The role of socio-economic factors responsible for non-compliance of directly observed treatment short-course among tuberculosis patients

Introduction: Tuberculosis has gone out of control in many parts of world. The therapeutic regimens given under direct observation as recommended by world Health Organization have been shown to be highly effective for both preventing and treating tuberculosis but poor compliance to anti-tuberculosis medication (ATT) is a major barrier to its global controls. Objectives: The role of socio-economic factors responsible for non-compliance of directly observed treatment short-course (DOTS) among Tuberculosis Patients. Materials and Methods: Cross sectional study, interviewed using a set of questionnaire. Participants were enrolled at DOTS centre of Lala Ram Sarup Institute of tuberculosis and Respiratory disease New Delhi, India. Patients who interrupted treatment for more than 2 months consecutively were classified as non-compliance cases. T-test was used for comparing the means of the control and case groups. Results: Majority of study population (45%) was in age group of 31 -45 years, which is the productive age. 85 % noncompliant patients believed anti TB drugs to be bad and 15 % accepted as good for them. Similarly 87 % believed to stop ATT very prematurely as soon as symptoms disappear while 13 % believed to continue as per DOTS volunteer advice. The study revealed that the non-compliance of DOTS was significantly high among those who were less educated, unskilled worker, low family income and upper lower class family. Conclusion: In Delhi region, the treatment of Tuberculosis was good and did not have much involvement of socio-economical role in the non-compliance of treatment. But still major hurdle is the inadequate education, occupation, and income

Identification of Hot and Cold spots in genome of Mycobacterium tuberculosis using Shewhart Control Charts

The organization of genomic sequences is dynamic and undergoes change during the process of evolution. Many of the variations arise spontaneously and the observed genomic changes can either be distributed uniformly throughout the genome or be preferentially localized to some regions (hot spots) compared to others. Conversely cold spots may tend to accumulate very few variations or none at all. In order to identify such regions statistically, we have developed a method based on Shewhart Control Chart. The method was used for identification of hot and cold spots of single-nucleotide variations (SNVs) in Mycobacterium tuberculosis genomes. The predictions have been validated by sequencing some of these regions derived from clinical isolates. This method can be used for analysis of other genome sequences particularly infectious microbes

Voltage Gated Calcium Channels Negatively Regulate Protective Immunity to Mycobacterium tuberculosis

Mycobacterium tuberculosis modulates levels and activity of key intracellular second messengers to evade protective immune responses. Calcium release from voltage gated calcium channels (VGCC) regulates immune responses to pathogens. In this study, we investigated the roles of VGCC in regulating protective immunity to mycobacteria in vitro and in vivo. Inhibiting L-type or R-type VGCC in dendritic cells (DCs) either using antibodies or by siRNA increased calcium influx in an inositol 1,4,5-phosphate and calcium release calcium activated channel dependent mechanism that resulted in increased expression of genes favoring pro-inflammatory responses. Further, VGCC-blocked DCs activated T cells that in turn mediated killing of M. tuberculosis inside macrophages. Likewise, inhibiting VGCC in infected macrophages and PBMCs induced calcium influx, upregulated the expression of pro-inflammatory genes and resulted in enhanced killing of intracellular M. tuberculosis. Importantly, compared to healthy controls, PBMCs of tuberculosis patients expressed higher levels of both VGCC, which were significantly reduced following chemotherapy. Finally, blocking VGCC in vivo in M. tuberculosis infected mice using specific antibodies increased intracellular calcium and significantly reduced bacterial loads. These results indicate that L-type and R-type VGCC play a negative role in M. tuberculosis infection by regulating calcium mobilization in cells that determine protective immunity

Prevalence, Distribution and Functional Significance of the −237C to T Polymorphism in the IL-12Rβ2 Promoter in Indian Tuberculosis Patients

Cytokine/cytokine receptor gene polymorphisms related to structure/expression could impact immune response. Hence, the −237 polymorphic site in the 5′ promoter region of the IL-12Rβ2 (SNP ID: rs11810249) gene associated with the AP-4 transcription motif GAGCTG, was examined. Amplicons encompassing the polymorphism were generated from 46 pulmonary tuberculosis patients, 35 family contacts and 28 miscellaneous volunteers and sequenced. The C allele predominated among patients, (93.4%, 43/46), and in all volunteers and contacts screened, but the T allele was exclusively limited to patients, (6.5%, 3/46). The functional impact of this polymorphism on transcriptional activity was assessed by Luciferase-reporter and electrophoretic mobility shift assays (EMSA). Luciferase-reporter assays showed a significant reduction in transcriptional efficiency with T compared to C allele. The reduction in transcriptional efficiency with the T allele construct (pGIL-12Rb2-T), in U-87MG, THP-1 and Jurkat cell lines, were 53, 37.6, and 49.8% respectively, compared to the C allele construct (pGIL-12Rb2-C). Similarly, densitometric analysis of the EMSA assay showed reduced binding of the AP-4 transcription factor, to T compared to the C nucleotide probe. Reduced mRNA expression in all patients (3/3) harboring the T allele was seen, whereas individuals with the C allele exhibited high mRNA expression (17/25; 68%, p = 0.05). These observations were in agreement with the in vitro assessment of the promoter activity by Luciferase-reporter and EMSA assays. The reduced expression of IL-12Rβ2 transcripts in 8 patients despite having the C allele was attributed to the predominant over expression of the suppressors (IL-4 and GATA-3) and reduced expression of enhancers (IFN-α) of IL-12Rβ2 transcripts. The 17 high IL-12Rβ2 mRNA expressers had significantly elevated IFN-α mRNA levels compared to low expressers and volunteers. Notwithstanding the presence of high levels of IL-12Rβ2 mRNA in these patients elevated IFN-α expression could modulate their immune responses to Mycobacterium tuberculosis

A 46-year-old man with tracheomegaly, tracheal diverticulosis, and bronchiectasis: Mounier-Kuhn syndrome

Lower respiratory tract infection is one of the common causes of morbidity in India which is occasionally undiagnosed. In this regard tracheobronchomegaly is one of those conditions which masquerade as chronic bronchitis and bronchiectasis and are usually undiagnosed. It is a well-defined clinical and radiologic entity characterized by marked dilatation of the trachea and the central bronchi and is frequently associated with recurrent lower respiratory tract infection. Tracheobronchomegaly has been described by a variety of names, including Mounier-Kuhn syndrome, tracheal diverticulosis, tracheobronchiectasis, tracheocele, tracheomalacia, and tracheobronchopathia malacia

Tuberculosis associated immune reconstitution inflammatory syndrome in patients infected with HIV: meningitis a potentially life threatening manifestation

Abstract Background Tuberculosis (TB) is the most common co infection in HIV-infected persons in India, requiring concomitant administration of anti TB and antiretroviral therapies. Paradoxical worsening of tuberculosis after anti-retroviral therapy (ART) initiation is frequently seen. Objective To study the frequency, clinical presentation and outcome of paradoxical tuberculosis associated immune reconstitution inflammatory syndrome (TB-IRIS) in HIV infected patients in a TB hospital in North India. Design A retrospective chart review of HIV-infected TB patients on anti-tubercular treatment (ATT) at time of ART initiation over a 3 year period. Medical records were reviewed for clinical manifestations and outcome in patients who developed TB-IRIS. Results 514 HIV-infected patients were enrolled between January 2006 and December 2008. Thirteen (12.6%) of 103 patients who had received ART and ATT simultaneously developed paradoxical TB-IRIS. Clinical presentations of paradoxical TB-IRIS included new lymphadenopathy (n = 3), increase in size of existing lymphadenopathy (n = 3), worsening of existing pulmonary lesions (n = 2), appearance of new pleural effusion (n = 1) and prolonged high grade fever (n = 2). Four patients developed new tubercular meningitis as manifestation of TB-IRIS. Our cases developed TB-IRIS a median of 15 days after starting ART (IQR 15–36). TB-IRIS patients were older (> 35 years) than those with no IRIS (P = 0.03), but were not distinguishable by CD4 T-cell count, duration of ATT before ART or the outcome of TB treatment. Eight (62%) patients had a complete recovery while 5 (38%) patients with TB-IRIS died, of which majority (n = 3) had meningitis. Conclusions Paradoxical TB-IRIS is a frequent problem during concomitant ATT and ART in HIV-TB co infected patients in north India. Meningitis is a potentially life threatening manifestation of TB-IRIS.</p

Phenol-ammonium sulfate microscopy method for diagnosis of pulmonary tuberculosis

Background: Collection and processing of sputum samples for the detection of acid fast bacilli (AFB) is hazardous for health-workers in developing countries with limited facilities. The phenol ammonium sulfate (PhAS) method involves smear microscopy and Ziehl-Neelson (ZN) staining of precipitates/ floccules formed in sputum samples when PhAS is added. The present study has been designed to assess the performance and safety of this method. Materials and methods: The study was conducted from January 2011 to March 2011 at the Department of Microbiology, Lala Ram Sarup Institute of Tuberculosis and Respiratory Diseases, New Delhi. A total of 1038 sputum samples were subjected to ZN staining before and after treatment with PhAS. The smear microscopy results of the PhAS treated and untreated samples were compared. In addition, 200 representative samples were inoculated after processing by petroff's method directly for culture and after treatment with PhAS. Result: The sensitivity, specificity, positive predictive value and negative predictive value of the PhAS solution treated ZN smear microscopy method were found to be 98.8%, 88.5%, 98.0% and 92.7% respectively in comparison with direct smear microscopy. The overall correlation between the two methods was found to be 97.3%. None of the PhAS treated samples grew Mycobacterium tuberculosis on culture. Conclusion: Sputum microscopy with PhAS solution is a safe, reliable and inexpensive alternative for direct microscopy. This method can be conveniently applied for usage in microscopy centers with limited bio-safety facilities

Predominance of Beijing genotype in extensively drug resistant Mycobacterium tuberculosis isolates from a tertiary care hospital in New Delhi, India

Out of a total of 311 Mycobacterium tuberculosis isolates from sputum specimens subjected to first- and second-line drug-susceptibility testing (DST) at a hospital serving as a referral center for chronic tuberculosis (TB) cases in New Delhi, 232/311 (74.6%) isolates were found to be resistant to isoniazid and rifampicin. Among multidrug-resistant (MDR) isolates, 119/232 (51.3%) were resistant to four first-line drugs (streptomycin, isoniazid, rifampicin and ethambutol). Mono-resistance to isoniazid was observed in 18 (5.7%) isolates, while none of the isolates tested showed mono-resistance to rifampicin. 50/232 (21.5%) isolates met the definition of extensively drug resistant (XDR) TB, i.e., additional resistance to a fluoroquinolone and at least one of the three injectable second-line drugs: kanamycin, capreomycin, or amikacin. Spoligotyping of the XDR-TB isolates revealed 14 patterns; 39/50 (78%) isolates being grouped in three clusters vs. 11/50 (22%) isolates being unique. SIT1/Beijing represented the largest cluster (n = 21, 42%), followed by SIT26/CAS1-Delhi (n = 10, 20%) and SIT 53/T1 (n = 8 isolates; 16%). This study corroborates recent observations from North India suggesting that both Beijing and CAS1-Delhi lineages constitute the bulk of XDR-TB isolates that are disseminating rapidly across a large geographical region in and around the capital city of India

Endoscopic ultrasound-guided fine-needle aspiration with an echobronchoscope in undiagnosed mediastinal lymphadenopathy: First experience from India

Background and Aim: Several studies have reported the performance of combining endoscopic ultrasound fine-needle aspiration with an echobronchoscope (EUS-B-FNA), with endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA), for the diagnosis of mediastinal lymphadenopathy. Herein we report our initial experience with this technique. Patients and Methods: In this retrospective study, we report the outcome of patients who underwent EUS-B-FNA with or without EBUS-TBNA during the same session. Details of the lymph nodes sampled, number of passes, duration of the procedure, results of the cytological examination, and the final diagnosis of all the patients are presented. Results: Eleven patients underwent EUS-B-FNA during the study period. EUS-B-FNA and EBUS-TBNA together yielded a sensitivity and specificity of 72.7 and 100%, respectively. The number of lymph nodes sampled per patient and the number of passes per lymph node were significantly higher with EBUS-TBNA compared to EUS-B-FNA. The mean duration of procedure was also significantly higher in the EBUS-TBNA group. The most common reason (five patients (45.5%)) for resorting to EUS-B-FNA was the patient being unfit for EBUS-TBNA or the inability of the operator to complete the procedure. Sarcoidosis was the most frequent final diagnosis in the study patients (four cases), followed by bronchogenic carcinoma (three cases). Conclusions: EUS-B-FNA is a useful additional tool for the diagnosis of mediastinal lymphadenopathy. In combination with EBUS-TBNA, it has a fairly good diagnostic yield and is a good alternative in situations where EBUS-TBNA is not feasible