Antibacterial activity of peptides and bio-safety evaluation: in vitro and in vivo studies against bacterial and fungal pathogens.

BACKGROUND: Antimicrobial peptides are promising alternatives to antibiotics to treat bacterial and fungal infections, especially drug-resistant clinical pathogens. METHODS: Antimicrobial peptides (AMPs) were synthesized and antimicrobial activity was assayed. The antibacterial mechanism, ATP production, ROS generation and molecular mechanism were determined. Biofilm inhibition assay was performed in planktonic bacterial cells and biofilm degradation assay was performed using mature biofilm. The synthesized AMP2 was subjected to in vitro and in vivo analysis to analyze the safety. RESULTS: The synthesized peptides AMP1, AMP2, AMP3 and AMP4 exhibited antimicrobial activity against Gram-positive and Gram-negative bacteria. The MIC values ranged from 1.5 ± 0.25-12.5 ± 1.25 µM and the MFC values range from 2.25 ± 0.12-25 ± 1.25 µM. F. solani showed fewer MFC values than other fungal strains. Time kill assay was performed and the AMP2 killed about 70 % of Acinetobacter baumannii at 1 × MIC concentration within 10 min incubation and killed 97 % of bacteria at 1 × MBC concentration within 15 min. The antimicrobial peptide AMP2 was highly effective against planktonic A. baumannii and L. monocytogenes. The tested AMP2 showed less toxicity to cell lines and Zebrafish. CONCLUSIONS: Antimicrobial peptides have potential antimicrobial properties against Gram-positive and Gram-negative bacteria. The in silico studies of these antimicrobial peptides are useful for eradicating drug-resistant bacteria.

PS9, Derived from an Aquatic Fungus Virulent Protein, Glycosyl Hydrolase, Arrests MCF-7 Proliferation by Regulating Intracellular Reactive Oxygen Species and Apoptotic Pathways.

One of the most common diseases in women is breast cancer, which has the highest death globally. Surgery, chemotherapy, hormone treatments, and radiation are the current treatment options for breast cancer. However, these options have several adverse side effects. Recently, peptide-based drugs have gained attention as anticancer therapy. Studies report that peptides from biological toxins such as venom and virulent pathogenic molecules have potential therapeutic effects against multiple diseases, including cancers. This study reports on the in vitro anticancer effect of a short peptide, PS9, derived from a virulent protein, glycosyl hydrolase, of an aquatic fungus, Aphanomyces invadans. This peptide arrests MCF-7 proliferation by regulating intercellular reactive oxygen species (ROS) and apoptotic pathways. Based on the potential for the anticancer effect of PS9, from the in silico analysis, in vitro analyses using MCF-7 cells were executed. PS9 showed a dose-dependent activity; its IC50 value was 25.27-43.28 µM at 24 h. The acridine orange/ethidium bromide (AO/EtBr) staining, to establish the status of apoptosis in MCF-7 cells, showed morphologies for early and late apoptosis and necrotic cell death. The 2,7-dichlorodihydrofluorescein diacetate (DCFDA) staining and biochemical analyses showed a significant increase in reactive oxygen species (ROS). Besides, PS9 has been shown to regulate the caspase-mediated apoptotic pathway. PS9 is nontoxic, in vitro, and in vivo zebrafish larvae. Together, PS9 may have an anticancer effect in vitro.

Bacterial clearance and anti-inflammatory effect of Withaferin A against human pathogen of Staphylococcus aureus in infected zebrafish.

The emergence of antibiotic resistance is the most challenging factor for developing a proper drug to treat S. aureus infection. These bacterial pathogens can survive in fresh water and spread to various environments. Plant sources, especially pure compounds, are the material of interest amongst researchers for developing drugs of therapeutic value. Here, we report the bacterial clearance and anti-inflammatory potential of the plant compound Withaferin A, using the zebrafish infection model. The minimum inhibitory concentration of the Withaferin A was calculated as 80 µM against S. aureus. The DAPI/PI staining and scanning electron microscopy analysis showed the pore-forming mechanism of Withaferin A on the bacterial membrane. Along with the antibacterial activity, the results from the tube adherence test reveal the antibiofilm property of Withaferin A. In vivo studies were demonstrated to determine the effect of Withaferin A on survival, inflammatory response and behavioural changes during S. aureus infection. Staining zebrafish larvae with neutral red and Sudan black indicates a substantial decrease in the number of localized macrophages and neutrophils. The gene expression analysis showed the downregulation of inflammatory marker genes. Additionally, we observed the improvement in locomotory behaviour among Withaferin A treatment adult zebrafish. In conclusion, S. aureus can infect zebrafish and induces toxicological effect. In comparison, the results from in vitro and in vivo experiments suggest that Withaferin A can be used for synergistic antibacterial, antibiofilm and anti-inflammatory activity to treat infections due S. aureus.

Biocompatibility of Veratric Acid-Encapsulated Chitosan/Methylcellulose Hydrogel: Biological Characterization, Osteogenic Efficiency with In Silico Molecular Modeling.

The limitations of graft material, and surgical sites for autografts in bone defects treatment have become a significant challenge in bone tissue engineering. Phytocompounds markedly affect bone metabolism by activating the osteogenic signaling pathways. The present study investigated the biocompatibility of the bio-composite thermo-responsive hydrogels consisting of chitosan (CS), and methylcellulose (MC) encapsulated with veratric acid (VA) as a restorative agent for bone defect treatment. The spectroscopy analyses confirmed the formation of CS/MC hydrogels and VA encapsulated CS/MC hydrogels (CS/MC-VA). Molecular analysis of the CS-specific MC decamer unit with VA complex exhibited a stable integration in the system. Further, Runx2 (runt-related transcription factor 2) was found in the docking mechanism with VA, indicating a high binding affinity towards the functional site of the Runx2 protein. The formulated CS/MC-VA hydrogels exhibited biocompatibility with the mouse mesenchymal stem cells, while VA promoted osteogenic differentiation in the stem cells, which was verified by calcium phosphate deposition through the von Kossa staining. The study results suggest that CS/MC-VA could be a potential therapeutic alternative source for bone regeneration.

Nimbin analogs N5 and N7 regulate the expression of lipid metabolic genes and inhibit lipid accumulation in high-fat diet-induced zebrafish larvae: An antihyperlipidemic study.

BACKGROUND: Excess accumulation of lipids leads to obesity. Triterpenoids are a group of plant compounds which poses various biological activities. The biological activities of Nimbin analogs N5 and N7 were addressed in this study on inhibiting lipid aggregation and underlying the derivatives molecular mechanisms for a therapeutical approach. AIM: This study aims to evaluate the anti-adipogenic activity of semi-natural Nimbin analogs, N5 and N7, on zebrafish larvae induced with oxidative stress due to a high-fat diet (HFD) and adipogenesis using specific fluorescent stains. MATERIALS AND METHODS: Zebrafish at 4 days post fertilized (dpf) larvae were divided into groups for the HFD diet along with exposure to various concentrations of N5 and N7. HFD induced accumulation of neutral lipids and triglycerides (Oil Red O and Nile red staining, respectively) with weight gain, which generated intracellular ROS (DCFH-DA staining) and superoxide anion production (DHE staining) with depleted glutathione levels (NDA staining) were assayed. HFD exposure promoted the accumulation of inflammatory macrophages (Neutral red staining) and impaired glucose metabolism (2NBDG staining). The ability of N5 and N7 to reduce total regulating lipogenic specific genes C/EBP-α, SREBP-1 and FAS were evaluated using relative gene expression. KEY FINDINGS: The Nimbin analogues N5 and N7 suppressed adipogenesis, forming intracellular ROS and superoxide anion while simultaneously restoring glutathione levels. The analogues significantly lowered total TC and TG levels, prevented inflammatory macrophage build-up and boosted glucose absorption. Also, N5 and N7 down-regulate the lipogenic-specific genes. SIGNIFICANCE: Nimbin analogs N5 and N7 enhance lipolysis and inhibit adipogenesis in in-vivo zebrafish larvae model.

Effective degradation of Chlortetracycline using dual bio catalyst.

Chlortetracycline (CTC) degradation using potential microbial consortia or individual bacterial strains was useful method for improving bioremediation potential. The co-culture (Klebsiella pneumoniae CH3 and Bacillus amyloliquefaciens CS1) of bacterial strains have the ability to degrade chlortetracycline (91.8 ± 1.7%), followed by sulfamethoxazole (62.1 ± 1.2%) and amoxicillin (73.9 ± 3.3%). It was observed that the degradation potential was maximum after 10 days incubation, 8-10% inoculum, pH 7.5, and antibiotic concentration ranged from 150 to 200 mg/L. The initial concentrations of CTC significantly affected CTC degradation. In strain CH3, maximum biodegradation of CTC (99.4 ± 2.3%) was observed at 200 mg/L initial CTC concentrations. In CS1, maximum biodegradation of CTC was obtained at 150 mg/L concentration (80.5 ± 3.2%) after 10 days of culture. Alkaline pH was found to be suitable for the degradation of antibiotic than acidic range. After initial optimization by one factor at a time approach in free cells, the bacterial strains (CH3 and CS1) were co-immobilized. The co-immobilized bacterial cells showed improved degradation potential than free cells. To determine the biodegradation potential of immobilized cells, the selected strains were immobilized in polymer beads and treated with CTC with 175 mg/L initial concentration. The experimental results revealed that after 3 days of treatment the residual CTC concentration was 150.1 ± 3.2 mg/L and it decreased as 1.28 ± 0.01 mg/L after 10 days of treatment. The present study confirmed the effectiveness and feasibility of biodegradation ability of K. pneumoniae CH3 and B. amyloliquefaciens CS1 immobilized for CTC degradation in wastewater.

Biosorption of hazardous waste from the municipal wastewater by marine algal biomass.

Lead is one of the highly toxic heavy metals causes various diseases even at very lower concentrations to human and affects eco-system. It is mainly released into the water through industrial activities. Phytoremediation is useful to degrade, reduce, metabolize and assimilate lead from wastewater. In this study, Turbinaria ornata was collected from the sea and dried biomass was used for biosorption of heavy metals. Adsorption of heavy metal was maximum after 100 min incubation with alga powder at acidic pH (4.5). The interactive effects of lead concentration, contact times, pH, biomass concentration and agitation speed was evaluated by a two-level full factorial design. Initial lead concentration, agitation speed and biomass concentration were the most important variables affecting lead removal (p < 0.001) were selected for optimization using central composite rotatable design. Lead removal was found to be maximum (99.8%) in optimized conditions: initial lead 99.8 mg/L, 250 rpm agitation speed and 16.2 g/L biomass concentrations. Municipal wastewater was collected and lead concentration (0.013 mg/L) and physiochemical factors were analyzed. Algal biomass removed >98.5% lead form the wastewater within 10 min in an optimized condition. The present study confirmed the potential application of T. ornata for the removal of lead from contaminated environment.

Assessment on In Vitro Probiotic Attributes of Lactobacillus plantarum Isolated From Horse Feces.

This study was designed to assess in vitro probiotic attributes of potent bacterium isolated from the feces of healthy horse. Initially, a total of eight bacteria were isolated from the feces and evaluated their antibacterial activities against indicator bacterial pathogens using agar well diffusion assay. Results showed significant (P < .05) antibacterial property of Lactobacillus plantarum strain LF4 against pathogens tested with maximum growth inhibitory activity of 320.16 ± 3.4 AU/mL against Staphylococcus aureus. Further, in vitro probiotic properties of strain LF4 were determined using standard methodologies. Strain LF4 maintained its viability towards acidic condition (pH 2.0) and simulated gastric juice (pH 2.0) with total cell counts of 1.6 ± 0.18 and 1.7 ± 0.18 log cfu/mL, respectively. Moreover, the strain was observed resistant to oxgall (0.5% w/v) up to 36 hours. The isolate showed significant (P < .05) hydrophobicity property (60.3 ± 1.6%), auto-aggregation trait (41.31 ± 1.5%), and moderate proteolytic activity. Strain LF4 revealed significant (P < .05) rate of DPPH scavenging (15.3 ± 1.3-69.7 ± 1.3%) and hydroxyl radical scavenging (11.3 ± 1.3 to 56.4 ± 1.3%) in a concentration dependent manner. Additionally, the isolate was observed susceptible to all the conventional antibiotics tested, thereby indicating its safer utilization. In conclusion, findings suggested the colossal applications of L. plantarum strain LF4 as an ideal probiotic bacterium in equine industries.

GR15 peptide of S-adenosylmethionine synthase (SAMe) from Arthrospira platensis demonstrated antioxidant mechanism against H2O2 induced oxidative stress in in-vitro MDCK cells and in-vivo zebrafish larvae model.

GR15 is a short molecule or peptide composed of aliphatic amino acids and possesses to have antioxidant properties. The GR15, 1GGGAFSGKDPTKVDR15 was identified from the protein S-adenosylmethionine synthase (SAMe) expressed during the sulfur departed state of Arthrospira platensis (spirulina or cyanobacteria). The in-silico assessment and the structural features of GR15 showed its antioxidant potency. Real-time PCR analysis found the up-regulation of ApSAMe expression on day 15 against oxidative stress due to 10 mM H2O2 treatment in A. platensis (Ap). The antioxidant activity of GR15 was accessed by the cell-free antioxidant assays such as ABTS, SARS, HRAS and NO; the results showed dose-dependent antioxidant activity. The toxicity assay was performed in both in vitro and in vivo models, in which peptide does not exhibit any toxicity in MDCK cell and zebrafish embryos. The intercellular ROS reduction potential of GR15 peptide was also investigated in both in vitro and in vivo models including LDH assay, antioxidant enzymes (SOD and CAT), and fluorescent staining assay (DCFDA, Hochest and Acridine orange sting) was performed; the results showed that the GR15 peptide was effectively reduced the ROS level. Further, RT-PCR demonstrated that GR15 enhanced the antioxidant property and also up-regulated the antioxidant gene, thus reduced the ROS level in both in vitro and in vivo models. Based on the results obtained from this study, we propose that GR15 has the potential antioxidant ability; hence further research can be directed towards the therapeutic product or drug development against disease caused by oxidative stress.

Cholecalciferol and metformin protect against lipopolysaccharide-induced endothelial dysfunction and senescence by modulating sirtuin-1 and protein arginine methyltransferase-1.

Endothelial cell activation through nuclear factor-kappa-B (NFkB) and mitogen-activated protein kinases leads to increased biosynthesis of pro-inflammatory mediators, cellular injury and vascular inflammation under lipopolysaccharide (LPS) exposure. Recent studies report that LPS up-regulated global methyltransferase activity. In this study, we observed that a combination treatment with metformin (MET) and cholecalciferol (VD) blocked the LPS-induced S-adenosylmethionine (SAM)-dependent methyltransferase (SDM) activity in Eahy926 cells. We found that LPS challenge (i) increased arginine methylation through up-regulated protein arginine methyltransferase-1 (PRMT1) mRNA, intracellular concentrations of asymmetric dimethylarginine (ADMA) and homocysteine (HCY); (ii) up-regulated cell senescence through mitigated sirtuin-1 (SIRT1) mRNA, nicotinamide adenine dinucleotide (NAD+) concentration, telomerase activity and total antioxidant capacity; and (iii) lead to endothelial dysfunction through compromised nitric oxide (NOx) production. However, these LPS-mediated cellular events in Eahy926 cells were restored by the synergistic effect of MET and VD. Taken together, this study identified that the dual compound effect inhibits LPS-induced protein arginine methylation, endothelial senescence and dysfunction through the components of epigenetic machinery, SIRT1 and PRMT1, which is a previously unidentified function of the test compounds. In silico results identified the presence of vitamin D response element (VDRE) sequence on PRMT1 suggesting that VDR could regulate PRMT1 gene expression. Further characterization of the cellular events associated with the dual compound challenge, using gene silencing approach or adenoviral constructs for SIRT1 and/or PRMT1 under inflammatory stress, could identify therapeutic strategies to address the endothelial consequences in vascular inflammation-mediated atherosclerosis.

Isolation and characterization of novel Streptomyces strain from Algeria and its in-vitro antimicrobial properties against microbial pathogens.

BACKGROUND: The constant development of microbial resistance to the traditional antimicrobial agents and the emergence of new infectious diseases justify the urgent need for new effective antimicrobial molecules. However, the irrational use of antibiotics increases microbial resistance dramatically and along with that the frequency of mortality associated with infections is higher. Therefore, to combat the antimicrobial resistance, the screening of compounds with novel chemical structures is essential. This study intended to determine the antimicrobial potential of Streptomyces GLD22 strain isolated from Algeria. METHODS: The characterization of Streptomyces strain GLD22 was performed by physiological, biochemical and molecular tests. The antimicrobial activity was tested by the well diffusion method and the minimum inhibitory concentration value calculation were performed using broth micro dilution technique. The extracellular metabolites profiling was done using GC-MS. RESULTS: Physiological, biochemical and phylogenetic analysis confirmed that the strain GLD22 showed maximum identity towards Streptomyces species. The extra cellular metabolites revealed their antimicrobial activity at 1 mg/ml for Klebsiella pneumoniae, Pseudomonas aeruginosa and Escherichia coli, whereas Staphylococcus aureus, Bacillus cereus and Bacillus subtilis documented 0.5, 1 and 1 mg/ml respectively. GC-MS analysis confirmed that 2-tert-butyl-4,6-bis(3,5-di-tert-butyl-4-hydroxybenzyl) phenol, Dibutyl phthalate and Cyclo(leucyloprolyl) were the major drug molecules present in the extract. CONCLUSION: The novel Streptomyces strain GLD22 recovered from the Gueldaman cave of Algeria showed better antimicrobial activity towards both Gram positive and Gram negative pathogens. Interestingly, the MIC values were comparable with the standard drug molecules. In addition, the identification of active metabolites present in the crude extracts was an advantage.

Oxidative stress induced antioxidant and neurotoxicity demonstrated in vivo zebrafish embryo or larval model and their normalization due to morin showing therapeutic implications.

AIMS: The study examined that morin as possible antioxidant and neuroprotective due to oxidative stress (H2O2) in zebrafish larval model. MATERIALS AND METHODS: Zebrafish larvae were induced with oxidative stress using H2O2 at 1 mM; their behavioural changes were assessed through partition preference and horizontal compartment test. The head section without eyes and yolk sac of zebrafish larvae were employed for enzyme assays such as SOD, CAT, Thiobarbituric acid reactive substances assay, reduced glutathione, glutathione peroxidase activity, glutathione S transferase, Acetylcholinesterase activity and nitrate levels. Also, intracellular ROS and apoptosis in larval head was detected by DCFDA and acridine orange staining followed by gene expression studies. KEY FINDINGS: Morin exposure was not harmful to the larvae at concentration between 20 and 60 µM, but it caused non-lethal deformity between 80 and 100 µM. In the partition test, zebrafish embryos treated with H2O2 showed cognitive impairment, whereas the morin-treated groups showed an improved behavioural activity. The study also found that restoring antioxidant enzymes and reduced lipid peroxidation which had a neuroprotective impact. Inhibition of NO overproduction and increased AChE activity were also shown to reduce the neuronal damage. Apoptosis and intracellular ROS levels were reduced in larvae when it was co-incubated with morin. Morin treatment up regulated the antioxidant enzymes against oxidative stress. SIGNIFICANCE: Morin provides protection against H2O2 induced oxidative stress through a cellular antioxidant defence mechanism by up-regulating gene expression, thus increasing the antioxidant activity at cellular or organismal stage.

Effective removal of pharmaceutical impurities and nutrients using biocatalyst from the municipal wastewater with moving bed packed reactor.

The presence of antibiotics in the wastewater is one of the important issues related to environmental management. In this study, antibiotics-degrading bacteria were screened from the enriched sewage sludge sample. Among the isolated bacterial strains, Bacillus subtilis AQ03 showed maximum antibiotic tolerance (>2000 ppm). The characterized strain B. subtilis AQ03 degraded sulfamethaoxazole and sulfamethoxine and the optimum nutrient and physical-factors were analyzed. B. subtilis AQ03 degraded 99.8 ± 1.3 % sulfamethaoxazole, and 93.3 ± 6.2 % sulfamethoxine. Sodium nitrate and ammonium chloride were improved antibiotics degradation (<90 %). The optimized conditions were maintained in a moving bed bioreactor for the removal of antibiotics and nutrients from the wastewater. The selected strain considerably produced proteases (109.4 U/mL), amylases (55.1 U/mL), cellulase (9.6 U/mL) and laccases (15.2). In moving bed reactor, sulfamethaoxazole degradation was maximum after 8 days (100 ± 1.5 %) and sulfamethoxazole (100 ± 0) was removed completely from wastewater after 10 days. In moving bed reactor, biological oxygen demand (92.1 ± 2.8 %), chemical oxygen demand (79.6 ± 1.2 %), nitrate (89.4 ± 3.9 %) and phosphate (91.8 ± 1.2) were removed from the wastewater along with antibiotics after 10 days of treatment. The findings indicate that the indigenous bacterial communities and the ability to survive in the presence of high antibiotic concentrations and xenobiotics. Moving bed bioreactor is useful for the removal of nutrients and antibiotics from wastewater.

Environmental photochemistry in Solanum trilobatum mediated plasmonic nanoparticles as a probe for the detection of Cd2+ ions in water.

Nowadays world deals with a lot of environmental troubles out of which water pollution is very dangerous. Water gets contaminated by heavy metal ions is a universal problem which needs suitable consideration to keep up the quality of the water. It will be advantageous that an easy device can be detecting the concentration of heavy metal ions in water. Here, a contaminant, cadmium from industrial affluent into water is considered and focused. Gold nanoparticles (AuNPs) have been synthesized by Solanum trilobatum leaf extract and its applications of antifungal and sensing activity was reported here. The influences of different concentration of these reducing agent on the synthesis of AuNPs (G5 and G10) have been evaluated. The structural, optical, vibrational, morphological and compositional properties of the AuNPs were studied through XRD, UV-vis spectra, FTIR, HRTEM and EDAX analysis. The optical studies showed surface plasmon absorbance peak at 526 nm. It shows that the absorbance of the peak becomes narrow with a higher concentration of leaf extract. XRD results showed the average size of the AuNPs was 8 nm. It also confirmed the high crystallinity of nanoparticles. FTIR exposes that amine and carboxyl groups may be involved in the stabilization and reduction mechanism. TEM pictures of both G10 and G5 demonstrate merely spherical nanoparticles. This morphology control is taken place owing to the adsorbed amine and carboxyl groups onto the gold nanoparticles cap the particles and improve the stability. The presence of gold elements in the sample was identified with the help of EDAX. The sensitivity of the system towards various Cd2+ concentrations was measured as 0.058/mM for G5 and 0.095/mM for G10. The prepared nanoparticles produced highest zone of inhibition (ZOI) of 17.5 mm and 19 mm against human being pathogenic fungi Aspergillus Flavus and Candida albicans respectively. Here, small sized spherical nanoparticles showed good antifungal activity.

Synthesis and characterization of magnetite carbon nanocomposite from agro waste as chromium adsorbent for effluent treatment.

The waste water released from industries which contain pollutants like heavy metals, dyes and other toxic chemicals brings numerous harms to the ecosystem and humans. Nowadays the nanocomposites based technologies are effectively used for environmental remediation. In the present study, hexavalent chromium was removed from the industrial effluent using magnetite carbon nanocomposite. The nanocomposite composed of highly porous carbon and iron oxide nanoparticles prepared by using agrowastes (sugarcane bagasse and orange peel extract). Iron oxide nanoparticles (FeONPs) formation was confirmed by UV-visible spectroscopy; incorporation of magnetite with highly porous carbon was established by Fourier Transforms Infrared Spectroscopy and X-ray Diffraction Spectroscopy. Morphological features of magnetite nanoparticles and highly porous carbon were analyzed using Scanning Electron Microscope and Transmission Electron Microscope. Magnetic properties analyzed by Vibrating Sample Magnetometer revealed magnetite carbon nanocomposite exhibited better Ms value than highly porous carbon. The concentration of Cr6+ in treated effluent was determined using Atomic Absorption Spectroscopy. Pseudo-second order equation fitted with kinetics and the Langmuir monolayer favors for isotherm. This study reveals efficiency in Cr6+ removal from effluent using magnetite carbon nanocomposites which extends their application in waste water treatment.

Insulin signaling pathway assessment by enhancing antioxidant activity due to morin using in vitro rat skeletal muscle L6 myotubes cells.

BACKGROUND: Plant-derived phytochemicals such as flavonoids have been explored to be powerful antioxidants that protect against oxidative stress-related diseases. In the present study, Morin, a flavonoid compound was studied for its antioxidant and antidiabetic properties in relation to oxidative stress in insulin resistant models conducted in rat skeletal muscle L6 cell line model. METHODS: Evaluation of antioxidant property of morin was assayed using in vitro methods such as cell viability by MTT assay, estimation of SOD and CAT activity and NO scavenging activity. The anti-oxidative nature of morin on L6 cell line was conducted by the DCF-DA fluorescent activity. Glucose uptake in morin treated L6 myotubes are accessed by 2-NBDG assay in the presence or absence of IRTK and PI3K inhibitors. Further glycogen content estimation due to the morin treatment in L6 myotubes was performed. Antioxidant and insulin signaling pathway gene expression was examined over RT-PCR analysis. RESULTS: Morin has a negligible cytotoxic effect at doses of 20, 40, 60, 80, and 100 µM concentration according to cell viability assay. Morin revealed that the levels of the antioxidant enzymes SOD and CAT in L6 myotubes had increased. When the cells were subjected to the nitro blue tetrazolium assay, morin lowered reactive oxygen species (ROS) formation at 60 µM concentration displaying 39% ROS generation in oxidative stress condition. Lesser NO activity and a drop in green fluorescence emission in the DCFDA assay, demonstrating its anti-oxidative nature by reducing ROS formation in vitro. Glucose uptake by the L6 myotube cells using 2-NBDG, and with IRTK and PI3K inhibitors (genistein and wortmannin) showed a significant increase in glucose uptake by the cells which shows the up regulated GLUT-4 movement from intracellular pool to the plasma membrane. Morin (60 µM) significantly enhanced the expression of antioxidant genes GPx, GST and GCS as well as insulin signalling genes IRTK, IRS-1, PI3K, GLUT-4, GSK-3ß and GS in L6 myotubes treated cells. CONCLUSION: Morin has the ability to act as an anti-oxidant by lowering ROS levels and demonstrating insulin mimetic activity by reversing insulin resistance associated with oxidative stress.

NV14 from serine O-acetyltransferase of cyanobacteria influences the antioxidant enzymes in vitro cells, gene expression against H2 O2 and other responses in vivo zebrafish larval model.

In this study, we have identified a novel peptide NV14 with antioxidative functions from serine O-acetyltransferase (SAT) of Artrospira platensis (Ap). The full sequence of ApSAT and its derived NV14 peptide "NVRIGAGSVVLRDV" (141-154) was characterized using bioinformatics tools. To address the transcriptional activity of ApSAT in response to induce generic oxidative stress, the spirulina culture was exposed to H2 O2 (10 mM). The ApSAT expression was studied using RT-PCR across various time points and it was found that the expression of the ApSAT was significantly upregulated on Day 15. The in vitro cytotoxicity assay against NV14 was performed in human dermal fibroblast cells and human blood leukocytes. Results showed that NV14 treatment was non-cytotoxic to the cells. Besides, in vivo treatment of NV14 in zebrafish larvae did not exhibit the signs of developmental toxicity. Further, the in vitro antioxidant assays enhanced the activity of the antioxidant enzymes, such as SOD and CAT, due to NV14 treatment; and also significantly reduced the MDA levels, while increasing the superoxide radical and H2 O2 scavenging activity. The expression of antioxidant enzyme genes glutathione peroxidase, γ-glutamyl cysteine synthase, and glutathione S-transferase were found to be upregulated in the NV14 peptide pretreated zebrafish larvae when induced with generic oxidative stress, H2 O2 . Overall, the study showed that NV14 peptide possessed potent antioxidant properties, which were demonstrated over both in vitro and in vivo assays. NV14 enhanced the expression of antioxidant enzyme genes at the molecular level, thereby modulating and reversing the cellular antioxidant balance disrupted due to the H2 O2 -induced oxidative stress.

Sequestration of toxic Pb(II) ions using ultrasonic modified agro waste: Adsorption mechanism and modelling study.

Treating the effluents from industries by using biological and agricultural wastes is an emerging field of research. In this study, three different biosorbents are prepared from tamarind seeds such as; raw, sulphuric acid-modified and ultrasonic-assisted surface-modified tamarind seed powder has been utilized to expel the Pb (II) ions from synthetic solution. The surface characteristics of the newly synthesized raw and surface modified agro-waste biomass were studied by FTIR and SEM. An experimental study was carried out to investigate the effect of different parameters on adsorption of Pb(II) ions using raw, sulphuric acid-modified and ultrasonic-assisted surface-modified tamarind seeds. The maximum Pb(II) ions adsorption was found at pH - 6.0, temperature - 303 K, biosorbent dosage - 3.5 g/L and contact time - 60 min for raw tamarind seeds and 30 min for sulphuric acid-modified and ultrasonic-assisted surface-modified tamarind seeds. The adsorption mechanism was described by Langmuir isotherm and pseudo-first order kinetic model. Among the three biosorbents, ultrasonic-assisted surface-modified tamarind seeds show higher adsorption capacity (18.86 mg/g) of Pb(II) ions removal from the synthetic solution. The thermodynamic study declared that the present Pb(II) ions adsorption onto the prepared biosorbents was spontaneous, exothermic and followed physical adsorption process. Results have shown that tamarind seed was found to be the best adsorbent in the expulsion of Pb(II) ions from the wastewater environment.

Medicinal Plants, Phytochemicals, and Herbs to Combat Viral Pathogens Including SARS-CoV-2.

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome corona virus-2 (SARS-CoV-2), is the most important health issue, internationally. With no specific and effective antiviral therapy for COVID-19, new or repurposed antiviral are urgently needed. Phytochemicals pose a ray of hope for human health during this pandemic, and a great deal of research is concentrated on it. Phytochemicals have been used as antiviral agents against several viruses since they could inhibit several viruses via different mechanisms of direct inhibition either at the viral entry point or the replication stages and via immunomodulation potentials. Recent evidence also suggests that some plants and its components have shown promising antiviral properties against SARS-CoV-2. This review summarizes certain phytochemical agents along with their mode of actions and potential antiviral activities against important viral pathogens. A special focus has been given on medicinal plants and their extracts as well as herbs which have shown promising results to combat SARS-CoV-2 infection and can be useful in treating patients with COVID-19 as alternatives for treatment under phytotherapy approaches during this devastating pandemic situation.

Green synthesis of lignin nanorods/g-C3N4 nanocomposite materials for efficient photocatalytic degradation of triclosan in environmental water.

Triclosan (TCS) is a common anti-microbial ingredient in pharmaceutical and personal care products. The usage of TCS was banned by the United States Food and Drug Administration (in 2016) due to its potential health risks. However, TCS has been frequently detected in the aquatic environment. Therefore, it is vital to design low-cost and highly efficient photocatalysts to enhance TCS's photocatalytic degradation in wastewater treatment to eliminate its toxicity to environmental health. In this study, we developed a highly efficient catalyst by incorporating lignin nanorods (LNRs) into graphitic carbon nitride (GCN) nanomaterials as green LNRs/GCN-based nanocomposite photocatalysts for the effective degradation of TCS in waters. LNRs/GCN nanosheets (NSs) and LNRs/GCN-NRs based nanocomposite materials were prepared using a simple wet-impregnation method. The surface morphology and optical properties of as-synthesized materials were well-characterized using FE-SEM, XRD, XPS, and UV-DRS. The photocatalyst (LNRs/GCN-NRs) material showed maximum TCS degradation efficiency of 99.9% and a high rate constant of 0.0661 min-1 under pH-10 with crucial reactive spices (OH and O2-), and excellent cycling performance (over five cycles) within 90 min of UV-light illumination. LNRs/GCN-NRs nanocomposite indicated enhanced photocatalytic performances for TCS degradation due to its strong synergistic effect between LNRs and GCN-NRs as bifunctional catalyst substrate morphology with efficient bandgap energy and accessible active sites compared to LNRs/GCN-NSs. Therefore, LNRs/GCN-NRs nanocomposite was observed to be a highly-active, low-cost, stable, eco-friendly, and efficient photocatalyst for complete degradation of TCS under UV-light irradiation.