AN ASSESSMENT OF POTENTIAL DRUG-DRUG INTERACTIONS IN HYPERTENSIVE PATIENTS IN A TERTIARY CARE HOSPITAL

Objective: To an assessment of potential drug-drug interactions in hypertensive patients in a tertiary care hospital. Methods: A prospective, observational study was conducted at a tertiary care hospital, Erode for a period of 8 mo. A sample of 480 patients was assessed for PDDIs using drug checker in Micromedex®-2.7. Results: A total of 430 patients were analyzed and it was found to be 396 (82.50%) hypertensive patients had PDDIs, and a sum total of 1160 PDDIs were observed. Potential drug-drug interactions (PDDIs) higher in female hypertensive patients [255 (64.39%)] compared to males. Incidences of PDDIs were found to be higher in the age group of 60-70 y were [177 (44.69%)] and incidences of interactions based on the duration of (4-6 d) hospital stays were 272 (68.68%). Moreover, 49.24% of patients were found to be prescribed with more than 7 drugs, with higher incidences of PDDIs. Some of the most common drug interacting pair was between aspirin and clopidogrel combination observed in 325 PDDIs in the major, with pharmacodynamics in nature. Conclusion: Clinical pharmacist ought to have the role of regular monitoring of drug therapy in identifying and preventing the medications that have the potential to cause drug-drug interactions, thereby minimizing the undesirable outcomes in drug medical care and improving the quality of care

Rapid and Efficient Generation of Recombinant Human Pluripotent Stem Cells by Recombinase-mediated Cassette Exchange in the AAVS1 Locus

Even with the revolution of gene-targeting technologies led by CRISPR-Cas9, genetic modification of human pluripotent stem cells (hPSCs) is still time consuming. Comparative studies that use recombinant lines with transgenes integrated into safe harbor loci could benefit from approaches that use site-specific targeted recombinases, like Cre or FLPe, which are more rapid and less prone to off-target effects. Such methods have been described, although they do not significantly outperform gene targeting in most aspects. Using Zinc-finger nucleases, we previously created a master cell line in the AAVS1 locus of hPSCs that contains a GFP-Hygromycin-tk expressing cassette, flanked by heterotypic FRT sequences. Here, we describe the procedures to perform FLPe recombinase-mediated cassette exchange (RMCE) using this line. The master cell line is transfected with a RMCE donor vector, which contains a promoterless Puromycin resistance, and with FLPe recombinase. Application of both a positive (Puromycin) and negative (FIAU) selection program leads to the selection of RMCE without random integrations. RMCE generates fully characterized pluripotent polyclonal transgenic lines in 15 d with 100% efficiency. Despite the recently described limitations of the AAVS1 locus, the ease of the system paves the way for hPSC transgenesis in isogenic settings, is necessary for comparative studies, and enables semi-high-throughput genetic screens for gain/loss of function analysis that would otherwise be highly time consuming

Epigenetic Induction of Definitive and Pancreatic Endoderm Cell Fate in Human Fibroblasts

Reprogramming can occur by the introduction of key transcription factors (TFs) as well as by epigenetic changes. We demonstrated that histone deacetylase inhibitor (HDACi) Trichostatin A (TSA) combined with a chromatin remodeling medium (CRM) induced expression of a number of definitive endoderm and early and late pancreatic marker genes. When CRM was omitted, endoderm/pancreatic marker genes were not induced. Furthermore, treatment with DNA methyltransferase inhibitor (DNMTi) 5-azacytidine (5AZA) CRM did not affect gene expression changes, and when 5AZA was combined with TSA, no further increase in gene expression of endoderm, pancreatic endoderm, and endocrine markers was seen over levels induced with TSA alone. Interestingly, TSA-CRM did not affect expression of pluripotency and hepatocyte genes but induced some mesoderm transcripts. Upon removal of TSA-CRM, the endoderm/pancreatic gene expression profile returned to baseline. Our findings underscore the role epigenetic modification in transdifferentiation of one somatic cell into another. However, full reprogramming of fibroblasts to β-cells will require combination of this approach with TF overexpression and/or culture of the partially reprogrammed cells under β-cell specific conditions

Generation of hepatocyte- and endocrine pancreatic-like cells from human induced endodermal progenitor cells

Multipotent Adult Progenitor Cells (MAPCs) are one potential stem cell source to generate functional hepatocytes or β-cells. However, human MAPCs have less plasticity than pluripotent stem cells (PSCs), as their ability to generate endodermal cells is not robust. Here we studied the role of 14 transcription factors (TFs) in reprogramming MAPCs to induced endodermal progenitor cells (iENDO cells), defined as cells that can be long-term expanded and differentiated to both hepatocyte- and endocrine pancreatic-like cells. We demonstrated that 14 TF-iENDO cells can be expanded for at least 20 passages, differentiate spontaneously to hepatocyte-, endocrine pancreatic-, gut tube-like cells as well as endodermal tumor formation when grafted in immunodeficient mice. Furthermore, iENDO cells can be differentiated in vitro into hepatocyte- and endocrine pancreatic-like cells. However, the pluripotency TF OCT4, which is not silenced in iENDO cells, may contribute to the incomplete differentiation to mature cells in vitro and to endodermal tumor formation in vivo. Nevertheless, the studies presented here provide evidence that reprogramming of adult stem cells to an endodermal intermediate progenitor, which can be expanded and differentiate to multiple endodermal cell types, might be a valid alternative for the use of PSCs for creation of endodermal cell types

Efficient Recombinase-Mediated Cassette Exchange in hPSCs to Study the Hepatocyte Lineage Reveals AAVS1 Locus-Mediated Transgene Inhibition

Tools for rapid and efficient transgenesis in "safe harbor" loci in an isogenic context remain important to exploit the possibilities of human pluripotent stem cells (hPSCs). We created hPSC master cell lines suitable for FLPe recombinase-mediated cassette exchange (RMCE) in the AAVS1 locus that allow generation of transgenic lines within 15 days with 100% efficiency and without random integrations. Using RMCE, we successfully incorporated several transgenes useful for lineage identification, cell toxicity studies, and gene overexpression to study the hepatocyte lineage. However, we observed unexpected and variable transgene expression inhibition in vitro, due to DNA methylation and other unknown mechanisms, both in undifferentiated hESC and differentiating hepatocytes. Therefore, the AAVS1 locus cannot be considered a universally safe harbor locus for reliable transgene expression in vitro, and using it for transgenesis in hPSC will require careful assessment of the function of individual transgenes

cis-Bromidobis(ethylene-1,2-diamine)(2-methylpropan-1-amine)cobalt(III) dibromide

In the title compound, [CoBr(C2H8N2)2(C4H11N)]Br2, the cobalt(III) ion has a distorted octahedral coordination environment and is ligated by three N atoms and a bromine ion in the equatorial plane, and by two N atoms occupying the axial positions. In the crystal, the complex cation and the two Br− counter-anions are linked by N—H...Br hydrogen bonds, forming a supramolecular framework

Enhancement of Fracture Toughness Characteristics of Woven Jute Fabric Mat Reinforced Epoxy Composites with SiC Fillers

Natural fiber reinforced composites are attracting the attention of industries toward a sustainable and environment-friendly material for the future demands. The main drawback pertaining to the use of natural fiber composites lies in its inferior strength in comparison to its counterpart synthetic fibers. This research focuses on improving the fracture toughness of woven jute fabric mat with different orientations (0°, 30° and 45°) in epoxy resin by incorporating fillers and tested in universal tensile testing machine. Silicon carbide (SiC) particles of three sizes 26 µm, 54 µm and 72 µm were incorporated in the matrix in different weight ratios (5%, 10% and 15%). The fracture toughness of the jute/epoxy composites increases with the increase in the amount of SiC particles up to 10% addition and then it shows decreasing trend with further addition. The fracture toughness values show upward trend when the fiber orientation changed from 0° to 45°. Fractography of the tested specimen captured by scanning electron microscope which reveals the fracture of fibers and micro cracks in the epoxy matrix due to the applied load. Analysis of variance (ANOVA) is also conducted for predicting the most influencing parameters on the fracture toughness

Spectroscopic (FT-IR, FT-Raman and NMR) and NBO analysis of 3, 4-dimethylanisole by density functional method

236-249Combined experimental and theoretical studies have been conducted on the molecular structure and vibrational, spectra of 3, 4-dimethyl anisole (DMA). The FT-IR and FT-Raman spectra of DMA have been recorded in the solid phase. The molecular geometry and vibrational frequencies of DMA in the ground state have been calculated by using the ab-initio Hartree-Fock (HF) and density functional methods (B3LYP) invoking 6-31+G (d,p) basis set. The optimized geometric bond lengths and bond angles obtained by HF method shows best agreement with the experimental values. Comparison of the observed fundamental vibrational frequencies of DMA with calculated results by HF and density functional methods indicates that B3LYP is superior to the scaled HF approach for molecular vibrational problems. The difference between the observed and scaled wave number values of most of the fundamental is very small. The thermodynamic functions and atomic change of the title compound has also been performed at HF/B3LYP/6-31+G(d,p) level of theories. A detailed interpretation of the FT-IR, FT-Raman, NMR spectra of DMA has also been reported. The theoretical spectrograms for infrared and Raman spectra of the title molecule have been constructed. The thermodynamic function of the title compound has also been performed at HF/6-31+G (d,p) and B3LYP/6-31+G (d,p) level of theories. Natural bond orbital analysis has been carried out to explain the change transfer or delocalization of change due to the intra-molecular interactions. Energy of the highest occupied molecular (HOMO) orbital and lowest unoccupied (LUMO) molecular orbital have been predicted

Mechanical and Corrosion Behavior of Al7075 (Hybrid) Metal Matrix Composites by Two Step Stir Casting Process

Abstract This paper investigates the mechanical properties and corrosion behavior of Metal Matrix Composites prepared using Al7075 alloy as a matrix, Silicon Carbide and Titanium Carbide as reinforcement particles. Two step stir casting process was used to fabricate the composites by varying volume fractions of Silicon Carbide and Titanium Carbide (0 to 15 vol. %). Microstructural analysis, mechanical and corrosion behavior were used to evaluate the performance of the composites. Uniform distribution of reinforcement particle was observed through optical photomicrographs. Vickers micro hardness tests were performed and the hardness values were increased with an increase in reinforcement from 0 to 15 vol. %. The tensile strength of the 10 vol. % of aluminum hybrid matrix composite was better than that of the base alloy. In 3.5% NaCl solution, it was observed that the 15 vol. % of the aluminum hybrid matrix composite have higher corrosion resistance in comparison the base alloy