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Find video protocols related to scientific articles indexed in Pubmed.
Development of a POC test for TB based on multiple immunodominant epitopes of M. tuberculosis specific cell-wall proteins.
PLoS ONE
PUBLISHED: 01-01-2014
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The need for an accurate, rapid, simple and affordable point-of-care (POC) test for Tuberculosis (TB) that can be implemented in microscopy centers and other peripheral health-care settings in the TB-endemic countries remains unmet. This manuscript describes preliminary results of a new prototype rapid lateral flow TB test based on detection of antibodies to immunodominant epitopes (peptides) derived from carefully selected, highly immunogenic M. tuberculosis cell-wall proteins. Peptide selection was initially based on recognition by antibodies in sera from TB patients but not in PPD-/PPD+/BCG-vaccinated individuals from TB-endemic settings. The peptides were conjugated to BSA; the purified peptide-BSA conjugates striped onto nitrocellulose membrane and adsorbed onto colloidal gold particles to devise the prototype test, and evaluated for reactivity with sera from 3 PPD-, 29 PPD+, 15 PPD-unknown healthy subjects, 10 patients with non-TB lung disease and 124 smear-positive TB patients. The assay parameters were adjusted to determine positive/negative status within 15 minutes via visual or instrumented assessment. There was minimal or no reactivity of sera from non-TB subjects with the striped BSA-peptides demonstrating the lack of anti-peptide antibodies in subjects with latent TB and/or BCG vaccination. Sera from most TB patients demonstrated reactivity with one or more peptides. The sensitivity of antibody detection ranged from 28-85% with the 9 BSA-peptides. Three peptides were further evaluated with sera from 400 subjects, including additional PPD-/PPD+/PPD-unknown healthy contacts, close hospital contacts and household contacts of untreated TB patients, patients with non-TB lung disease, and HIV+TB- patients. Combination of the 3 peptides provided sensitivity and specificity>90%. While the final fully optimized lateral flow POC test for TB is under development, these preliminary results demonstrate that an antibody-detection based rapid POC lateral flow test based on select combinations of immunodominant M. tb-specific epitopes may potentially replace microscopy for TB diagnosis in TB-endemic settings.
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Impact of introducing the line probe assay on time to treatment initiation of MDR-TB in Delhi, India.
PLoS ONE
PUBLISHED: 01-01-2014
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National Institute of Tuberculosis and Respiratory Diseases (erstwhile Lala Ram Sarup Institute) in Delhi, India.
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Direct drug susceptibility testing of Mycobacterium tuberculosis for rapid detection of multidrug resistance using the Bactec MGIT 960 system: a multicenter study.
J. Clin. Microbiol.
PUBLISHED: 12-07-2011
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Conventional indirect drug susceptibility testing of Mycobacterium tuberculosis with liquid medium is well established and offers time-saving and reliable results. This multicenter study was carried out to evaluate if drug susceptibility testing (DST) can be successfully carried out directly from processed smear-positive specimens (direct DST) and if this approach could offer substantial time savings. Sputum specimens were digested, decontaminated, and concentrated by the laboratory routine procedure and were inoculated in Bactec MGIT 960 as well as Lowenstein-Jensen (LJ) medium for primary isolation. All the processed specimens which were acid-fast bacterium (AFB) smear positive were used for setting up direct DST for isoniazid (INH) and rifampin (RIF). After the antimicrobial mixture of polymyxin B, amphotericin B, nalidixic acid, trimethoprim, and azlocillin (PANTA) was added, the tubes were entered in the MGIT 960 instrument using the 21-day protocol (Bactec 960 pyrazinamide [PZA] protocol). Results obtained by direct DST were compared with those obtained by indirect DST to establish accuracy and time savings by this approach. Of a total of 360 AFB smear-positive sputum specimens set up for direct DST at four sites in three different countries, 307 (85%) specimens yielded reportable results. Average reporting time for direct DST was 11 days (range, 10 to 12 days). The average time savings by direct DST compared to indirect DST, which included time to isolate a culture and perform DST, was 8 days (range, 6 to 9 days). When results of direct DST were compared with those of indirect DST, there was 95.1% concordance with INH and 96.1% with rifampin. These findings indicate that direct DST with the Bactec MGIT 960 system offers further time savings and is a quick method to reliably detect multidrug resistance (MDR) cases.
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Unilateral and painless development of isoniazid induced gynecomastia during re-treatment of pulmonary tuberculosis.
J Assoc Physicians India
PUBLISHED: 09-14-2011
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A case of isoniazid induced gynecomastia is being reported in an 18-year old male, who received a re-treatment regimen for the relapse of pulmonary tuberculosis (TB). At the end of two months of the treatment, the patient developed a painless unilateral gynecomastia, which completely disappeared after a month of the cessation of isoniazid. A review of literature on isoniazid induced gynecomastia is discussed.
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Reasons for interruption of anti-tubercular treatment as reported by patients with tuberculosis admitted in a tertiary care institute.
Indian J Tuberc
PUBLISHED: 03-26-2011
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Department of Tuberculosis and Chest Diseases, Tertiary Level tuberculosis (TB) institute in Delhi, India.
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A prospective randomized controlled trial on the efficacy of noninvasive ventilation in severe acute asthma.
Respir Care
PUBLISHED: 04-28-2010
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Noninvasive ventilation (NIV) is an emerging modality in the management of patients with acute respiratory failure. However, its role in severe acute asthma is not well defined.
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Disparity in circulating peripheral blood dendritic cell subsets and cytokine profile of pulmonary tuberculosis patients compared with healthy family contacts.
Hum. Immunol.
PUBLISHED: 03-25-2010
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Dendritic cell (DC) subsets, myeloid DCs (mDCs), and plasmacytoid DCs (pDCs) play a fundamental role in immune response to Mycobacterium tuberculosis (M. tuberculosis). Flow-cytometric estimation of DC subsets showed differences in the ratio of these subsets in untreated, smear-positive pulmonary tuberculosis patients compared with healthy family contacts (HFC, p < 0.05). The percentage of pDCs (0.14 +/- 0.01) was higher than mDCs (0.12 +/- 0.01) in patients, whereas in HFC, mDCs (0.15 +/- 0.01) was higher than pDCs (0.1 +/- 0.01). The percentage of mDCs (0.15 +/- 0.01) and pDCs (0.11 +/- 0.01) was restored in treated patients. Alteration in the DC subsets before and after chemotherapy was confirmed in the follow-up of acid-fast bacilli (AFB)-positive patients. This reversal in the percentage of mDC vs pDCs implicates the influence of active disease on circulating DC subsets. The cytokine bead array revealed an inverse relationship in the circulating levels of IL-12 and IFN-gamma. High IL-12 (37.9 +/- 15.2) and low IFN-gamma (11.09 +/- 3.6) was seen in HFCs derived serum samples compared with that of patients (p < 0.05). The higher percentage of mDCs and elevated IL-12 levels was found to be associated with high risk HFCs investigated. Furthermore CpG/LPS-stimulated whole-blood culture of untreated patients expressed high IFN-alpha in pDCs and less IL-12 in mDCs compared with those of treated patients.
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Evaluation of nitrate reductase assay for rapid detection of resistance to second-line drugs in Mycobacterium tuberculosis in a tertiary care hospital.
Diagn. Microbiol. Infect. Dis.
PUBLISHED: 03-11-2010
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The present study evaluated nitrate reductase assay (NRA) for rapid detection of resistance to 5 second-line drugs: kanamycin, ethionamide, ofloxacin, cycloserine, and para-aminosalicylic acid. Eighty-six multidrug-resistant clinical isolates of Mycobacterium tuberculosis were used to compare NRA with proportion method for rapid detection of resistance to second-line drugs. The sensitivity of the NRA for different drugs ranged from 86.4% to 100%, whereas the specificity ranged from 98.4% to 100%. Excellent agreement was found between the 2 tests with respect to their ? values. For 63% of the isolates, the results were available in 14 days (turnaround time). The NRA is a rapid, accurate, simple, and inexpensive method for detection resistance to second-line drugs. This method may become an appropriate alternative method, especially for the resource poor settings.
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Ocular and systemic morbidity profile in mass formic acid injuries.
Ophthalmic Surg Lasers Imaging
PUBLISHED: 02-05-2010
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A report on formic acid-induced ocular and systemic injuries.
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Outcomes of noninvasive ventilation in acute hypoxemic respiratory failure in a respiratory intensive care unit in north India.
Respir Care
PUBLISHED: 12-08-2009
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To determine the outcomes of noninvasive ventilation (NIV) and the factors associated with NIV failure in patients with acute hypoxemic respiratory failure (AHRF).
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Toll-like receptor 2 and DC-SIGNR1 differentially regulate suppressors of cytokine signaling 1 in dendritic cells during Mycobacterium tuberculosis infection.
J. Biol. Chem.
PUBLISHED: 07-17-2009
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A hallmark of protective immunity during Mycobacterium tuberculosis (M. tb) infection is the regulated secretion of pro-inflammatory and regulatory cytokines. Suppressors of Cytokine Signaling (SOCS) are key regulators of cytokine secretion and function. In this study we investigated regulation of Toll-like receptor 2 (TLR2) and dendritic cell-specific ICAM-3 grabbing non-integrin receptor 1 (DC-SIGNR1)-mediated SOCS1 expression in DCs during M. tb infection. We show that, compared with TLR2, stimulating DC-SIGNR1 on DCs induces higher SOCS1 expression and lower interleukin-12 production. Co-stimulating DC-SIGNR1 and TLR2 differentially regulates SOCS1 expression depending on the relative concentration of their ligands. Stimulating DC-SIGNR1 with M. tb infection increases SOCS1 expression, while stimulating TLR2 with M. tb infection reduces SOCS1 expression. Knockdown of SOCS1 in DCs by siRNA enhances interleukin-12 transcription and protein expression upon DC-SIGNR1 stimulation. Raf-1 and Syk differentially regulate TLR2- and DC-SIGNR1-mediated SOCS1 expression. In addition, DC-SIGNR1 shows greater association with SOCS1 when compared with TLR2. Interestingly, compared with healthy asymptomatic individuals, peripheral blood mononuclear cells of patients with active tuberculosis disease showed higher expression of SOCS1, which was reduced following chemotherapy. Similarly, stimulating DC-SIGNR1 on DCs from M. tb-infected TLR2(-/-) mice enhanced SOCS1 expression that was reduced following chemotherapy. Further, knockdown of SOCS1 in mouse DCs or human peripheral blood mononuclear cells resulted in increased killing of virulent M. tb. These results indicate that TLR2 and DC-SIGNR1 differentially regulate SOCS1 expression during M. tb infection. This in turn regulates M. tb survival by governing key cytokine expression.
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Post treatment sequelae of multi-drug resistant tuberculosis patients.
Indian J Tuberc
PUBLISHED: 02-13-2009
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Sequelae of patients treated for multi-drug resistance tuberculosis (MDR-TB) remain unknown.
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Voltage gated calcium channels negatively regulate protective immunity to Mycobacterium tuberculosis.
PLoS ONE
PUBLISHED: 01-16-2009
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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.
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Profile of HIV associated tuberculosis at a tertiary institute in setting of free anti-retroviral therapy.
J Assoc Physicians India
PUBLISHED: 01-01-2009
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With the rollout of the national program, free of cost antiretroviral therapy is available to the Indian population since 2004, resulting in sea change in outcome of HIV associated tuberculosis.
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Fate of sputum samples transported in bottles containing cetylpyridinium chloride and sodium chloride: a national reference laboratory study.
Indian J Tuberc
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The transportation of sputum samples may sometimes take more than one week which results in an increased contamination rate and loss of positive cultures. The current study was planned to analyze the recovery rate of mycobacteria from transported samples with and without Cetylpyridinium chloride (CPC). Addition of CPC is useful for isolation of M. tuberculosis from sputum subjected to long-term storage.
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A 46-year-old man with tracheomegaly, tracheal diverticulosis, and bronchiectasis: Mounier-Kuhn syndrome.
Lung India
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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.
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Tuberculosis associated immune reconstitution inflammatory syndrome in patients infected with HIV: meningitis a potentially life threatening manifestation.
AIDS Res Ther
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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.
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Prevalence, distribution and functional significance of the -237C to T polymorphism in the IL-12R?2 promoter in Indian tuberculosis patients.
PLoS ONE
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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-12R?2 (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-12R?2 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-?) of IL-12R?2 transcripts. The 17 high IL-12R?2 mRNA expressers had significantly elevated IFN-? mRNA levels compared to low expressers and volunteers. Notwithstanding the presence of high levels of IL-12R?2 mRNA in these patients elevated IFN-? expression could modulate their immune responses to Mycobacterium tuberculosis.
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Identification of hot and cold spots in genome of Mycobacterium tuberculosis using Shewhart Control Charts.
Sci Rep
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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.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.