Polyhydroxybutyrate degradation by biocatalyst of municipal sludge water and degradation efficacy in sequencing batch biofilm reactor.

The main aim of the study was to degrade poly-ß-hydroxybutyrate (P(3HB)) in the sequencing batch biofilm reactor (SBBR) using biocatalyst. Enrichment method was used for the isolation of P(3HB) degrading bacteria. These bacterial strains were isolated from the wastewater sludge sample treated with P(3HB) sheets. A total of 75 bacteria were isolated after 60 days of incubation. The zone of clearance varied between 12 ± 1 mm and 19 ± 2 mm. Two bacterial strains (Nitrobacter vulgaris SW1 and Pseudomonas aeruginosa KS10) showed rapid PHB degradation activity on agar plates. Plate screening experiments confirmed PHB degrading ability of P. aeruginosa KS10 and N. vulgaris SW1. Biodegrading potential improved after 72 h fermentation period. The bacteria produced depolymerase and enzyme activity was maximum after 72 h. The sequencing batch biofilm reactor (SBBR) co-cultured with N. vulgaris SW1 and P. aeruginosa KS10 was operated to remove PHB from the wastewater. Biofilm in the reactor degraded PHB and the production of polyhydroxybutyrate depolymerase influenced on PHB degradation. Polyhydroxybutyrate degradation improved continuously and maximum degradation (95.6%) was achieved after 8 days. The degradation of biopolymers help to reduce environmental pollution associated with the petroleum based polymers.

Anti-Obesity Efficacy of Pediococcus acidilactici MNL5 in Canorhabditis elegans Gut Model.

In the present study, thirty two lactic acid bacteria (LAB) were isolated from fermented Indian herbal medicine. In comparison to other strains, MNL5 had stronger bile salt hydrolase (BSH) and cholesterol-lowering properties. Furthermore, it can withstand the extreme conditions found in the GI tract, due to, e.g., pepsin, bile salts, pancreatin, and acids. Pediococcus acidilactici MNL5 was identified as a probiotic candidate after sequencing the 16S rRNA gene. The antibacterial activity of P. acidilactici MNL5 cell-free supernatants (CFS) against Escherichia coli, Staphylococcus aureus, Helicobacter pylori, Bacillus cereus, and Candida albicans was moderate. A Caenorhabditis elegans experiment was also performed to assess the effectiveness of P. acidilactici MNL5 supplementation to increase life span compared to E. coli supplementation (DAF-2 and LIU1 models) (p < 0.05). An immense reduction of the lipid droplets of C. elegans was identified through a fluorescent microscope. The drastic alteration of the expression of fat genes is related to obesity phenotypes. Hence, several paths are evolutionary for C. elegans; the results of our work highlight the nematode as an important model for obesity.

Antibacterial, Antioxidant, Larvicidal and Anticancer Activities of Silver Nanoparticles Synthesized Using Extracts from Fruits of Lagerstroemia speciose and Flowers of Couroupita guianensis.

The present study aimed to analyze the in vitro antibacterial, antioxidant, larvicidal and cytotoxicity properties of green synthesized silver nanoparticles (Ag NPs) using aqueous extracts from fruits of Lagerstroemia speciosa and flowers of Couropita guinensis. Synthesized Ag NPs were characterized using UV-DRS, FTIR, XRD, DLS, and High-Resolution SEM and TEM analyses. Absorption wavelength was observed at 386 nm by UV-DRS analysis and energy band gap was calculated as 3.24 eV. FTIR analysis showed the existence of various functional groups in the aqueous extract and in the NPs. DLS analysis showed the stability and particle size of the synthesized Ag NPs. SEM analysis revealed that Ag NPs are in a face centered cubic symmetry and spherical shape with a size of 23.9 nm. TEM analysis showed particle size as 29.90 nm. Ag NPs showed antibacterial activity against both Gram-positive and Gram-negative bacteria. DPPH scavenging trait of Ag NPs was ranging from 20.0 ± 0.2% to 62.4 ± 0.3% and observed significant larvicidal activity (LC50 at 0.742 ppm and LC90 at 6.061 ppm) against Culex quinquefasciatus. In vitro cytotoxicity activity of Ag NPs was also tested against human breast cancer (MCF-7) and fibroblast cells (L-929) and found that cells viabilities are ranging (500 to 25 µg/mL) from 52.5 ± 0.4 to 94.0 ± 0.7% and 53.6 ± 0.5 to 90.1 ± 0.8%, respectively. The synthesized Ag NPs have the potential to be used in the various biomedical applications.

Anti-tubercular and probiotic properties of coagulase-negative staphylococci isolated from Koozh, a traditional fermented food of South India.

In the last few years, the demand for the tremendous therapeutic applications of indigenous probiotic bacteria from diversified fermented food products has surged. In view of this, the present study was documented to evaluate the anti-tubercular and probiotic properties of coagulase-negative staphylococci (CNS) indigenous to Koozh, a traditional fermented food product of South India. A total of 18 isolates were purified from Koozh, and tested for anti-tubercular activity against Mycobacterium tuberculosis H37Rv using luciferase reporter phage (LRP) assay. Among them, six isolates revealed higher percentage (>90%) of relative light unit (RLU) reduction. These six isolates were further evaluated for their in vitro probiotic attributes using standard protocols. All six staphylococci strains disclosed good probiotic properties. Moreover, Staphylococcus hominis strain MANF2 showed high cell survival percentage (92.2%) at pH 2.0 as well as towards simulated gastric juice (88.51%). Furthermore, strain MANF2 was found to be resistant to bile salt after 24 h of incubation with maximal viability of 5.71 ± 0.02 log cfu/mL, and depicted the deconjugation of bile salt as well. All the isolates exhibited strong auto-aggregation capacity (44.4 ± 1.2-68.1 ± 1.5%), and hydrophobicity against toluene (55.0 ± 1.2-72.0 ± 1.1%). Additionally, strain MANF2 was observed to be highly resistant to phenol (6.27 ± 0.01 log cfu/mL) and lysozyme (81.1 ± 1.6% viability). Most importantly, all six isolates depicted good hypocholesterolemic effect, slight ß-galactosidase activity, and moderate proteolytic property. The strains were sensitive to all the tested conventional antibiotics, except Nalidixic acid. In addition to this, all staphylococci strains demonstrated significant DPPH (2,2-Diphenyl-1-picrylhydrazyl) scavenging, hydrogen peroxide tolerance, and hydroxyl radical scavenging activity in a dose dependent manner, thereby exhibiting the potent antioxidative properties of isolates. The negative results obtained from haemolytic, DNase, and gelatinase tests revealed the non-pathogenicity and safety aspect of these strains. In a nutshell, the present investigation divulges the persuasive anti-tubercular and probiotic properties of staphylococci, particularly strain MANF2, and recommended the further exploitation of Koozh associated CNS in pharmaceutics.

Enhancement of anti-tubercular activity and biomass of fermented food associated Staphylococcus hominis strain MANF2 using Taguchi orthogonal array and Box-Behnken design.

The prime focus of the present investigation was to optimize statistically the anti-tubercular activity and biomass of fermented food associated Staphylococcus hominis strain MANF2 using Taguchi orthogonal array (OA) and Box-Behnken design (BBD). The anti-tubercular activity of strain MANF2 was determined against Mycobacterium tuberculosis H37Rv using luciferase reporter phase assay. Among varied media examined, the isolate exhibited impressive anti-tubercular activity with paramount relative light unit reduction of >90% in de Man Rogose Sharpe (MRS) broth. Primarily, the anti-tubercular activity and biomass of strain MANF2 were estimated in MRS broth by optimizing eight diversified parameters using one factor at a time (OFAT) method after working out a series of experiments. The most significant contributing factors selected through OFAT tool were optimized using Taguchi approach with a standard OA layout of L18 (22 × 36). Results demonstrated the significant (P ≤ 0.05) influence of pH, temperature, yeast extract, magnesium sulphate, and glycerol on response variables. These controlled variables were further optimized using BBD matrix at N = 46 by second-order polynomial equation. The fermentation medium of pH 6.5 constituting yeast extract (0.5% w/v), magnesium sulphate (0.1% w/v), and glycerol (1.5% v/v), being further incubated at 30 °C showed enhanced anti-tubercular activity (98.7%) and approximately 4 fold increment in the bacterial biomass yield (8.3 mg/mL) with respect to traditional OFAT method. Three-dimensional response plots of the quadratic model showed interdependent interaction between the significant variables. In conclusion, the present study revealed the first report on the optimization of anti-tubercular activity and biomass of S. hominis via Taguchi OA as well as BBD design, and thus, paved a path for its proficient applications in pharmaceutical industries as dynamic mycobactericidal agent in future.

Carboxymethyl cellulase production optimization from Glutamicibacter arilaitensis strain ALA4 and its application in lignocellulosic waste biomass saccharification.

In this context, carboxymethyl cellulase (CMCase) production from Glutamicibacter arilaitensis strain ALA4 was initially optimized by one factor at a time (OFAT) method using goat dung as proficient feedstock. Two-level full factorial design (25 factorial matrix) using first-order polynomial model revealed the significant (p < 0.05) influence of pH, moisture, and peptone on CMCase activity. Central composite design at N = 20 was further taken into account using a second-order polynomial equation, and thereby liberated maximum CMCase activity of 4925.56 ± 31.61 U/g in the goat dung medium of pH 8.0 and 100% moisture containing 1% (w/w) peptone, which was approximately two fold increment with respect to OFAT method. Furthermore, the partially purified CMCase exhibited stability not only at high pH and temperature but also in the presence of varied metal ions, organic solvents, surfactants, and inhibitors with pronounced residual activities. The enzymatic hydrolysis using partially purified CMCase depicted the maximum liberation of fermentable sugars from alkali pretreated lignocellulosic wastes biomass in the order of paddy straw (13.8 ± 0.15 mg/g) > pomegranate peel (9.1 ± 0.18 mg/g) > sweet lime peel (8.37 ± 0.16 mg/g), with saccharification efficiency of 62.1 ± 0.8, 40.95 ± 0.4, and 37.66 ± 0.4%, respectively after 72 hr of treatment.

Removal of SDS from biological protein digests for proteomic analysis by mass spectrometry.

BACKGROUND: Metal-organic frameworks (MOFs - MIL-101) are the most exciting, high profiled developments in nanotechnology in the last ten years, and it attracted considerable attention owing to their uniform nanoporosity, large surface area, outer-surface modification and in-pore functionality for tailoring the chemical properties of the material for anchoring specific guest moieties. MOF's have been particularly highlighted for their excellent gas storage and separation properties. Recently biomolecules-based MOF's were used as nanoencapsulators for antitumor and antiretroviral controlled drug delivery studies. However, usage of MOF material for removal of ionic detergent-SDS from biological samples has not been reported to date. Here, first time we demonstrate its novel applications in biological sample preparation for mass spectrometry analysis. METHODS: SDS removal using MIL-101 was assessed for proteomic analysis by mass spectrometry. We analysed removal of SDS from 0.5 % SDS solution alone, BSA mixture and HMEC cells lysate protein mixture. The removal of SDS by MIL-101 was confirmed by MALDI-TOF-MS and LC-MS techniques. RESULTS: In an initial demonstration, SDS has removed effectively from 0.5 % SDS solution by MIL-101via its binding attraction with SDS. Further, the experiment also confirmed that MIL-101 strongly removed the SDS from BSA and cell lysate mixtures. CONCLUSIONS: These results suggest that SDS removal by the MIL-101 method is a practical, simple and broad applicable in proteomic sample processing for MALDI-TOF-MS and LC-MS analysis.

Chemical analysis of Punica granatum fruit peel and its in vitro and in vivo biological properties.

BACKGROUND: The medical application of pomegranate fruits and its peel is attracted human beings. The aim of the present study was to evaluate the in vitro α-Glucosidase inhibition, antimicrobial, antioxidant property and in vivo anti-hyperglycemic activity of Punica granatum (pomegranate) fruit peel extract using Caenorhabditis elegans. METHODS: Various invitro antioxidant activity of fruit peel extracts was determined by standard protocol. Antibacterial and antifungal activities were determined using disc diffusion and microdilution method respectively. Anti-hyperglycemic activity of fruit peel was observed using fluorescence microscope for in vivo study. RESULTS: The ethyl acetate extract of P. granatum fruit peel (PGPEa) showed α-Glucosidase inhibition upto 50 % at the concentration of IC50 285.21 ± 1.9 µg/ml compared to hexane and methanol extracts. The total phenolic content was highest (218.152 ± 1.73 mg of catechol equivalents/g) in ethyl acetate extract. PGPEa showed more scavenging activity on 2,2-diphenyl-picrylhydrazyl (DPPH) with IC50 value 302.43 ± 1.9 µg/ml and total antioxidant activity with IC50 294.35 ± 1.68 µg/ml. PGPEa also showed a significant effecton lipid peroxidation IC50 208.62 ± 1.68 µg/ml, as well as high reducing power. Among the solvents extracts tested, ethyl acetate extract of fruit peel showed broad spectrum of antimicrobial activity. Ethyl acetate extract supplemented C.elegans worms showed inhibition of lipid accumulation similar to acarbose indicating good hypoglycemic activity. The normal worms compared to test (ethyl acetate extract supplemented) showed the highest hypoglycaemic activity by increasing the lifespan of the worms. GC-MS analysis of PGPEa showed maximum amount of 5-hydroxymethylfurfural and 4-fluorobenzyl alcohol (48.59 %). CONCLUSION: In the present investigation we observed various biological properties of pomegranate fruit peel. The results clearly indicated that pomegranate peel extract could be used in preventing the incidence of long term complication of diabetics.

Acetaminophen Induced Hepatotoxicity in Wistar Rats--A Proteomic Approach.

Understanding the mechanism of chemical toxicity, which is essential for cross-species and dose extrapolations, is a major challenge for toxicologists. Standard mechanistic studies in animals for examining the toxic and pathological changes associated with the chemical exposure have often been limited to the single end point or pathways. Toxicoproteomics represents a potential aid to the toxicologist to understand the multiple pathways involved in the mechanism of toxicity and also determine the biomarkers that are possible to predictive the toxicological response. We performed an acute toxicity study in Wistar rats with the prototype liver toxin; the acetaminophen (APAP) effects on protein profiles in the liver and its correlation with the plasma biochemical markers for liver injury were analyzed. Three separate groups--control, nontoxic (150 mg/kg) and toxic dose (1500 mg/kg) of APAP--were studied. The proteins extracted from the liver were separated by 2-DE and analyzed by MALDI-TOF. The differential proteins in the gels were analyzed by BIORAD's PDQuest software and identified by feeding the peptide mass fingerprint data to various public domain programs like Mascot and MS-Fit. The identified proteins in toxicity-induced rats were classified based on their putative protein functions, which are oxidative stress (31%), immunity (14%), neurological related (12%) and transporter proteins (2%), whereas in non-toxic dose-induced rats they were oxidative stress (9%), immunity (6%), neurological (14%) and transporter proteins (9%). It is evident that the percentages of oxidative stress and immunity-related proteins were up-regulated in toxicity-induced rats as compared with nontoxic and control rats. Some of the liver drug metabolizing and detoxifying enzymes were depleted under toxic conditions compared with non-toxic rats. Several other proteins were identified as a first step in developing an in-house rodent liver toxicoproteomics database.

Some biological activities of Epaltes divaricata L. - an in vitro study.

BACKGROUND: Novel chemical molecules recovered from endangered medicinal plants have wide applications and have the potential to cure different diseases caused by microorganisms. The aim of this study was to investigate In vitro antimicrobial, α-glucosidase inhibition and antioxidant activity of different solvent extracts of Epaltes divaricata L. METHODS: Antimicrobial activity of hexane, ethyl acetate and methanol extract of Epaltes divaricata was determined against bacteria and fungi using disc diffusion and microdilution method respectively. α-glucosidase inhibition, Total phenolic content (TPC), Reducing power activity, DPPH radical scavenging assay, hydroxyl radical scavenging activity, nitric oxide scavenging activity, superoxide scavenging activity and lipid peroxidation assay of plant extracts were performed according to standard protocol. Compound detection from the potential solvent extract was done through GC-MS analysis. RESULTS: Epaltes divaricata ethyl acetate extracts (EDEa) (1.25 mg/disc) showed significant inhibition for E. lentum (23 mm), E. aerogenes (18 mm), P. fluorescence (15 mm) and A. baumanii (15 mm). Minimum inhibitory concentration (MIC) of EDEa was found to be 31.25 µg/ml, 62.5 µg/ml and 62.5 µg/ml against A. flavus, A. niger and T. rubrum respectively. EDEa showed more α-glucosidase inhibition and antioxidant activity compared to hexane and methanol. EDEa showed 50% α-glucosidase inhibition at the concentration of 525.20 ± 2.37 µg/ml. The TPC of EDEa was 412.0 ± 2.21 mg of catechol equivalents/g extract. EDEa showed great scavenging activity on 2,2-diphenyl-picrylhydrazyl (DPPH) (IC50 560 ± 2.02 µg/ml), hydroxyl (IC50 314.75 ± 2.56 µg/ml), nitric oxide (IC50 648.20 ± 2.09 µg/ml) and superoxide (IC50 361.14 ± 1.45 µg/ml) radicals, as well as high reducing power. EDEa also showed a more suppressive effect on lipid peroxidation. Using Antioxidant ß-carotene linoleate method, the scavenging values of EDEa was significantly lower than BHT. GC-MS analysis of EDEa showed maximum amount of 2-butenamide, N-(4-fluorophenyl)-3-methyl trans-cinnamyl tiglate silane and trichlorocyclohexyl silane (36.86%). CONCLUSION: The results obtained in this study clearly indicate that EDEa can be used as a natural antimicrobial, α-glucosidase inhibition and antioxidant agent.

In-vitro antimicrobial, antibiofilm, cytotoxic, antifeedant and larvicidal properties of novel quinone isolated from Aegle marmelos (Linn.) Correa.

BACKGROUND: Plant metabolites have wide applications and have the potential to cure different diseases caused by microorganisms. The aim of the study was to evaluate the antimicrobial, antibiofilm, cytotoxic, antifeedant and larvicidal properties of novel quinine isolated from Aegle marmelos (Linn.) Correa. METHODS: A compound was obtained by eluting the crude extract, using varying concentrations of the solvents by the chromatographic purification. Broth micro dilution method was used to assess the antimicrobial activity and anticancer study was evaluated using MTT assay. Larvicidal activity was studied using leaf disc no-choice method. RESULTS: Based on the IR, 13C NMR and 1H NMR spectral data, the compounds were identified as quinone related antibiotic. It exhibited significant activity against Gram positive and Gram negative bacteria. The lowest Minimum Inhibitory Concentration (MIC) of the compound against Bacillus subtilis and Staphylococcus aureus was 100 and 75 µg mL(-1) respectively. Against Escherichia coli and Pseudomonas aeruginosa it exhibited MIC value of 25 µg mL(-1). The MIC of the compound against Aspergillus niger, A. clavatus, Penicillium roqueforti was 20 µg mL(-1) and that against Fusarium oxysporum (20 µg mL(-1)), A. oryzae (40 µg mL(-1)), and Candida albicans (60 µg mL(-1)), respectively. It showed effective antibiofilm activity against E. coli, S. typhii and P. aeroginosa at 8 µg mL(-1) and did not exhibit considerable cytotoxic activity against Vero and HEP2 cell lines. Additionally, the compound documented significant antifeedant and larvicidal activities against Helicoverpa armigera and Spodoptera litura at 125, 250, 500 and 1000 ppm concentrations. CONCLUSION: The results concluded that the compound can be evaluated further in industrial applications and also an agent to prepare botanical new pesticide formulations.

Untargeted metabolomics of fermented onion (Allium cepa L) using UHPLC Q-TOF MS/MS reveals anti-obesity metabolites and in vivo efficacy in Caenorhabditis elegans.

The bioconversion of onion extracts with P. acidilactici MNL5 enhances the metabolites and has a synergistic lipid-reduction impact that is beneficial for anti-obesity studies. The 48 h fermented onion extracts (FOE) demonstrated an enhanced inhibitory activity against pancreatic lipase (89.5 ± 1.25 %) as compared to the raw onion extract (ROE) (33.4 ± 0.86 %). The antioxidant properties of FOE significantly increased compared to the ROE inhibitory effect on DPPH (99.5 ± 2.40 mg vitamin C equiv./mg, DW FOE), and ABTS (104.5 ± 2.32 mg vitamin C equiv./mg, DW FOE). Based on FOE's higher antioxidant activity, UHPLC-Q-TOF-MS/MS demonstrated dramatic changes in the untargeted metabolite profile as compared to ROE. Moreover, C. elegans supplemented with FOE and quercetin exhibited an enhanced lifespan activity, lipid reduction, and decreased triglycerides. FOE can lower cholesterol and enhance quercetin to promote pancreatic lipase activity for synergistic anti-obesity effects.

Emerging paradigms of viral diseases and paramount role of natural resources as antiviral agents.

In the current scenario, the increasing prevalence of diverse microbial infections as well as emergence and re-emergence of viral epidemics with high morbidity and mortality rates are major public health threat. Despite the persistent production of antiviral drugs and vaccines in the global market, viruses still remain as one of the leading causes of deadly human diseases. Effective control of viral diseases, particularly Zika virus disease, Nipah virus disease, Severe acute respiratory syndrome, Coronavirus disease, Herpes simplex virus infection, Acquired immunodeficiency syndrome, and Ebola virus disease remain promising goal amidst the mutating viral strains. Current trends in the development of antiviral drugs focus solely on testing novel drugs or repurposing drugs against potential targets of the viruses. Compared to synthetic drugs, medicines from natural resources offer less side-effect to humans and are often cost-effective in the productivity approaches. This review intends not only to emphasize on the major viral disease outbreaks in the past few decades and but also explores the potentialities of natural substances as antiviral traits to combat viral pathogens. Here, we spotlighted a comprehensive overview of antiviral components present in varied natural sources, including plants, fungi, and microorganisms in order to identify potent antiviral agents for developing alternative therapy in future.

Carbapenemases producing Klebsiella pneumoniae from the pus of hospitalized patients: In-vitro antibiotic properties of Streptomyces against multidrug resistant infectious bacteria.

BACKGROUND: Klebsiella pneumoniae is predominantly exists in the pus of the human wounds and it creates massive infections in the skin and causes serious health associated infections. Modern antibiotics are highly active in the treatment of wound infections. In this study was aimed to determine resistance of K. pneumoniae screened from wound specimens of patients. Sample was collected from the pus of the patients associated with secondary infection. METHODS: Samples were serially diluted and the isolated bacterial strains were characterized by biochemical tests, colony morphology and Gram's staining methods. Resistance of K. pneumoniae was tested using antibiotics such as, Gentamycin, Ampicillin, Tetracycline, Cefurooxime, Oxacillin, Ofloxacin, Erythromycin, Nalidic acid, Cefepine, Piperacillin, Norfloxacin, Imipenem, Nitrofurantoin, Amikacin, Ciprofloxacin, Vancomycin, Meropeneum and Cefotaxime with Kirby-Bauer disc diffusion method. RESULTS: Among the 73 K. pneumoniae strains, four strains produced AmpC and ESBLs, 42 strains produced ESBLs and 7 bacterial strains synthesized only AmpC enzyme. Four stains produced ESBLs and showed multidrug resistance against various antibiotics. Most of the strains synthesized extracellular polysaccharides and mediated biofilm formation. Among the K. pneumoniae strains, K. pneumoniae PS02 showed multidrug resistant against most of the tested antibiotics. It produced ESBLs and AmpC enzyme. To produce secondary metabolites, actinomycetes were isolated and characterized as Streptomyces sp. AC14. The secondary metabolite was effective against Klebsiella strains. CONCLUSIONS: To conclude, secondary metabolites extracted from Streptomyces sp. AC14 was found to be effective against multidrug resistant bacterium. Further studies are warranted to analyze the drug hydrolyzing pathways of bacteria and to identify the mechanism of action of secondary metabolites from Streptomyces sp. AC14.

Hepatoprotective effect of selected isoandrographolide derivatives on steatotic HepG2 cells and High Fat Diet fed rats.

Non-alcoholic Fatty Liver Disease (NAFLD) is one of the growing epidemics of the globe. This study was aimed to evaluate the anti-NAFLD effect of selected IAN derivatives using in silico, in vitro and in vivo models. In silico tools viz., DataWarrior, SwissADME and Gaussian 09 were used to predict the pharmacokinetic properties and electronic distribution patterns of the derivatives; docking analysis was done with Autodock against PPARα. Toxicities of the derivatives were assessed in HepG2 cells using MTT assay. Anti-NAFLD efficacies of the derivatives were assessed in free fatty acid induced steatotic HepG2 cells. In vivo anti-NAFLD effect of active isoandrographolide (IAN) derivative, 19-propionyl isoandrographolide (IAN-19P) was assessed in High Fat Diet fed rats. In silico and in vitro studies indicated that IAN-19P showed improved drug-likeness and drug score. The toxicity of IAN-19P to HepG2 cells was comparatively less than IAN and other derivatives. In free fatty acid induced steatotic HepG2 cells, treatment with IAN-19P significantly lowered intracellular triglyceride content and leakage of LDH and transaminases. Treating High Fat Diet fed animals with IAN-19P significantly lowered plasma lipids, transaminases, LDH and GGT levels. The treatment with IAN-19P upregulated the expressions of PPARα and CPT-1. IAN-19P did not produce any noticeable adverse effect till 2 g/kg concentration in acute and 250 mg/kg concentration in subacute toxicity studies. This study indicated the beneficial effect of IAN-19P for the treatment of NAFLD; however robust investigations are needed to establish the potential of IAN-19P to treat NAFLD.

Molecular diversity and hydrolytic enzymes production abilities of soil bacteria.

Soil is an integral part of ecosystem which is niche for varieties of microflora. The present study was investigated to isolate varied strains of bacteria from soil samples of three different geographical regions of Tamil Nadu (India) and evaluate their hydrolytic enzymes (amylase, cellulase, and inulinase) producing potentialities. Among 72 bacterial cultures isolated from Ambattur Industrial Estate, Neyveli Lignite Corporation, and Arignar Anna Zoological Park regions, 41.66, 38.88, and 36.11% of isolates were observed amylase, cellulase, and inulinase producers, respectively. On the other hand, 20.83% of total bacteria isolated from all three regions exhibited concurrent production of amylase, cellulase, and inulinase. Potent isolates depicting maximum enzyme activities were identified as Bacillus anthracis strain ALA1, Bacillus cereus strain ALA3, Glutamicibacter arilaitensis strain ALA4, and Bacillus thuringiensis strain ALA5 based on molecular characterization tools. Further, the thermodynamics parameters, open reading frames (ORFs) regions, and guanine-cytosine (GC) content were determined by distinct bioinformatics tools using 16S rRNA sequences of strains. Minimum free energy values for strain ALA1, strain ALA3, strain ALA4, and strain ALA5 were calculated as -480.73, -478.76, -496.63, and -479.03 kcal/mol, respectively. Mountain plot and entropy predicted the hierarchical representation of RNA secondary structure. The GC content of sequence for strain ALA1, strain ALA3, strain ALA4, and strain ALA5 was calculated as 53.06, 52.94, 56.78, and 53.06%, respectively. Nine ORFs were obtained for strain ALA1, strain ALA3, and strain ALA5 while 10 ORFs were observed for strain ALA4. Additionally, bootstrap tree demonstrated close resemblance of strains with existing bacteria of similar genus. Findings showed higher variability of bacterial diversity as hydrolytic enzymes producers in the investigated geographical regions.

Green synthesis and characterization of gold nanoparticles using endophytic fungi Fusarium solani and its in-vitro anticancer and biomedical applications.

The present study aimed to explore the anticancer potentials of the gold nanoparticles (NPs) obtained by green synthesis method using an endophytic strain Fusarium solani ATLOY - 8 has been isolated from the plant Chonemorpha fragrans. The formation of the NPs was analyzed by UV, FTIR, SEM and XRD. The synthesized NPs showed pink-ruby red colors and high peak plasmon band was observed between 510 and 560 nm. It is observed that intensity of absorption steadily increases the wavelength and band stabilizes at 551 nm. The XRD pattern revealed the angles at 19, 38.32, 46.16, 57.50, and 76.81° respectively. Interestingly, the FTIR band absorption noted at 1413 cm-1, 1041 cm-1 and 690 cm-1 ascribed the presence of amine II bands of protein, C-N and C-H stretching vibrations of the nanoparticles. SEM analysis indicated that the average diameter of the synthesized nanoparticles was between 40 and 45 nm. These NPs showed cytotoxicity on cervical cancer cells (He La) and against human breast cancer cells (MCF-7) and the NPs exhibited dose dependent cytotoxic effect. IC50 value was 0.8 ± 0.5 µg/mL on MCF-7 cell line and was found to be 1.3 ± 0.5 µg/mL on MCF-7 cell lines. The synthesized NPs induced apoptosis on these cancer cell lines. The accumulation of apoptotic cells decreased in sub G0 and G1 phase of cell cycle in the MCF-7 cancer cells were found to be 55.13%, 52.11% and 51.10% after 12 h exposure to different concentrations. The results altogether provide an apparent and versatile biomedical application for safer chemotherapeutic agent with little systemic toxicity.

Purification and characterization of anti-tubercular and anticancer protein from Staphylococcus hominis strain MANF2: In silico structural and functional insight of peptide.

The present context was investigated to purify and characterize anti-tubercular as well as anticancer protein from fermented food associated Staphylococcus hominis strain MANF2. Initially, the anti-tubercular potency of strain MANF2 was assessed against Mycobacterium tuberculosis H37Rv using luciferase reporter phase assay which revealed pronounced relative light unit (RLU) reduction of 92.5 ± 1.2%. The anticancer property of strain MANF2 was demonstrated against lung cancer (A549) and colon cancer (HT-29) cell lines using MTT assay which showed reduced viabilities. Anti-tubercular activities of the purified protein were observed to be increased significantly (P < 0.05) ranging from 34.6 ± 0.3 to 71.4 ± 0.4% of RLU reduction. Likewise, the purified protein showed significantly (P < 0.05) reduced viabilities of A549 and HT-29 cancer cells with IC50 values of 46.6 and 48.9 µg/mL, respectively. The nominal mass of the purified protein was found to be 7712.3 Da as obtained from MALDI-TOF MS/MS spectrum. The protein showed the sequence homology with 1-336 amino acids of Glyceraldehyde-3-phosphate dehydrogenase from Staphylococcus sp., thus, categorizing as a new class of Glyceraldehyde-3-phosphate dehydrogenase-like protein. The amino acid sequence of the most abundant peptide (m/z = 1922.12) in the purified protein was obtained as 'KAIGLVIPEIDGKLDGGAQRV' and it was identified as peptide NMANF2. In silico tools predicted significant stereo-chemical, physiochemical, and functional characteristics of peptide NMANF2. In a nutshell, protein purified from strain MANF2 can certainly be used as an ideal therapeutic agent against tuberculosis and cancer (lung and colon).

Chemopreventive potential of Epoxy clerodane diterpene from Tinospora cordifolia against diethylnitrosamine-induced hepatocellular carcinoma.

Medicinal plants are a promising source for identification of lead molecules for cancer therapy. In our continuous search to discover bioactive compounds from natural products, we isolated (5R, 10R)-4R, 8R-dihydroxy-2S, 3R:15, 16-diepoxycleroda-13(16), 17, 12S:18,1S-dilactone (ECD), a diterpenoid from Tinospora cordifolia and studied its chemopreventive potential in diethylnitrosamine (DEN) induced hepatocellular carcinoma (HCC) rats. Fifty male Wistar rats were divided into five groups. Group I served as normal control. Group II-IV were given DEN (0.01% in drinking water) for twenty weeks. In addition, Group III (preventive treatment) received ECD (10 mg/kg body weight) throughout the study. Group IV (curative treatment) received ECD (10 mg/kg body weight) for the last 8 weeks. Group V received ECD alone (10 mg/kg body weight) throughout the experimental period. At the end of the experimental period all the animals were sacrificed and analyzed for biochemical end points to assess the effect of ECD treatment in DEN induced HCC. The animals treated with DEN showed a decrease in the activities of antioxidant (SOD, CAT) and detoxification enzymes (GSH, GPx) with increase in the activities of the hepatic markers (SGOT, SGPT, LDH). Treatment of ECD in both preventive and curative DEN induced animals increased the level of antioxidants and detoxification enzymes, and decreased serum transaminase level and hepatic marker enzymes to near normal. Histopathological and nodular incidence also confirmed that ECD remarkably reduced tumor incidence and reversed damaged hepatocytes to normal. Our findings confirm that ECD exhibits preventive effect against chemically induced HCC in rats. ECD can be a potent chemopreventive drug for HCC.

Isolation and characterization of Aspergillus sp. for the production of extracellular polysaccharides by response surface methodology.

In this study, Aspergillus sp. was isolated for the production of extracellular polysaccharide. The process parameters were initially optimized by traditional methods. The cheap substrate, wheat bran was used for the production of extracellular polysaccharide in solid state fermentation. Supplementation of (1%, w/w) maltose, gelatin enhanced EPS production (5.36 mg/g). The salts such as, Cu2+ (4.9 mg/g), Ca2+ (3.5 mg/g), Zn2+ (2.9 mg/g), Mn2+ (3.4 mg/g) and Mg2+ (1.8 mg/g) stimulated EPS production. In two level full factorial experimental designs, the EPS yield varied from 3.18 to 11.65 mg/g wheat bran substrate with various combinations of the components supplemented with wheat bran substrate. Among these selected factors in central composite design, maltose significantly influenced on extracellular polysaccharide production.