LC-QTOF-MS/MS metabolic profiling and hepatoprotective effects of Litsea monopetala bark methanol extract against liver injury in rats and HepG2 cells.

Fresh stem bark decoction of Litsea monopetala has been practiced for the treatment of jaundice and other liver disorders by the tribal communities of Thakht-e-Sulaiman hills from West Pakistan. As per the folkloric claim, this study aims to identify the phytoconstituents and evaluate the hepatoprotective action of stem bark methanol extract of L. monopetala (LMME). The in-vitro hepatoprotective effect of L. monopetala was performed by H2O2-induced toxicity in the HepG2 cell line and in-vivo by cclt;sub>4-induced hepatotoxicity in Wistar albino rats taking Silymarin as standard drug. Phytoconstituents were identified using LC-QTOF-MS analysis followed by in-silico docking and validation. Molecular docking interactions between identified compounds of L. monopetala and two target proteins, namely 1VJY and 5HYK were presented. In this study, treatment with LMME at 100 µg/mL showed 67.73 % cell viability as compared to H2O2 (100 µM) treated alone i.e., 18.55 % in the HepG2 cell line. In-vivo treatment of LMME reversed the altered serum biochemical parameters and reduced the inflammatory response similar to that of the Silymarin-treated group supported by histopathological investigation. This research reveals that L. monopetala is a rich source of flavonoids and phenols which supports its hepatoprotective effects and is proposed for its usage as a promising hepatoprotective agent after controlled trials.

Cancer Cell-Selective Inhibition of Migration by Styrenic Catiomer Emulsions.

Cancer metastasis remains the most formidable cause of mortality and morbidity in cancer patients. Developing an effective and economical method toward cancer antimetastatic strategy demands immediate attention in anticancer therapy. Herein, we followed a cost-effective greener method for preparing a small family of amphiphilic catiomers with varied styrene content (45, 63, and 83%), which revealed the unique potential of promoting normal cell migration while retarding cancer metastasis. The styrenic polymers formed micellar self-assembly in aqueous phase and exhibited a cationic charge. Polymers were quite nontoxic up to 200 µg/mL concentration toward human embryonic kidney cell HEK293 as well as human, triple negative breast cancer cell MDAMB-231, mouse melanoma cell B16F10, and human oral squamous carcinoma cell FaDu. Confocal imaging and fluorescence activated cell sorting (FACS) showed effective incorporation of polymers within cells. Interestingly, the polymer-treated HEK293 cells underwent prominent wound healing in scratch assay. However, the as-synthesized polymer-treated cancer cells resisted migration as analyzed from the scratch assay. A mechanistic study using immunoblotting assay established upregulation of migratory proteins vimentin and TGF-ß and downregulation of E-cadherin in normal HEK293 cells. Remarkably, this trend was completely reversed in cancer cell MDAMB-231. This study describes the extraordinary potential of styrenic catiomers as wound healers for normal cells while inhibiting cancer metastasis.

Structural Phase Transition in 0D (3,5-DMP)2Bi1-xSbxCl5 Metal Halides: Expression of the Lone Pair Effect and Polyhedral Distortion.

Low-dimensional Bi/Sb-based organic-inorganic metal halides (OIMHs) have attracted immense attention from the research community because of their structural diversity and efficient luminescence properties. Further understanding of the relationship between the structure and luminescence properties of these materials is of utmost importance for tuning the luminescence properties for various practical applications. Herein, we have synthesized two lead-free Bi/Sb-based novel OIMHs, (3,5-DMP)2BiCl5 and (3,5-DMP)2SbCl5 [(3,5-DMP) = 3,5-dimethylpiperidine], with zero-dimensional (0D) structures and crystallizing in triclinic (P1¯ space group) and monoclinic (P21/c space group) crystal systems, respectively. Both the compounds behave as typical semiconductors with indirect optical band gaps of 3.34 and 3.36 eV for pristine Bi and Sb compounds. These compounds exhibit higher environmental and thermal stability at ambient conditions. Gradual substitution of Sb at the Bi site in (3,5-DMP)2Bi1-xSbxCl5 resulted in the introduction of structural strain due to the significant expression of the lone pair effect, thus leading to a structural transition from the triclinic to monoclinic phase. The effect of the structural phase transition on the optical properties is also studied in (3,5-DMP)2Bi1-xSbxCl5. This work may offer new direction and guidance for exploring various 0D hybrid metal halides and tuning the structures for improvement in the luminescence properties.

Hepatoprotective potency of Litsea glutinosa (L.) C.B. Rob. leaf methanol extract on H2O2-induced toxicity in HepG2 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaves of Litsea glutinosa (L.) (Lauraceae) are traditionally used to treat hepatitis and liver injury by Bangladeshi folks. However, the hepatoprotective study of leaves of L. glutinosa has not been supported by any research. AIM OF THE STUDY: To evaluate the antioxidant and hepatoprotective effects of leaves of methanol extract of L. glutinosa using the HepG2 cell line. Phytochemicals were identified with the help of a GC-MS study followed by In-silico docking of the promising compounds to justify our hepatoprotective effect. MATERIALS & METHODS: The dried leaves of L. glutinosa (LGAO) were extracted by Soxhlet using methanol as solvent. Antioxidant effects were investigated using Superoxide dismutase (SOD), Reduced glutathione (GSH), Glutathione peroxidase (GPx), and Malondialdehyde (MDA) in HepG2 cells against H2O2 intoxicated group. The In-vitro hepatoprotective effect of LGAO (100 µg/ml) was determined in HepG2 cells as compared with the Silymarin-treated standard group (100 µg/ml) along with morphological changes of cells. Twelve numbers of phytochemicals were identified by GC-MS study. In-silico studies are performed for their inhibitory effects against Peroxisome proliferator-activated receptor alpha (PPAR-α) and Transforming growth factor-beta1 (TGF-ß1) using AUTODOCK Tools-1.5.6 and Discovery studio 4.0. RESULTS: Methanol extract of L. glutinosa possesses (LGAO) significant (p < 0.0001) increase in SOD, GSH, and GPx levels and a decrease in MDA as compared with the control one. MTT assay in HepG2 cells showed a significant (p < 0.0001) increase in the percentage of cell viability in LGAO and Silymarin-treated group i.e., 71.98%, 88.59% respectively as compared with the H2O2 intoxicated group alone i.e., 22.74%. Restoration of cell architecture in HepG2 cells was obtained by the LGAO and Silymarin-treated group treated with H2O2. Further, the In-silico study of Neophytadiene compound showed the highest docking score -10.2 and -8.6 towards receptors. CONCLUSION: Methanol extract of leaves of L. glutinosa showed potential hepatoprotective effect In-vitro which justified our traditional claim. The presence of phytochemical Neophytadiene may be responsible for the said effect. Furthermore, molecular docking scores were consistent with the In-vitro results. They targeted the substantial inhibitory effects of Litsea glutinosa against receptors to establish the correlation between experimental and theoretical results.

Cholate-conjugated cationic polymers for regulation of actin dynamics.

Cytoskeletal movement is a compulsory necessity for proper cell functioning and is largely controlled by actin filament dynamics. The actin dynamics can be fine-tuned by various natural and artificial materials including cationic proteins, polymers, liposomes, and lipids, although most of the synthetic substrates have toxicity issues. Herein, we show actin nucleation and stabilization with a synthetic family of cholic acid (CA)-conjugated cationic macromolecules. Architectural conjugation of CA is designed by attaching it to the polymer chain end, as well as to the side chain of the polymer. The side-chain cholate content is also varied in the copolymer, which results in self-aggregation in aqueous media above a certain critical aggregation concentration (CAC). Below the CAC, the in vitro actin dynamics modulation behaviour is studied using a pyrene actin fluorescence assay, actin co-sedimentation assay, dynamic light scattering (DLS), and transmission electron microscopy (TEM). These polymers are nontoxic to HeLa cells, and the 2% cholate conjugated cationic copolymer showed maximum enhancement of G-actin nucleation, as well as F-actin stabilization. We further develop a theoretical model to elucidate the underlying dynamics of the actin polymerization process under the influence of cationic copolymers with cholate pendants. Finally, we proposed macromolecular self-aggregation as a unique tool for modulating actin dynamics, as revealed from the experimental findings and theoretical modelling.

Modulating Insulin Aggregation with Charge Variable Cholic Acid-Derived Polymers.

To understand the effect of cholic acid (CA)-based charge variable polymeric architectures on modulating the insulin aggregation process, herein, we have designed side-chain cholate-containing charge variable polymers. Three different types of copolymers from 2-(methacryloyloxy)ethyl cholate with anionic or cationic or neutral units have been synthesized by reversible addition-fragmentation chain transfer polymerization. The effects of these copolymers on the insulin fibrillation process was studied by multiple biophysical approaches including different types of spectroscopic and microscopic analyses. Interestingly, the CA-based cationic polymer (CP-10) was observed to inhibit the insulin fibrillation process in a dose-dependent manner and to act as an effective anti-amyloidogenic agent. Corresponding anionic (AP-10) and neutral (NP-10) copolymers with cholate pendants remained insignificant in controlling the aggregation process. Tyrosine fluorescence assays and Nile red fluorescence measurements demonstrate the role of hydrophobic interaction to explain the inhibitory potencies of CP-10. Furthermore, circular dichroism spectroscopic measurements were carried out to explore the secondary structural changes of insulin fibrils in the presence of cationic polymers with and without cholate moieties. Isothermal titration calorimetry measurements revealed the involvement of electrostatic polar interaction between the CA-based cationic polymer and insulin at different stages of fibrillation. Overall, this work demonstrates the efficacy of the CA-based cationic polymer in controlling the insulin aggregation process and provides a novel dimension to the studies on protein aggregation.

Efficacy of a novel bone preprocessing method for better DNA yield.

In cases involving identification of missing persons, mass disasters and ancient DNA investigations, bone and teeth samples are often the only, and almost always the best, biological material available for DNA profiling. Standard methods for extraction of DNA from such samples involve grinding of the bone and teeth samples. Here, we present an extremely efficient protocol for recovery of DNA from bone samples by a method of scrapping. The study was carried out on 25 samples and it was found that the quantity of DNA isolated by the scrapping method was up to 1.131 ng/µl with a success rate of 93% as compared to a much lower yield of 0.359 ng/µl DNA isolated with a success rate of 28% through the grinding method. The scrapping method of DNA extraction has been proven to be extremely useful in forensic examination of challenging samples that had multiple failures using the traditional grinding method.

Cholate Conjugated Polymeric Amphiphiles as Efficient Artificial Ionophores.

A family of amphiphilic copolymers containing hydrophobic cholate pendants has been prepared by copolymerization of cholic acid-based monomer 2-(methacryloxy)-ethyl cholate (MAECA) with polyethylene glycol methyl ether methacrylate (PEGMA). The polymers differ for the content of MAECA that increases from 0 to 35%. The copolymers partition within liposomes and display potent ionophoric activity forming large pores in the membrane and allowing the leakage of small inorganic ions (H+, Na+) and of large polar organic molecules (calcein). Their activity is strictly correlated to the content of cholic acid subunits, increasing as the fraction of cholate moiety increases.

Genomic insight into Y-STR diversity in the population of Odisha, India.

This study evaluated the haplotype diversity of 17 Y chromosomal genetic markers among 202 unrelated males who were randomly selected in the population of Odisha, India. Out of total 196 haplotypes observed in this study, 190 were unique haplotypes. Forensic relevant parameters, viz., gene diversity (GD) and discrimination capacity (DC), were calculated as 0.999999998 and 0.970 respectively, for the studied population. The highest genetic diversity was observed at the locus DYS385a/b (0.9541) and lowest at the locus DYS437 (0.3326) among all the studied Y chromosomal loci. The polymorphic information content (PIC), power of discrimination (PD), and matching probability (PM) was found to be 0.999999965, 0.999999998, and 1.6×10-9 for the tested Y STR loci. The genetic data observed in this study would enrich the existing Y STR data of the Indian population and would also be useful for forensic application.

Genomic portrait of Odisha, India drawn by using 21 autosomal STR markers.

In order to find out the genetic diversity in the eastern Indian population of Odisha consisting of various linguistic and ethnic groups, we undertook a study on 508 unrelated healthy individuals belonging to Odisha, India. We assessed genetic variation and compared the data with published literature of Indian population consisting of different ethnic groups from different geographical areas using 21 autosomal STR markers. The most polymorphic and discriminatory STR locus in the studied population was found to be SE33 with the calculated values of 0.94 and 0.991 respectively for both the parameters. The combined power of discrimination (CPD) and combined power of exclusion (CPE) were found to be 1 and 0.999999999704865 respectively. The combined probability of match (CPm) and combined paternity index (CPI) for all 21 autosomal STR loci were found to be 8.01 × 10-26 and 3.45 × 109 respectively. Though, the studied eastern Indian population of Odisha shared its closest genetic affinity with nearest Indian geographical regions, i.e., the population of Jharkhand which is geographically located in eastern India as well as the Central Indian population.