How Do Patients Who Fail First-Line TB Treatment but Who Are Not Placed on an MDR-TB Regimen Fare in South India?

SETTING: Seven districts in Andhra Pradesh, South India. OBJECTIVES: To a) determine treatment outcomes of patients who fail first line anti-TB treatment and are not placed on an multi-drug resistant TB (MDR-TB) regimen, and b) relate the treatment outcomes to culture and drug susceptibility patterns (C&DST). DESIGN: Retrospective cohort study using routine programme data and Mycobacterium TB Culture C&DST between July 2008 and December 2009. RESULTS: There were 202 individuals given a re-treatment regimen and included in the study. Overall treatment outcomes were: 68 (34%) with treatment success, 84 (42%) failed, 36 (18%) died, 13 (6.5%) defaulted and 1 transferred out. Treatment success for category I and II failures was low at 37%. In those with positive cultures, 81 had pan-sensitive strains with 31 (38%) showing treatment success, while 61 had drug-resistance strains with 9 (15%) showing treatment success. In 58 patients with negative cultures, 28 (48%) showed treatment success. CONCLUSION: Treatment outcomes of patients who fail a first-line anti-TB treatment and who are not placed on an MDR-TB regimen are unacceptably poor. The worst outcomes are seen among category II failures and those with negative cultures or drug-resistance. There are important programmatic implications which need to be addressed

Microsatellite Genotyping of Plasmodium vivax Isolates from Pregnant Women in Four Malaria Endemic Countries

Plasmodium vivax is the most widely distributed human parasite and the main cause of human malaria outside the African continent. However, the knowledge about the genetic variability of P. vivax is limited when compared to the information available for P. falciparum. We present the results of a study aimed at characterizing the genetic structure of P. vivax populations obtained from pregnant women from different malaria endemic settings. Between June 2008 and October 2011 nearly 2000 pregnant women were recruited during routine antenatal care at each site and followed up until delivery. A capillary blood sample from the study participants was collected for genotyping at different time points. Seven P. vivax microsatellite markers were used for genotypic characterization on a total of 229 P. vivax isolates obtained from Brazil, Colombia, India and Papua New Guinea. In each population, the number of alleles per locus, the expected heterozygosity and the levels of multilocus linkage disequilibrium were assessed. The extent of genetic differentiation among populations was also estimated. Six microsatellite loci on 137 P. falciparum isolates from three countries were screened for comparison. The mean value of expected heterozygosity per country ranged from 0.839 to 0.874 for P. vivax and from 0.578 to 0.758 for P. falciparum. P. vivax populations were more diverse than those of P. falciparum. In some of the studied countries, the diversity of P. vivax population was very high compared to the respective level of endemicity. The level of inter-population differentiation was moderate to high in all P. vivax and P. falciparum populations studied

Proteins identified by MALDI-TOF/TOF-MS/MS analysis after 2-DE gel separation of proteins from healthy and diseased leaf spot samples of <i>W</i>. <i>somnifera</i>.

<p>Proteins identified by MALDI-TOF/TOF-MS/MS analysis after 2-DE gel separation of proteins from healthy and diseased leaf spot samples of <i>W</i>. <i>somnifera</i>.</p

Features associated with the protein yield and significance of protein spots selected for the identification on 2-DE.

<p>Features associated with the protein yield and significance of protein spots selected for the identification on 2-DE.</p

2-DE analysis of proteins from <i>W</i>. <i>somnifera</i> leaves extracted using the three protocols under evaluation.

<p>(A) Tris buffer (B) Phenol and (C) TCA-acetone. Upper panel showed the proteins were separated on a 3–10 nonlinear pH strips. Lower panels are the gel photographs showing better resolution after re-analysis of the same proteins within the pH 5–8 range. Regions of the gels that show particularly different distributions of protein spots when comparing extraction methods are highlighted by rectangular dashed boxes.</p

Differential spot distribution of proteins extracted by tris based, phenol based and TCA-acetone based method.

<p>According to (A) molecular weight ranges (Mr) and (B) isoelectric point (pI). Bars with different letters within each range correspond to statistically significant differences (ANOVA, p<0.05).</p

Gene expression levels of selected genes as determined by quantitative RT-PCR in healthy (H) and diseased (D) plant samples.

<p>Here 14-3-3, Argonaute, CASP, COM, Expansin, GST, Ketoacyl, NADPH oxidase and SIN3 are 14-3-3 like protein, Argonaute protein, Casparian strip membrane domain proteins, Caffeoyl-CoA O-methyltransferase, Expansin, Glutathione-S-transferase, 3-ketoacyl-CoA synthase, NADPH oxidase, Sin3-like protein. Gene expression data was normalized against expression of cyclophilin (<i>CYP</i>) internal control.</p

Pie chart showing distribution according to biological function of proteins identified in <i>W</i>. <i>somnifera</i> in response to fungal infection.

<p>This classification is based on KEGG (<a href="http://www.kegg.jp/kegg/pathway.html" target="_blank">http://www.kegg.jp/kegg/pathway.html</a>) and on the literature.</p

Changes in the leaf proteome profile of <i>Withania somnifera</i> (L.) Dunal in response to <i>Alternaria alternata</i> infection

<div><p><i>Withania somnifera</i> is a high value medicinal plant which is used against large number of ailments. The medicinal properties of the plant attributes to a wide array of important secondary metabolites. The plant is predominantly infected with leaf spot pathogen <i>Alternaria alternata</i>, which leads to substantial biodeterioration of pharmaceutically important metabolites. To develop an effective strategy to combat this disease, proteomics based approach could be useful. Hence, in the present study, three different protein extraction methods tris-buffer based, phenol based and trichloroacetic acid-acetone (TCA-acetone) based method were comparatively evaluated for two-dimensional electrophoresis (2-DE) analysis of <i>W</i>. <i>somnifera</i>. TCA-acetone method was found to be most effective and was further used to identify differentially expressed proteins in response to fungal infection. Thirty-eight differentially expressed proteins were identified by matrix assisted laser desorption/ionization time of flight-mass spectrometry (MALDI TOF/TOF MS/MS). The known proteins were categorized into eight different groups based on their function and maximum proteins belonged to energy and metabolism, cell structure, stress and defense and RNA/DNA categories. Differential expression of some key proteins were also crosschecked at transcriptomic level by using qRT-PCR and were found to be consistent with the 2-DE data. These outcomes enable us to evaluate modifications that take place at the proteomic level during a compatible host pathogen interaction. The comparative proteome analysis conducted in this paper revealed the involvement of many key proteins in the process of pathogenesis and further investigation of these identified proteins could assist in the discovery of new strategies for the development of pathogen resistance in the plant.</p></div

Feasibility and effectiveness of provider initiated HIV testing and counseling of TB suspects in Vizianagaram district, South India.

BACKGROUND: Though internationally recommended, provider initiated HIV testing and counseling (PITC) of persons suspected of tuberculosis (TB) is not a policy in India; HIV seroprevalence among TB suspects has never been reported. The current policy of PITC for diagnosed TB cases may limit opportunities of early HIV diagnosis and treatment. We determined HIV seroprevalence among persons suspected of TB and assessed feasibility and effectiveness of PITC implementation at this earlier stage in the TB diagnostic pathway. METHODS: All adults examined for diagnostic sputum microscopy (TB suspects) in Vizianagaram district (population 2.5 million), in November-December 2010, were offered voluntary HIV counseling and testing (VCT) and assessed for TB diagnosis. RESULTS: Of 2918 eligible TB suspects, 2465(85%) consented to VCT. Among these, 246(10%) were HIV-positive. Of the 246, 84(34%) were newly diagnosed as HIV (HIV status not known previously). To detect a new case of HIV infection, the number needed to screen (NNS) was 26 among 'TB suspects', comparable to that among 'TB patients'. Among suspects aged 25-54 years, not diagnosed as TB, the NNS was 17. CONCLUSION: The seroprevalence of HIV among 'TB suspects' was as high as that among 'TB patients'. Implementation of PITC among TB suspects was feasible and effective, detecting a large number of new HIV cases with minimal additional workload on staff of HIV testing centre. HIV testing of TB suspects aged 25-54 years demonstrated higher yield for a given effort, and should be considered by policy makers at least in settings with high HIV prevalence