assess the effectiveness of the green chemistry technique in producing silver nanoparticles using an liquid extract from broccoli florets (Brassica oleracea) under ideal environment. The successful production of silver nanoparticles was achieved through silver nitrate (AgNO₃) biological reduction with the help of an aqueous broccoli florets extract at a slightly acidic pH of 6–7. The silver nanoparticles occurrence was shown by a change of color that moved from colorless to reddish-brown. To characterize the green-produced nanoparticles, various analytical techniques such as Ultraviolet–Visible Spectroscopy (UV-VIS), Fourier Transform Infrared Spectroscopy (FT-IR), X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM), and Energy-Dispersive X-ray Spectroscopy (EDAX) were employed. The antioxidant properties of the formed silver nanoparticles (AgNPs) were examined in vitro using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Ferric Reducing Antioxidant Power (FRAP) tests. Additionally, the antibacterial properties of AgNPs against various pathogenic bacteria was evaluated. The reduction procedure was easy and simple manageable, with UV–Vis spectroscopy indicating the surface plasmon resonance (SPR) presence at 425 nm. FTIR was utilized to identify active chemical groups in the biomass before and after reduction. SEM and X-ray diffraction analyses indicated that the silver nanoparticles had an average the size of individual particles of 33 nm and exhibited a face-centered cubic (FCC) structure. EDAX analysis confirmed the occurrence of elemental silver in the nanoparticles. The study demonstrated that the biosynthesis of AgNPs led to significant variations in antioxidant activity, which was dose-dependent and showed a similar pattern to the testing of the scarfing action of the ascorbic acid against free radicals using DPPH and FRAP. The AgNPs also dispalyed firm deep-spectrum antibacterial action observed against the tested pathogenic bacteria, outperforming certain medications. Interestingly, the silver nanoparticles remained stable at ambient temperature for 25 days without precipitation, retaining their antioxidant and antibacterial properties. In conclusion, the research findings suggest that an aqueous extract of fresh broccoli florets can serve as a viable and environmentally friendly method for producing stable silver nanoparticles with beneficial antioxidant and antibacterial characteristics

Design of a novel multi-epitopes based vaccine against brucellosis

Brucella melitensis is a gram-negative coccobacillus that causes brucellosis in humans. The lack of effective treatment and increasing antibiotic-resistant patterns shown by B. melitensis, warrant the search for novel therapeutics. In this study, comprehensive bioinformatics, reverse vaccinology, and biophysics techniques were employed to design a novel multi-epitopes based vaccine (MEBV) against B. melitensis. Core proteomics, subtractive proteomics and immunoinformatic studies revealed three core proteins: Flagellar hook protein (FlgE), TonB-dependent receptor, and Porin family protein as promising vaccine targets. The proteins have exposed topology, and are antigenic, and adhesive. Furthermore, B and T cell epitopes were predicted from these proteins. Highly antigenic, immunogenic, non-toxic and non-allergenic epitopes were shortlisted and used in the MEBV design. The designed MEBV also showed stable docked conformation with different immune receptors such as MHC-I, MHC-II, and TLR-4. The global energy of selected docked complexes was as; solution 4 of MEBV-TLR-4 (−45.73 kJ/mol), solution 5 of MEBV-MHC-I (−20.94 kJ/mol), and solution 1 of MEBV-MHC-II (−3.45 kJ/mol). Molecular dynamics simulation studies unveiled a steady root mean square deviation (RMSD) pattern for the systems. However, all of them were stable in terms of intermolecular binding conformation and chemical interactions. Further, the systems showed robust binding energies with net binding energy < −300 kcal/mol. The van der Waals and electrostatic energies were the dominating energies and were found as intermolecular stabilizing factors. The vaccine was also predicted to generate promising immunological responses and thus could be an attractive candidate to be evaluated in experimental studies

Whole Proteome-Based Therapeutic Targets Annotation and Designing of Multi-Epitope-Based Vaccines against the Gram-Negative XDR-Alcaligenes faecalis Bacterium

This study involved therapeutic targets mining for the extremely drug-resistant bacterial species called Alcaligenes faecalis, which is known to infect humans. The infections caused by this species in different parts of the human body have been linked with a higher degree of resistance to several classes of antibiotics. Meanwhile, alternate therapeutic options are needed to treat these bacterial infections in clinical settings. In the current study, a subtractive proteomics approach was adapted to annotate the whole proteome of Alcaligenes faecalis and prioritize target proteins for vaccine-related therapeutics design. This was followed by targeted protein-specific immune epitope prediction and prioritization. The shortlisted epitopes were further subjected to structural design and in silico validation of putative vaccines against Alcaligenes faecalis. The final vaccine designs were also evaluated for potential interaction analysis with human TLR-2 through molecular docking. Finally, the putative vaccines were subjected to in silico cloning and immune simulation approaches to ensure the feasibility of the target-specific vaccine constructs in further experimental designs

Integrated Mechanisms of Polarity&ndash;Based Extracts of Cucumis melo L. Seed Kernels for Airway Smooth Muscle Relaxation via Key Signaling Pathways Based on WGCNA, In Vivo, and In Vitro Analyses

The present study aimed to determine the mechanisms responsible for calcium&ndash;mediated smooth muscle contractions in C. melo seeds. The phytochemicals of C. melo were identified and quantified with the aid of Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometric (LC/ESI&ndash;MS/MS) and high&ndash;performance liquid chromatography (HPLC), and then tested in&ndash;vitro and in vivo to confirm involvement in smooth muscle relaxation. Allergic asthma gene datasets were acquired from the NCBI gene expression omnibus (GEO) and differentially expressed gene (DEG) analysis, weighted gene co&ndash;expression network analysis (WGCNA), and functional enrichment analysis were conducted. Additionally, molecular docking of key genes was carried out. Kaempferol, rutin, and quercetin are identified as phytochemical constituents of C. melo seeds. Results indicated that C. melo seeds exhibit a dose&ndash;dependent relaxant effect for potassium chloride (80 mM)&ndash; induced spastic contraction and calcium antagonistic response in calcium dose&ndash;response curves. The functional enrichment of WGCNA and DEG asthma&ndash;associated pathogenic genes showed cytokine&ndash;mediated pathways and inflammatory responses. Furthermore, CACNA1A, IL2RB, and NOS2 were identified as key genes with greater binding affinity with rutin, quercitrin, and kaempferol in molecular docking. These results show that the bronchodilator and antidiarrheal effects of C. melo were produced by altering the regulatory genes of calcium&ndash;mediated smooth muscle contraction