Structural and functional analysis of chitinase gene family in wheat (Triticum aestivum).

Chitinases are the hydrolytic enzymes which protect plants against pathogen attack. However, the precise role of chitinases in disease resistance has not been explored in wheat. In the present study, in silico approach, including secondary structure analysis, detailed signature pattern study, cis-acting regulatory elements survey, evolutionary trends and three-dimensional molecular modeling was used for different chitinase classes of wheat (Triticum aestivum). Homology modeling of class I, II, IV and 3 chitinase proteins was performed using the template crystal structure. The model structures were further refined by molecular mechanics methods using different tools, such as Procheck, ProSA and Verify3D. Secondary structure studies revealed greater percentage of residues forming α helix conformation with specific signature pattern, similar to casein kinase II phosphorylation site, amidation site, N-myristoylation (N-MYR) site and protein kinase C phoshorylation site. The expression profile suggested that wheat chitinase gene was highly expressed in cell culture and callus. We found that wheat chitinases showed more functional similarity with rice and barley. The results provide insight into the evolution of the chitinase family, constituting a diverse array of pathogenesis-related proteins. The study also provides insight into the possible binding sites of chitinase proteins and may further enhance our knowledge of fungal resistance mechanism in plants.

In silico structural and functional analysis of protein encoded by wheat early salt-stress response gene (WESR3).

Salt stress is one of the major abiotic stresses limiting grain yield in wheat (Triticum aestivum L.). Wheat early salt-stress response gene (WESR3) is one of the major salt stress genes, which is affected in the first phase of salt stress. In this study, sequence and structural analysis of protein coded by WESR3 gene was carried out using various bioinformatics tools. Sequence analysis of WESR3 protein revealed the presence of highly conserved regions of Mlo gene family. Three-dimensional modeling was carried out to elucidate its structure and its active site. The sequence analysis revealed that WESR3 protein might be involved in fungal pathogen attack pathway. Thus, in addition to its involvement in abiotic stresses, it also seemed to play an important part in biotic stress pathways. Out of the three modeled protein structures obtained from I-TASSER, HHPred and QUARK, the I-TASSER protein model was the best model based on high confidence score and lesser number of bad contacts. The Ramchandran plot analysis also showed that all amino acid residues of I-TASSER model lie in the allowed region and thus indicating towards the overall good quality of the predicted model. Seventeen active sites were predicted in the protein bearing resemblance to the Mlo family conserved regions. In conclusion, a detailed analysis of WESR3 protein suggested an important role of WESR3 in biotic and abiotic stress. These results aid to the experimental data and help to build up a complete view of WESR3 proteins and their role in plant stress response.

A study on factors influencing farmer’s satisfaction level towards Agricultural produce marketing committee (Rajnandgaon district).

Agriculture is an important element of Indian economy. The vast majority of this country’s population depends on agriculture for its livelihood. So it becomes very important to offer farmers a fair market for his produce at fair price .Governments agricultural policies and Agri-marketing schemes should be huge,effective, organized and systematic. For fulfilling this purpose the Agricultural produce marketing committees were established. They work under district regulated market set up by the different state governments to sell the agricultural produce directly from farmer(as seller) to purchasers . There is no middleman in between the market transactions. The present study is focused on the factors influencing farmers satisfaction level towards Agricultural produce marketing committee of Rajnandgaon district .

Determinants of infant mortality in a developing region in rural Andhra Pradesh.

Introduction: Background & Objectives: Despite several interventions implemented for the improvement of infant survival, the pace of decline in infant mortality in the State of Andhra Pradesh during the last two decades has been rather slow. This paper examines determinants of infant mortality in a rural population of about 45000 in Medchal region of Andhra Pradesh, India. Methods: Data obtained through continuous (longitudinal) enumeration of household population and also from the cross-sectional survey of households carried out during 2008-09 in the study area were analyzed. A univariate analysis was carried out and followed by an application of binary logistic regression model to identify significant factors associated with infant mortality. Results: Despite a substantial increase in institutional deliveries in the study area, infant mortality rate remained relatively constant at 43±1. Socioeconomic variables such as education and household economic status and environmental variables such as access to safe drinking water and sanitation facility and use of clean cooking fuel emerged as significant predictors of infant survival. Interpretation & Conclusions: Promoting institutional deliveries by providing cash assistance to women by itself cannot be expected to bring down infant mortality to a low level in the absence of an improvement in socioeconomic and environmental conditions of the households.

Effectiveness of interventions among HIV reactive pregnant women (in Andhra Pradesh).

Background: Over the years, single dose Nevirapine has been the cornerstone of the PPTCT program in India. However after successful scale up of ART program, initiation of ART for the pregnant was also considered another means of reducing the risk of transmission of HIV. As there were no specific studies to demonstrate the advantages of full-course ART during pregnancy compared to single dose Nevirapine, the current study was undertaken. Objectives: To study the socio-demographic variables, effectiveness of the different interventions and factors affecting the interventions among the PPTCT program beneficiaries. Materials and Methods: As part of the retrospective cohort study the details of the HIV positive pregnant women registered at four ICTCs of Andhra Pradesh were analyzed in two groups (Nevirapine and ART). Results: Total 145 pregnant women were studied and among them, 3.4% opted for MTP, 66.2% had live-birth and 60% had normal vaginal delivery. Upon testing for HIV at 18 months, 35% babies were HIV reactive in the Nevirapine group, 17.6% babies were HIV reactive in the ART group. Interpretation & Conclusion: The findings from the present study showed higher protection among babies with mothers on ART before delivery, compared to those with single dose Nevirapine.

Profile of HIV positive clients in an ICTC of a private medical college, Andhra Pradesh: A situational analysis.

Background: HIV/AIDS has turned out into global pandemic. In India under National AIDS Control Program (NACP) –III Integrated Counseling and Testing Centers (ICTC) were established throughout the country for providing prevention services through a holistic approach for all segments of the population. Objectives: To study the sociodemographic profile and risk behavior pattern of clients diagnosed HIV positive in the ICTC. Material & method: Study Design: Cross-sectional study (record based) Setting: Mediciti Institute of Medical Sciences, Andhra Pradesh. Subjects- All the 448 seropositive attendees during the year 2007-2010. Data pertaining to total 8845 clients, who availed the services from the ICTC during the above period was assessed for the time trend of HIV infection .Study Variables- Year of reporting, in-referral of the clients, age, sex, educational status, marital status, occupation and pattern of risk behavior in relation to HIV/AIDS. Statistical analysis- Data was analyzed in SPSS version 17.0 . Results: HIV prevalence was found to be declining over the last 4 years, except in 2008. Among the seropositives 62.7% were males and 37.3% were females. 85% were in the reproductive age group 78.1% were married;53.1% were illiterates. Majority of them are labourers from the agricultural background and housewives. In 83.9% of the individuals it is acquired through unprotected heterosexual intercourse. Conclusion- These findings should call for greater introspection among the program managers and facilitate for more customized local planning involving various stakeholders.

Growth, biomass production and photosynthesis of Cenchrus ciliaris L. under Acacia tortilis (Forssk.) Hayne based silvopastoral systems in semi arid tropics.

The growth, biomass production and photosynthesis of Cenchrus ciliaris was studied under the canopies of 17 yr old Acacia tortilis trees in semi arid tropical environment. On an average the full grown canopy of A. tortilis at the spacing of 4x4 m allowed 55% of total Photosynthetically Active Radiation (PAR) which in turn increased Relative Humidity (RH) and reduced under canopy temperature to -1.75oC over the open air temperature. C. ciliaris attained higher height under the shade of A. tortilis. The tiller production and leaf area index decreased marginally under the shade of tree canopies as compared to the open grown grasses. C. ciliaris accumulated higher chlorophyll a and b under the shade of tree canopies indicating its shade adaptation potential. The assimilatory functions such as rate of photosynthesis, transpiration, stomatal conductance, photosynthetic water use efficiency (PN/TR) and carboxylation efficiency (PN/CINT) decreased under the tree canopies due to low availability of PAR. The total biomass production in term of fresh and dry weight decreased under the tree canopies. On average of 2 yr C. ciliaris had produced 12.78 t ha-1 green and 3.72 t ha-1 dry biomass under the tree canopies of A. tortilis. The dry matter yield reduced to 38% under the tree canopies over the open grown grasses. The A. tortilis + C. ciliaris maintained higher soil moisture, organic carbon content and available N P K for sustainable biomass production for the longer period. The higher accumulation of crude protein, starch, sugar and nitrogen in leaves and stem of C. ciliaris indicates that this grass species also maintained its quality under A. tortilis based silvopastoral system. The photosynthesis and dry matter accumulation are closely associated with available PAR indicating that for sustainable production of this grass species in the silvopasture systems for longer period about 55% or more PAR is required.

Solitary rectal ulcer syndrome: uncommon cause of rectal bleeding in children.

Rectal polyp and hemorrhoids are common causes of bleeding per rectum in pediatric age group. However, there are some other causes which should be considered in differential diagnosis. We have acquainted a case of rectal bleeding due to solitary rectal ulcer in a child of 10 year. Colonoscopic examination was required for diagnosis as proctoscopic examination and digital rectal examination missed the diagnosis, probably due to poor co-operation by the patient and rare nature of the disease. Although well recognized in the adult population, the pediatric experience with this condition is limited.

Hepatitis E virus infection in chronic liver disease causes rapid decompensation.

Hepatitis E previously known as enterically transmitted non-A, non-B hepatitis, is a self limiting infectious viral disease of developing countries. Various issues regarding the pathogenesis of liver injury and its natural history remain unanswered after two decades of its discovery. A small proportion of patients develop fulminant hepatic failure. Mortality is very high if it is associated with pregnancy, especially during third trimester. After establishment of hepatitis A virus as a cause of decompensation of chronic liver disease, now there are reports that hepatitis E viruses also does the same. Acute hepatitis E in these patients has a protracted course with high morbidity and mortality. Many patients develop hepatorenal syndrome, hepatic encephalopathy and even liver failure after co-infection with hepatitis E virus. Now time has come to institute hepatitis E virus superinfection as one of the cause of acute on chronic liver failure. Hepatitis E is a problem of developing countries and Nepal is in the endemic zone. Sudden decompensation in chronic liver disease patient, who were otherwise stable and under regular follow up, should be carefully dealt with. Patient statistics at our unit shows that 7 cases of chronic liver diseases with superinfection with hepatitis E virus were dealt from April 2004 to August 2005. Two patients (29%) died and 5 recovered. In patients with recovery, there was deterioration of Child-Pugh grading and the duration of hospital stay was longer. Thus, hepatitis E in diagnosed chronic liver disease case should be taken apprehensively. Similarly patients of chronic liver disease traveling to endemic zone should take precaution. If vaccine against hepatitis E virus is developed, chronic liver disease patient would be the eligible candidate for vaccination beside pregnant ladies.

Enhancement of radionuclide induced cytotoxicity by 2-deoxy-D-glucose in human tumor cell lines.

The efficacy of targeted radiotherapy can be enhanced by selective delivery of radionuclide to the tumors and/or by differentially enhancing the manifestation of radiation damage in tumors. Our earlier studies have shown that the 2-deoxy-D-glucose (2-DG), an inhibitor of glucose transport and glycolytic ATP production, selectively enhances the cytotoxicity of external beam radiation in tumor cells. Therefore, it is suggested that 2-DG may also enhance the cytotoxic effects of radionuclides selectively in tumor cells, thereby improving the efficacy of radionuclide therapy. In vitro studies on breast carcinoma (MDA-MB-468) and glioma (U-87) cell lines, has been carried out to verify this proposition. Clonogenicity (macrocolony assay), cell proliferation, cytogenetic damage (micronuclei formation) and apoptosis were investigated as parameters of radiation response. Mean inactivation dose D (dose required to reduce the survival from 1 to 0.37), was 48 MBq/ml and 96 MBq/ml for 99 mTc, treated MDA-MB-468 and U-87, respectively. The dose response of growth inhibition, induction of micronuclei formation and apoptosis observed under these conditions, were correlated well with the changes in cell survival. Presence of 2-DG (5 mM) during radionuclide exposure (24 hrs), reduced the survival by nearly 2 folds in MDA-MB-468 (from 48.5 MBq to 18.5 MBq) and by 1.6 folds in U-87 cells (from 96 MBq to 66 Mbq). These results clearly show that the presence of 2-DG during radionuclide exposure, significantly enhances the cytotoxicity, by increasing mitotic as well as interphase death. Further studies to understand the mechanisms of radio-sensitization by 2-DG and preclinical studies using tumor-bearing animals, are required for optimizing the treatment schedule.

The paradox of pyrazinamide: an update on the molecular mechanisms of pyrazinamide resistance in Mycobacteria.

Pyrazinamide (PZA) is an important front line anti-tuberculosis drug because of its sterilizing activity against semi-dormant tubercle bacilli. In spite of its remarkable role in shortening the treatment duration from 9 months to 6 months when used in combination with Rifampicin and Isoniazid, PZA remains a difficult paradox because of its incompletely understood mode of action and mechanism of resistance. PZA is a nicotinamide analog prodrug which is converted into the active bactericidal form pyrazinoic acid by the bacterial enzyme pyrazinamidase (PZase). PZA does not appear to have a specific cellular target and instead, exerts its bactericidal effect by disrupting the membrane energetics and acidification of cytoplasm. Majority (72-97%) of PZA-resistant isolates of M. tuberculosis exhibit mutations in their pncA gene or upstream area leading to loss of PZase activity. A wide diversity of pncA mutations scattered along the entire length of pncA gene is unique to PZA resistance. However, PZA resistant isolates with normal PZase activity and wild type pncA sequences have also been reported in several studies which indicate that alternate mechanisms of PZA resistance exist. Investigations into these mechanisms would be useful in developing alternative diagnostic/therapeutic measures. This review presents the update of various mechanisms of PZA resistance in different mycobacteria with special emphasis on mode of action of PZA and mechanisms of resistance in Mycobacterium tuberculosis.

Byssinosis among male textile workers in Pondicherry: a case-control study.

BACKGROUND: In India, 20 million workers are involved in the manufacturing of textiles. However, there are few epidemiological studies from India that have assessed the magnitude or the risk factors associated with byssinosis. In Pondicherry, textile mills have been in existence for over a century. This case-control study aimed to find the factors associated with the development of byssinosis in textile workers. METHODS: The sample consisted of 761 men above the age of 30 years who had worked for at least 10 years in a textile factory. All the respondents were interviewed by a pretested questionnaire to gather information regarding the symptoms of byssinosis, certain personal characteristics and occupational history. Byssinosis was identified using the classification proposed by the World Health Organization. Two age-matched controls were selected for each case. RESULTS: Univariate analysis of the factors for symptomatic byssinosis showed that dusty worksites, heavy smoking and duration of service > or = 30 years were significant. Logistic regression analysis showed that working in the spinning (odds ratio 6.1) and weaving sections (odds ratio 1.9), heavy smoking (odds ratio 3.9) and > or = 30 years of service (odds ratio 2.0) were independent significant risk factors. CONCLUSION: Efforts to reduce dust levels in the working environment and to discourage smoking among textile workers need to be strengthened to minimize the risk of developing byssinosis.

Reversal of catecholamine induced cardiomyopathy in a patient with bilateral malignant pheochromocytoma.

A patient of catecholamine induced cardiomyopathy (CMP) due to bilateral malignant adrenal pheochromocytoma (PCC), successfully reversed on treatment is described here. To the best of our knowledge, this is first report of catecholamine induced CMP due to bilateral malignant adrenal PCC, which could be reversed after surgery.

Deconvolution of the fluorescence spectra into its component Gaussians: a method to analyze the binding of coumarin to bovine serum albumin.

7-N,N-Diethylamino-4-methylcoumarin, (cou-1), a readily available laser dye binding to bovine serum albumin (BSA), at room temperature has been studied by steady state fluorescence spectroscopy. Existing methods of analysis of the binding to obtain the binding parameters are based on the change in fluorescence intensity at a particular wavelength. These methods are not convenient when there is a gradual shift in the emission maxima for increasing protein concentration. In this paper we present a method to obtain the binding constants of cou-1 to BSA using a Windows '95 based package to deconvolute the asymmetrical spectrum (fluorescence intensity versus wave number curve) into two Gaussians, each corresponding to the binding of the fluorophore to a particular site. This method is convenient to analyze the binding constant data and obtain the binding parameters of each binding site, and can also provide information about the microenvironment of each site, relating micropolarity and microviscosity.

Efficacy of light-traps in sampling malaria vectors in different ecological zones in central India.

This preliminary field study was designed chiefly to test the efficiency of the light-trap as a tool for sampling malaria vectors, in tribal villages located in different ecological settings in comparison with indoor resting collections as an alternative method. Anopheles culicifacies, a known malaria vector, was the most prevalent species in the study villages and more than 80% of trap catches were obtained before midnight with peak activity during dusk. Reproductive status of trapped specimens revealed proportional representations of unfed, freshly fed, and gravid females. Another vector, An. fluviatilis was found in small numbers by both the methods. Thus the trap could give a reliable and unbiased sample of vector population. Seven species were abundant in the light-trap catches while only four in the indoor resting collections indicates the usefulness of the light-trap for sampling exophilic species. The study revealed that light-traps did not have any bias in favor of any particular species. The method may be useful for assessing the night time densities of different species or the fluctuation of a species at different dates and village to village variations. Light-traps could be used for sampling both endophilic and exophilic anophelines.

Fungitoxic properties of Prunus persica oil.

During screening of leaves of 15 higher plant species, the leaves of Prunus persica exhibited absolute inhibition of mycelial growth of Aspergillus flavus. The active fraction was isolated in form of an essential oil which exhibited fungistatic nature at its minimum inhibitory concentration of 0.1% and showed a broad antifungal range. The oil was found non-phytotoxic to the seedling growth and seed germination of wheat.

Action of 2,4-dichlorophenoxyacetic acid and rifampicin on heterocyst differentiation in the blue-green alga, Nostoc linckia.

2,4-Dichlorophenoxyacetic acid, a commonly used herbicide, increased the growth of the filamentous blue-green alga, Nostoc linckia at doses upto 100 μg /ml. The herbicidetreated N2-cultures showed enhanced heterocyst frequency and N2-growth. Thus, the herbicide stimulated algal growth at the expense of molecular nitrogen under aerobic growth conditions. Rifampicin caused chain formation of heterocysts. This was effectively counteracted by 2,4-dichlorophenoxyacetic acid, suggesting a biological interaction between them at the level of the heterocyst spacing control mechanism.