Isolation of potentially pathogenic Escherichia coli O157:H7 from the Ganges River

Escherichia coli serotype O157:H7 was detected among bacteria collected from the Ganges River. O157:H7 isolates tested positive for stx(1), stx(2), and eae gene sequences. Identification of potentially pathogenic isolates from extensively used source water indicates that O157:H7 may be a significant but as yet underacknowledged public health concern in India.     

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.     

KATP channel knockout worsens myocardial calcium stress load in vivo and impairs recovery in stunned heart

Gene knockout of the KCNJ11-encoded Kir6.2 ATP-sensitive K(+) (K(ATP)) channel implicates this stress-response element in the safeguard of cardiac homeostasis under imposed demand. K(ATP) channels are abundant in ventricular sarcolemma, where subunit expression appears to vary between the sexes. A limitation, however, in establishing the full significance of K(ATP) channels in the intact organism has been the inability to monitor in vivo the contribution of the channel to intracellular calcium handling and the superimposed effect of sex that ultimately defines heart function. Here, in vivo manganese-enhanced cardiac magnetic resonance imaging revealed, under dobutamine stress, a significantly greater accumulation of calcium in both male and female K(ATP) channel knockout (Kir6.2-KO) mice compared with sex- and age-matched wild-type (WT) counterparts, with greatest calcium load in Kir6.2-KO females. This translated, poststress, into a sustained contracture manifested by reduced end-diastolic volumes in K(ATP) channel-deficient mice. In response to ischemia-induced stunning, male and female Kir6.2-KO hearts demonstrated accelerated time to contracture and increased peak contracture compared with WT. The outcome on reperfusion, in both male and female Kir6.2-KO hearts, was a transient reduction in systolic performance, measured as rate-pressure product compared with WT, with protracted increase in left ventricular end-diastolic pressure, exaggerated in female knockout hearts, despite comparable leakage of creatine kinase across groups. Kir6.2-KO hearts were rescued from diastolic dysfunction by agents that target alternative pathways of calcium handling. Thus K(ATP) channel deficit confers a greater susceptibility to calcium overload in vivo, accentuated in female hearts, impairing contractile recovery under various conditions of high metabolic demand.     

TGF-beta-regulated collagen type I accumulation: role of Src-based signals

Transforming growth factor- (TGF-) stimulates myofibroblast transdifferentiation, leading to type I collagen accumulation and fibrosis. We investigated the function of Src in TGF--induced collagen I accumulation. In human mesangial cells, PTyr416 Src (activated Src) was 3.3-fold higher in TGF--treated cells than in controls. Src activation by TGF- was blocked by rottlerin and by a dominant negative mutant of protein kinase C (PKC), showing that TGF- activates Src by a PKC-based mechanism. Pharmacological inhibitors and a dominant negative Src mutant prevented the increase in collagen type I secretion in cells exposed to TGF-. Similarly, on-target Src small interference RNA (siRNA) prevented type I collagen secretion in response to TGF-, but off-target siRNA complexes had no effect. It is well established in mesangial cells that upregulation of type I collagen by TGF- requires extracellular signal-regulated kinase 1/2 (ERK1/2), and we found that activation of ERK1/2 by TGF- requires Src. In conclusion, these results suggest that stimulation of collagen type I secretion by TGF- requires a PKC-Src-ERK1/2 signaling motif. mesangial cells; fibrosis; glomerulus; transforming growth factor-     

Sperm motion characteristics of GarolexMalpura sheep evolved in a semi-arid tropical environment through introgression of FecB gene

The arid and semi-arid tropical climates of India are endowed with vast diversity of non-prolific sheep breeds. The GarolexMalpura sheep has been evolved in a semi-arid tropical environment by introgression of FecB gene via artificial insemination of Malpura ewes using diluted semen of prolific microsheep Garole and subsequently multiplied by inter se mating among GarolexMalpura halfbreds. The aim of the present study was to identify FecB mutation in sexually mature GarolexMalpura rams by forced RFLP-PCR of BMPR-1B gene and evaluate: (i) semen production and sperm motion characteristics of GM rams and (ii) influence of age and FecB genotype on their semen attributes. Semen was collected during autumn season from 12 donor rams by artificial vagina on 8 occasions at weekly interval. The overall means of traits which did not differed significantly with age or FecB genotyping were volume (0.72 ml), mass motility (4.44), sperm concentration (2721.56 x 10(6)ml(-1)), curvilinear velocity (134.51 microm/s), motility (81.3%), amplitude of lateral head displacement (6.24 microm), beat frequency (44.43 Hz), sperm head elongation (48.9%) and sperm head area (10.01 microm(2)). The FecB genotyping had a significant effect (P<0.05) on percent linearity and rapid motile sperms, which did not vary significantly with age. Although sperm concentration was higher in FecB(BB) and FecB(B+), compared to FecB(++) genotypes but the effect was non-significant. The age and FecB genotyping had significant effects (P<0.05) on straightness, average path velocity, straight-line velocity and percentage of medium or slow motile sperms. It is concluded that GarolexMalpura rams with introgressed FecB gene are capable of producing good quality semen in a semi-arid tropical climate.     

Effect of Allium sativum on the immunity and survival of Labeo rohita infected with Aeromonas hydrophila

The aim of this study was to evaluate dietary dosages of garlic on the immune response and disease resistance against infections due to the opportunistic pathogen Aeromonas hydrophila in Labeo rohita fingerlings. Garlic, Allium sativum, was incorporated into the diets (0%, 0.1%, 0.5% and 1.0%) of rohu, L. rohita fingerlings (10 ± 2 g). Every 20 days, different biochemical (serum total protein, albumin, globulin, albumin : globulin ratio, blood glucose), haematological (WBC, RBC and haemoglobin content) and immunological (superoxide anion production, lysozyme activity and serum bactericidal activity) parameters were evaluated. Superoxide anion production, lysozyme, serum bactericidal, serum protein and albumin were enhanced in garlic treated groups compared with the control group. After 60 days, fish were challenged with A. hydrophila and mortality (%) was recorded up to day 10 post‐challenge. Survival decreased in control group (57%) up to day 10 after infection. However, this was increased in the garlic treatment group, i.e. 85% survivability in the 1 g garlic kg^?1 (B group) and 5 g garlic kg^?1 (C group), and 71% survivability in the 10 g garlic kg^?1 (D group), respectively. These results indicate that Allium sativum stimulates the immunity and makes L. rohita more resistant to infection by A. hydrophila.     

Negative regulation of ASK1 by p21Cip1 involves a small domain that includes Serine 98 that is phosphorylated by ASK1 in vivo

The cyclin-dependent kinase inhibitor p21(Cip1) regulates multiple cellular functions and protects cells from genotoxic and other cellular stresses. Activation of apoptosis signal-regulating kinase 1 (ASK1) induced by inhibition of mTOR signaling leads to sustained phospho-c-Jun that is suppressed in cells with functional p53 or by forced expression of p21(Cip1). Here we show that small deletions of p21(Cip1) around S98 abrogate its association with ASK1 but do not affect binding to Cdk1, hence distinguishing between the cell cycle-regulating functions of p21(Cip1) and its ability to suppress activation of the ASK1/Jun N-terminal protein kinase (JNK) pathway. p21(Cip1) is phosphorylated in vitro by both ASK1 and JNK1 at S98. In vivo phosphorylation of p21(Cip1), predominantly carried out by ASK1, is associated with binding to ASK1 and inactivation of ASK1 kinase function. Binding of p21(Cip1) to ASK1 requires ASK1 kinase function and may involve phosphorylation of S98.     

Vibrational dynamics and heat capacity in polyglycine II

The earlier works on the vibrational dynamics of polyglycine II (PG II) suffer from several infirmities, which not only suppress the dynamical nature of normal modes, but also lead to several incorrect assignments and interactive constants of the potential field. In this study, we have re-examined the phonon dispersion profiles of PG II using Higgs method for evaluation of phase-related normal modes and have attempted to remove the infirmities, as far as possible. The Wilson's GF matrix method combined with the Urey-Bradley force field has been used for normal mode analysis. This potential field leads to correct assignments of Raman, infrared and inelastic neutron scattering frequencies. Characteristic features of the dispersion curve (v versus delta/pi plot), such as repulsion and regions of high density-of-states have been interpreted. In addition, the heat capacity as a function of temperature has been obtained via density-of-states. It agrees well with the experimental data and is being reported for the first time.