DOTS Awareness and the Myths and Misconceptions about DOTS among Medical Practitioners in Mysore

Background: Annually 2 million people in India develop Tuberculosis and 330,000 die. WHO-recommended DOTS strategy was pilot-tested in 1993 and launched as Revised National Tuberculosis Control Program (RNTCP) in 1997. Awareness of DOTS among the doctors in the private sector was appalling although nationwide coverage was attained by 2006. OBJECTIVE: To study awareness of DOTS among Medical Practitioners of urban and rural Mysore. Methodology: 401 Medical practitioners in hospitals and nursing homes of urban and rural areas of Mysore who treated Tuberculosis patients (private and public sector) were approached. They were grouped under different specialties as per the year of graduation (before or after introduction of DOTS). Results: 38 % doctors who graduated before the introduction of DOTS didn’t follow DOTS compared to 14.9% doctors who graduated later. 100% doctors working in Government sector felt that DOTS was better than daily regimen while 85% from the private sector felt so. Only 47.9% of the doctors in the private sector practiced DOTS compared to 95.1 % in the Govt. Sector. Hence, the number of doctors practicing DOTS in Private Sector was less than 50 % of that in the Govt. Sector. Both of these comparisons were found to be statistically highly significant (p<0.001). Awareness of DOTS was alarmingly low among Orthopedic Surgeons, Gynecologists and Pediatricians when compared to Physicians and General Practitioners. Conclusions: DOTS awareness is still low among doctors who graduated before the introduction of DOTS. Private practitioners harbored myths and misconceptions about DOTS

Control of transcription initiation

Mechanism of control of transcription initiation have expanded far beyond the classical operon concept. Control elements are multipartite and well separated from each other. Thetrans-factors bound to these sites make contacts with RNA polymerase: promoter complexes by DNA bending or looping to influence the initiation event. Activators and repressors are like two faces of the same coin and their function depends on the site of action, mode of interaction with DNA and also the nutritional status of the cell

Regulation of DNA topology in mycobacteria

DNA topoisomerases catalyse essential DNA transaction processes in order to attain the balanced topology of the genome. Contrasting activities of DNA topoisomerase I and DNA gyrase result in the maintenance of topological homeostasis. The regulation of expression of different topoisomerases ensure steady state optimum level expression of the enzymes. Many aspects of their organization and regulation seem to be different in mycobacteria when compared with that of Escherichia coli. Here we present several aspects of the regulation of mycobacterial topoisomerases and discuss the significance

A comparative study of frequency of postnatal depression among subjects with normal and caesarean deliveries

Background: Prevalence of postnatal depression (PND) is 12-15%. Recent studies are equivocal about the earlier inference that PND is higher among caesarian than normal delivery. Objective: The aim of this study is to investigate the frequency of PND among the Indian women and the association between the mode of delivery and PND. Material and method: Fifty subjects each; having delivered normally and by caesarian section was chosen. All the women were within 3 months post delivery and could understand Kannada language. Those who consented were asked to complete the Edinburgh Postnatal Depression Scale (EPDS). Those found to have scores suggestive of depression on EPDS were assessed for depression according to ICD-10. The data was analyzed using paired t test and chi square test. Result and conclusion: Among Post caesarean subjects, depression was diagnosed in 20% (n=10) as compared to 16% (n=8) in subjects that delivered normally. However there was no significant difference in the frequency of depression among the two groups. Due to the small sample size the results cannot be generalized

An orphangyr B in the Mycobacterium smegmatis genome uncovered by comparative genomics

DNA gyrase is an essential topoisomerase found in all bacteria. It is encoded by gyrB and gyrA genes. These genes are organized differently in different bacteria. Direct comparison of Mycobacterium tuberculosis and Mycobacterium smegmatis genomes reveals presence of an additionalgyr B in M. smegmatis flanked by novel genes. Analysis of the amino acid sequence of GyrB from different organisms suggests that the orphan GyrB in M. smegmatis may have an important cellular role

Alignment and phylogenetic analysis of type II DNA topoisomerases

DNA topoisomerases have been evolved to solve the topological problems of DNA during replication, transcription, recombination and segregation. Discovery of several new enzymes and their characterization has necessitated this compilation. This analysis shows the distinct evolutionary relatedness of type II DNA topoisomerases. A striking feature is the absence of a contiguous stretch of about 160 amino acids in one of the subunits of prokaryotic type II enzymes, which might have important implications to their structure and function

Transcriptional specificity after mycobacteriophage I3 infection

Transcriptional regulation following mycobacteriophage I3 infection has been investigated. For this purpose, RNA polymerase mutants (rif) of host bacterium, Mycobacterium smegmatis have been isolated and characterised. Phage growth inrifs and rifr cells in presence of rifampicin revealed the involvement of host RNA polymerase in phage genome transcription. This was confirmed by studies onin vivo RNA synthesis as well as by direct RNA polymerase assay after phage infection. Significant stimulation in RNA polymerase activity was seen following phage infection. The maximal levels were attained in about 60 min post infection and maintained throughout the phage development period. The stimulation of polymerase activity was most pronounced when the phage DNA was used as the template. RNA polymerases from uninfected and phage-infected Mycobacterium smegmatis have been purified to homogeniety. The enzyme purification was accomplished by a rapid procedure utilising affinity chromatography on rifampicin-Sepharose columns. Subunit structure analysis of the purified RNA polymerase from uninfected and phage-infected cells showed the presence of α ,β , β ' and σ subunits similar to the other prokaryotic RNA polymerases. In addition, a polypeptide of 79, 000 daltons was associated with the enzyme after phage infection. The enzymes differed in their properties with respect to template specificity. Phage 13 DNA was the preferred template for the modified RNA polymerase isolated from infected cells which may account for the transcriptional switch required for phage development

Requirement for calcium ions in mycobacteriophage I3 DNA injection and propagation

Ca2+ ions are absolutely necessary for the propagation of mycobacteriophage I3 in synthetic medium. These ions are required for successful infection of the host and during the entire span of the intracellular development of the phage. A direct assay of the phage DNA injection using 32[p] labelled phage, shows that Ca2+ ions are necessary for the injection process. The injection itself is a slow process and takes 15 min to complete at 37&#176;. The bacteria infected in presence of Ca2+ tend to abort if the ions are subsequently withdrawn from the growth medium. The effect of calcium withdrawal is maximally felt during the early part of the latent period; however, later supplementation of Ca2+ ions salvage phage production and the mature phage progeny appear after a delayed interval, proportional to the time of addition of Ca2+

Use of protein A gene fusions for the analysis of structure-function relationship of the transactivator protein C of bacteriophage Mu

A sensitive dimerization assay for DNA binding proteins has been developed using gene fusion technology. For this purpose, we have engineered a gene fusion using protein A gene of Staphylococcus aureus and C gene, the late gene transactivator of bacteriophage Mu. The C gene was fused to the 3' end of the gene for protein A to generate an A-C fusion. The overexpressed fusion protein was purified in a single step using immunoglobulin affinity chromatography. Purified fusion protein exhibits DNA binding activity as demonstrated by electrophoretic mobility shift assays. When the fusion protein A-C was mixed with C and analyzed for DNA binding, in addition to C and A-C specific complexes, a single intermediate complex comprising of a heterodimer of C and A-C fusion proteins was observed. Further, the protein A moiety in the fusion protein A-C does not contribute to DNA binding as demonstrated by proteolytic cleavage and circular dichroism (CD) analysis. The assay has also been applied to analyze the DNA binding domain of C protein by generating fusions between protein A and N- and C-terminal deletion mutants of C. The results indicate a role for the region towards the carboxy terminal of the protein in DNA binding. The general applicability of this method is discussed