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1.
Environ Res ; 260: 119610, 2024 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-39004393

RESUMO

Photocatalysis has been widely used as one of the most promising approaches to remove various pollutants in liquid or gas phases during the last decade. The main emphasis of the study is on the synergy of vacancy engineering and heterojunction formation, two widely used modifying approaches, to significantly alter photocatalytic performance. The vacancy-induced Ag2CO3/BiOBr/WO3-x heterojunction system has been fabricated using a co-precipitation technique to efficiently abate methylene blue (MB) dye and doxycycline (DC) antibiotic. The as-fabricated Ag2CO3/BiOBr/WO3-x heterojunction system displayed improved optoelectronic characteristic features because of the rational combination of dual charge transferal route and defect modulation. The Ag2CO3/BiOBr/WO3-x system possessed 97% and 74% photodegradation efficacy for MB and DC, respectively, with better charge isolation and migration efficacy. The ternary photocatalyst possessed a multi-fold increase in the reaction rate for both MB and DC, i.e., 0.021 and 0.0078 min-1, respectively, compared to pristine counterparts. Additionally, more insightful deductions about the photodegradation routes were made possible by the structural investigations of MB and DC using density functional theory (DFT) simulations. This study advances the understanding of the mechanisms forming visible light active dual Z-scheme heterojunction for effective environmental remediation.

2.
Ann Diagn Pathol ; 73: 152355, 2024 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-38878689

RESUMO

Long noncoding RNAs (lncRNAs) participate in transcriptional, epigenetic, and post-transcriptional regulation of gene expression and may influence carcinogenesis. MALAT1 is a lncRNA that is expressed in endocrine and many other neoplasms and it has been shown to have oncogenic and/or tumor suppressor effects in tumor development. Olfactory neuroblastomas arise in the nasal cavity while sympathetic neuroblastomas are present mainly in the adrenal and periadrenal regions. These neoplasms have overlapping histopathological features. Rare cases of sympathetic neuroblastomas metastatic to the nasal cavity have been reported. PHOX2B has been shown to be relatively specific for sympathetic neuroblastomas, but only a limited number of cases of olfactory neuroblastomas have been examined for PHOX2B expression. This study aimed to explore the potential utilization of MALAT1 and PHOX2B in distinguishing these two entities. Tissue microarrays (TMA) were created for olfactory neuroblastomas (n = 26) and sympathetic neuroblastomas (n = 52). MALAT1 lncRNA expression was assessed by in situ hybridization using RNAScope technology. TMA slides were scanned by Vectra multispectral imaging system and image analysis and quantification were performed with inForm software. PHOX2B expression was analyzed by immunohistochemistry. MALAT1 showed predominantly nuclear expression in both tumor types and MALAT1 expression was 2-fold higher in olfactory neuroblastomas compared to sympathetic neuroblastomas (p < 0.0001). PHOX2B showed nuclear staining in most sympathetic neuroblastomas (51/52, 98 %) while only 1 olfactory neuroblastoma (3.8 %) was focally positive for this marker. These findings suggest immunostaining of PHOX2B could be an excellent marker in distinguishing between these two tumor types.

3.
J Gastroenterol Hepatol ; 38(4): 619-624, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36652396

RESUMO

BACKGROUND AND AIM: Emergence of drug resistance, especially to second-line drugs, hampers tuberculosis elimination efforts. The present study aimed to evaluate MTBDRplus and MTBDRsl assays for detecting first-line and second-line drug resistance, respectively, in gastrointestinal tuberculosis (GITB). METHODS: Thirty ileocecal biopsy specimens, processed in the Department of Microbiology between 2012 and 2022, that showed growth of Mycobacterium tuberculosis on culture were included in the study. DNA, extracted from culture, was subjected to MTBDRplus and MTBDRsl (Hain Lifescience GmbH, Nehren, Germany), following manufacturer's instructions. Their performance was compared against phenotypic drug susceptibility testing (pDST) and gene sequencing. RESULTS: Out of the 30 specimens, 4 (13.33%) were mono-isoniazid resistant, 4 (13.33%) were multidrug resistant (MDR), 2 (6.67%) were pre-extensively drug resistant (pre-XDR), and 2 (6.67%) were mono-fluoroquinolone resistant. The results were 100% concordant with pDST and gene sequencing. CONCLUSIONS: In the wake of growing drug resistance in all forms of extrapulmonary tuberculosis, including GITB, MTBDRplus and MTBDRsl are reliable tools for screening of resistance to both first-line and second-line drugs.


Assuntos
Mycobacterium tuberculosis , Tuberculose Gastrointestinal , Humanos , Antituberculosos/farmacologia , Mycobacterium tuberculosis/genética , Tuberculose Gastrointestinal/diagnóstico , Tuberculose Gastrointestinal/tratamento farmacológico , Testes de Sensibilidade Microbiana , Isoniazida , Genótipo , Sensibilidade e Especificidade
4.
Indian J Clin Biochem ; 38(3): 393-399, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37234181

RESUMO

Human immunodeficiency virus (HIV) infection associated with weakened immune system due to decreased CD4 T cell count favors development of tuberculosis. Effector immune responses are also associated with micronutrient status due to their prominent role in maintaining immune functions. Micronutrient deficiencies are quite common among HIV patients that further result into compromised immunity thus making the conditions even more favorable for mycobacteria to establish disease. So, current study was designed to assess association of different micronutrients with development of TB in HIV patients. Micronutrient levels were measured in asymptomatic HIV patients who were monitored for the development of TB during follow up period (incident TB) within one month to one year and also in symptomatic microbiologically confirmed HIV-TB patients. Among various micronutrients assessed, levels of ferritin were found to be significantly increased (p < 0.05) with significant decreased zinc (p < 0.05) and selenium (p < 0.05) levels in incident TB group as well as in HIV-TB subjects compared to asymptomatic HIV patients who did not develop TB in the follow up period. Importantly, increased levels of ferritin and decreased levels of selenium were significantly associated with development of tuberculosis in HIV patients.

5.
J Gastroenterol Hepatol ; 37(8): 1571-1578, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35501293

RESUMO

BACKGROUND AND AIM: Prompt and accurate diagnosis of gastrointestinal tuberculosis (GITB) along with simultaneous detection of drug resistance is inevitable for tuberculosis elimination. Truenat MTB Plus (TruPlus), a chip-based real-time polymerase chain reaction assay, was evaluated for the first time for diagnosing GITB and detecting rifampicin resistance. METHODS: Fifty ileocecal biopsy specimens (5 microbiologically confirmed GITB [culture-positive], 25 clinically confirmed GITB [culture-negative], and 20 control patients) processed in the Department of Microbiology between 2011 and 2021 were subjected to TruPlus assay, Xpert MTB RIF assay multiplex polymerase chain reaction. Their performance was evaluated against both culture and composite reference standard. RESULTS: The overall sensitivity and specificity of TruPlus in diagnosing GITB was 70% (21/30) and 100%, respectively. The sensitivity was 60% (3/5) for microbiologically confirmed cases and 72% (18/25) for clinically confirmed cases. Performance of TruPlus was superior to Xpert (sensitivity = 30%; P = 0.001) and comparable with MPCR (sensitivity = 83.33%; P = 0.13). Both TruPlus and MPCR had moderate agreement with reference standards, and MPCR detected additional three cases. Both TruPlus and Xpert correctly reported Rifampicin resistance in three cases. CONCLUSIONS: TruPlus, with its greater portability and higher sensitivity than Xpert, could serve as an important tool for diagnosing GITB and rifampicin resistance at outreach endemic areas.


Assuntos
Antibióticos Antituberculose , Mycobacterium tuberculosis , Tuberculose Gastrointestinal , Tuberculose Pulmonar , Farmacorresistência Bacteriana , Humanos , Mycobacterium tuberculosis/genética , Rifampina/farmacologia , Sensibilidade e Especificidade , Tuberculose Gastrointestinal/diagnóstico
6.
Eur Radiol ; 31(5): 3297-3305, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33146793

RESUMO

OBJECTIVE: Comparison of virtual CT enteroscopy (VCTE) using carbon dioxide with small-bowel enteroclysis (SBE) and capsule endoscopy (CE) in small-bowel tuberculosis (SBTB). METHODS: This prospective study comprised consecutive patients suspected to have SBTB. VCTE and SBE were performed on the same day and evaluated by independent radiologists. CE was performed within 2 weeks. VCTE was performed following insufflation of carbon dioxide via catheters in the jejunum and anorectum. A contrast-enhanced CT was followed by a delayed non-contrast CT. Image processing was done using virtual colonoscopy software. Findings on VCTE, SBE, and CE were compared. The final diagnosis of SBTB was based on either histopathological or cytological findings, response to antitubercular treatment, or a combination of these. RESULTS: Of the 55 patients in whom VCTE was performed, complete data was available in 52 patients. A final diagnosis of SBTB was established in 37 patients. All patients had VCTE and SBE. CE was performed in 34 patients. Adequate luminal distension was achieved in all patients with SBE and 35 patients with VCTE. SBE showed more strictures in jejunum (10.8%) and ileum (75.7%) compared with VCTE (jejunum, 8.1%, and ileum, 64.9%) and CE (jejunum, 5.9%, and ileum, 61.8%). However, difference was not statistically significant. VCTE revealed a greater length of strictures in both the jejunum and ileum compared with SBE and CE. CONCLUSION: VCTE allows adequate evaluation of the bowel in most patients with SBTB. It allows detection of greater length of abnormality in jejunum and ileum compared with SBE and CE. KEY POINTS: • The use of VCTE using CO2 bowel insufflation in patients with SBTB should be considered. • VCTE allows detection of a greater length of abnormality in the jejunum and ileum.


Assuntos
Endoscopia por Cápsula , Tuberculose , Dióxido de Carbono , Endoscopia Gastrointestinal , Humanos , Estudos Prospectivos , Tomografia Computadorizada por Raios X
7.
Retina ; 41(4): 793-803, 2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-32833411

RESUMO

PURPOSE: To analyze and describe serial follow-up of choriocapillaris alterations in tubercular serpiginouslike choroiditis (SLC) using optical coherence tomography angiography (OCTA) and to compare it with multimodal imaging. METHODS: In this prospective cohort study, patients with active tubercular SLC underwent OCTA using Optovue RTVue XR Avanti and other imaging techniques including enhanced-depth imaging OCT (EDI-OCT) (Heidelberg Spectralis; Heidelberg Engineering, Heidelberg, Germany), fundus autofluorescence, fluorescein angiography, and indocyanine green angiography. Serial imaging was performed for a total follow-up of 3 months. Choriocapillaris alterations at the site of lesions were analyzed on OCTA imaging, and their mean lesion areas were calculated. RESULTS: Twenty-six eyes (26 patients; 20 males; mean age: 32.68 ± 10.56 years) were included. Fourteen eyes had active multifocal lesions (n = 39), whereas 12 eyes had active placoid lesions (n = 12). At baseline, OCTA showed hyporeflective flow deficit lesions corresponding to the hypofluorescent lesions on indocyanine green angiography in all eyes. In the multifocal type of SLC, the mean lesion area decreased in all eyes compared with baseline, and lesions with a lesion area less than 0.1 mm2 on OCTA showed near-complete resolution with minimal choriocapillaris atrophy. In comparison, all eyes with a placoid type of SLC showed no significant reduction in the lesion area and showed extensive choriocapillaris atrophy. CONCLUSION: Optical coherence tomography angiography has the unique ability to demonstrate pathological flow impairment at the level of choriocapillaris in active tubercular SLC. Serial OCTA analysis reveals that large tubercular SLC lesions result in choriocapillaris atrophy as the lesions heal, whereas smaller multifocal lesions show resolution of choriocapillaris hypoperfusion with minimal atrophy.


Assuntos
Corioide/irrigação sanguínea , Corioidite/diagnóstico por imagem , Tuberculose Ocular/diagnóstico por imagem , Adulto , Antituberculosos/uso terapêutico , Corioidite/tratamento farmacológico , Corioidite/fisiopatologia , Corantes/administração & dosagem , Feminino , Angiofluoresceinografia , Seguimentos , Humanos , Aumento da Imagem , Verde de Indocianina/administração & dosagem , Masculino , Pessoa de Meia-Idade , Imagem Multimodal , Estudos Prospectivos , Tomografia de Coerência Óptica , Tuberculose Ocular/tratamento farmacológico , Tuberculose Ocular/fisiopatologia , Acuidade Visual/fisiologia , Adulto Jovem
8.
Mol Cell Probes ; 53: 101595, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32437884

RESUMO

BACKGROUND: Despite several recent advances in detection techniques, there is still an unmet need for simple tests for the diagnosis of tuberculous meningitis (TBM). Therefore, in an effort towards developing a simple and rapid diagnostic test for resource-poor settings, we designed an assay in which magnetic bead flocculation test (MBF) was used to detect the amplified DNA. Multi-targeted (using two multicopy gene targets IS6110 and IS1081) loop-mediated isothermal amplification (MLAMP) was used for amplification. METHODS: MLAMP-MBF assay was performed on CSF samples of 600 patients, out of which 120 were definite TBM (culture confirmed), 280 were probable TBM and 200 were non-TB controls, based on Marais's criteria. The performance of assay was evaluated by comparing the result of definite TBM with culture and that of probable TBM with composite reference standard consisting of clinical, microbiological(smear/culture) and radiological parameters. RESULTS: The overall sensitivity of MLAMP-MBF (using any of the two gene targets) was 89.5% and specificity was 100%. The sensitivity was 96.6% (116/120) in diagnosing definite TBM and 86.4% (242/280) in diagnosing probable TBM. The sensitivity of IS1081 was 88% and that of IS6110 was 83% in diagnosing TBM. Specificity of both the gene targets was 100%. There were 20 cases positive only by IS1081 LAMP and 6 cases positive only by IS6110; thus 26 of 400 (6.5%) TBM cases could be additionally detected following multi-targeted approach. CONCLUSION: MLAMP-MBF is a sensitive, robust, cost-effective and promising technique for diagnosis of TBM in low-resource high-endemic settings.


Assuntos
Testes de Floculação/métodos , Tipagem de Sequências Multilocus/métodos , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose Meníngea/diagnóstico , Técnicas de Tipagem Bacteriana , Líquido Cefalorraquidiano/microbiologia , Diagnóstico Precoce , Feminino , Humanos , Fenômenos Magnéticos , Masculino , Mycobacterium tuberculosis/genética , Pobreza , Sensibilidade e Especificidade , Tuberculose Meníngea/líquido cefalorraquidiano
9.
Can J Microbiol ; 66(12): 723-732, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32762636

RESUMO

CcpN is a transcriptional repressor in Bacillus subtilis that binds to the promoter region of gapB and pckA, downregulating their expression in the presence of glucose. CcpN also represses sr1, which encodes a small noncoding regulatory RNA that suppresses the arginine biosynthesis gene cluster. CcpN has homologues in other Gram-positive bacteria, including Enterococcus faecalis. We report the interaction of CcpN with DivIVA of B. subtilis as determined using bacterial two-hybrid and glutathione S-transferase pull-down assays. Insertional inactivation of CcpN leads to cell elongation and formation of straight chains of cells. These findings suggest that CcpN is a moonlighting protein involved in both gluconeogenesis and cell elongation.


Assuntos
Bacillus subtilis/citologia , Bacillus subtilis/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Ciclo Celular/metabolismo , Bacillus subtilis/genética , Proteínas de Bactérias/genética , Repressão Catabólica/genética , Regulação Bacteriana da Expressão Gênica/genética , Gluconeogênese/genética
10.
Retina ; 39(8): 1595-1606, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29689028

RESUMO

PURPOSE: To study optical coherence tomography angiography (OCTA) and multimodal imaging features of Type 1 inflammatory choroidal neovascularization (CNV) in tubercular serpiginous-like choroiditis and response to anti-vascular endothelial growth factor therapy. METHODS: In this study, multimodal imaging was performed using OCTA, enhanced-depth imaging optical coherence tomography, fluorescein angiography, and indocyanine green angiography. Correlation of OCTA with other imaging modalities in the detection of CNV was performed. The changes in CNV configuration after anti-vascular endothelial growth factor therapy were assessed. RESULTS: In this study, nine eyes (8 patients; 5 females; mean age: 32.5 ± 11.57 years) with diagnosis of tubercular serpiginous-like choroiditis were included. All the eyes had presence of low-lying pigment epithelial detachments on enhanced-depth imaging optical coherence tomography. Using OCTA, it was possible to detect Type 1 CNV in all eyes. Type 1 CNV networks comprised fine anastomotic network of vessels, some of which had a hairpin loop configuration. After anti-vascular endothelial growth factor therapy, there was a decrease in branching and anastomosis. The visual acuity significantly improved from 0.49 ± 0.26 (20/60 Snellen equivalent) at baseline to 0.26 ± 0.17 (20/36 Snellen equivalent) (P = 0.03) in all eyes. CONCLUSION: Type 1 CNV can occur among patients with tubercular serpiginous-like choroiditis, leading to significant visual loss even in the healed stages of the disease. Optical coherence tomography angiography can help in the detection of Type 1 CNV where conventional multimodal imaging, including fluorescein angiography and OCT, fails to make a definitive diagnosis and thereby guide the initiation of anti-vascular endothelial growth factor therapy.


Assuntos
Neovascularização de Coroide/diagnóstico , Corioidite/diagnóstico , Tuberculose Ocular/diagnóstico , Adolescente , Adulto , Inibidores da Angiogênese/uso terapêutico , Corioide/irrigação sanguínea , Corioidite/microbiologia , Corantes/administração & dosagem , Feminino , Angiofluoresceinografia , Humanos , Verde de Indocianina/administração & dosagem , Masculino , Pessoa de Meia-Idade , Imagem Multimodal , Tomografia de Coerência Óptica , Tuberculose Ocular/microbiologia , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Acuidade Visual , Adulto Jovem
11.
J Assoc Physicians India ; 67(7): 22-24, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31559763

RESUMO

BACKGROUND: Cardiovascular disease (CVD) is the leading cause of mortality in patients with rheumatoid arthritis (RA). Along with traditional cardiovascular risk factors and systemic inflammation, metabolic syndrome (MetS) contributes to CVD and increased mortality in patients with RA. In this study we determine the prevalence of MetS in RA patients presenting to a tertiary care centre in north India. METHODS: This is a case control study involving 114 patients of RA with disease duration of ≥1 year and 114 healthy controls who are age and sex matched. Components of MetS were assessed in all the subjects and disease activity of RA was determined by DAS28-ESR. MetS was defined according to modified ATP-III criteria and consensus definition of metabolic syndrome for adult Asian Indians. RESULTS: Women constituted 81.6% in RA group and 80.5% in control group. Mean age of subjects was 44.81±12.7 years in RA group and 43.27±12.6 years in control group. According to modified ATP-III criteria, 36 (31.6%) RA subjects and 17 (14.9%) controls had MetS (p=0.03). According to the consensus definition of metabolic syndrome for adult Asian Indian criteria, 40 (35.1%) RA subjects and 18 (15.8%) controls had MetS (P=0.01). There was no significant difference in disease activity between subjects of RA with or without MetS (p=0.276). CONCLUSION: The prevalence of MetS was higher in RA subjects compared to controls. There is no association of MetS with disease activity in our cohort. Larger studies are needed to determine the relation between MetS and disease activity.


Assuntos
Artrite Reumatoide/epidemiologia , Síndrome Metabólica/epidemiologia , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Índia/epidemiologia , Pessoa de Meia-Idade , Prevalência , Fatores de Risco , Centros de Atenção Terciária
12.
J Assoc Physicians India ; 67(12): 21-24, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31801325

RESUMO

BACKGROUND: Female genital tract tuberculosis (FGTB) is a very common disease in developing countries. Rapid and specific diagnosis is of paramount importance. PURPOSE: To evaluate Multiplex PCR using MPB 64 and IS6110 primers directed against M. tuberculosis for the diagnosis of FGTB and to compare the different methods available for diagnosis like histopathology, smear microscopy and TB culture. MATERIALS AND METHODS: Multiplex PCR was performed on endometrial biopsy samples of 21FGTB confirmed cases, 49 clinically suspected FGTB cases and 25 Non TB (control group) patients. RESULTS: : Multiplex PCR had sensitivity of 95.23% for confirmed cases and specificity of 100% for confirmed FGTB cases. In 49 clinically diagnosed, but unconfirmed FGTB cases multiplex PCR was positive in 61.22% cases. The overall sensitivity of microscopy, culture, Histopathology and multiplex PCR were 1.42%, 8.57%, 21.42%, 72.85% and specificity was 100%, 100%, 100% and 100% respectively. CONCLUSION: Multiplex PCR using MPB 64 and IS6110 primers has a high sensitivity and specificity in diagnosis of FGTB.


Assuntos
Mycobacterium tuberculosis , Tuberculose dos Genitais Femininos/diagnóstico , Tuberculose , Antígenos de Bactérias , Feminino , Humanos , Reação em Cadeia da Polimerase Multiplex , Sensibilidade e Especificidade
13.
Immunol Invest ; 47(6): 615-631, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29733740

RESUMO

PURPOSE: To assess cellular composition and local cytokine response in vitreous humor of tubercular uveitis. METHODS: Cells were collected from vitreous cassettes and peripheral blood of 8 tubercular uveitis and 5 control subjects, undergoing vitrectomy and analyzed by flow cytometry for cellular composition, activation status, proinflammatory cytokine expression, and uptake of TLR9 ligand, CpG ODN 2216. RESULTS: CD3 + T cells with equal proportion of CD4+ and CD8 + T cells formed major fraction of infiltrating cells. The vitreous humor showed higher expression of recent activation marker, CD69, and proinflammatory cytokines, IFN-γ and IL-17A, in CD4 + T cells as compared to peripheral blood. Lastly, intraocular CD4 + T cells showed reduced uptake of ODN 2216 than peripheral blood. CONCLUSIONS: Our results indicate that local antigenic stimuli trigger T cell infiltration and activation of CD4 + T cells that are hyporesponsive to TLR9 stimulation. These infiltrating T cells might be responsible in further aggravating ocular inflammation.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Ativação Linfocitária/imunologia , Tuberculose Ocular/imunologia , Uveíte/imunologia , Corpo Vítreo/citologia , Corpo Vítreo/imunologia , Adulto , Idoso , Antígenos CD/metabolismo , Antígenos de Diferenciação de Linfócitos T/metabolismo , Feminino , Humanos , Interferon gama/metabolismo , Interleucina-17/metabolismo , Lectinas Tipo C/metabolismo , Masculino , Pessoa de Meia-Idade , Oligonucleotídeos/metabolismo , Receptor Toll-Like 9/imunologia , Tuberculose Ocular/microbiologia , Uveíte/microbiologia , Corpo Vítreo/microbiologia , Adulto Jovem
14.
Neurol India ; 66(6): 1550-1571, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30504541

RESUMO

It has been approximately five decades since Dastur et al., published their seminal work on pathology of tuberculous meningitis (TBM). Though most of their findings find relevance in today's era, there is an important difference; these findings can now be replicated during life using modern day technology. In this article, we review the seminal words of Professor Dastur and colleagues, analyse their findings, interpret how these revolutionized our understanding of TBM and highlight their relevance in today's era. We also discuss challenges in the management of TBM, which we continue to face today and the various options required to overcome these challenges.


Assuntos
Antituberculosos/uso terapêutico , Tuberculose Meníngea/história , História do Século XX , Humanos , Tuberculose Meníngea/diagnóstico , Tuberculose Meníngea/tratamento farmacológico
15.
J Proteome Res ; 16(4): 1632-1645, 2017 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-28241730

RESUMO

Mycobacterium tuberculosis, the causative agent of tuberculosis, accounts for 1.5 million human deaths annually worldwide. Despite efforts to eradicate tuberculosis, it still remains a deadly disease. The two best characterized strains of M. tuberculosis, virulent H37Rv and avirulent H37Ra, provide a unique platform to investigate biochemical and signaling pathways associated with pathogenicity. To delineate the biomolecular dynamics that may account for pathogenicity and attenuation of virulence in M. tuberculosis, we compared the proteome and phosphoproteome profiles of H37Rv and H37Ra strains. Quantitative phosphoproteomic analysis was performed using high-resolution Fourier transform mass spectrometry. Analysis of exponential and stationary phases of these strains resulted in identification and quantitation of 2709 proteins along with 512 phosphorylation sites derived from 257 proteins. In addition to confirming the presence of previously described M. tuberculosis phosphorylated proteins, we identified 265 novel phosphorylation sites. Quantitative proteomic analysis revealed more than five-fold upregulation of proteins belonging to virulence associated type VII bacterial secretion system in H37Rv when compared to those in H37Ra. We also identified 84 proteins, which exhibited changes in phosphorylation levels between the virulent and avirulent strains. Bioinformatics analysis of the proteins altered in their level of expression or phosphorylation revealed enrichment of pathways involved in fatty acid biosynthesis and two-component regulatory system. Our data provides a resource for further exploration of functional differences at molecular level between H37Rv and H37Ra, which will ultimately explain the molecular underpinnings that determine virulence in tuberculosis.


Assuntos
Mycobacterium tuberculosis/genética , Fosfoproteínas/biossíntese , Proteoma/genética , Tuberculose/microbiologia , Humanos , Espectrometria de Massas , Mycobacterium tuberculosis/patogenicidade , Fosfoproteínas/genética , Fosforilação/genética , Proteômica/métodos , Transdução de Sinais/genética , Tuberculose/genética , Tuberculose/patologia
17.
Acta Neurochir (Wien) ; 159(3): 503-507, 2017 03.
Artigo em Inglês | MEDLINE | ID: mdl-28110400

RESUMO

BACKGROUND: Establishment of a reliable and rapid diagnosis is of paramount importance in spinal tuberculosis. The available gadgetry of investigations, such as AFB smear, culture of Mycobacterium tuberculosis, and Uniplex PCR, suffers from a lack of adequate sensitivity and/or a lack of rapidity. Therefore, many times a diagnosis is made either very late in the disease process or sometimes empirical therapy has to be started because a definite diagnosis could not be made. All of these are not ideal situations for a clinician. The present study was done with the aim to establish a rapid and reliable diagnosis of M. tuberculosis infection. This was established by identifying M. tuberculosis genes. METHODS: The study was done on nine consecutive patients who presented with non-traumatic spontaneous vertebral compression collapse. CT-guided aspirate from the involved vertebra was subjected to Multiplex PCR (MPCR) using three primers: IS6110, protein b, and MPB 64. The aspirate was also subjected to smear and culture. The results of MPCR were compared with the final diagnosis. RESULTS: Seven out of nine patients had a final diagnosis of tuberculosis. MPCR was positive in six of these seven patients, thus showing sensitivity of 85.7% and specificity of 100%. Results of MPCR were obtained within 24 h. CONCLUSIONS: MPCR using IS6110, protein b, and MPB64 primers has a high sensitivity and specificity in rapid diagnosis of spinal tuberculosis. To the best of our knowledge, this has not been attempted before in spinal tuberculosis. This is particularly useful for paucibacillary infections like spinal tuberculosis. However, further studies using large sample sizes are needed to confirm the practical applicability of this technique.


Assuntos
Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Tuberculose da Coluna Vertebral/diagnóstico , Feminino , Humanos , Masculino , Técnicas de Diagnóstico Molecular/normas , Reação em Cadeia da Polimerase Multiplex/normas , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/isolamento & purificação , Projetos Piloto , Sensibilidade e Especificidade , Tuberculose da Coluna Vertebral/microbiologia
18.
J Assoc Physicians India ; 65(12): 40-47, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31556271

RESUMO

BACKGROUND AND AIMS: There is paucity of data on pulmonary and ENT involvement in ANCA associatd vasculitis from India. We aimed to review the pattern of lung and upper respiratory tract involvement in patients with AAV diagnosed at our centre. METHODS: A retrospective review of all AAV patients between January 2007 and June 2014 was done. A complete clinical evaluation for Pulmonary and ENT involvement was done. Advanced investigations including computed tomography (CT) bronchoscopy and nasal endoscopy were done when indicated. Proportion of involvement was noted and different variables among patient groups were compared. RESULTS: 92 patients (median age 42 years; 60% female) of AAV were included. Clinical and/or radiological evidence of lung involvement was seen in 70 (76.1%) patients. Diffuse alveolar haemorrhage was present in 6 (60%) patients with MPA and 7 (10.1%) patients with GPA (p=0.002). ENT involvement was present in 55 (59.8%) patients and was more in GPA (p=0.000). Absence of renal involvement [p=0.047] and absence of GI involvement [p=0.012] were associated with ENT involvement in GPA. CONCLUSION: Pulmonary involvement was common in GPA, MPA and CSS, ENT involvement was almost characteristic of GPA. DAH was common in MPA. Population based and multicentre studies are needed to assess the true burden of organ involvement in AAV in the Indian population.

20.
Ophthalmology ; 122(4): 840-50, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25578256

RESUMO

PURPOSE: To compare 3 different molecular techniques to detect the Mycobacterium tuberculosis genome in vitreous fluid of eyes with multifocal serpiginoid choroiditis (MSC). DESIGN: Prospective, interventional case series. PARTICIPANTS: Eleven patients (11 eyes) with active MSC in at least 1 eye underwent diagnostic pars plana vitrectomy (PPV) between October 2012 and December 2013. METHODS: Vitreous fluid samples were subjected to multitargeted polymerase chain reaction (PCR) for a M. tuberculosis assay, the Gene Xpert MTB/RIF assay (Cepheid, Sunnyvale, CA), and a line probe assay (GenoType MTBDRplus; Hain Lifescience, GmbH, Nehren, Germany). The samples with positive results were subjected to rpoB gene sequencing to demonstrate rifampicin resistance. The clinical details, digital fundus imaging, and treatment details and outcomes also were noted. MAIN OUTCOME MEASURES: Detection of the M. tuberculosis genome and rifampicin resistance in the vitreous samples. RESULTS: Of the 11 eyes subjected to PPV, the multitargeted PCR results for tuberculosis were positive for 10 eyes, the MTBDRplus assay results were positive in 6 eyes, and the Gene Xpert MTB/RIF assay results were positive in 4 eyes. Rifampicin resistance was detected in 3 eyes by rpoB gene sequencing, in 3 eyes by the MTBDRplus assay, and in 1 eye by the Gene Xpert MTB/RIF assay. CONCLUSIONS: We detected the M. tuberculosis genome in the vitreous fluid of eyes with MSC using 3 different molecular techniques. Rifampicin resistance was detected for the first time in eyes with MSC.


Assuntos
Corioidite/microbiologia , Genoma Bacteriano/genética , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose Resistente a Múltiplos Medicamentos/microbiologia , Tuberculose Ocular/microbiologia , Corpo Vítreo/microbiologia , Adolescente , Adulto , Antibióticos Antituberculose/farmacologia , Antituberculosos/uso terapêutico , Corioidite/diagnóstico , Corioidite/tratamento farmacológico , Farmacorresistência Bacteriana , Quimioterapia Combinada , Feminino , Angiofluoresceinografia , Humanos , Masculino , Pessoa de Meia-Idade , Técnicas de Diagnóstico Molecular , Coroidite Multifocal , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/genética , Reação em Cadeia da Polimerase , Estudos Prospectivos , Rifampina/farmacologia , Tuberculose Resistente a Múltiplos Medicamentos/diagnóstico , Tuberculose Resistente a Múltiplos Medicamentos/tratamento farmacológico , Tuberculose Ocular/diagnóstico , Tuberculose Ocular/tratamento farmacológico , Vitrectomia , Adulto Jovem
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