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.

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 ±â€¯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 burn wounds exhibited significant acceleration in BW-healing on the 14th day (CGH-SGO: 96 ±â€¯2.1 %) in contrast with the control (Gauze: 71 ±â€¯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.

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.

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.

A comprehensive review on ayurvedic herb Leptadenia reticulata (Jeevanti): a phytochemistry and pharmacological perspective.

Leptadenia reticulata is a vital Ayurvedic medicinal herb, commonly known as Jivanti or Jiv, and contains revitalising, rejuvenating, and lactogenic activities. It has been used in traditional medicine for treating respiratory disorders, wounds, inflammation, cough, dehydration, tuberculosis, colitis, chickenpox, dysentery, eye diseases, night blindness, fever, and snake bites. It is a perennial herb of Indian origin belonging to the Asclepiadaceae family and has been utilised for its therapeutic properties since ancient times. It is a key ingredient in several marketed herbal drugs, including chyawanprash, speman, and leptaden. Several potent compounds, including ß-sitosterol, γ-sitosterol, phytol, α-amyrin, ß-amyrin, apigenin, reticulin, deniculatin, leptaculatin, diosmetin, and rutin are present in this herb and attributed various pharmacological activities, including antidiabetic, antimicrobial, antioxidant, anti-abortifacient, anticancer, antipyretic, analgesic, anti-inflammatory, and antiulcer properties. This review provides an in-depth analysis of the distribution, ethnobotanical use, botanical description, phytocompounds, and pharmacological activities of Leptadenia reticulata.

Leptadenia reticulata is a vital Ayurvedic herb containing revitalising, rejuvenating, and lactogenic activities.It is present in polyherbal formulations chyawanprash, speman, and leptaden.Its secondary metabolites have anticancer, anticholesterol, antidiabetic, antiabortifacient, and anti-inflammatory potential.This review shows the distribution, morphology, and therapeutic potential of Leptadenia reticulata.A summary of the phytochemicals present in Leptadenia reticulata will also be provided.

Exploring plant-based alpha-glucosidase inhibitors: promising contenders for combatting type-2 diabetes.

Objective: This systematic review aimed to provide comprehensive details on the α-G inhibitory potential of various bioactive compounds derived from natural sources.Methods: A comprehensive literature search was conducted using various databases and search engines, including Science Direct, Google Scholar, SciFinder, Web of Science, and PubMed until May, 2023.Results and conclusions: The enzyme alpha-glucosidase (α-G) is found in the brush border epithelium of the small intestine and consists of duplicated glycoside hydrolase (GH31) domain. It involves the conversion of disaccharides and oligosaccharides into monosaccharides by acting on alpha (1 → 4) and (1 → 6) linked glucose residue. Once absorbed, glucose enters the bloodstream and elevates postprandial glucose, which is associated with the development of type 2 Diabetes (T2D). Epidemic obesity, cardiovascular disease, and nephropathy are linked to T2D. Traditional medicinal plants with α-G inhibitory potential are commonly used to treat T2D due to the adverse effects of currently used α-G inhibitors miglitol, acarbose, and voglibose. Various bioactive compounds derived from natural sources, including lupenone, Wilforlide A, Baicalein, Betulinic acid, Ursolic acid, Oleanolic acid, Katononic acid, Carnosol, Hypericin, Astilbin, lupeol, betulonic acid, Fagomine, Lactucaxanthin, Erythritol, GP90-1B, Procyanidins, Galangin, and vomifoliol retain α-G inhibitory potential for regulating hyperglycaemia.

Exploring the potential of Diplosphaera mucosa VSPA for the treatment of petroleum effluent with simultaneous lipid production.

The discovery of unexplored, robust microalgal strains will assist in treating highly polluted industrial effluent, including petroleum effluent. In the current analysis, a newly isolated microalgal strain, Diplosphaera mucosa VSPA, was used to treat petroleum effluent in a lab-scale raceway bioreactor. Its treatment efficiency was compared with a well-known species, Chlorella pyrenoidosa. The D. mucosa VSPA strain proliferated in petroleum effluent at a high growth rate, with final biomass, and lipid concentrations reaching 6.93 g/L and 2.72 g/L, respectively. Treatment efficiency was calculated based on the final removal efficiency of ammonium nitrogen, phosphate phosphorus, and chemical oxygen demand, which was more than 90%. Control experiments suggested that the maximum removal of pollutants from petroleum effluent was due to microalgae growth. Some growth models, including the Gompertz, Logistic, Stannard, Richard, and Schnute, were used to simulate the experimental data, verifying the results. Good fitting of all models was obtained, with the R2 value reaching more than 0.90. The development of a suitable model can help in decreasing the efforts required for the scale-up of the process.

Phytochemical profiling and evaluation of the antidiabetic potential of Ichnocarpus frutescens (Krishna Sariva): kinetic study, molecular modelling, and free energy approach.

This research explored novel antidiabetic drugs from natural sources using the Ayurvedic Rasayana herb Ichnocarpus frutescens through invitro enzyme assay, kinetics study, and computational approaches. Invitro enzyme inhibition assay demonstrated the promising inhibitory activity of root extract against alpha-amylase (α-A) and alpha-glucosidase (α-G) enzyme with IC50 value 7.34 ± 0.22 mg/ml and 4.40 ± 0.25 mg/ml respectively. Enzyme kinetic study revealed the competitive inhibition of both proteins by Ichnocarpus frutescens extract. High-Resolution Liquid Chromatography Mass Spectrometer and Docking study revealed the better binding energy of phytoconstituents 23-Acetoxysoladulcidine, Atrovirinone, Bismurrayaquinone A, Lamprolobine, Zygadenine, and Gambiriin A3 than standard drug acarbose. Molecular modelling showed stable protein-ligands binding interaction during the 100 ns simulation. It revealed comparable Root Mean Square Deviation, Radius of Gyration, and Solvent Accessible Surface Area of these compounds with acarbose. The active site residues of both proteins remained stable and showed significantly less Root Mean Square Fluctuation. Molecular Mechanics with Generalised Bonn Surface Area analysis has illustrated the similar inhibitory activity of Zygadenine for α-A, 23-Acetoxysoladulcidine, and Gambiriin A3 for α-G protein, compared to the FDA-approved drug acarbose. Thus, the study suggested that the root of Ichnocarpus frutescens can be used as α-A and α-G inhibitors and be considered a compelling lead for the medication of type 2 diabetes.Communicated by Ramaswamy H. Sarma.

Atrovirinone, 23-Acetoxysoladulcidine, Bismurrayaquinone A, Gambiriin A3, Zygadenine, and Lamprolobine are the major phytoconstituents identified from Ichnocarpus frutescens.The computational study suggested that this compound possesses α-A and α-G inhibition potential.Invitro study confirmed the competitive mode of inhibition of both enzymes by root extract of Ichnocarpus frutescens.The antidiabetic potential of Ichnocarpus frutescens was investigated for the first time by kinetic study and insilico approach.

Treatment of carpet and textile industry effluents using Diplosphaera mucosa VSPA: A multiple input optimisation study using artificial neural network-genetic algorithms.

The wastewater treatment efficiency of Diplosphaera mucosa VSPA was enhanced by optimising five input parameters and increasing the biomass yield. pH, temperature, light intensity, wastewater percentage (pollutant concentration), and N/P ratio were optimised, and their effects were studied. Two competitive techniques, response surface methodology (RSM) and artificial neural network (ANN), were applied for constructing predictive models using experimental data generated according to central composite design. Both MATLAB and Python were used for constructing ANN models. ANN models predicted the experimental data with high accuracy and less error than RSM models. Generated models were hybridised with a genetic algorithm (GA) to determine the optimised values of input parameters leading to high biomass productivity. ANN-GA hybridisation approach performed in Python presented optimisation results with less error (0.45%), which were 7.8 pH, 28.8 °C temperature, 105.20 µmol m-2 s-1 light intensity, 93.10 wastewater % (COD) and 23.5 N/P ratio.

Downstream process intensification for biotechnologically generated hyaluronic acid: Purification and characterization.

Hyaluronic acid (HA), an anionic, non-sulfated glycosaminoglycan, has several clinical applications. This study examines several downstream methods for purifying HA with maximum recovery and purity. Following the fermentation of Streptococcus zooepidemicus MTCC 3523 to produce HA, the broth was thoroughly purified to separate cell debris and insoluble impurities using a filtration procedure and a variety of adsorbents for soluble impurities. Nucleic acids, proteins with high molecular weight, were successfully removed from the broth using activated carbons and XAD-7 resins. In contrast, insoluble and low molecular weight impurities were removed using diafiltration, with HA recovery of 79.16% and purity close to 90%. Different analytical and characterization procedures such as Fourier transform-infrared spectroscopy, X-ray diffraction, nuclear magnetic resonance, and scanning electron microscopy validated the presence, purity, and structure of HA. Microbial HA showed activity in tests for 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) radical-scavenging (4.87 ± 0.45 kmol TE/g), total antioxidant capacity (13.32 ± 0.52%), hydroxyl radical-scavenging (32.03 ± 0.12%), and reducing power (24.85 ± 0.45%). The outcomes showed that the precipitation, adsorption, and diafiltration processes are suitable for extracting HA from a fermented broth under the chosen operating conditions. The HA produced was of pharmaceutical grade for non-injectable applications.

Glypican1: A potential cancer biomarker for nanotargeted therapy.

Glypicans (GPCs) are generally involved in cellular signaling, growth and proliferation. Previous studies reported their roles in cancer proliferation. GPC1 is a co-receptor for a variety of growth-related ligands, thereby stimulating the tumor microenvironment by promoting angiogenesis and epithelial-mesenchymal transition (EMT). This work reviews GPC1-biomarker-assisted drug discovery by the application of nanostructured materials, creating nanotheragnostics for targeted delivery and application in liquid biopsies. The review includes details of GPC1 as a potential biomarker in cancer progression as well as a potential candidate for nano-mediated drug discovery.

HR-LCMS and evaluation of anti-diabetic activity of Hemidesmus indicus (anantmool): Kinetic study, and molecular modelling approach.

This study delved into the exploration of novel antidiabetic medications acquired from natural resources, utilizing the Ayurvedic Rasayana herb Hemidesmus indicus through cutting-edge chemoprofiling and molecular modelling techniques. The methanolic extract of Hemidesmus indicus root exhibited the highest extractive yield (24.70 ± 0.08 %) and contained substantial levels of total phenolic and flavonoid content as 154.15 ± 1.24 mg Gallic Acid Equivalent/g extract and 70.61 ± 0.35 Quercetin Equivalent/g extract respectively. Invitro study revealed the potent inhibitory potential of methanolic extract of the herb against essential carbohydrate hydrolytic enzymes α-amylase (IC50 = 4.19 ± 0.04 mg/ml) and α-glucosidase (IC50 = 5.78 ± 0.10 mg/ml). Further, the enzyme kinetic study demonstrated the competitive mode of inhibition of both enzymes. HR-LCMS analysis identified the major phytoconstituents present in the extracts, including Solanocapsine, Cyclovirobuxine C, Lucidine B, Zygadenine, Aspidospermidine, silychristin, 3beta-3-Hydroxy-18-lupen-21-one, Manglupenone, and 19-Noretiocholanolone. Molecular docking, molecular dynamic simulation, and MM/GBSA analysis have proved stable, rigid, compact, and folded form of complexes during the entire 100 ns simulation, illustrating Zygadenine, Solanocapsine, and Cyclovirobuxine C as the superior inhibitors of α-A protein, while Zygadenine, Plumieride, and Phlegmarine exhibited greater inhibitory behaviour towards α-G protein than the FDA-approved drug acarbose. Collectively, our findings indicate that the Hemidesmus indicus could be a promising source of α-A and α-G inhibitors, potentially serving as a lead in order to develop medications for type-2 diabetes.

Discovery of the allosteric inhibitor from actinomyces metabolites to target EGFRCSTMLR mutant protein: molecular modeling and free energy approach.

EGFR (epidermal growth factor receptor), a surface protein on the cell, belongs to the tyrosine kinase family, responsible for cell growth and proliferation. Overexpression or mutation in the EGFR gene leads to various types of cancer, i.e., non-small cell lung cancer, breast, and pancreatic cancer. Bioactive molecules identified in this genre were also an essential source of encouragement for researchers who accomplished the design and synthesis of novel compounds with anticancer properties. World Health Organization (WHO) report states that antibiotic resistance is one of the most severe risks to global well-being, food safety, and development. The world needs to take steps to lessen this danger, such as developing new antibiotics and regulating their use. In this study, 6524 compounds derived from Streptomyces sp. were subjected to drug-likeness filters, molecular docking, and molecular dynamic simulation for 1000 ns to find new triple mutant EGFRCSTMLR (EGFR-L858R/T790M/C797S) inhibitors. Docking outcomes revealed that five compounds showed better binding affinity (- 9.074 to - 9.3 kcal/mol) than both reference drug CH7233163 (- 6.11 kcal/mol) and co-crystallized ligand Osimertinib (- 8.07 kcal/mol). Further, molecular dynamic simulation confirmed that ligand C_42 exhibited the best interaction at the active site of EGFR protein and comprised a better average radius of gyration (3.87 Å) and average SASA (Solvent Accessible Surface Area) (82.91 Å2) value than co-crystallized ligand (4.49 Å, 222.38 Å2). Additionally, its average RMSD (Root Mean Square Deviation) (3.25 Å) and RMSF (Root Mean Square Fluctuation) (1.54 Å) values were highly similar to co-crystallized ligand (3.07 Å, 1.54 Å). Compared to the reference ligand, it also demonstrated conserved H-bond interactions with the residues MET_793 and GLN_791 with strong interaction probability. In conclusion, we have found a potential drug with no violation of the rule of three, Lipinski's rule of five, and 26 other vital parameters having great potential in medicinal and pharmaceutical industries applications and can overcome synthetic drug issues.

Design and modelling of photobioreactor for the treatment of carpet and textile effluent using Diplosphaera mucosa VSPA.

The current study investigated the potential of one less explored microalgae species, Diplosphaera mucosa VSPA, for treating carpet and textile effluent in a conventionally designed 10 L bubble column photobioreactor. To the best of our knowledge, this is the first study to evaluate COD (chemical oxygen demand) removal efficiency by microalgae in carpet effluent. To evaluate D. mucosa VSPA's potential, its growth and bioremediation efficacy were compared to those of a well-known strain, Chlorella pyrenoidosa. D. mucosa VSPA outperformed C. pyrenoidosa in both effluents, with the highest biomass concentration reaching 4.26 and 3.98 g/L in carpet and textile effluent, respectively. D. mucosa VSPA also remediated 94.0% of ammonium nitrogen, 71.6% of phosphate phosphorus, and 91.9% of chemical oxygen demand in carpet effluent, approximately 10% greater than that of C. pyrenoidosa. Both species also removed more than 65% of colour from both effluents, meeting the standard set by governing bodies. Microalgae growth and substrate removal patterns in the photobioreactor were simulated using photobiotreatment and the Gompertz model. Simulation results revealed that photobiotreatment was the better-fit model, concluded based on the coefficient of regression value and the second-order Akaike information criterion test. Modelling studies can assist in increasing the performance and scale-up of the photobioreactor. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03655-3.

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.

Tapping on the Potential of Hyaluronic Acid: from Production to Application.

The manufacture, purification, and applications of hyaluronic acid (HA) are discussed in this article. Concerning the growing need for affordable, high-quality HA, it is essential to consider diverse production techniques using renewable resources that pose little risk of cross-contamination. Many microorganisms can now be used to produce HA without limiting the availability of raw materials and in an environmentally friendly manner. The production of HA has been associated with Streptococci A and C, explicitly S. zooepidemicus and S. equi. Different fermentation techniques, including the continuous, batch, fed-batch, and repeated batch culture, have been explored to increase the formation of HA, particularly from S. zooepidemicus. The topic of current interest also involves a complex broth rich in metabolites and residual substrates, intensifying downstream processes to achieve high recovery rates and purity. Although there are already established methods for commercial HA production, the anticipated growth in trade and the diversification of application opportunities necessitate the development of new procedures to produce HA with escalated productivity, specified molecular weights, and purity. In this report, we have enacted the advancement of HA technical research by analyzing bacterial biomanufacturing elements, upstream and downstream methodologies, and commercial-scale HA scenarios.

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.

In Vitro and In Vivo Scalp Retention and Penetration of 99mTc-Minoxidil Solution.

The present study assessed the effect of retention on ex vivo skin and in vivo scalp penetration of radiolabeled minoxidil formulations (5% w/v). Minoxidil was radiolabeled with technetium (99mTc) with an efficiency of 99.1% using 0.2% stannous chloride as reducing agent at pH 6 and incubation temperature of 40 °C. Three different 99mTc-minoxidil formulations were prepared using aqueous ethanolic solution as vehicle. Formulation A contains 99mTc-minoxidil dissolved in vehicle, formulation B contains 10% propylene glycol (PG) and formulation C contains 10% hydroxypropyl cellulose (HPC), in addition. Results showed that addition of HPC resulted in enhanced viscosity (400 mPa.s) and significantly higher ex vivo retention (p < 0.05) and permeation (0.75±0.12%, 8 h). PG does not improve the permeation and the results (0.44±0.05%, 8 h) were not significantly different from vehicle alone (0.40±0.05%, 8 h). The results of the in vivo human scalp studies corroborated with the ex vivo results and addition of hydroxypropyl cellulose (HPH) showed significantly higher (p < 0.05) scalp retention. Post 8 h application, scalp penetration in group treated with formulation C was nearly 2.8-fold and 2.2-fold higher than those treated with formulation A and B, respectively. Further, absence of minoxidil in systemic circulation during study duration indicates safety. In conclusion, our results showed that increasing contact time of minoxidil with scalp by modifying viscosity results in reduced frequency of application and improved efficacy.