PURIFICATION AND BIOCHEMICAL CHARACTERIZATION OF L-AMINO ACID OXIDASE FROM WESTERN REGION INDIAN COBRA (NAJA NAJA) VENOM

Objective: Purification and biochemical characterization of LAAO from western region Indian Cobra (Naja naja) venom.Methods: LAAO was purified from Indian cobra (Naja naja) venom using sequential chromatography on Sephadex G-75 gel filtration followed by Ion exchange on CM-Sephadex C-25 column. Biochemical characterization viz., pH, Temperature, Km and Vmax were determined. Molecular weight of LAAO was determined by electrophoresis. Inhibition of LAAO from cold water extracts of Curcuma zedoria, Curcuma ceasia, Curcuma aromatic, Curcuma longa, Curcuma amada, Cucumis sativus and Benincasa hispida was done.Results: Purified LAAO showed the single band on non reducing SDS-PAGE with approximate molecular weight of 65 kDa. Further biochemical characterization revealed that, LAAO from Naja naja (western region) has an optimum pH of 7.0 and is stable at room Temperature upto 37 °C and showed an optimum enzyme concentration of 4µg/ml and Km 134.1µM and Vmax is 21.87 U/min. Cold water extract of Curcuma ceasia, Curcuma aromatic and Benincasa hispida showed comparatively significant inhibition of LAAO.Conclusion: LAAO has promising therapeutic prospects because of its effects on various biological functions. Variation in snake species and their geographical distribution also contributes to the venom properties such as composition, toxicity level, pharmacological and biological activities. The significant difference in protein profiling leads to substantial lethality among different geographical regions. Thus in this study Indian Cobra (Naja naja) venom from the western region of India was subjected to purification of LAAO and biochemical characterization. Â

Serine protease from Indian Cobra venom: its anticoagulant property and effect on human fibrinogen

An unusual low molecular weight serine protease (NnP28) has been purified from Indian Cobra (Naja naja) venom from the western region of the Indian sub-continent. We reported the purification and characterization of low molecular NnP28, emphasizing its role on human fibrinogen and anticoagulant property. NnP28 was purified using gel filtration column chromatography followed by ion exchange chromatography. Protein gel electrophoresis revealed its molecular weight approximate to 28 kDa. The exact molecular mass of NnP28 was found to be 27.12 kDa by mass spectrometry, hydrolyzing casein specifically, inhibited by PMSF suggesting it has a serine protease. NnP28 prolonged the clotting time of re-calcified human citrated plasma and activated partial thromboplastin time (APTT) exhibiting anticoagulant property. NnP28 exhibited fibrinogenolytic activity. Thus, the present study demonstrates the presence of unusual low molecular weight serine protease, emphasizing its importance of region-specific Indian cobra species

Purification and biochemical characterization of a novel secretory dipeptidyl peptidase IV from porcine serum

Purification of DPP-IV enzyme from porcine serum, is presented in this study for the first time. The high molecular weight DPP-IV from porcine serum was fractioned using Sephadex G-75 gel filtration followed by DEAE Sephadex anion exchange and Sephadex G-100 gel filtration chromatography columns with a final yield of 11.25%. The SDS-PAGE of the purified sample showed a single band of molecular mass nearing 160 kDa. Distinct single band was observed after PAS staining confirmed it to be a glycoprotein. The purified enzyme showed an optimum pH and temperature of 8 and 37 degrees C, respectively. The enzyme effectively cleaved fluorogenic substrate Gly-Pro-AMC with Km and Vmax of 4.578 mu M and 90.84 nmoles/min, respectively. Purified DPP-IV activity was inhibited by Diprotin A with an IC(50)value of 8.473 mu M. Among the three plant extracts used to study DPP-IV inhibition, the aqueous hot extract ofTerminalia chebulashowed the highest inhibition of 87.19%, followed by the aqueous cold extract ofMomordica carantia, ( 31.6%) andAzadirachta indica(34.16%) at the concentration of 25 mu g

Prevalence of high blood pressure and associated factors among adolescents and young people in Tanzania and Uganda.

We conducted a cross-sectional study among school/college students in Tanzania and Uganda to determine the prevalence of high blood pressure (BP) and associated factors. Participants were classified to have high BP if they had pre-hypertension or hypertension. Interviews were done using the WHO STEPS instrument. Using data from both countries (n = 1596), the overall prevalence of high BP was 40% (95% CI: 37-42). The prevalence of pre-hypertension was 29% (95% CI: 26-31) and that of hypertension was 11% (95% CI: 10-13). High BP was independently associated with obesity (aOR = 6.7, 95% CI: 2.2-20.0), male sex (aOR = 3.2, 95% CI: 2.4-4.4), and among males aged above 20 years (aOR = 5.5, 95% CI: 2.9-10.5). Consumption of fruits/vegetables was associated with decreased odds for high BP (aOR = 0.7, 95% CI: 0.50-0.98). The increasing burden of pre-hypertension across age groups could explain the early onset of hypertension and cardiovascular diseases (CVDs) among young African adults. There is a need for longitudinal studies to explore the drivers of pre-hypertension in East African adolescents

Postnatal depression across countries and cultures : a qualitative study

Background: Postnatal depression seems to be a universal condition with similar rates in different countries. However, anthropologists question the cross-cultural equivalence of depression, particularly at a life stage so influenced by cultural factors. Aims: To develop a qualitative method to explore whether postnatal depression is universally recognised, attributed and described and to enquire into people’s perceptions of remedies and services for morbid states of unhappiness within the context of local services. Method: The study took place in 15 centres in 11 countries and drew on three groups of informants: focus groups with new mothers, interviews with fathers and grandmothers, and interviews with health professionals.Textual analysis of these three groups was conducted separately in each centre and emergent themes compared across centres. Results: All centres described morbid unhappiness after childbirth comparable to postnatal depression but not all saw this as an illness remediable by health interventions. Conclusions: Although the findings of this study support the universality of a morbid state of unhappiness following childbirth, they also support concerns about the cross-cultural equivalence of postnatal depression as an illness requiring the intervention of health professionals; this has implications for future research

On the limits of sexual health literacy: Insights from Ugandan schoolgirls

This article makes the case that current conceptions of sexual health literacy have limited relevance to the Ugandan context because they assume that knowledge of unsafe sexual practices will lead to changes in behavior and lifestyle. Drawing on a longitudinal case study with 15 Ugandan schoolgirls in rural Uganda from August 2004 to September 2006, this study argues that despite being well-informed about the risks and responsibilities of sexual activity, poverty and sexual abuse severely constrained options for these young women. Although many believed in the value of abstaining from sexual activity until marriage, they engaged in transactional sex to pay for school fees, supplies, clothing, and food. Further, fear of sexual abuse, early pregnancy, and HIV–AIDS compromised attempts to embrace sexuality. The article concludes with implications of the study for research and policy on sexual health literacy in Uganda and other poorly resourced regions of the world

Loss of p53 results in protracted electrographic seizures and development of an aggravated epileptic phenotype following status epilepticus

The p53 tumor suppressor is a multifunctional protein, which regulates cell cycle, differentiation, DNA repair and apoptosis. Experimental seizures up-regulate p53 in the brain, and acute seizure-induced neuronal death can be reduced by genetic deletion or pharmacologic inhibition of p53. However, few long-term functional consequences of p53 deficiency have been explored. Here, we investigated the development of epilepsy triggered by status epilepticus in wild-type and p53-deficient mice. Analysis of electroencephalogram (EEG) recordings during status epilepticus induced by intra-amygdala kainic acid (KA) showed that seizures lasted significantly longer in p53-deficient mice compared with wild-type animals. Nevertheless, neuronal death in the hippocampal CA3 subfield and the neocortex was significantly reduced at 72 h in p53-deficient mice. Long-term continuous EEG telemetry recordings after status epilepticus determined that the sum duration of spontaneous seizures was significantly longer in p53-deficient compared with wild-type mice. Hippocampal damage and neuropeptide Y distribution at the end of chronic recordings was found to be similar between p53-deficient and wild-type mice. The present study identifies protracted KA-induced electrographic status as a novel outcome of p53 deficiency and shows that the absence of p53 leads to an exacerbated epileptic phenotype. Accordingly, targeting p53 to protect against status epilepticus or related neurologic insults may be offset by deleterious consequences of reduced p53 function during epileptogenesis or in chronic epilepsy

Investigating the metabolic capabilities of Mycobacterium tuberculosis H37Rv using the in silico strain iNJ661 and proposing alternative drug targets

<p>Abstract</p> <p>Background:</p> <p><it>Mycobacterium tuberculosis </it>continues to be a major pathogen in the third world, killing almost 2 million people a year by the most recent estimates. Even in industrialized countries, the emergence of multi-drug resistant (MDR) strains of tuberculosis hails the need to develop additional medications for treatment. Many of the drugs used for treatment of tuberculosis target metabolic enzymes. Genome-scale models can be used for analysis, discovery, and as hypothesis generating tools, which will hopefully assist the rational drug development process. These models need to be able to assimilate data from large datasets and analyze them.</p> <p>Results:</p> <p>We completed a bottom up reconstruction of the metabolic network of <it>Mycobacterium tuberculosis </it>H37Rv. This functional <it>in silico </it>bacterium, <it>iNJ</it>661, contains 661 genes and 939 reactions and can produce many of the complex compounds characteristic to tuberculosis, such as mycolic acids and mycocerosates. We grew this bacterium <it>in silico </it>on various media, analyzed the model in the context of multiple high-throughput data sets, and finally we analyzed the network in an 'unbiased' manner by calculating the Hard Coupled Reaction (HCR) sets, groups of reactions that are forced to operate in unison due to mass conservation and connectivity constraints.</p> <p>Conclusion:</p> <p>Although we observed growth rates comparable to experimental observations (doubling times ranging from about 12 to 24 hours) in different media, comparisons of gene essentiality with experimental data were less encouraging (generally about 55%). The reasons for the often conflicting results were multi-fold, including gene expression variability under different conditions and lack of complete biological knowledge. Some of the inconsistencies between <it>in vitro </it>and <it>in silico </it>or <it>in vivo </it>and <it>in silico </it>results highlight specific loci that are worth further experimental investigations. Finally, by considering the HCR sets in the context of known drug targets for tuberculosis treatment we proposed new alternative, but equivalent drug targets.</p