Structure-function studies of HNF1A (MODY3) gene mutations in South Indian patients with monogenic diabetes.

Maturity-onset diabetes of the young (MODY) is a genetically heterogeneous monogenic form of diabetes characterized by onset of diabetes below 25 years of age, autosomal dominant mode of inheritance and primary defect in insulin secretion. Mutations in the gene (HNF1A) encoding transcription factor hepatocyte nuclear factor 1A (HNF-1A) results in one of the most common forms of MODY (MODY3). HNF-1A is mainly enriched in pancreatic ß-cells and hepatocytes and important for organ development and normal pancreatic function. We here report on the functional interrogation of eight missense HNF1A mutations associated with MODY3 in South Indian subjects, and the contributing effect of common variant (S487N) within HNF1A. Of the eight mutations, three mutations (p.R171G, p.G245R and p.R263H), in particular, affected HNF-1A function in transfected HeLa cells by reducing both transcriptional activity and nuclear localization, possibly due to disruption of the integrity of the three dimensional structure. The common variant p.S487N contributed further to the loss-of-function of p.R271Q (p.R271Q+p.S487N double mutant), in vitro, on both activity and localization. Our data on the first functional study of HNF1A mutations in South India subjects confers that the defect of the HNF-1A mutant proteins are responsible for MODY3 diabetes in these patients.

Caenorhabditis elegans-based in vivo screening of bioactives from marine sponge-associated bacteria against Vibrio alginolyticus.

AIM: To establish Caenorhabditis elegans based in vivo method for screening bioactives from marine sponge associated bacteria (SAB) against Vibrio species. METHODS AND RESULTS: About 256 SAB isolates were screened for their ability to rescue C. elegans infected with Vibrio species. The chloroform extract of the positive isolate was subjected to column fractionation and purity of the active fraction was analysed using HPLC. Further, the components were elucidated using GC/MS. The active fraction was tested for its in vivo rescue activity, antibacterial and anti-QS activity. In vivo colonization reduction and biofilm inhibition efficiency were assessed using GFP-tagged V. alginolyticus using confocal laser scanning microscopy (CLSM). The ability of the active fraction in modulating expression of V. alginolyticus quorum sensing (QS) regulators luxT and lafK was measured using real-time PCR. The results indicated that the chloroform extract of SAB4.2 displayed significant rescue activity against V. alginolyticus by inhibiting the QS pathway. HPLC analysis of the active fraction revealed a single major peak and GC/MS analysis suggested Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl) as the major constituent. The potent bacterial isolate was identified as Alcaligenes faecalis. CONCLUSIONS: In vivo screening using C. elegans identified a marine isolate that inhibits the virulence of V. alginolyticus by interrupting the QS pathway. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides a C. elegans based in vivo screening method for identifying bioactives from natural resources by overcoming the disadvantages of traditional in vitro plate assays.

Cloning of the human aspartoacylase cDNA and a common missense mutation in Canavan disease.

Canavan disease, an autosomal recessive leukodystrophy, is caused by deficiency of aspartoacylase and accumulation of N-acetylaspartic acid in brain. We have cloned the human aspartoacylase (ASP) cDNA spanning 1,435 basepairs, and show that the isolated cDNA expresses aspartoacylase activity in bacteria. Furthermore, an A to C base change, at nucleotide 854, has been found in 85% of the 34 Canavan alleles tested so far. This base change results in a missense Glu285Ala mutation that is predicted to be part of the catalytic domain of aspartoacylase. The data suggest that the catalytic centre of aspartoacylase involves a triad of Ser, His and Glu residues. Our findings have implications for diagnosis and screening of Canavan disease.

Performance of magnetic dipole contribution on ferromagnetic non-Newtonian radiative MHD blood flow: An application of biotechnology and medical sciences.

Casson flow ferromagnetic liquid blood flow over stretching region is studied numerically. The domain is influence by radiation and blood flow velocity and thermal slip conditions. Blood acts an impenetrable magneto-dynamic liquid yields governing equations. The conservative governing nonlinear partial differential equations, reduced to ODEs by the help of similarity translation technique. The transport equations were transformed into first order ODEs and the resultant system are solved with help of 4th order R-K scheme. Performing a magnetic dipole with a Casson flow across a stretched region with Brownian motion and Thermophoresis is novelty of the problem. Significant applications of the study in some spheres are metallurgy, extrusion of polymers, production in papers and rubber manufactured sheets. Electronics, analytical instruments, medicine, friction reduction, angular momentum shift, heat transmission, etc. are only few of the many uses for ferromagnetic fluids. As ferromagnetic interaction parameter value improves, the skin-friction, Sherwood and Nusselt numbers depreciates. A comparative study of the present numerical scheme for specific situations reveals a splendid correlation with earlier published work. A change in blood flow velocity magnitude has been noted due to Casson parameter. Increasing change in blood flow temperature noted due to Casson parameter. Skin-friction strengthened and Nusselt number is declined with Casson parameter. The limitation of current work is a non-invasive magnetic blood flow collection system using commercially available magnetic sensors instead of SQUID or electrodes.

On the perturbation of the H-bonding interaction in ethylene glycol clusters upon hydration.

Ab initio and density functional methods have been employed to study the structure, stability, and spectral properties of various ethylene glycol (EG(m)) and ethylene glycol-water (EG(m)W(n)) (m = 1-3, n = 1-4) clusters. The effective fragment potential (EFP) approach was used to explore various possible EG(m)W(n) clusters. Calculated interaction energies of EG(m)W(n) clusters confirm that the hydrogen-bonding interaction between EG molecules is perturbed by the presence of water molecules and vice versa. Further, energy decomposition analysis shows that both electrostatic and polarization interactions predominantly contribute to the stability of these clusters. It was found from the same analysis that ethylene glycol-water interaction is predominant over the ethylene glycol-ethylene glycol and water-water interactions. Overall, the results clearly illustrate that the presence of water disrupts the ethylene glycol-ethylene glycol hydrogen bonds.

Evaluation of Luteolin in the Prevention of N-nitrosodiethylamine-induced Hepatocellular Carcinoma Using Animal Model System.

Hepatocellular carcinoma (HCC) is one of the commonest tumors worldwide. The treatment of HCC is vital for disease diagnosis and prognosis, as the liver is the most important organ controlling metabolic functions. Now-a-days, western folklore medicines are largely dependent on the phyto compounds which are highly effective in therapy and with low side effects. Luteolin is a flavonoid (3,4,5,7-Tetrahydro flavones) possess anti-inflammatory, anticancer and anti allergic property. The present study evaluates the efficacy of luteolin against N-nitrosodiethylamine (DEN) induced HCC in albino rats. In the highlight of the above, luteolin was evaluated for its efficacy against DEN induced HCC in male Wistar albino rats. The Biochemical parameters such as tissue damaging enzymes viz., AST, ALP, LDH and γ-GT, enzymatic antioxidants viz., SOD, CAT, GSH and GPx and histopathological changes have been estimated. The tissue damaging enzymes were found to be high in DEN alone treated group whereas the enzymatic antioxidants decreased destructively. Severe lesions and cirrhosis were observed in the toxin (DEN alone) treated group. The luteolin treated DEN group altered the tissue damaging enzymes and the enzymatic antioxidants. The damaged lesion in the histoarchitecture of DEN treated rat liver was almost completely restored. Finally this study strongly demonstrates that luteolin has potent curative property against HCC in albino rats.

Surface alloying of FeCoCrNiMn particles on Inconel-718 using plasma-transferred arc technique: microstructure and wear characteristics.

Inconel-718 (IN-718) is a commonly used nickel-based superalloy in various fields, such as gas turbine and power generation applications. However, the lower wear and oxidation resistance hinder their wide usage. In this work, FeCoCrNiMn particles were mechanically ball-milled and preplaced on the IN-718 substrate. Then, the preplaced FeCoCrNiMn particles were scanned by heat source using plasma-transferred arc (PTA) technique. The effect of PTA alloying on the phase changes, microstructure, nanohardness and wear resistance has been investigated. The result showed that the PTA region contained different phases, such as FCC, BCC and intermetallic. No cracks were observed in the PTA alloyed region. Moreover, the porous free structure was viewed in the alloyed region, which revealed that the PTA alloying process was effectively used to perform the alloying process. More hard phases, such as NiFe, CoMn, Cr9Mn25Ni21, MnNi, FeCo, FeMn and MnCo, were formed on the PTA-alloyed region. The obtained wear rate of the substrate specimen at 30 N applied load is 2.45 × 10-3 mm3 m-1 and 1.79 × 10-3 mm3 m-1 for the PTA specimen. Similarly, the wear rate of the substrate specimen at 50 N is 5.38 × 10-3 mm3 m-1 and for the PTA sample, it is 2.29 × 10-3 mm3 m-1. The PTA specimen showed lower CoF than the substrate specimen due to increased surface hardness and minimum deformation of asperities. The primary wear type was mildly abrasive, accompanied by slight oxidative wear. Oxygen reacted with the surface alloying elements and formed different oxides, such as CoO, Cr2O3, MnO2, Mn2O3, Mn3O4, FeO and Fe2O3. These dense oxidation films covered the working surface and enhanced the wear resistance. The worn-out PTA surface showed that the wear scar depths were shallow and lower than the substrate, and reduced the roughness.

Nonprescription sale of schedule H1 antibiotics in a city of South India.

CONTEXT: The acquisition of antibiotic without a prescription by the general population is a typical practice found in community pharmacies across India, which is a notable contributor of antimicrobial resistance. According to the present regulation in India, sale of certain antimicrobials included in schedule H1 without prescription is unlawful. In this contest, a program was organized by the Drug Control Administration, Government of Andhra Pradesh, to educate pharmacists regarding schedule H1. AIMS: The aim of our study is to assess the impact of the program on the rate of antibiotics dispensed at community pharmacies. SETTINGS AND DESIGN: A cross-sectional study was designed to investigate the nonprescription sale of antibiotics, from September to December 2018 through 200 community pharmacies located in and around Guntur city located in the state of Andhra Pradesh in India. SUBJECTS AND METHODS: A simulated client methodology was used in this study. A total of 3 female actors including an author of this present study are prior trained to present a standardized simulation of clinical conditions (sore throat, urinary tract infection, cold, and fever) to the pharmacist at the community pharmacies. STATISTICAL ANALYSIS USED: Microsoft excel sheet was used for data analysis. RESULTS: The simulated patients successfully obtained antibiotic from 78% pharmacies with the highest rate of urinary tract infection when compared to other conditions. Pharmacists who objected to dispense antibiotics (22%) are found in developed locations in the city and appeared well qualified. CONCLUSIONS: The present study revealed that the antibiotics are continued to be sold without prescription even after the education program on schedule H1. The deregulation of the act is definitely the problem to be addressed by the government.

2-(2,4-Dichloro-phen-yl)-9-phenyl-2,3-di-hydro-thieno[3,2-b]quinoline.

In the title compound, C(23)H(15)Cl(2)NS, the quinoline system is almost planar [r.m.s. deviation = 0.013 (2) Å]. The phenyl group is disordered over two positions with site occupancies of 0.55 and 0.45, and is oriented in a nearly perpendicular configuration to the quinoline ring [the dihedral angles between the quinoline ring and the major and minor disordered components of the phenyl ring are 81.8 (2) and 71.6 (2)°, respectively]. The dihydro-thiene ring adopts an envelope conformation. The dihedral angle between the chloro-phenyl ring and the quinoline system is 79.32 (1)°. In the crystal weak C-H⋯π inter-actions occur.

Studies on the antifungal and serotonin receptor agonist activities of the secondary metabolites from piezotolerant deep-sea fungus Ascotricha sp.

The potent antifungal agent sesquiterpenes and serotonin 5-HT2C agonist ascotricin were produced by a newly isolated deep-sea fungus Ascotricha sp. This fungus was isolated from deep-sea sediment collected at a depth of 1235 m and characterized. Piezotolerance was successfully tested under high pressure-low temperature (100 bar pressure and 20ºC) microbial cultivation system. Production of secondary metabolites was enhanced at optimized culture conditions. The in-vivo antifungal activity of sesquiterpenes was studied using the Caenorhabditis elegans - Candida albicans model system. The sesquiterpenes affected the virulence of C. albicans and prolonged the life of the host C. elegans. These findings suggest that sesquiterpenes are attractive antifungal drug candidates. The 5-HT2C receptor agonist is a potential target for the development of drugs for a range of central nervous system disorders. The interaction of 5-HT2C agonist ascotricin with the receptor was studied through bioinformatic analysis. The in silico molecular docking and molecular dynamic simulation studies demonstrated that they fit into the serotonin 5-HT2C active site and the crucial amino acid residues involved in the interactions were identified. To our knowledge, this is first report of in vivo antifungal analysis of sesquiterpenes and in silico studies of serotonin 5-HT2C receptor-ascotricin complex.

The C/EBPδ protein is stabilized by estrogen receptor α activity, inhibits SNAI2 expression and associates with good prognosis in breast cancer.

Hypoxia and inflammatory cytokines like interleukin-6 (IL-6, IL6) are strongly linked to cancer progression, and signal in part through the transcription factor Ccaat/enhancer-binding protein δ (C/EBPδ, CEBPD), which has been shown to promote mesenchymal features and malignant progression of glioblastoma. Here we report a different role for C/EBPδ in breast cancer. We found that the C/EBPδ protein is expressed in normal breast epithelial cells and in low-grade cancers. C/EBPδ protein (but not mRNA) expression correlates with estrogen receptor (ER+) and progesterone receptor (PGR) expression and longer progression-free survival of breast cancer patients. Specifically in ER+ breast cancers, CEBPD-but not the related CEBPB-mRNA in combination with IL6 correlated with lower risk of progression. Functional studies in cell lines showed that ERα promotes C/EBPδ expression at the level of protein stability by inhibition of the FBXW7 pathway. Furthermore, we found that C/EBPδ attenuates cell growth, motility and invasiveness by inhibiting expression of the SNAI2 (Slug) transcriptional repressor, which leads to expression of the cyclin-dependent kinase inhibitor CDKN1A (p21CIP1/WAF1). These findings identify a molecular mechanism by which ERα signaling reduces the aggressiveness of cancer cells, and demonstrate that C/EBPδ can have different functions in different types of cancer. Furthermore, our results support a potentially beneficial role for the IL-6 pathway specifically in ER+ breast cancer and call for further evaluation of the role of intra-tumoral IL-6 expression and of which cancers might benefit from current attempts to target the IL-6 pathway as a therapeutic strategy.

Hydroxopentaamminechromium(III) promoted phosphorylation of bovine serum albumin: its potential implications in understanding biotoxicity of chromium.

Evidence for chromium(III) induced phosphorylation of a biomarker protein bovine serum albumin (BSA) is presented. Radiolabelled adenosine 5'-triphosphate (ATP) was reacted with BSA in the presence of various Cr(III) salts. While [Cr(NH3)5(H2O)]3+ brought about phosphorylation of BSA, several Cr(III) complexes, viz. [Cr(bpy)3]3+, [Cr(phen)3]3+, [Cr(en)3]3+, [Cr(salen)(H2O)2]+ and [Cr(salprn)(H2O)2]+, did not phosphorylate BSA. The Cr(III) mediated the transfer of gamma- and alpha-phosphates but not the adenine and the sugar moieties of the ATP molecule to BSA. The observed stoichiometry was 0.75 mol Pi to mol BSA for the gamma-phosphate and 0.5 mol Pi to mol BSA for the alpha-phosphate of ATP. The presence of serine phosphate and threonine phosphate was detected in the hydrolysate of phosphorylated BSA by means of comparison of Rf values with authentic samples of phosphoserine and phosphothreonine after chromatographic separation and autoradiography. [Cr(NH3)5(H2O)]3+ at pH 7.4 is known to exist as the conjugate base [Cr(NH3)5(OH)]2+ and is capable of ligand substitution involving metal-oxygen bond retention. Such anation reaction of [Cr(NH3)5(OH)]2+ with ATP subsequently leads to the esterification of alcoholic hydroxyl groups of serine and threonine of BSA. Possible consequences of chromium(III) induced in vivo phosphorylation of proteins are discussed.

Synthesis and characterization of polyethylene glycol (PEG) coated Fe3O4 nanoparticles by chemical co-precipitation method for biomedical applications.

Polyethylene glycol (PEG) coated Fe3O4 nanoparticles were synthesized by chemical co-precipitation method. With polyethylene glycol (PEG) as a stabilizer and dispersant. The X-ray diffraction and selected area electron diffraction (SAED) results show that the cubic inverse spinel structure of pure phase polycrystalline Fe3O4 was obtained. The scanning electron microscopy (SEM) and field emission transmission electron microscopy (FE-TEM) results exhibited that the resulted Fe3O4 nanoparticles were roughly spherical in shape with narrow size distribution and homogenous shape. Fourier transform infrared spectroscopy (FT-IR) results suggested that PEG indicated with Fe3O4 via its carbonyl groups. Results of vibrating sample magnetometer (VSM) indicated that the prepared Fe3O4 nanoparticles exhibit superparamagnetic behavior and high saturation magnetization at room temperature. Such Fe3O4 nanoparticles with favorable size and tunable magnetic properties are promising biomedical applications.

Spectroscopic investigation on the production of clay bricks with SCBA waste.

In this paper, the effect of sugarcane bagasse ash (SCBA) addition to the brick making clay has been analyzed using spectroscopic techniques. For that, mixtures of brick making clay (BMC) with sugarcane bagasse ash (SCBA) in proportions of 0-20 wt.% were hydraulic uniaxially pressed and sintered at temperatures of 800-1100 °C. The partial replacement of the brick making clay with SCBA was studied with chemical and mineralogical analyzes (XRF and X-ray diffraction). The quantitative estimation of minerals was made by FTIR analysis. The results of FT-IR reveal that kaolinite, quartz, and lignin are predominant, whereas, cellulose and calcite are in moderate levels. In addition, magnetite and hematite are found in trace level. The overall results reveal that the brick making clay substituted with 15 wt.% of SCBA can open up a new path for the fabrication of quality bricks at low cost.

Allelic distribution of nine short tandem repeat (STR), HLA-DQA1, and polymarker loci in an Omani sample population.

Allele frequency distributions of nine short tandem repeat (STR) loci, D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D7S820, and D13S317, HLA-DQA1 and polymarker (PM) loci were studied in a sample population from Sultanate of Oman, Middle East. Blood samples were collected from 162 unrelated individuals. For all STR, HLA-DQA1 and PM loci, no deviations from Hardy-Weinberg equilibrium, based on the exact test, were observed. The most discriminating loci were D18S51 (PD=0.966) and FGA (PD=0.964), while the least informative locus is GYPA (PD=0.585). The allele frequency data may be useful in forensic case work.

Typing of eight short tandem repeat (STR) loci in a Saudi Arabian population.

Allele frequency data for eight short tandem repeat (STR) loci, HUMF13A01, HUMFESFPS, HUMF13B, HUMLPL, HUMCSF1PO, HUMTPOX, HUMTHO1 and HUMvWA, were obtained for unrelated individuals in a Saudi Arabian population. All loci, except F13B (P = 0.037) and LPL (P = 0.035), meet Hardy-Weinberg expectations, based on the exact test. The most informative locus is HUMvWA (PD = 0.936) and the least discriminating is the HUMTPOX locus (PD = 0.820). There was only one observation of a departure from expectation from pairwise locus comparisons. These data can be used for estimating the frequency of STR profiles in a Saudi Arabian population.

Partial purification of maturation-promoting factor from catfish, Clarias batrachus: identification as the histone H1 kinase and its periodic activation.

Maturation-promoting factor (MPF) has been demonstrated in the 100,000 g supernatant of 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one (17 alpha, 20 beta-DP)-induced catfish, Clarias batrachus oocytes using DEAE-cellulose and sephadex G-200 chromatography. Partially purified MPF molecule eluted as a single peak on sephadex G-200 with molecular mass of approximately 200 kDa in native PAGE. SDS-PAGE analysis showed the presence of five proteins of 32, 34, 45, 46 and 48 kDa. Antibody against the PSTAIR sequence of p34cdc2 recognized 32 and 34 kDa proteins, whereas rabbit anti-cyclin B1 and B2 crossreacted with 46 and 48 kDa proteins, respectively. Cyclin B was absent in immature oocytes and appeared after 7 h of 17 alpha, 20 beta-DP stimulation, coinciding with the histone H1 kinase (HH1K) activity and start of germinal vesicle breakdown (GVBD). Our data indicate that C. batrachus MPF is a complex of cdc2 kinase and cyclin B molecules. A close relationship between HH1K activity and catfish oocyte maturation has been demonstrated using cycloheximide, cytochalasin B and colchicine. HH1K activation was inhibited by cycloheximide, while cytochalasin B and colchicine were ineffective. These finding suggests that the activation of HH1K depends on protein synthesis, whereas disruption of microfilaments influences only nucleus migration without effect on GVBD or HH1K activation. An increase of phosphorylated proteins after activation of catfish oocytes with 17 alpha, 20 beta-DP has also been observed.

Distribution of D1S80 alleles in the Bahrainian population.

This study demonstrates that the locus D1S80 is highly polymorphic in the Bahrainian population. There were 24 different D1S80 alleles and 51 distinct genotypes observed in 198 Bahrainians. There was one allele observed that was smaller than the 14 repeat allele. This data set meets the Hardy-Weinberg expectations (HWE) and could be a useful marker for parentage testing and forensic applications.

Ellagic acid normalizes mitochondrial outer membrane permeabilization and attenuates inflammation-mediated cell proliferation in experimental liver cancer.

Despite great advances in our understanding of the molecular causes of liver cancer, significant gaps still remain in our knowledge of the disease pathogenesis and development of effective strategies for early diagnosis and treatment. The present study was conducted to evaluate the chemopreventive activity of ellagic acid (EA) against experimental liver cancer in rats. This is the first report that implies a possible role of EA in controlling liver cancer through activation of mitochondrial outer membrane permeability via activating proteins such as Bax, bcl-2, cyt-C, and caspase-9, which play important roles in apoptosis. Downregulation of NF-κB, cyclin D1, cyclin E1, matrix metalloproteinases (MMP)-2, MMP-9, and proliferating cell nuclear antigen (PCNA) were noted in EA-treated experimental rats and controlled inflammation mediated liver cancer when compared to the diethylnitrosamine (DEN)-induced group. Transmission electron microscopy (TEM) analysis of the livers of experimental rats demonstrated that EA treatment renovated its internal architecture. Overall, these results demonstrate the value of molecular approaches in identifying the potential role of EA as an effective chemopreventive agent.

Molecular dynamics simulations of retinoblastoma protein.

Tumor suppressor proteins play a crucial role in cell cycle regulation. Retinoblastoma protein (pRB) is one among them which regulates G1-S transition by binding with transcription factors. The activity of pRB is deregulated by cyclin dependent kinases-mediated hyper-phosphorylation and also due to cancer-derived mutations. In addition, it is also deactivated by binding of viral onco-proteins such as large T antigen, E1A, and E7. These viral proteins initially recognize pRB through their conserved LxCxE motif and facilitate dissociation of preexisting pRB-E2F complex. Based on these features, molecular dynamics (MD) simulation is performed for four different states of pRB for which the crystal structure is available. The unliganded/apo form and complex forms with E2F and E7 peptides reveal the molecular mechanism behind the activation and inactivation of pRB. In addition, the ternary complex of pRB with both E7 and E2F (for which no crystal structure is available) is modeled and simulated to understand the influence of binding of one ligand on the other. The variations in the three major factors such as conformational changes, inter- and intra-molecular interactions, and binding free energies between the apo and complex forms confirm the possibility for designing a small molecule inhibitor to inhibit pRB-E7 interactions without altering the prebound E2F. The present study deals with the molecular modeling and MD simulations of pRB in free and ligand-bound forms and confirms that pRB could be a valid target for the anticancer drug design when the cancer is induced by the viral onco-proteins and forms a clear base for designing E7 antagonists.