COLON SPECIFIC DELIVERY OF COMBINATION OF 5-FLUOROURACIL AND CELECOXIB: PREPARATION, CHARACTERIZATION, AND IN VITRO CYTOTOXICITY ASSAY

Objective: 5-Fluorouracil (5-FU) and celecoxib (Cel) combination offered additive effect in the treatment of colon cancer. However, physicochemical and biopharmaceutical attributes of both drugs deliver suboptimal concentration at the site of action. The objective of the current study is the development of a microparticulate drug delivery system loaded with a combination of 5-FU and Cel to achieve prolonged drug delivery in colon cancer. Methods: 5-FU and Cel combination were loaded in Eudragit coated chitosan (CH) microspheres (MSs) and characterized. Results: The average particle size of the MSs was in the range of 2.7±0.9μm to 4.8±1.1μm. A substantial drug encapsulation efficiency of 71.30±2.3% as obtained for 5-FU as compared to 35.20±1.9% of Cel in the tailored microparticles. The drug loading capacity of 6.5 mg/10 mg and 2.3 mg/10 mg was obtained for 5-FU and Cel, respectively. By Eudragit S 100 (Ed) coating, significant pH-dependent release profile was achieved, and no drug release was observed in simulated gastric and intestinal fluids. The developed MSs exhibited the release of 92.1±2.9% of 5-FU in 8h whereas 18.9±0.7% Cel was found to be released from the developed MSs. The drug-loaded MSs exhibited appreciable potency against HT-29 cells with an IC50 value of 35.9 μM. Conclusion: The results indicated that these microparticles are a promising vehicle for selectively targeting drugs to the colon in the chemotherapy of colon cancer

Short-hairpin RNA library: identification of therapeutic partners for gefitinib-resistant non-small cell lung cancer.

Somatic mutations of the epidermal growth factor receptor often cause resistance to therapy with tyrosine kinase inhibitor in non-small cell lung cancer (NSCLC). In this study, we aimed to identify partner drugs and pathways that can induce cell death in combination with gefitinib in NSCLC cells. We undertook a genome-wide RNAi screen to identify synthetic lethality with gefitinib in tyrosine kinase inhibitor resistant cells. The screening data were utilized in different approaches. Firstly, we identified PRKCSH as a candidate gene, silencing of which induces apoptosis of NSCLC cells treated with gefitinib. Next, in an in silico gene signature pathway analysis of shRNA library data, a strong correlation of genes involved in the CD27 signaling cascade was observed. We showed that the combination of dasatinib (NF-κB pathway inhibitor) with gefitinib synergistically inhibited the growth of NSCLC cells. Lastly, utilizing the Connectivity Map, thioridazine was identified as a top pharmaceutical perturbagen. In our experiments, it synergized with gefitinib to reduce p-Akt levels and to induce apoptosis in NSCLC cells. Taken together, a pooled short-hairpin library screen identified several potential pathways and drugs that can be therapeutic targets for gefitinib resistant NSCLC

Clinical profile of snake bite patients in tertiary care hospital in Himachal Pradesh: a prospective study

Background: To study clinical profile of snake bite patients in tertiary care hospital in Indra Gandhi Medical College at Shimla, Himachal Pradesh of North India.Methods: Hospital based prospective observational study was conducted in the Department of Medicine, for the duration of one year from 1st June2013 to 31st May 2014.Results: A total of 78 patients were admitted with mean age of 38.46 years with male to female ratio of 1:1.6. Seasonal variation with peak incidence during rainy season was seen. Most common snake identified was green coloured and peak timing of snake bite was between 07:00am-04:00pm. There was delay in admission of more than 6 hours in 66.67% of cases. Hemotoxicity was predominant manifestation seen in 62.82% of cases and persistence of coagulopathy was most common complication (51.02%) despite giving optimal ASV. There was paucity in ASV administration seen in only 59.46% of referred patients. Mean ASV vials used were 23.41 vials ±8.72 vials.Conclusions: Mass education is required at both general population and health professional levels to improve snake bite management and monovalent ASV against Green pit viper is more practical option to manage cases in this region

Role of norepinephrine in the regulation of rapid eye movement sleep

Sleep and wakefulness are instinctive behaviours that are present across the animal species. Rapid eye movement (REM) sleep is a unique biological phenomenon expressed during sleep. It evolved about 300 million years ago and is noticed in the more evolved animal species. Although it has been objectively identified in its present characteristic form about half a century ago, the mechanics of how REM is generated, and what happens upon its loss are not known. Nevertheless, extensive research has shown that norepinephrine plays a crucial role in its regulation. The present knowledge that has been reviewed in this manuscript suggests that neurons in the brain stem are responsible for controlling this state and presence of excess norepinephrine in the brain does not allow its generation. Furthermore, REM sleep loss increases levels of norepinephrine in the brain that affects several factors including an increase in Na-K ATPase activity. It has been argued that such increased norepinephrine is ultimately responsible for REM sleep deprivation, associated disturbances in at least some of the physiological conditions leading to alteration in behavioural expression and settling into pathological conditions

In vitro antifilarial activity, antioxidant potential and phenolic constituents of Quisqualis indica L.

648-654Quisqualis indica L., commonly known as ‘Rangoon-ki-bel’ or ‘Madhumalti’, has been used by the traditional healers as it is active against some of the commonly occurring diseases like boils, fevers diarrhea and helminthiasis. However, no systematic and scientifically validated studies on antifilarial activity of Q. indica are available. In the present study, we report in vitro antifilarial activity of ethanolic and hydroethanolic extracts of the leaves (QILE and QILEW) and flowers (QIFE and QIFEW) of this plant on microfilariae (mf) and female adult worms of human lymphatic filariid Brugia malayi using motility and or 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT)-reduction assays. The hydroalcoholic extract of flowers (QIFEW) was found effective as it killed adult female worms (LC100: 62.5 µg/mL) and mf (LC100: 125 µg/mL); IC50 values for the respective parasite stages were 34.50 and 31.88 µg/mL. SI values recorded with respect to motility of female parasite and mf was more than 20. The active principle(s) responsible for antifilarial activity may thus be present in QIFEW. The antioxidant activity results also indicated QIFEW to possess better antioxidant potential than the other extracts studied. HPLC analysis showed that the 02 keyphenolics present in hydroalcoholic extract of the flowers (QIFEW) were gallic acid and ellagic acid. In the different extracts, the concentration of gallic acid was found to vary from 26.9 mg/g to 2.50 mg/g while ellagic acid ranged between 11.5 mg/g to 6.77 mg/g. It was also observed that the leaves were rich in flavonoids whereas the flowers were rich in phenolics. The findings indicate that active molecule (s) of hydroalcoholic extractfrom Q. indica flowers may help in providing new leads for developing antifilarial agents. We believe that this is the first systematically studied report on the in vitro antifilarial activity of the hydroalcoholic extract of Q. indica flowers

Aberrant splicing of U12-type introns is the hallmark of ZRSR2 mutant myelodysplastic syndrome.

Somatic mutations in the spliceosome gene ZRSR2-located on the X chromosome-are associated with myelodysplastic syndrome (MDS). ZRSR2 is involved in the recognition of 3'-splice site during the early stages of spliceosome assembly; however, its precise role in RNA splicing has remained unclear. Here we characterize ZRSR2 as an essential component of the minor spliceosome (U12 dependent) assembly. shRNA-mediated knockdown of ZRSR2 leads to impaired splicing of the U12-type introns and RNA-sequencing of MDS bone marrow reveals that loss of ZRSR2 activity causes increased mis-splicing. These splicing defects involve retention of the U12-type introns, while splicing of the U2-type introns remain mostly unaffected. ZRSR2-deficient cells also exhibit reduced proliferation potential and distinct alterations in myeloid and erythroid differentiation in vitro. These data identify a specific role for ZRSR2 in RNA splicing and highlight dysregulated splicing of U12-type introns as a characteristic feature of ZRSR2 mutations in MDS

In vitro antifilarial activity, antioxidant potential and phenolic constituents of Quisqualis indica L.

Quisqualis indica L., commonly known as ‘Rangoon-ki-bel’ or ‘Madhumalti’, has been used by the traditional healers as it is active against some of the commonly occurring diseases like boils, fevers diarrhea and helminthiasis. However, no systematic and scientifically validated studies on antifilarial activity of Q. indica are available. In the present study, we report in vitro antifilarial activity of ethanolic and hydroethanolic extracts of the leaves (QILE and QILEW) and flowers (QIFE and QIFEW) of this plant on microfilariae (mf) and female adult worms of human lymphatic filariid Brugia malayi using motility and or 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT)-reduction assays. The hydroalcoholic extract of flowers (QIFEW) was found effective as it killed adult female worms (LC100: 62.5 μg/mL) and mf (LC100: 125 μg/mL); IC50 values for the respective parasite stages were 34.50 and 31.88 μg/mL. SI values recorded with respect to motility of female parasite and mf was more than 20. The active principle(s) responsible for antifilarial activity may thus be present in QIFEW. The antioxidant activity results also indicated QIFEW to possess better antioxidant potential than the other extracts studied. HPLC analysis showed that the 02 keyphenolics present in hydroalcoholic extract of the flowers (QIFEW) were gallic acid and ellagic acid. In the different extracts, the concentration of gallic acid was found to vary from 26.9 mg/g to 2.50 mg/g while ellagic acid ranged between 11.5 mg/g to 6.77 mg/g. It was also observedthat the leaves were rich in flavonoids whereas the flowers were rich in phenolics. The findings indicate that active molecule (s) of hydroalcoholic extractfrom Q. indica flowers may help in providing new leads for developing antifilarial agents. We believe that this is the first systematically studied report on the in vitro antifilarial activity of the hydroalcoholicextract of Q. indica flowers