Neurogenin-2 induced neuronal differentiation in co-cultured bone marrow stromal and neonatal cortical cells.

Objective: To explore a novel and dynamic role for neurogenin-2 in promoting cortical neurogenesis in cells produced from co-culturing neonatal cortical neural progenitor cells with bone marrow stromal cells. METHODS: The experimental study was conducted from June 2016 to January 2019 at the neuropharmacology laboratory of the Hussein Ebrahim Jamal Research Institute of Chemistry, International Centre for Chemical and Biological Sciences, Karachi. The growth of cells at different stages in harvested cells was determined by 3-(4, 5- dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay. Immunocytochemistry was used to evaluate the protein expressions of neuronal markers and transcription factors. Data was analysed using SPSS 20. RESULTS: Data showed significant generation of neuronal cells and this was also verified by increased expression of nesting in cortical co-cultures with bone marrow stromal cells. Immunoreactive outcomes showed over expressions in co-cultured chlorotoxin cells. Subsequently, neurogenin-2 was found intermixed with induced expressions of transcriptional factor NeuroD1 and reduced glial fibrillary acidic protein-labelled cells. Conclusion: Better understanding of the mechanisms underlying transcriptional modulation of neurogenic events hold the key for emerging treatment approaches towards neurodegeneration.

Memory and cognition enhancing effects of combination of Melissa officinalis and Panax ginseng.

Melissa officinalis and Panax ginseng extracts were investigated to determine combinatorial effects on cognitive behaviors' of albino-rats. The study was prospective-experimental; lasted from June-2022 to March-2023. Learning and memory measurements were done by animal-models. Data analyzed by 22nd version of SPSS. In Passive-avoidance-test both doses of Melissa officinalis and Panax ginseng (100/100mg/kg and 200/200mg/kg) showed significant differences in number of acquisition-trial between groups (p<0.001); drug treated groups showed longer latency-period compared to control and scopolamine (p<0.001). In time-spent-in-dark-chamber treated groups spent less-time in dark-chamber as compared to control and scopolamine (p<0.001). In Morris-water-maze-task treatment groups (100/100mg/kg and 200/200mg/kg) showed significant (p<0.001) decrease in escape-latency compared with control and scopolamine. Spatial-memory-probe showed significant interaction between drugs and days (p<0.001); time-spent in platform region is significantly increased (p<0.001) in both treatment groups compared with control and scopolamine. 8-arm-radial-maze-test showed the significant increase (p<0.05) in total number of correct responses in treatment groups (100/100mg/kg and 200/200mg/kg) compared to control and scopolamine. In-vitro studies revealed acetyl-choline-esterase inhibition by 36.40% from Melissa officinalis and Panax ginseng combination. Study concluded that combination of M. officinalis and P. ginseng extracts may significantly improve the effects on memory and cognition.

Unravelling the enigma of siRNA and aptamer mediated therapies against pancreatic cancer.

Pancreatic cancer (PC) is a fatal disease that has a poor 5-year survival rate. The poor prognosis can be attributed to both troublesome detections at the initial stage, which makes the majority of the treatment options largely unsuccessful and leads to extensive metastasis, as well as to its distinct pathophysiological characteristics, such as rich desmoplastic tumours bounded by dysplastic and hypo perfused vessels restricting the mobility of therapeutic agents. Continued attempts have been made to utilise innovative measures for battling PC to increase the therapeutic effectiveness of therapies and overcome their cytotoxicity. Combined cancer targeting and gene silencing approach has shown improved outcomes in patients' survival rates and quality of life, offering a potential solution to therapeutic complications. It particularly targets various barriers to alleviate delivery problems and diminish tumour recurrence and metastasis. While aptamers, a type of single-stranded nucleic acids with strong binding affinity and specificity to target molecules, have recently surfaced as a viable PC strategy, siRNA can interfere with the expression of certain genes. By concurrently suppressing genes and boosting targeted approach, the cocktail of siRNA/Aptamer and other therapeutic drugs can circumvent the multi-drug resistance phenomena. Additionally, combination therapy with additive or synergistic effects can considerably increase the therapeutic efficacy of anti-cancer medications. This study outlines the primary difficulties in treating PC, along with recent developments in siRNA/Aptamer mediated drug delivery to solve the major hiccup of oncology field.

Highly potent anti-inflammatory, analgesic and antioxidant activities of 3,5-disubstituted tetrahydro-2H-1,3,5-thiadiazine thiones.

Four series of tetrahydro-2H-1,3,5-thiadiazine-2-thiones (series A and B including two novel enantiopure isomers), tetrahydro-2H-1,3,5-thiadiazine-6-thiones (series C) and N-3 ester derivatives of tetrahydro-2H-1,3,5-thiadiazine-6-thiones (series D) were synthesized and evaluated for their anti-inflammatory, analgesic and anti-oxidant activities. These THTT analogues specially series D were first time examined for their in vitro anti-inflammatory, in vivo analgesic and anti-oxidant activities. Among them lipophilic compounds (series B and D) were found to be highly active for anti-inflammatory evaluation with IC50 values between 5.1-16.9 and 4.1-32.4 µM, respectively when compared with the standard drug ibuprofen IC50 = 11.2 µM. The structure-activity relationship exposed the importance of lipophilic substituents especially ester and n-propyl group for inhibition of inflammation. The molecular docking studies demonstrated that all the active analogues of THTT have notable binding relations with Arg120 of the active sites of COX-1 enzyme either through CS moiety of the THTT nucleus or with COO attached at N-3 of THTT nucleus. In vivo analgesic activity of the selected THTT compounds 14, 17, 18, 19 (series B) and 28 (series D) were also carried out by acetic acid-induced writhing procedure. The compound 28 showed significant anti-nociceptive/analgesic activity at the oral dose of 5 mg/kg body weight with the percent protection (32.05 %) when compared with standard indomethacin at 10 mg/kg (48.83 %). Additionally, these compounds demonstrated the moderate level of antioxidant potential with IC50 values in the range of 60.9 to 93.6 µM (standard butylated hyroxyanisole; IC50 = 44.2 µM). These results indicated that this class of heterocyclic compounds may be a template specially to design better anti-inflammatory and analgesic agents.

Lipid-based nanocarrier mediated CRISPR/Cas9 delivery for cancer therapy.

CRISPR/Cas mediated gene-editing has opened new avenues for therapies that show great potential for treating or curing cancers, genetic disorders, and microbial infections such as HIV. CRISPR/Cas9 tool is highly efficacious in revolutionizing the advent of genome editing; however, its efficient and safe delivery is a major hurdle due to its cellular impermeability and instability. Nano vectors could be explored to scale up the safe and effective delivery of CRISPR/Cas9. This review highlights the importance of CRISPR/Cas9 genome editing system in cancer treatment along with the effect of lipid-based nanoparticles in its safe delivery to cancer cells. The solid-lipid nanoparticles, nanostructured lipid carrier, lipid nanoparticles and niosomes have shown great effect in the delivery of CRISPR compounds to the cancer cells. The design and genome editing application in cancer therapy has been discussed along with the future concern and prospects of lipid nanoparticle based CRISPR/Cas9 has been focused toward the end.

Neuronal transcription program induced in hippocampal cells cocultured with bone marrow derived mesenchymal cells.

Several approaches have been applied to harvest bone marrow stromal cells (BMSCs) and to differentiate into neurons or neuronal-like cells through chemical stimulation or exposing to growth factors. To date, the data regarding induction or regulation of neuronal transcription program in neuronal-like cells derived from BMSCs is yet unknown. The objective of this study is to co-culture BMSCs with neonatal hippocampal cells and generate neuronal-like cells by direct cell-to-cell contact system without using neuronal growth factors or neurobasal medium. Here, we proposed a role for NeuroD1 and Neurogenin -2 bHLH family of transcription factors implicated in onset of neurogenesis and differentiation of cells into neurons in promoting the interaction of hippocampal cells with BMSCs and their differentiation in to neurons. The proliferation of the cells was assessed with MTT assay and the role of neuronal induction and differentiation transcription regulators NeuroD1 and Neurogenin-2 in cocultured cells was determined through immunocytochemical analysis. We observed activation and expression of the neurogenic transcription factors NeuroD1 and NGN-2 associated with neuronal activation program to initiate the onset of neurogenesis in cocultured cells. Further, our results have shown a significant expression of neuronal progenitor and immature neuronal marker i.e., nestin and tubulin respectively in cocultured cells endorsing the initiation of neuronal activation.

Chitosan­zinc sulfide nanoparticles, characterization and their photocatalytic degradation efficiency for azo dyes.

Herein, chitosan­zinc sulfide nanoparticles (CS-ZnS-NPs) were developed as an efficient photocatalyst for the degradation of toxic dyes. The as-synthesized CS-ZnS-NPs were analyzed using XRD, FTIR, SEM, and EDS. The functional groups of CS-ZnS-NPs were validated with FTIR spectroscopy. The SEM envisaged the average particle size as 40 nm, whereas EDS interpreted the compositional analysis of the nanocomposite. XRD analysis illustrated the crystallinity and hexagonal crystal structure of the CS-ZnS-NPs. The photocatalytic efficiency of CS-ZnS-NPs was evaluated using two carcinogenic azo dyes, Acid Brown 98 and Acid Black 234. A UV lamp (254 nm) was used as an irradiation source during the photocatalytic degradation of dyes. At the optimum conditions, the synthesized CS-ZnS-NPs showed 96.7% degradation for Acid Black 234 in 100 min and 92.6% for Acid Brown 98 in 165 min. The degradation phenomena followed pseudo-first-order kinetics. The values of rate constant (k) were 0.01464 and 0.04096 min-1 with correlation coefficient (R2) of 0.98891 and 0.99406 for Acid Brown 98 and Acid Black 234, respectively. The CS-ZnS-NPs were easily recovered and recycled for four successive batches. The results showed that CS-ZnS-NPs are considered as highly productive, cost-effective and promising photocatalyst in degrading pollutants in several consecutive cycles.

N-(2-hydroxyphenyl) acetamide (NA-2) elicits potent antitumor effect against human breast cancer cell line (MCF-7).

Breast cancer is the most dominating malignancy in females worldwide. Treatment with conventional chemotherapeutics is associated with severe adverse effects. Thus need of new compounds, with better therapeutic potential and lesser side effects still exist. In this context the present study is planned to investigate therapeutic potential of anti-inflammatory compound N-(2- hydroxyphenyl) acetamide (NA-2) against breast cancer cells (MCF-7). The compound was selected on the basis of its reported anti-inflammatory, anti-arthritic and anti-glioblastoma activities in our previous studies. MTT, Annexin-V-FITC and wound healing assays were used to analyze the effect of compound on growth inhibition, apoptosis and metastasis. While flow cytometry, RT-PCR and immunocytochemistry techniques were used to assess the effect of NA-2 on cell cycle arrest, and expression of apoptotic markers (Bax and Bcl-2) at both mRNA and protein level respectively. Data analysis revealed that NA-2 significantly inhibits growth of MCF-7 cells after 48 h treatment (IC50 = 1.65 mM). NA-2 also delayed the wound healing process, arrested cell cycle at G0/G1 phase and induced apoptosis by enhancing Bax/Bcl-2 ratio. We concluded that NA-2 possesses strong anticancer activity against MCF-7 cells, which is mediated through different mechanisms, making it a useful molecule for the development of new antitumor drugs.

Design and synthesis of mixed micellar system for enhanced anticancer efficacy of Paclitaxel through its co-delivery with Naringin.

Emergence of multidrug resistance (MDR) has limited the success of chemotherapeutic agents. Reversal of drugs efflux systems through combination therapy has got wider attention for increasing anticancer drugs efficacy. This study aims at co-encapsulation of Paclitaxel with Naringin in mixed polymeric micelles for enhanced anticancer activity of the drug. Drug-loaded micelles were prepared using two different amphiphilic block co-polymers and were characterized for morphology, size, zeta potential, drug encapsulation, in vitro release and stability using atomic force microscope (AFM), zetasizer, UV spectrophotometer, and FT-IR. MTT assay and fluorescence microscopy were used for in vitro cytotoxicity and cellular uptake studies. Nano-size micelles with spherical morphology and negative charge encapsulated 76.52 ± 0.94% and 32.87 0.61% Paclitaxel and Naringin, respectively. The micelles were thermally stable and retained 87.05 ± 0.69% and 92.88 ± 2.17% Paclitaxel and Naringin upon one-month storage. Maximum drug release was achieved at fourth hour of the study for both the loaded drugs. Paclitaxel co-encapsulation with Naringin synergistically improved its intracellular uptake and 65% in vitro cytotoxicity against breast cancer cells was achieved at its lower dose of 15 µg/mL. Results suggest that co-encapsulation of Paclitaxel with Naringin in mixed micelles is an effective strategy for achieving its higher anticancer activity.