From nature to cancer therapy: Evaluating the Streptomyces clavuligerus secondary metabolites for potential protein kinase inhibitors.

The study aimed to evaluate the antioxidant, protein kinase inhibitory (PKIs) potential, cytotoxicity activity of Streptomyces clavuligerus extract. DPPH assay revealed a robust free radical scavenging capacity (IC50 28.90 ± 0.24 µg/mL) of organic extract with a maximum inhibition percentage of 61 ± 1.04%. PKIs assay revealed the formation of a whitish bald zone by S. clavuligerus extracts which indicates the presence of PKIs. The cytotoxicity activity of organic fraction of extract through Sulforhodamine B assay on MCF-7, Hop-62, SiHa, and PC-3 cell lines demonstrated the lowest GI50 value against the MCF-7 cell line followed by the PC-3 cell line, showing potent growth inhibitory potential against human breast cancer and human prostate cancer cell line. HR-LCMS analysis identified multiple secondary metabolites from the organic and aqueous extracts of S. clavuligerus when incubated at 30°C under 200 rpm for 3 days. All the secondary metabolites were elucidated for their potential to inhibit RTKs by molecular docking, molecular dynamic simulation, MM/GBSA calculations, and free energy approach. It revealed the superior inhibitory potential of epirubicin (Epi) and dodecaprenyl phosphate-galacturonic acid (DPGA) against fibroblast growth factors receptor (FGFR). Epi also exhibited excellent inhibitory activity against the platelet-derived growth factor receptor (PDGFR), while DPGA effectively inhibited the vascular endothelial growth factor receptor. Additionally, the presence Epi in S. clavuligerus extract was validated through the HPLC technique. Thus, our findings highlight a superior inhibitory potential of Epi against FGFR and PDGFR RTKs than the FDA-approved drug.

On the potential activity of hyaluronic acid as an antimicrobial agent: experimental and computational validations.

This century has seen the rise of antibiotic resistance as a significant public health problem. In addition, oxidative stress may also be a factor in selecting resistant strains of bacteria. The current study analyzed microbially produced hyaluronic acid's antibacterial activity and antioxidant activity. It had significant antibacterial action against strains of Staphylococcus aureus and Escherichia coli, with the IC50 value obtained being 487.65 µg mL-1 for antioxidant assay. Our molecular docking investigations of hyaluronic acid on tyrosyl-tRNA synthetase (Staphylococcus aureus: -6.13 kcal/mol, Escherichia coli: -5.79 kcal/mol) and topoisomerase II DNA gyrase (Staphylococcus aureus: -5.02 kcal/mol, Escherichia coli: -4.90 kcal/mol) confirmed the ligands' possible binding mode to the appropriate targets' sites. We also employed molecular dynamics simulation and showed that HA binds more strongly with 1JIL (-85.455 ± 12.623 kJ/mol) compared to 2YXN (-49.907 ± 64.191 kJ/mol), 5CDP (-47.285 ± 13.925 kJ/mol), and 6RKS (-45.306 ± 21.338 kJ/mol). We also report that the ligand forms several hydrogen bonds in molecular simulation, implying regular interaction with key residues of the enzymes. The results in this study indicate the potential use of HA in the vast field of applications having both asthetic and medicinal values.

Effects of monochromatic lights on the melanophores arrangement in the spotted snakehead fish Channa punctata (Bloch, 1793).

Some freshwater teleost fish have pigment cells whose arrangement and shape are affected by the environment. Natural light has a wide range of light intensity. Fish are sensitive to the background and exposed light colour. Fish body colour is a significant criterion in fixing its market value, whether it is ornamental or edible. By favourable light exposure, a culturist may get a good market value of fish on most ethical grounds. In this study, we recorded the changes in melanophore response with the changes in light colour on Channa punctata. Adult fish were treated with monochromatic lights (darkness, white, blue and red light) for 5 and 28 days. After treatment, their body colour and melanophore size, number, length and the number of dendrites were studied. The results showed a significant influence of monochromatic light on melanophore arrangement in fish skin. The data showed that blue light is appropriate for the overall species colour of photic C. punctata. Continuous black or white light caused severe damage to the fish's appearance.

L-asparaginase production in solid-state fermentation using Aspergillus niger: process modeling by artificial neural network approach.

L-asparaginase has proven itself as a potential anti-cancer drug and in the mitigation of acrylamide formation in the food industry. In the present investigation, a novel utilization of niger (Guizotia abyssinica) de-oiled cake as the sole source for the cost-effective production of L-asparaginase was evaluated and compared with different agro-substrates in solid-state fermentation. The substrate provided a favorable C/N content for the L-asparaginase production as evident from the chemical composition (CHNS analysis) of the substrate. The influential process parameters viz; autoclaving time, moisture content, temperature and pH were optimized and modeled using machine-learning based artificial neural network (ANN) and statistical-based response surface methodology (RSM). The maximum enzyme activity of 34.65 ± 2.18 IU/gds was observed at 30.3 min of autoclaving time, 62% moisture content, 30 °C temperature and 6.2 pH in 96 h. A 1.36 fold improvement in enzyme activity was observed on utilizing optimized parameters. In comparison with RSM, the ANN model showed superior prediction with a low mean squared error of 0.072, low root mean squared error of 0.268 and 0.99 value of regression coefficient. The present study demonstrates the novel utilization of inexpensive and readily available agro-industrial waste for the development of cost-effective L-asparaginase production process.

Biocomputational Prediction Approach Targeting FimH by Natural SGLT2 Inhibitors: A Possible Way to Overcome the Uropathogenic Effect of SGLT2 Inhibitor Drugs.

The Food and Drug Administration (FDA) approved a new class of anti-diabetic medication (a sodium-glucose co-transporter 2 (SGLT2) inhibitor) in 2013. However, SGLT2 inhibitor drugs are under evaluation due to their associative side effects, such as urinary tract and genital infection, urinary discomfort, diabetic ketosis, and kidney problems. Even clinicians have difficulty in recommending it to diabetic patients due to the increased probability of urinary tract infection. In our study, we selected natural SGLT2 inhibitors, namely acerogenin B, formononetin, (-)-kurarinone, (+)-pteryxin, and quinidine, to explore their potential against an emerging uropathogenic bacterial therapeutic target, i.e., FimH. FimH plays a critical role in the colonization of uropathogenic bacteria on the urinary tract surface. Thus, FimH antagonists show promising effects against uropathogenic bacterial strains via their targeting of FimH's adherence mechanism with less chance of resistance. The molecular docking results showed that, among natural SGLT2 inhibitors, formononetin, (+)-pteryxin, and quinidine have a strong interaction with FimH proteins, with binding energy (∆G) and inhibition constant (ki) values of -5.65 kcal/mol and 71.95 µM, -5.50 kcal/mol and 92.97 µM, and -5.70 kcal/mol and 66.40 µM, respectively. These interactions were better than those of the positive control heptyl α-d-mannopyranoside and far better than those of the SGLT2 inhibitor drug canagliflozin. Furthermore, a 50 ns molecular dynamics simulation was conducted to optimize the interaction, and the resulting complexes were found to be stable. Physicochemical property assessments predicted little toxicity and good drug-likeness properties for these three compounds. Therefore, formononetin, (+)-pteryxin, and quinidine can be proposed as promising SGLT2 inhibitors drugs, with add-on FimH inhibition potential that might reduce the probability of uropathogenic side effects.

Characterization, inhibitory activity and mechanism of polyphenols from faba bean (gallic-acid and catechin) on α-glucosidase: insights from molecular docking and simulation study.

The chemo-profiling of ethanolic extract of faba beans seeds was performed and explored as an α-glucosidase inhibitor. The inhibition of α-glucosidase is one of the alternatives approach to control postprandial hyperglycemia by, resulting in the delay of the carbohydrate digestion of absorbable monosaccharides. Ethanolic seed extract showed phenolic compounds, flavonoid such as gallic acid (m/z [M- H] = 169.0124,C7H6O5) ellagic acid derivatives epigallocatechin (m/z [M- H = 305.0644,C15H14O7),catechin (m/z [M- H] = 289.0656,C15H14O6), epigallocatechin gallate (m/z [M- H] = 457.0578,C22H18O11) and epicatechin monogallate (m/z [M- H] = 441.081, C22H18O10). The extract was found to exert inhibitory activity (88.28 ± 2.67%) (IC50 value of 2.30 ± 0.032 mg/mL) with a mixed mode of inhibition (Km, apparent = 0.54 ± 0.020 mM and Vmax, apparent 0.136 ± 0.04 mM/min). Molecular docking studies of gallic acid and catechin on α-glucosidase proposed productive binding modes having binding energy (-6.58 kcal/mol and -7.25 kcal/mol) with an effective number of hydrogen bonds and binding energy. Tyr63, Arg197, Asp198, Glu 233, Asn324, Asp 326 of α-glucosidase participated in binding events with gallic acid and catechin. Molecular dynamics simulation studies were performed for both complexes i.e. gal:α-glucosidase and cat:α-glucosidase along with apo state of α-glucosidase, which revealed stable systems during the simulation. These findings of the present study may give an insight into the further development of the novel antidiabetic drug from the seeds of faba beans.

Protective effect of a protease inhibitor from Agaricus bisporus on Saccharomyces cerevisiae cells against oxidative stress.

Protease inhibitors are known to resist damage to host organisms against external threats, hence form a part of their defense system. This property of protease inhibitors was studied on protecting oxidatively stressed Saccharomyces cerevisiae yeast cells. The protease inhibitor was extracted from Agaricus bisporus, an edible mushroom. The inhibitor showed the presence of antioxidant activity as the purified inhibitor fraction gave an IC50 value of 45.13 ± 0.88 µg/mL and 33.30 ± 1.5 µg/mL when checked, respectively, by 2, 2-diphenyl-1-picrylhydrazyl, DPPH and 2, 2'-azo-bis(3-ethylbenzthiazoline-6- sulfonic acid), ABTS•+ scavenging activity. The yeast cells' survival rate (%), was determined through 3-(4, 5-dimethylthiazol-2-yl) - 2, 5-diphenyltetrazolium bromide, MTT assay, and it was found that in the presence of 2 mM H2O2 cell survival decreased to 26.33%, whereas when the experiment was conducted in the presence of protease inhibitor and 2 mM H2O2 cell survival percentage rose to 74%. The protease inhibitor's effect on the oxidatively stressed yeast cells was further studied by using Scanning Electron Microscopy (SEM), Atomic Force Microscopy (AFM) and Confocal Microscopy to understand the morphological changes. The viable and non-viable cell populations were quantified using Fluorescence Assorted Cell Sorting (FACS) using propidium iodide, PI, 4', 6-diamidino-2-phenylindole, DAPI and 2', 7'-dichlorofluorescein, DCF dyes.

In vitro investigation of hypoglycemic and oxidative stress properties of fava bean (Vicia faba L.) seed extract in Saccharomyces cerevisiae 2376.

Imbalance of free radicals over antioxidants in human body may result in oxidative damage to biomolecules (lipids, proteins, DNA) that causes severe chronic diseases. The aim of this proposed research was to examine the hypoglycemic and oxidative stress potential of fava bean (Vicia faba L.) seed extract. Acetone extract showed a significant effect on glucose uptake rate (77.28 ± 2.42%) in yeast cells at 25 mM glucose concentration. Minimum glucose uptake rate was found to be 52.36 ± 2.06% % by chloroform seed extract. Atomic force microscopy revealed that 3% hydrogen peroxide concentration results in roughness was found to be maximum (441 ± 6.7 nm) and along with extract treatment showed a significant reduction in roughness (251 ± 6.2 nm). Propidium iodide and DAPI staining showed apoptotic ratio as 0.40 (40 ± 1.18%,) and 0.42 (42 ± 1.16%) in hydrogen peroxide treated cell only as compared to other treatments. MTT assay showed that acetone extract had maximum survival rate (82.067%) and least survival rate was found in chloroform extract (70.48%). Hypoglycaemic potential and oxidative stress might be polyphenols (phenolics, flavonoids) present in seed extract or synergistic effect.

Alpha-amylase inhibitors from mycelium of an oyster mushroom.

The α-Amylase and α-glucosidase are two main enzymes involved in carbohydrate metabolism. This study was aimed at detecting alpha-amylase inhibitory activity from edible mushroom mycelia. Oyster mushroom was collected from a natural source, from Indian Institute of Technology (Banaras Hindu University) campus and was maintained in vitro in mycelial form. Chloroform, acetone, methanol, and water were used separately for extraction of an active constituent from mycelial cells grown, for 7 days, in potato dextrose broth. The extracts were tested for alpha-amylase inhibitory activity. Chloroform, acetone, and methanol extracts were found to have alpha-amylase inhibitory activity, with IC50 values of 1.71, 224, and 383 µg/mL, respectively. Aqueous extract had no enzyme inhibitory activity. The acetone extract inhibited α-amylase non-competitively whereas chloroform extract showed competitive inhibition. Acetone extraction yielded highest total phenolic content (TPC) of 0.524 mM of gallic acid equivalent, whereas chloroform extraction resulted in lowest TPC of 0.006 mM. The HPLC and absorbance maxima of acetone and chloroform extracts suggest that the bioactive component responsible for enzyme inhibition could be glycoproteins in chloroform extract and catechins (flavonoids) in acetone extract. Thus, the mushroom mycelia under study may be exploited for production and purification of a lead compound for the development of the α-amylase inhibitory drug.

Production of a protease inhibitor from edible mushroom Agaricus bisporus and its statistical optimization by response surface methodology.

The production of a protease inhibitor from Agaricus bisporus through solid-state fermentation was studied. The purpose was to produce protease inhibitor from natural, cheap, and readily available carbon and nitrogen sources. Solid-state fermentation enhanced the mycelia growth and also gave a higher yield of the product. Further, fungal growth and other production parameters were statistically optimized. The specificity of the inhibitor was tested and was effective against trypsin. Screening of significant factors (wheat bran, cyanobacterial biomass, initial pH, temperature, incubation period, and moisture content and inoculum size) was performed using Plackett-Burman design. Central composite design was used to determine the optimized values of the significant variables which were found to be temperature (27.5°C), incubation time (156 hr), cyanobacterial biomass (1 g), and moisture content (50%) and gave a statistical yield of 980 PIU/g which was 25.6% higher than experimental yield (780 PIU/g). The inhibitor was purified by ammonium sulfate precipitation and diethylaminoethyl (DEAE) cellulose chromatography (yield 43.89% and 0.21%, respectively) and subjected to reversed-phase HPLC to validate its identity. Since protease inhibitors act against proteases, finding ample therapeutic roles; the isolated protease inhibitor from A. bisporus can also be a probable medicinal agent after its further characterization.

The effect of fulvic acid on pre- and postaggregation state of Aß(17-42): molecular dynamics simulation studies.

Alzheimer's disease (AD), a neurodegenerative disorder, is directly related to the aggregation of Aß peptides. These peptides can self-assemble from monomers to higher oligomeric or fibrillar structures in a highly ordered and efficient manner. This self-assembly process is accompanied by a structural transition of the aggregated proteins from their normal fold into a predominantly ß-sheet secondary structure. 14ns molecular dynamics simulation revealed that fulvic acid interrupted the dimer formation of Aß(17-42) peptide while in its absence Aß(17-42) dimer formation occurred at ~12ns. Additionally, fulvic acid disrupted the preformed Aß(17-42) trimer in a very short time interval (12ns). These results may provide an insight in the drug design against Aß(17-42) peptide aggregation using fulvic acid as lead molecule against Aß(17-42) mediated cytotoxicity and neurodegeneration.

Synergistic effects of ternary mixture formulation and process parameters optimization in a sequential approach for enhanced L-asparaginase production using agro-industrial wastes.

A novel ternary mixture of inexpensive and nutrient-rich agro-substrates comprising groundnut de-oiled cake, corn gluten meal, and soybean meal has been explored to enhance the L-asparaginase production in solid-state fermentation. To achieve the aim, a hybrid strategy was implemented by utilizing a combination of a mixture design and artificial neural networks. The study initiated with the judicious selection of the agro-substrates based on their low C/N content in comparison to the control using the CHNS elemental analysis. The mixture composition of soybean meal (49.0%), groundnut de-oiled cake (31.5%), and corn gluten meal (19.5%) were found optimum using the simplex lattice mixture design. The agro-industrial substrates mix revealed synergistic effects on the L-asparaginase production than either of the substrates alone. The maximum L-asparaginase activity of 141.45 ± 5.24 IU/gds was observed under the physical process conditions of 70% moisture content, autoclaving period of 30 min and 6.0 pH by adopting the machine learning-derived artificial neural network (ANN) methodology. The ANN modeling showed excellent prediction ability with a low mean squared error of 0.7, a low root mean squared error of 0.84, and a high value of 0.99 for regression coefficient. Moisture content (%) was assessed to be the most sensitive process parameter in the global sensitivity analysis. The net outcome from the two sequential optimization designs is the selection of the ideal mixture composition followed by the optimum physical process parameters. The application of the enzyme demonstrated significant cytotoxicity against leukemia cell line and therefore exhibited an anti-cancer effect. The present study reports a novel mixture combination and methodology that can be used to lower the cost and enhance the production of L-asparaginase using an agro-industrial substrate mixture.

Development of Daruharidra (Berberis aristata) Based Biogenic Cadmium Sulfide Nanoparticles: Their Implementation as Antibacterial and Novel Therapeutic Agents against Human Breast and Ovarian Cancer.

BACKGROUND: This article presents a new and environmentally friendly method for generating DH-CdSNPs (cadmium sulfide nanoparticles) ranging from 5-10 nm in size. A green synthesis method for the development of inorganic nanoparticles was developed a few years back for their applications in diverse fields, such as medicine, bioimaging and remediation. The biogenic synthesis of these nanoparticles containing daruharidra (Berberis aristata) and cadmium sulfide is an effective alternative. AIMS: By employing Daruharidra extract as a herbal analog, we aim to minimize the risks and adverse effects that come along with the use of other chemically synthesized nanoparticles. This study's main goal was to investigate the potential of these nanoparticles as powerful antibacterial and anticancer agents. METHODS: We used a crude powdered daruharidra extract as a stabilizer ingredient to create CdSbased nanoformulations in an environmentally responsible way. By exposing the breast cancer cell line (MDAMB-231) and ovarian teratocarcinoma cell line (PA1) to these nanoformulations, we were able to evaluate their anticancer activities. Additionally, flow cytometry analysis was conducted to scrutinize the process of cell cycle arrest and apoptosis in reference to anticancer studies. Furthermore, DH-CdSNPs were applied on different gram-positive as well as gramnegative bacteria in a disc diffusion assay to ascertain their antibacterial activity. Nanoparticles were tested on bacterial strains to check if they were resistant after the MIC or minimum inhibitory concentration. RESULTS: The cytotoxicity of nanoparticles was tested by MTT assay. The impact of increasing concentrations of NPs on cell lines was tested, revealing a cytotoxic effect. The half-maximal inhibitory concentration values for a 24-hour treatment were determined to be 95.74µg/ml for ovarian cancer cells and 796.25 µg/ml for breast cancer cells. Treatment with DH-CdSNP resulted in a noteworthy increase in early apoptotic cells, with percentages rising from approximately 3% to 14.5% in ovarian cancer cell lines and from 4% to 13.6% in breast cancer cell lines. Furthermore, the NPs induced arrest of the cell cycle, specifically in the interphase of G2 and mitosis phase, with DNA damage observed in sub G1 in ovarian cancer cells and G0/G1 arrest observed in breast cancer cells. Additionally, the NPs exhibited exceptional potency against both gram-positive as well as gram-negative bacteria. CONCLUSION: Less research has been done on using bioinspired DH-CdSNP to deliver anticancer medications. The amalgamation of plant extract and the DH-CdSNP could cause a paradigm shift in the cancer therapy approach. The findings revealed that the biosynthesized DH-CdSNP limited the growth of human breast and ovarian cancer cells. This property can be further investigated against a variety of additional cell lines to determine whether this property makes the DH-CdSNP a promising treatment alternative. The results obtained from these nanoformulations exhibit faster efficacy compared to traditional medications.

Graphene-silymarin-loaded chitosan/gelatin/hyaluronic acid hybrid constructs for advanced full-thickness burn wound management.

Burn wounds (BWs) with extensive blood loss, along with bacterial infections and poor healing, may become detrimental and pose significant rehabilitation obstacles in medical facilities. Therefore, the freeze-drying method synthesized novel hemocompatible chitosan, gelatin, and hyaluronic acid infused with graphene oxide-silymarin (CGH-SGO) hybrid constructs for application as a BW patch. Most significantly, synthesized hybrid constructs exhibited an interconnected-porous framework with precise pore sizes (≈118.52 µm) conducive to biological functions. Furthermore, the FTIR and XRD analyses document the constructs' physiochemical interactions. Similarly, enhanced swelling ratios, adequate WVTR (736 ± 78 g m-2 hr-1), and bio-degradation rates were seen during the physiological examination of constructs. Following the in vitro investigations, SMN-GO added to constructs improved their anti-bacterial (against E.coli and S. aureus), anti-oxidant, hemocompatible, and bio-compatible characteristics in conjunction with prolonged drug release. Furthermore, in vivo, implanting constructs on wounds exhibited significant acceleration in full-thickness burn wound (FT-BW) healing on the 14th day (CGH-SGO: 95 ± 2.1 %) in contrast with the control (Gauze: 71 ± 4.2 %). Additionally, contrary to gauze, the in vivo rat tail excision model administered with constructs assured immediate blood clotting. Therefore, CGH-SGO constructs with an improved porous framework, anti-bacterial activity, hemocompatibility, and biocompatibility could represent an attractive option for healing FT-BWs.

Folate-Mediated Targeting and Controlled Release: PLGA-Encapsulated Mesoporous Silica Nanoparticles Delivering Capecitabine to Pancreatic Tumor.

The discovery of specifically tailored therapeutic delivery systems has sparked the interest of pharmaceutical researchers considering improved therapeutic effectiveness and fewer adverse effects. The current study concentrates on the design and characterization of PLGA (polylactic-co-glycolic acid) capped mesoporous silica nanoparticles (MSN)-based systems for drug delivery for pH-sensitive controlled drug release in order to achieve a targeted drug release inside the acidic tumor microenvironment. The physicochemical properties of the nanoformulations were analyzed using TEM, zeta potential, AFM, TGA, FTIR, and BET analyses in addition to DLS size. The final formed PLGA-FoA-MSN-CAP and pure MSN had sizes within the therapeutic ranges of 164.5 ± 1.8 and 110.7 ± 2.2, respectively. Morphological characterization (TEM and AFM) and elemental analysis (FTIR and XPS) confirmed the proper capping and tagging of PLGA and folic acid (FoA). The PLGA-coated FoA-MSN exhibited a pH-dependent controlled release of the CAP (capecitabine) drug, showing efficient release at pH 6.8. Furthermore, the in vitro MTT test on PANC1 and MIAPaCa-2 resulted in an IC50 value of 146.37 µg/ml and 105.90 µg/ml, respectively. Mitochondrial-mediated apoptosis was confirmed from the caspase-3 and annexin V/PI flow cytometry assay, which displayed a cell cycle arrest at the G1 phase. Overall, the results predicted that the designed nanoformulation is a potential therapeutic agent in treating pancreatic cancer.

DDX3X dynamics, glioblastoma's genetic landscape, therapeutic advances, and autophagic interplay.

Glioblastoma is one of the most aggressive and deadly forms of cancer, posing significant challenges for the medical community. This review focuses on key aspects of Glioblastoma, including its genetic differences between primary and secondary types. Temozolomide is a major first-line treatment for Glioblastoma, and this article explores its development, how it works, and the issue of resistance that limits its effectiveness, prompting the need for new treatment strategies. Gene expression profiling has greatly advanced cancer research by revealing the molecular mechanisms of tumors, which is essential for creating targeted therapies for Glioblastoma. One important protein in this context is DDX3X, which plays various roles in cancer, sometimes promoting it or otherwise suppressing it. Additionally, autophagy, a process that maintains cellular balance, has complex implications in cancer treatment. Understanding autophagy helps to identify resistance mechanisms and potential treatments, with Chloroquine showing promise in treating Glioblastoma. This review covers the interplay between Glioblastoma, DDX3X, and autophagy, highlighting the challenges and potential strategies in treating this severe disease.

In vitro and in vivo assessment of curcumin-quercetin loaded multi-layered 3D-nanofibroporous matrix prepared by solution blow-spinning for full-thickness burn wound healing.

Burn wounds (BWs) cause impairment of native skin tissue and may cause significant microbial infections that demand immediate care. Curcumin (Cur) and quercetin (Que) exhibit antimicrobial, hemocompatibility, ROS-scavenging, and anti-inflammatory properties. However, its instability, water insolubility, and low biological fluid absorption render it challenging to sustain local Cur and Que doses at the wound site. Therefore, to combat these limitations, we employed blow-spinning and freeze-drying to develop a multi-layered, Cur/Que-loaded gelatin/chitosan/PCL (GCP-Q/C) nanofibroporous (NFP) matrix. Morphological analysis of the NFP-matrix using SEM revealed a well-formed multi-layered structure. The FTIR and XRD plots demonstrated dual-bioactive incorporation and scaffold polymer interaction. Additionally, the GCP-Q/C matrix displayed high porosity (82.7 ± 2.07 %), adequate pore size (∼121 µm), enhanced water-uptake ability (∼675 % within 24 h), and satisfactory biodegradation. The scaffolds with bioactives had a long-term release, increased antioxidant activity, and were more effective against gram-positive (S. aureus) and gram-negative (E. coli) bacteria than the unloaded scaffolds. The in vitro findings of GCP-Q/C scaffolds showed promoted L929 cell growth and hemocompatibility. Additionally, an in vivo full-thickness BW investigation found that an implanted GCP-Q/C matrix stimulates rapid recuperation and tissue regeneration. In accordance with the findings, the Gel/Ch/PCL-Que/Cur NFP-matrix could represent an effective wound-healing dressing for BWs.

Apoptosis induction in leukemic cells by L-asparaginase preparation from Bacillus indicus: bench-scale production, purification and therapeutic application.

With the emergence of multiple side effects on the usage of commercial L-asparaginase formulations, keen interest is provoked to investigate new sources of L-asparaginases that possess antileukemic properties with minimal side effects. The present study reports the cost-effective bench-scale production, homogeneity purification and apoptosis induction potential of a new L-asparaginase preparation from Bacillus indicus against human leukemia cells. The enzyme is highly specific toward the natural substrate L-asparagine. The study initiated with the enzyme production using cost-effective substrates in which a 3.28-fold enhancement of enzyme activity was achieved in comparison with an unoptimized medium using the central composite experimental design approach. The scale-up of the process in a 3.7-L batch bioreactor resulted in 16.42 ± 0.17 IU/mL of L-asparaginase activity in 24 h. The crude extracellular enzyme was purified to homogeneity using anion exchange chromatography followed by gel filtration chromatography. A single band of approximately 35 kDa molecular weight was obtained on SDS-PAGE, while native PAGE analysis confirmed it to be a tetramer of four identical subunits. The circular dichroism spectroscopic study revealed the α + ß mixed type of secondary structure with 38.7% α-helices and 27.4% ß pleated sheets. The antitumor toxicity exhibited on the MOLT-4 leukemia cells by the new L-asparaginase was revealed using the MTT assay and acridine orange/propidium iodide dual staining for live/dead cells. The flow cytometry analysis established the potential of the purified L-asparaginase to induce the apoptotic cell death mechanism in MOLT-4 leukemia cells. Conclusively, the L-asparaginase of Bacillus indicus is a highly promising candidate that can be introduced as a new enzyme therapeutic against various leukemia disorders.

Molecular modelling study to discover novel JAK2 signaling pathway inhibitor.

The JAK2/STAT signaling cascades facilitates receptor signals which is responsible for cell growth, survival and homeostasis. Ligand binding to JAKs causes phosphorylation other proteins known as STATs, which translocate to the nucleus and regulate transcription of several important proteins. Growth hormone, prolactin and γ-interferon known agonists of JAK STAT receptors, signal to the nucleus by a more direct manner than the receptor tyrosine kinases. Mutations in JAKs may be responsible for immunodeficiency and myeloproliferative disorders because of its important role in cytokine signaling and making the pathway a therapeutic target for various disease. The present study screened Zinc database to find novel JAK2 inhibitors using virtual high throughput screening techniques. Selection of compound for further study was on the basis of docking score, free energy and binding pattern of the compound. Molecular simulation and MM/GBSA free energy was evaluated for the binding interactions and the stability of docked conformations. Several parameters which determine protein ligand interaction like RMSD, RMSF, Rg and binding pattern were observed. Hydrogen bonds (Glu 930, 932 and Asp 994) after 150 ns simulation were observed between identified compound INC000096136346 and it was similar to known inhibitor ruxolitinib. MM/GBSA free energy was comparable to known inhibitor ruxolitinib. ZINC000096136346 qualify Lipinski's rule of five, rule of three, WDI like rule and there is one violation in lead like rule.Communicated by Ramaswamy H. Sarma.

Investigation of molecular mechanism leading to gefitinib and osimertinib resistance against EGFR tyrosine kinase: molecular dynamics and binding free energy calculation.

Tyrosine kinase (TK) is an important protein responsible for phosphorylation of variety of proteins that helps in signal transduction process in transferring signal to regulate various physiological and biochemical processes. Drugs inhibiting signal transduction pathways can be a very rational approach to inhibit cellular physiological and biochemical process. Tyrosine kinase inhibitors are a wide family of drugs that have been used successfully in cancer chemotherapy. Certain mutations around the catalytic cleft may cause conformational changes at binding site and leads to decrease in inhibitor sensitivity to TK mutants. EGFRT790M mutation is the first recognized acquired resistance after tyrosine kinase inhibitor therapy that leads to resistant to first generation TKI in about 50% of non-small cell lung carcinoma patients. Third generation EGFR-TKIs bind irreversibly to the C797, which is present in the ATP-binding pocket. The present work provides a molecular mechanism for understanding the Gefitinib and Osimertinib sensitivities with the EGFRWILD, EGFRL858R, EGFRT790M, EGFRT790M+C797S mutants using molecular modelling techniques. Changes in response against Gefitinib and Osimertinib were observed with the change of amino acids at the tyrosine kinase domain of EGFRWILD and its mutants (EGFRL858R, EGFRT790M, EGFRT790M+C797S). RMSD, RMSF and binding energies calculation well correlates with the change in clinical observation.Communicated by Ramaswamy H. Sarma.