Science attitudes and knowledge among preclinical medical students in Pokhara, Nepal

Knowledge of science and the scientific method are important in learning about and using evidence-based medicine in practice. Courses in research methodology have been introduced for both medical students and practicing doctors. In Pokhara, the basic science subjects are taught in an integrated manner during the first four semesters of the undergraduate medical course. Studies on students' attitudes towards and knowledge of science are lacking in medical colleges in Nepal. Hence the study was carried out to obtain information on students' attitude towards and knowledge of science and scientific methodology among preclinical medical students and note the association, if any, of students' attitudes and their demographic characteristics. The study was carried out in March 2005 among the students of the first four semesters at the Manipal College of Medical Sciences, Pokhara, Nepal using a questionnaire developed by Hren and coworkers. Two hundred and twenty students (overall response rate 73.3%) successfully completed the questionnaire. Seventy-five respondents were Nepalese, 115 were Indians, 27 were Sri Lankans and 3 belonged to other nationalities. The X +/- SD total attitude score was 147.4 +/- 10.8 (neutral score 135). The X +/- SD scores on the subscales, value of science to humanity, value of scientific methodology and value of science to medicine were 51.3 +/- 5.4, 39.6 +/- 3.7 and 58.5 +/- 5.9 (neutral scores were 36, 51 and 48 respectively). The knowledge score measured using a set of 8 multiple choice questions was 3.3 +/- 1.4. The attitude scores were lower and the knowledge score was comparable to that reported previously in a study in Croatia but higher than that reported from Southeast Europe.     

An analog of BIX-01294 selectively inhibits a family of histone H3 lysine 9 Jumonji demethylases

BIX-01294 and its analogs were originally identified and subsequently designed as potent inhibitors against histone H3 lysine 9 (H3K9) methyltransferases G9a and G9a-like protein. Here, we show that BIX-01294 and its analog E67 can also inhibit H3K9 Jumonji demethylase KIAA1718 with half-maximal inhibitory concentrations in low micromolar range. Crystallographic analysis of KIAA1718 Jumonji domain in complex with E67 indicated that the benzylated six-membered piperidine ring was disordered and exposed to solvent. Removing the moiety (generating compound E67-2) has no effect on the potency against KIAA1718 but, unexpectedly, lost inhibition against G9a-like protein by a factor of 1500. Furthermore, E67 and E67-2 have no effect on the activity against histone H3 lysine 4 (H3K4) demethylase JARID1C. Thus, our study provides a new avenue for designing and improving the potency and selectivity of inhibitors against H3K9 Jumonji demethylases over H3K9 methyltransferases and H3K4 demethylases.     

Pathogenic and molecular characterisations of pigeonpea wilt pathogen Fusarium udum

Thirty two pathogenic isolates of Fusarium udum from different pigeonpea growing areas in India were studied for pathogenic and molecular variability. Pathogenic variability was tested on 12 pigeonpea differential genotypes, which revealed prevalence of five variants in F. udum. The amount of genetic variation was evaluated by Polymerase Chain Reaction (PCR) amplification with 20 random amplified polymorphic DNA (RAPD) markers and nine microsatellite markers. All amplifications revealed scorable polymorphisms among the isolates, and a total of 137 polymorphic fragments were scored for the RAPD markers and 16 alleles for the simple sequence repeat (SSR) markers. RAPD primers showed 86% polymorphism. Genetic similarity was calculated using Jaccard's similarity coefficient and cluster analysis was used to generate a dendrogram showing relationships between them. Isolates could be grouped into three subpopulations based on molecular analysis. Results indicated that there is high genetic variability among a subpopulation of F. udum as identified by RAPD and SSR markers and pathogenicity on differential genotypes.     

Spectroscopic characterization of the NO adduct of hydroxylamine oxidoreductase

Hydroxylamine oxidoreductase (HAO) from the autotrophic nitrifying bacterium Nitrosomonas europaea catalyzes the oxidation of NH2OH to NO2-. The enzyme contains eight hemes per subunit which participate in catalysis and electron transport. NO is found to bind to the enzyme and inhibit electron flow to the acceptor protein, cytochrome c554. NO is found to oxidize either partially or fully reduced HAO, but NO will not reduce ferric HAO. Since NO can be reduced but not oxidized to product by HAO, NO is not considered to be a long-lived intermediate in the catalytic mechanism. Substrate oxidation occurs in the presence of bound NO or cyanide, suggesting a second interaction site for substrate with HAO and providing a means for recovery of the NO-inhibited form of the enzyme. Upon addition of NO to oxidized HAO, the integer-spin EPR signal from the active site vanishes, an IR band from NO appears at 1920 cm(-1), and a diamagnetic quadrupole iron doublet appears in M?ssbauer spectroscopy with delta = 0.06 mm/s and DeltaEq = 2.1 mm/s. The NO stretching frequency and M?ssbauer parameters are characteristic of an [FeNO]6 heme complex. New M?ssbauer data on ferric myoglobin-NO are also presented for comparison. The results indicate that NO binds to heme P460 and that the loss of the integer-spin EPR signal is due to the conversion of heme P460 to a diamagnetic S = 0 state and concomitant loss of magnetic interaction with neighboring heme 6. In previous studies where the heme P460-heme 6 interaction was affected by substrate or cyanide binding, a signal attributable to heme 6 was not observable. In contrast, in this work, the NO-induced loss of the signal is accompanied by the appearance of a previously unobserved large g(max) (or HALS) low-spin EPR signal from heme 6.     

Variation in the Durations of the Photoperiod-sensitive and Photoperiod-insensitive Phases of Development to Flowering Among Eight Maturity Isolines of Soyabean [Glycine max (L.) Merrill]

In soyabean [Glycine max (L.) Merrill] the period between sowingand flowering is comprised of three successive developmentalphases—pre-inductive, inductive and post-inductive—inwhich the rate of development is affected, respectively, bytemperature only, by photoperiod and temperature, and then againby temperature only. A reciprocal-transfer experiment (carriedout at a mean temperature of 25°C) in which cohorts of plantswere transferred successively between short and long photoperiodsand vice-versa showed that eight combinations of three pairsof maturity alleles (E₁/e₁, E₂ /e₂, E₃ /e₃) had their greatesteffect on the duration of the inductive phase in long days.This phase was increased with the increasing photoperiod sensitivityinduced by the different gene combinations, and ranged fromabout 27 to 54 d according to genotype. In a short day regime(11·5 h d^-1), less than the critical photoperiod, theduration of the inductive phase was brief—requiring about11 photoperiodic cycles in the less photoperiod-sensitive genotypesand only about seven cycles in the more sensitive ones. Thematurity genes also affected the duration of the two photoperiod-insensitivephases; these durations were positively correlated with thephotoperiod-sensitivity potential of the gene combinations.The largest effect was on the pre-inductive phase which variedfrom 3 to 11 d, while the post-inductive phase varied from about13 to 18 d. As a consequence of these non-photoperiodic effectsof the maturity genes, even in the most inductive regimes (daylengthsless than the critical photoperiod) the time taken to flowerby the less photoperiod-sensitive combinations of maturity geneswas somewhat less than in the more sensitive combinations—rangingfrom about 28 to 34 d. The genetic and practical implicationsof these findings are discussed.Copyright 1994, 1999 AcademicPress Glycine max (L.) Merrill, soyabean, maturity genes, isolines, flowering, photoperiod     

Spectroscopic characterization and identification of Pseudomonas fluorescens mediated metabolic products of Acid Yellow-9

Pseudomonas flourescens NCIM 2100 was obtained from NCL, Pune, India, that was adapted to growth on 4 amino 1-1 azo benzene 3,4-Disulfonic acid. This strains was tested by UV, ¹H NMR, and IR spectroscopy for its ability to degrade the dye which resulted in to sulfonated analogs namely p-amino benzene sulfonic acid sodium salt and 2,4 diamino benzene sulfonic acid sodium salt. These compound further changed to either 2,4 dihydroxy benzene sulfonic acid sodium salt or 2 amino 4 hydroxy benzene sulfonic acid sodium salt, or 4 amino 2 hydroxy benzene sulfonic acid sodium salt. These breakdown compounds were non toxic in nature.     

Dependence of the response of an amperometric biosensor formed in a micro flow channel on structural and conditional parameters

Comprehensive analysis of the behavior of an amperometric biosensor incorporated in a micro flow channel was conducted by changing the structural and conditional parameters. The device used in the characterization consisted of a thin-film three-electrode system and a silicone rubber flow channel. An enzyme, glucose oxidase, was immobilized either at the bottom of the silicone rubber flow channel or on the electrode substrate. The flow rate, concentration, position of the immobilized enzyme, and channel height were changed, and the changes in the output current and the conversion efficiency were examined. When the flow rate and/or the channel height decreased, the output current and the conversion efficiency significantly increased. The conversion efficiency also increased by decreasing the concentration. The tendency of the flow dependence was reversed when the position of the immobilized enzyme was changed from the silicone rubber side to the electrode substrate. In addition, the influence of l-ascorbic acid was reduced by placing additional working electrodes in the upper stream. l-Ascorbic acid was eliminated more effectively as the flow rate decreased and the area of the working electrode for elimination increased.     

Impact of template overhang-binding region of HIV-1 RT on the binding and orientation of the duplex region of the template-primer

Fingers domain of HIV-1 RT is one of the constituents of the dNTP-binding pocket that is involved in binding of both dNTP and the template-primer. In the ternary complex of HIV-1 RT, two residues Trp-24 and Phe-61 located on the β1 and β3, respectively, are seen interacting with N + 1 to N + 3 nucleotides in the template overhang. We generated nonconservative and conservative mutant derivatives of these residues and examined their impact on the template-primer binding and polymerase function of the enzyme. We noted that W24A, F61A, and F61Y and the double mutant (W24A/F61A) were significantly affected in their ability to bind template-primer and also to catalyze the polymerase reaction while W24F remained unaffected. Using a specially designed template-primer with photoactivatable bromo-dU base in the duplex region at the penultimate position to the primer terminus, we demonstrated that F61A, W24A, F61Y as well as the double mutant were also affected in their cross-linking ability with the duplex region of the template-primer. We also isolated the E–TP covalent complexes of these mutants and examined their ability to catalyze single dNTP incorporation onto the immobilized primer terminus. The E–TP covalent complexes from W24F mutant displayed wild-type activity while those from W24A, F61A, F61Y, and the double mutant (W24A/F61A) were significantly impaired in their ability to catalyze dNTP incorporation onto the immobilized primer terminus. This unusual observation indicated that amino acid residues involved in the positioning of the template overhang may also influence the binding and orientation of the duplex region of the template-primer. Molecular modeling studies based on our biochemical results suggested that conformation of both W24 and F61 are interdependent on their interactions with each other, which together are required for proper positioning of the +1 template nucleotide in the binary and ternary complexes.     

SPARC Suppresses Apoptosis of Idiopathic Pulmonary Fibrosis Fibroblasts through Constitutive Activation of ��-Catenin

Idiopathic pulmonary fibrosis (IPF) is a poorly understood progressive disease characterized by the accumulation of scar tissue in the lung interstitium. A hallmark of the disease is areas of injury to type II alveolar epithelial cells with attendant accumulation of fibroblasts in areas called fibroblastic foci. In an effort to better characterize the lung fibroblast phenotype in IPF patients, we isolated fibroblasts from patients with IPF and looked for activation of signaling proteins, which could help explain the exaggerated fibrogenic response in IPF. We found that IPF fibroblasts constitutively expressed increased basal levels of SPARC, plasminogen activator inhibitor-1 (PAI-1), and active β-catenin compared with control cells. Control of basal PAI-1 expression in IPF fibroblasts was regulated by SPARC-mediated activation of Akt, leading to inhibition of glycogen synthase kinase-3β and activation of β-catenin. Additionally, IPF fibroblasts (but not control fibroblasts) were resistant to plasminogen-induced apoptosis and were sensitized to plasminogen-mediated apoptosis by inhibition of SPARC or β-catenin. These findings uncover a newly discovered regulatory pathway in IPF fibroblasts that is characterized by elevated SPARC, giving rise to activated β-catenin, which regulates expression of downstream genes, such as PAI-1, and confers an apoptosis-resistant phenotype. Disruption of this pathway may represent a novel therapeutic target in IPF.