Cell Line L6 Research Articles

The impact of bisphenol AF on skeletal muscle function and differentiation in vitro

Various environmental chemicals have been identified as contributors to metabolic diseases. Bisphenol AF (BPAF), a substitute for bisphenol A, has been associated with changes in glucose metabolism and incidence of type 2 diabetes mellitus in humans. However, its mode of action remains unclear. Considering that skeletal muscle is the primary tissue for glucose utilization and the development of insulin resistance, yet largely neglected in toxicological assessments, we investigated the impact of BPAF on skeletal muscle function and differentiation.We examined the effects of BPAF (0.01–10 μM) on glucose uptake, response to insulin, production of reactive oxygen species (ROS), intracellular calcium, and myocyte differentiation, during hyperglycemia, insulin stimulation, and muscle contraction. We used the rat myoblast cell line L6 differentiated into myotubes, and murine primary isolated muscle fibers.In myotubes and contracting adult fibers, BPAF increased mitochondrial ROS. Basal glucose uptake was increased in myotubes while cells' ability to respond to insulin was decreased. Additionally, in developing myotubes, differentiation markers were downregulated with BPAF, along with impaired formation of tube structures. These effects were primarily observed at 10 μM concentration, which is markedly higher than reported human exposure concentrations.The results provide an insight into potential hazards associated with BPAF in terms of metabolic disruption in skeletal muscle. The developed in vitro methods show promise for future usage in assessments of new chemicals and their mixtures.

DESIGN, SYNTHESIS, AND BIO PROFILING OF 2, 3-DIHYDROQUINAZOLIN-4(1H)-ONE DERIVATIVE AS TYPE II DIABETES AGENTS: A COMPREHENSIVE IN SILICO, IN VITRO, AND IN VIVO STUDY

Objective: Diabetes mellitus is a significant global health challenge, with Type 2 Diabetes Mellitus (T2DM) being a leading cause of mortality worldwide, demanding the need for effective interventions by developing innovative therapeutic strategies or novel antidiabetic agents. This study explores in silico, in vitro, and in vivo approaches to identify the most potent 2,3-Dihydroquinazolin-4(1H)-One derivative molecule with antidiabetic activity. Methods: Eleven new derivatives were designed, studied in silico to identify the most promising compounds, synthesized, studied spectrally to describe them, and evaluated for both in vitro and in vivo investigations. Alpha amylase and alpha-glucosidase inhibitory activities were investigated in vitro. The endogenous suppression of glucose synthesis in Hepatoblastoma cell line 2(HepG2) cells and the in vitro glucose absorption assay on cultivated L6 cell lines were conducted. To assess the ability of the newly synthesized compounds to prevent diabetes, in vivo investigations were conducted on Streptozotocin (STZ)-induced diabetic rats and the effects on various biochemical parameters were identified.Results: Leveraging computational methods, the QZ9 molecule was identified with stable interactions with key biomolecules associated with T2DM. Subsequent in vitro assays confirmed the inhibitory effects of QZ2, QZ8, and QZ9 on alpha-amylase and alpha-glucosidase activities, suggesting their potential as enzyme inhibitors. Additionally, QZ8 and QZ9 demonstrated enhanced glucose uptake and production inhibition in HepG2 cells, indicating their role in improving glucose homeostasis. In vitro, the top-ranked molecules QZ2, QZ8, and QZ9 were analyzed to validate the in silico findings and assess their potential as therapeutic agents for T2DM. The inhibition of α-amylase activity by QZ2, QZ8, and QZ9 was dose-dependent, with maximum inhibition observed at 1000 µg/ml: 57.33% for QZ2, 52.21% for QZ8, and 87.16% for QZ9. Similarly, α-glucosidase inhibition at 1000 µg/ml was 59.96% for QZ2, 53.50% for QZ8, and 81.51% for QZ9. Both QZ8 and QZ9 significantly increased glucose uptake and inhibited glucose production in HepG2 cells, with maximum glucose production inhibition at 100 µg/ml: 62.22% for QZ8 and 62.35% for QZ9. These findings suggest that QZ8 and QZ9 contribute to glucose homeostasis. QZ9 demonstrated superior enzyme inhibition compared to QZ2 and QZ8, with α-amylase and α-glucosidase inhibition up to 87.16% and 81.51%, respectively, at 1000 µg/ml. In vivo investigations in Diabetic rat models further confirmed the efficacy of these compounds by showing significant reductions in blood glucose levels. These results suggests the potentiality of QZ9 as a promising novel Antidiabetic agent.Conclusion: Combining computational predictions with experimental validations, this integrated approach highlights the promise of 2,3-Dihydroquinazolin-4(1H)-One derivative QZ9 as a novel antidiabetic agent, warranting further investigation for clinical translation.

Exploring the therapeutic potential of ruthenium(II)-bis(benzimidazol-2-yl)pyridine complex on MDA-MB-231, HCT-116 and normal L6 cell lines

The in vitro anticancer efficacy and cytotoxicity of the [Ru(bbp)2]2+ complex (bbp = 2,6-bis(benzimidazol-2-yl) pyridine) on human breast cancerous MDA-MB-231, colorectal HCT-116, and normal cell lines were investigated by direct microscopic and MTT assay methods. The synthesised [Ru(bbp)2]2+ complex was characterised by UV–visible, FTIR, 1H NMR, and MALDI-TOF-MS spectroscopic techniques. The complex showed a metal-to-ligand charge transfer (MLCT) absorption peak at 481 nm. The FTIR spectrum of the complex showed absorption bands in the range of 3458–573 cm−1. The 1H NMR chemical shift values resembled those of the bbp ligand. The molecular ion peak (M+) of the [Ru(bbp)2]2+ complex was obtained at m/z 1013.48. The IC50 values of the complex on MDA-MB-231, HCT-116 and normal L6 cell lines were calculated from the percentage cellular viability and are found to be 28.71, 9.06 and 153.82 µM respectively. The morphological changes in the cell lines at various concentrations of the complex were attributed to the generation of reactive oxygen species (ROS). The obtained result revealed that the anticancer activity of the synthesised complex depends on the concentration of the complex. The complex reduced the proportion of viable cells in both malignant cell lines, causing apoptosis. The results suggeseted that the synthesised complex showed good anticancer activity on the MDA-MB-231 and HCT-116 cell lines with low cytotoxicity. Thus, the present investigation paved the way for the development of novel anticancer drugs.

Development of probiotic functional food and evaluation of its functional properties

Probiotics are beneficial microorganisms that enhance human health by improving intestinal microbial balance. It helps in improving digestive health, enhances immunity, and improves psychiatric health, cardiac health and weight management. The present study explores the potential of using probiotic microorganisms from lotus (Nelumbo nucifera) and colocasia (Colocasia esculenta) leaves as sources for starter culture bacteria for curd preparation. Four types of curd were developed: cow's milk with colocasia leaf starter culture (MC), cow's milk with lotus leaf starter culture (ML), red sorghum and cow's milk mixture with colocasia leaf starter (MRC), and red sorghum and cow's milk mixture with lotus leaf starter (MRL). Metagenomic analysis showed that MC curd contained Acetobacter sp., (51%), Lactococcus sp., (26%), and Leuconostoc sp., (18%), while ML curd had Lactococcus sp., (67%), Leuconostoc sp. (20%), and Lactobacillus sp., (10%). Physicochemical study of MRC and MRL showed 2.94 ± 0.84 % and 2.84 ± 0.14 % of carbohydrate content, respectively. The analysis of MRC and MRL showed a significant increase in the color parameter “a*” probably due to the presence of anthocyanin in red sorghum milk. The MRC and MRL showed higher antioxidant and anti-inflammatory activities. The MRL curd showed the highest glucose uptake and cytotoxicity in the L6 cell line and also demonstrated potent α-amylase inhibition (IC50 = 2.383 ± 0.286 μg/ml) compared to MRC (IC50 = 13.02 ± 0.112 μg/ml). Thus, in-vitro anti-diabetic activity of red sorghum-fortified curds was found superior to MC and ML curds. This study suggests that red sorghum fortification enhances curd quality and provides notable health benefits.

Efficacy of Quercetin and Kaempferol from Methanolic Extracts of Adiantum lunulatum Burm. F. and Polystichum acrostichoides as Antidiabetic Activity

In present investigation, the aerial parts of two fern species viz. Polystichum acrostichoides (PA) and Adiantum lunulatum (AL) were used to evaluate the potent antidiabetic activity. Methanol solvent was used for extraction by soxhlation method followed by LC-MS analysis to identify the presence of compounds in both the extracts. Further, isolation of the major compounds was carried out and characterized by IR, mass and NMR techniques. Thereafter, in vitro α-amylase inhibitory activity for both the extracts was determined by comparing with acarbose as standard. Thereafter, α-glucosidase inhibition activity followed by glucose uptake was studied using L6 cell line study. Practical yield for both the fern extracts was calculated and observed A. lunulatum extract showed more yield (7 g) than P. acrostichoides (3 g). Based on the analytical data, isolated compound was identified as quercetin and kaempferol from the A. lunulatum and P. acrostichoides extracts, respectively. A. lunulatum extract even showed better dose dependent α-amylase inhibition and α-glucosidase inhibition activity than P. acrostichoides furthermore, L6 cell line study showed increased glucose uptake by A. lunulatum extract than P. acrostichoides extract when compared with standard insulin. Based on the findings, it appears that the both A. lunulatum and P. acrostichoides have the potential to be used as antidiabetic drugs due to presence of major isolated flavonoids i.e. quercetin and kaempferol as among the bioactive compounds. Antidiabetic efficacy was dosage dependent and A. lunulatum extract was more effective in inhibiting glucose uptake than P. acrostichoides extract.

Synthesis, characterization and biological evaluation of novel ruthenium(II)-polypyridine mixed ligand complexes bearing 1,2,4- triazole-thiol moiety

The novel mixed ligand complexes [Ru(bpy)2(pytrzSH)2](PF6)2 (complex A) and [Ru(phen)2(pytrzSH)2](PF6)2 (complex B) (bpy = 2,2′-bipyridine, phen = 1,10-phenanthroline and pytrzSH = 5-(3-pyridyl)-4H-1,2,4-triazole-3-thiol) has been synthesized and characterized by elemental analysis, UV, FTIR, 1H NMR, 13C NMR and MALDI-TOF mass spectral techniques. The spectral data confirms the formation of octahedral complexes. The lipophilic nature of the complex is determined from the partition coefficient (log P) values and it is found to be 1.20 ± 0.004 for complex A and 1.86 ± 0.004 for complex B. The synthesised complexes show slight antimicrobial activities on S. aureus, P. aeruginosa, E. coli, A. niger and C. albicans. The α-glucosidase inhibitory activity of the synthesised complexes A and B against the positive control acarbose display IC50 value at 490.32 and 223.01 µg/mL. In vitro antiproliferative and cytotoxic effects of complexes A and B are analysed on SK-MEL-28 and non-tumoral L6 cell lines by microscopic and MTT assay methods. The IC50 for complexes A and B on SK-MEL-28 cancerous cell line are found to be 27.444 and 40.721 µg/mL and for non-tumoral L6 cells are 25.869 and 38.425 µg/mL respectively. The apoptotic study suggests that the synthesized complexes show late apoptotic effect on SK-MEL-28 cancerous cells and early apoptotic effect on non-tumoral L6 cells. Hence the obtained results suggest that the biological activities of the synthesised complexes depend on the nature of the ligands present in the complexes.

Highly selective, sensitive and biocompatible rhodamine-based isomers for Al3+ detection: A comparative study

Selective detection of a metal ion with high selectivity is of great importance to understand its existence and its role in many chemical and biological processes. We report here the synthesis, characterization and Al3+ sensing properties of two rhodamine-based isomers, (E)-2-((2-(allyloxy)benzylidene)amino)ethyl)-3′,6′-bis(ethylamine)-2′,7′-dimethylspiro[isoindoline-1,9′-xanthen]-3-one (L-2-oxy) and (E)-2-((4-(allyloxy)benzylidene)amino)ethyl)-3′,6′-bis(ethylamine)-2′,7′-dimethylspiro[isoindoline-1,9′-xanthen]-3-one (L-4-oxy). L-2-oxyand L-4-oxy show pink coloration, significant enhancement in absorbance at 530 nm and fluorescence intensity at 553 nm in the presence of Al3+ among several cations. Quantum yield and lifetime of the probes increase in the presence of Al3+. LOD values have been determined as low as ∼1.0 nM for both the isomers. DFT study suggests that the cation induces opening of spirolactam ring resulting in the changes of the rhodamine dyes. Additional reason could be Chelation Enhanced Fluorescence (CHEF) effect due to the subsequent chelation of the metal ion. Between two isomers, L-2-oxy displays better sensing ability towards Al3+ in terms of fluorescence enhancement, limit of detection, lifetime enhancement. Both the probes have been utilized in cell imaging studies using rat skeletal myoblast cell line (L6 cell line).

Microbial mediated Synthesis and characterization of Ag Nanoparticles and its Synergistic impact on Actinobacterial, Anticancer and Antihelminthic activity

In this study, an endophytic actinomycetes strain of Streptomyces viridodiastaticus strain MGR23 was isolated from the roots of the medicinal plant Centella asiatica. The biosynthesized Ag-NPs were characterized using UV-Vis spectroscopy, Fourier transform infrared (FT-IR), Scanning electron microscopy (SEM), X-ray diffraction (XRD) and Zeta potential, which confirmed the successful formation of crystalline, spherical silver nanoparticles. Biosynthesized AgNPs significantly inhibited and reduced the growth of medically important survival microorganisms (Streptomyces viridodiastaticus MGR 23 against Streptomyces toxytricini D2, Acidothermus cellulolyticus CA16, Streptomyces sp. MGR 21, Aciditerrimonas ferrireducens SM7 and Streptomyces tritici D5). The biosynthesized Ag-NPs exhibited broad-spectrum antihelminthic activity. Our data elucidated that the biosynthesized Ag-NPs had a highly cytotoxic effect against the cancerous L6 cell line under in vitro conditions, revealing a maximum IC50 value of 1.0 μg/mL and no known anticancer drug was used as a positive regulation. AgNPs have a remarkable synergistic impact that points to their considerable potential in nanomedicine for future clinical use as a combination therapy.

Morroniside interaction with poly (ADP-ribose) polymerase accentuates metabolic mitigation of alloxan-induced genotoxicity and hyperglycaemia: a molecular docking based in vitro and in vivo experimental therapeutic insight

The present study tends to evaluate the possible potential of bio-active Morroniside (MOR), against alloxan (ALX)-induced genotoxicity and hyperglycaemia. In silico prediction revealed the interaction of MOR with Poly (ADP-ribose) polymerase (PARP) protein which corroborated well with experimental in vitro L6 cell line and in vivo mice models. Data revealed the efficacy of MOR in the selective activation of PARP protein and modulating other stress proteins NF-κB, and TNF-α to initiate protective potential against ALX-induced genotoxicity and hyperglycaemia. Further, the strong interaction of MOR with CT-DNA (calf thymus DNA) analyzed through CD spectroscopy, UV-Vis study and ITC data revealed the concerted action of bio-factors involved in inhibiting chromosomal aberration and micronucleus formation associated with DNA damage. Finally, MOR does not play any role in microbial growth inhibition which often occurs due to hyperglycemic dysbiosis. Thus, from the overall findings, we may conclude that MOR could be a potential drug candidate for the therapeutic management of induced-hyperglycaemia and genotoxicity. Communicated by Ramaswamy H. Sarma

LCMS Determination and Cytotoxicity of Abrus precatorius on L6 and SK-N-MC Cell Lines.

The present study aimed to investigate the cytotoxic effect of various extracts derived from Abrus precatorius Linn. leaves on rat L6 and human SK-N-MC neuroblastoma cell lines and determine the secondary metabolites responsible for the cytotoxicity of Abrus precatorius. Successive solvent extraction of A.precatorius leaves was carried out using the Soxhlet apparatus with solvents such as petroleum ether, chloroform, ethyl acetate, and ethanol. HPTLC fingerprinting and LC-MS studies were performed to assess the presence of secondary metabolites, such as flavonoids and phenols, in the ethyl acetate extract. Furthermore, the cytotoxic effect of extracts was tested on rat skeletal muscle cell line L6 and human neuroblastoma cell line SK-N-MC using MTT assay. The total phenolic content of ethyl acetate and ethanol extracts of A.precatorius were 72.67 and 60.73 mg, respectively, of GAE/g dry weight of the extract. The total flavonoid content of ethyl acetate and ethanol extract of A.precatorius were 107.33 and 40.66 mg of Quercetin equivalents/g dry weight of the extract. LCMS analysis demonstrated that the flavonoids in specific Naringenin, Diosmetin, Glycitin, and Genistein might play a prominent role in the cytotoxicity of A.precatorius. The cytotoxicity study revealed that the extracts of A.precatorius were non-toxic to rat L6 myotubes, and the IC50 values of the various extracts, such as APPE, APCH, APEA, and APET, were >100 µg/ml. The extracts exhibited cytotoxic activity against human neuroblastoma SK-N-MC cells, and the IC50 values of APPE, APCH, APEA, APET, and the standard drug "Cisplatin" were >100, >100, 64.88, >100, and 3.72 µg/ml, respectively. It was concluded from the study that the extracts of Abrus precatorius were cytotoxic to neuroblastoma cell lines but non-toxic to normal cell lines. HPTLC and LC-MS studies confirmed that flavonoids in the ethyl acetate extract could be responsible for the biological activity.

Barosmin against postprandial hyperglycemia: outputs from computational prediction to functional responses in vitro

Previously, barosmin has been demonstrated to possess anti-diabetic action. However, its effect to inhibit α-amylase and α-glucosidase, including glucose utilization efficacy, has yet to be revealed. Hence, the current study attempted to assess the efficiency of barosmin in inhibiting the α-amylase, α -glucosidase, and dipeptidyl peptidase 4 enzymes, including glucose uptake efficacy. Molecular docking and simulation were performed using AutoDock Vina and Gromacs respectively followed by gene ontology analysis using the database for annotation, visualization, and integrated discovery. Further, in vitro enzyme inhibitory activities and glucose uptake assay were performed in L6 cell lines. Density functional theory analysis detailed mechanistic insights into the crucial interaction sites of barosmin of which the electron-dense region was prone to nucleophilic attack (O-atoms) whereas hydroxyl groups (-OH) showed affinity for electrophilic attacks. Barosmin showed good binding affinity with α-amylase (-9.2 kcal/mol), α-glucosidase (-10.7 kcal/mol), and dipeptidyl peptidase 4 (-10.0 kcal/mol). Barosmin formed stable nonbonded contacts with active site residues of aforementioned enzymes throughout 200 ns molecular dynamics simulation. Further, it regulated pathway concerned with glucose homeostasis i.e. tumor necrosis factor signaling pathway. In addition, barosmin showed α-amylase (IC50= 95.77 ± 23.33 µg/mL), α-glucosidase (IC50= 68.13 ± 2.95 µg/mL), and dipeptidyl peptidase 4 (IC50= 13.27 ± 1.99 µg/mL) inhibitory activities including glucose uptake efficacy in L6 cell lines (EC50= 12.46 ± 0.90 µg/mL) in the presence of insulin. This study presents the efficacy of the barosmin to inhibit α-amylase and α-glucosidase and glucose uptake efficacy in L6 cell lines via the use of multiple system biology tools and in vitro techniques. Communicated by Ramaswamy H. Sarma

Bioresource-derived multifunctional carbon quantum dots as a fluorescence and electrochemical sensing platform for picric acid and noncytotoxic food storage application

This work deals with the synthesis of facile, water-soluble, and blue-emitting carbon quantum dots (CQDs) from Plumbago indica leaves by a hydrothermal method with a quantum yield of 16 %. The biomass Plumbago indica leaves exhibit several advances such as easy availability, cost-effectiveness, non-toxic, green source, and not requiring any sophisticated techniques to develop the sensor. The optical, morphological, structural, and compositional properties of the CQDs were confirmed by various characterization methods. As a fluorescent probe, CQDs exhibited good linear response to picric acid (PA) with a limit of detection (LOD) of 18 nM. The sensing mechanism involves a combination of static quenching and fluorescence resonance energy transfer (FRET). In the electrochemical sensing method, the differential pulse voltammetric (DPV) technique was performed using a CQD-modified gold electrode with a LOD of 0.35 nM for PA. Using fresh carrots as a model sample, the antioxidant activity of PVA, CQD, and a PVA-CQD mixture was qualitatively determined. The in vitro cytotoxicity analysis of CQDs by MTT assay using the L6 cell line confirmed its non-toxic behaviour. This work is noteworthy because it offers a new attempt to identify toxic PA from biomass-derived precursor material and finds future applications in the food industry.

Tangeretin alleviates Tunicamycin-induced endoplasmic reticulum stress and associated complications in skeletal muscle cells.

Endoplasmic reticulum (ER) stress and associated oxidative stress are involved in the genesis and progression of skeletal muscle diseases such as myositis and atrophy or muscle wasting. Targeting the ER stress and associated downstream pathways can aid in the development of better treatment strategies for these diseases with limited therapeutic approaches. There is a growing interest in identifying natural products against ER stress due to the lower toxicity and cost effectiveness. In the present study, we investigated the protective effect of Tangeretin, a citrus methoxyflavone found in citrus peels against Tunicamycin (pharmacological ER stress inducer)-induced ER stress and associated complications in rat skeletal muscle L6 cell lines. Treatment with Tunicamycin for a period of 24h resulted in the upregulation of ER stress marker proteins, ER resident oxidoreductases and cellular reactive oxygen species (ROS). Co-treatment with Tangeretin was effective in alleviating Tunicamycin-induced ER stress and associated redox-related complications by significantly downregulating the unfolded protein response (UPR), ER resident oxidoreductase proteins, cellular ROS and improving the antioxidant enzyme activity. Tunicamycin also induced upregulation of phosphorylated p38 MAP Kinase and loss of mitochondrial membrane potential. Tangeretin significantly reduced the levels of phosphorylated p38 MAP Kinase and improved the mitochondrial membrane potential. From the results, it is evident that Tangeretin can be explored further as a potential candidate for skeletal muscle diseases involving protein misfolding and ER stress.

Exploration of in vitro Antioxidant and Cytotoxic Activities of Whole Plant Extracts of Rhynchosia heynei Wight & Arn: An Endemic Medicinal Herbaceous Under Shrub of Eastern Ghats of India

This study reports the antioxidant and anticancer potential of Rhynchosia heynei, a widely used plant in traditional medicine, against various cancerous cell lines. The study found that R. heynei exhibited significant radical scavenging activity, indicating its potential as a natural antioxidant compound to guard against oxidative stress-induced diseases, including cancer. The plant extracts also displayed selective cytotoxicity towards various cell lines, with the most potent cytotoxicity exhibited by the ethanol extract towards the MCF-7 cell line and diethyl ether extract towards the HCT-116 cell line. The hexane extract exhibited substantial cytotoxic activity against HepG2, HCT-116 and L6 cell lines. However, the aqueous extract of R. heynei was the least cytotoxic of all the cell lines employed. Overall, the cytotoxic activity of R. heynei extracts was majorly selective towards MCF-7 and HCT-116. The findings suggest that R. heynei could be a potent natural product for developing anticancer drugs with fewer side effects. Further, in vitro and in vivo studies are required to determine the exact mechanism of action of the plant’s potential anticancer activity.

Design, synthesis, and anti-bacterial activities of piperazine based phthalimide derivatives against superbug-Methicillin-Resistant Staphylococcus aureus

A series of piperazine-based phthalimide derivatives 5 (a-l) were synthesized and extensively characterized using a variety of spectrum methods, including LC-MS, 1H-NMR, 13C-NMR, and FT-IR. All the derivatives were examined for their physicochemical, pharmacokinetic, bio-activity score, and PASS analysis. The 5e piperazine-based phthalimide derivative demonstrated promising antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) in the in vitro antibacterial studies. In comparison to streptomycin and bacitracin (10 µg/mL), the minimum inhibitory concentration of 5e against MRSA was discovered to be 45±0.15 micro g/ml. The anti-MRSA activity was validated with membrane damage studies by using SEM, and in silico docking studies were against 3VMT and 6FTB proteins of MRSA. In the toxicity study, 5e derivatives were evaluated against L6 cell lines. The results of the studies show the synthesized 2-(2-(4-((4-chlorophenyl) sulfonyl) piperazin-1-yl) ethyl) isoindoline-1,3-dione (5e) can be used for the development of anti-MRSA drugs.

Anti-hyperglycemic contours of Madhugrit are robustly translated in the Caenorhabditis elegans model of lipid accumulation by regulating oxidative stress and inflammatory response.

The prevalence of diabetes has considerably increased in recent years. In the long run, use of dual therapy of anti-diabetic agents becomes mandatory to attain euglycemia. Also, the incidences of diabetes-related co-morbidities have warranted the search for new therapeutic approaches for the management of the disease. Traditional herbo-mineral, anti-diabetic agents like Madhugrit are often prescribed to mitigate diabetes and related complications. The present study aimed to thoroughly characterize the pharmacological applications of Madhugrit. Phytometabolite characterization of Madhugrit was performed by ultra-high performance liquid chromatography. Evaluation of cell viability, α-amylase inhibition, glucose uptake, inflammation, and wound healing was performed by in vitro model systems using AR42J, L6, THP1, HaCaT cells, and reporter cell lines namely NF-κB, TNF-α, and IL-1β. The formation of advanced glycation end products was determined by cell-free assay. In addition, the therapeutic potential of Madhugrit was also analyzed in the in vivo Caenorhabditis elegans model system. Parameters like brood size, % curling, glucose and triglyceride accumulation, lipid deposition, ROS generation, and lipid peroxidation were determined under hyperglycemic conditions induced by the addition of supraphysiological glucose levels. Madhugrit treatment significantly reduced the α-amylase release, enhanced glucose uptake, decreased AGEs formation, reduced differentiation of monocyte to macrophage, lowered the pro-inflammatory cytokine release, and enhanced wound healing in the in vitro hyperglycemic (glucose; 25 mM) conditions. In C. elegans stimulated with 100 mM glucose, Madhugrit (30 µg/ml) treatment normalized brood size, reduced curling behavior, decreased accumulation of glucose, triglycerides, and lowered oxidative stress. Madhugrit showed multimodal approaches in combating hyperglycemia and related complications due to the presence of anti-diabetic, anti-inflammatory, anti-oxidant, wound healing, and lipid-lowering phytoconstituents in its arsenal. The study warrants the translational use of Madhugrit as an effective medicine for diabetes and associated co-morbidities.

Bioactive molecules and the antidiabetic efficacy of <em>Memecylon randerianum</em> — an ethnomedicinal plant from the Western Ghats

Diabetes is one of the most chronic diseases and a leading cause of death even though it is preventable. There are multiple therapeutic strategies like daily intake of drugs, insulin treatment and islet transplantation to manage severe diabetic complications. But the plant based therapy with regular exercise can improve diabetic control with reduced medication. The Malabar Memecylon, Memecylon randerianum S.M. Almeida & M.R. Almeida, is a medicinal shrub widely distributed in the Western Ghats of Kerala. Leaves are the most utilized plant part in traditional medicine. In vitro antidiabetic activity and HR-LCMS-QTOF analysis of the leaves, along with the screening of phytochemicals, has been executed in the present investigation. The methanolic extract showed the highest presence of phytochemicals such as flavonoids, tannins, saponins etc. The quantitative screening revealed that saponins (211.16±1.9 mg/g) and flavonoids (209.74±2.12 mg/g) were highly concentrated, and the leaf extract showed significant dose-dependent inhibition on α-amylase and α- glucosidase. The glucose uptake in L6 cell lines was found to be increased by 25% compared to untreated cells (control) at 100 μg/mL sample concentration. HR-LCMS-QTOF analysis detected nine major compounds and showed that leaves of the plant could be used as an effective therapeutic agent against the most common type 2 diabetes.

Poly lactide-co-glycolide encapsulated nano-curcumin promoting antagonistic interactions between HSP 90 and XRCC1 proteins to prevent cypermethrin-induced toxicity: An in silico predicted in vitro and in vivo approach

The present study describes the preparation and characterization of poly-lactide-co-glycolide encapsulated nano-curcumin (NCUR) drug, and its potential efficacy against the pesticide, such as cypermethrin-induced DNA damage and genotoxicity. Cypermethrin, the chosen pesticide, contaminates the aquatic environment after being washed off from the agricultural field to nearby water bodies leading to biomagnification-related perturbation of the ecological balance and overall environmental health by elevating adverse effects on non-target organisms producing toxic metabolites through biotransformation. The physico-chemical properties of NCUR were evaluated by employing the AFM, DLS and UV-Vis techniques. Sustainable release of NCUR, their bio-availability and ability to cross the blood-brain-barrier was assessed in the fish model. The in silico molecular docking study to identify the signalling proteins that interact with phyto-core-compound curcumin (CUR) was undertaken to predict the effectiveness of NCUR to combat pesticide-induced toxicity by modulating p53, PARP, HSP 90 and XRCC1 stress proteins, and other associated parameters in in vivo model using tilapia fish and in vitro model using L6 (mammalian skeletal muscle) cell line. Overall results revealed that negatively charged poly-lactide-co-glycolide (PLGA)-encapsulated NCUR (∼46 nm) showed hyperchromic binding with DNA and modulated the signalling cascades involved in stress and DNA repair mechanisms, corroborating well with the in silico prediction that would pave a new pathway in the arena of chemical and biological sciences to serve mankind.

Synthesis of vildagliptin loaded acrylamide-g-psyllium/alginate-based core-shell nanoparticles for diabetes treatment

Diabetes mellitus has become a major public health concern all over the world. Vildagliptin is one of the antidiabeticdrug that can overcome the existing problem of this prevalent disease. Present study aims to synthesize and investigate the role of vildagliptin-loaded core-shell nanoparticle of grafted psyllium and alginate (VG@P/A-NPs) in anti-diabetes application. FTIR, SEM, XRD, 13CNMR and zeta analyzer were used for characterization of the core-shell nanoparticles (VG@P/A-NPs). The synthesized acrylamide-grafted-psyllium was also optimized through varying grafting parameters such as acrylamide and ceric ammonium nitrate (CAN) concentration, time and temperature to obtain the maximum yield of acrylamide-grafted-psyllium. Rheological analysis of pure psyllium, grafted psyllium and alginate were also performed. For biological studies, the first cytotoxicity of grafted psyllium and VG@P/A-NPs were examined on human lung adenocarcinoma cell line A549 in which it was observed that VG@P/A-NPs did not exhibited any toxicity. The antidiabetic potential of VG@P/A-NPs was investigated by glucose uptake assay, using TNF-α induced insulin resistance skeletal cell model using mouse muscle L6 cell line. The insulin signaling impaired cell line displayed a highly significant (p < 0.0001) dose-dependent increase in glucose uptake after treatment with increasing doses of VG@P/A-NPs.The drug release behavior of VG@P/A-NPs was examined at various pH and the highest drug release (98 %) was obtained at pH (7.4). The drug release kinetic data was following the Higuchi (R2 = 0.9848) kinetic model, suggesting the release of drug from vildagliptin-loaded grafted psyllium-alginate core-shell nanoparticles (VG@P/A-NPs) as a square root of time-dependent process and diffusion controlled. This study provides an economical and environment-friendly approach towards the synthesis of VG@P/A-NPs with antidiabetes applications.

Establishing the potency of N-acyl amino acids versus conventional fatty acids as thermogenic uncouplers in cells and mitochondria from different tissues.

The possibility that N-acyl amino acids could function as brown or brite/beige adipose tissue-derived lipokines that could induce UCP1-independent thermogenesis by uncoupling mitochondrial respiration in several peripheral tissues is of significant physiological interest. To quantify the potency of N-acyl amino acids versus conventional fatty acids as thermogenic inducers, we have examined the affinity and efficacy of two pairs of such compounds: oleate versus N-oleoyl-leucine and arachidonate versus N-arachidonoyl-glycine in cells and mitochondria from different tissues. We found that in cultures of the muscle-derived L6 cell line, as well as in primary cultures of murine white, brite/beige and brown adipocytes, the N-acyl amino acids were proficient uncouplers but that they did not systematically display higher affinity or potency than the conventional fatty acids, and they were not as efficient uncouplers as classical protonophores (FCCP). Higher concentrations of the N-acyl amino acids (as well as of conventional fatty acids) were associated with signs of deleterious effects on the cells. In liver mitochondria, we found that the N-acyl amino acids uncoupled similarly to conventional fatty acids, thus apparently via activation of the adenine nucleotide transporter-2. In brown adipose tissue mitochondria, the N-acyl amino acids were able to activate UCP1, again similarly to conventional fatty acids. We thus conclude that the formation of the acyl-amino acid derivatives does not confer upon the corresponding fatty acids an enhanced ability to induce thermogenesis in peripheral tissues, and it is therefore unlikely that the N-acyl amino acids are of specific physiological relevance as UCP1-independent thermogenic compounds.