Identifying quantitative sncRNAs signature using global sequencing as a potential biomarker for tuberculosis diagnosis and their role in regulating host response.

OBJECTIVES: The study aimed to identify a quantitative signature of circulating small non-coding RNAs (sncRNAs) as a biomarker for pulmonary tuberculosis disease (active-TB/ATB) and explore their regulatory roles in host-pathogen interactions and disease progression. METHODS: We conducted a cross-sectional study recruiting subjects diagnosed with active-TB (drug-sensitive and drug-resistant) and healthy controls. Sera samples were collected and utilized for preparing small RNA libraries. Quantitative patterns of circulating sncRNAs (miRNAs, piRNAs and tRFs) were identified via high-throughput sequencing and DeSeq2 analysis and validated in independent active-TB cohorts. Functional knockdown for two selected miRNAs were also performed. RESULTS: A diagnostic signature of four sncRNAs for both drug-sensitive and drug-resistant active-TB cases was validated, exhibiting an AUC of 0.96 (95% CI: 0.937-0.996, p < 0.001) with 86.7% sensitivity (95% CI: 0.775-0.932) and 91.7% specificity (95% CI: 0.730-0.990) in ROC analysis. Functional knockdown demonstrated regulatory roles of hsa-miR-223-5p and hsa-miR-10b-5p in Mycobacterium tuberculosis (Mtb) growth and pro-inflammatory cytokine expression (IL-6 and IL-8). CONCLUSION: The study identified a diagnostic tool utilizing a signature of four sncRNAs with high specificity and sensitivity, enhancing our understanding of sncRNAs as ATB diagnostic biomarker. Additionally, hsa-miR-223-5p and hsa-miR-10b-5p demonstrated potential roles in Mtb pathogenesis and host-response to infection.

Evaluating the efficacy of stool sample on Xpert MTB/RIF Ultra and its comparison with other sample types by meta-analysis for TB diagnostics.

Precise and timely detection of tuberculosis (TB) is crucial to reduce transmission. This study aims to assess the accuracy of Xpert MTB/RIF Ultra on stool samples and systematically review the performance of Xpert MTB/RIF Ultra with different sample types by meta-analysis. Stool samples of smear-negative pulmonary TB (PTB), cervical lymph node TB, and abdominal TB patients were tested on the Xpert MTB/RIF Ultra system. Meta-analysis was performed on a set of 44 studies. Data were grouped by sample type, and the pooled sensitivity and specificity of Xpert MTB/RIF Ultra were calculated. The sensitivity of Xpert MTB/RIF Ultra with stool samples was 100% for smear-negative PTB, 27.27% for cervical lymph node TB, and 50% for abdominal TB patients, with 100% specificity for all included TB groups. The summary estimate for all PTB samples showed 84.2% sensitivity and 94.5% specificity, and EPTB samples showed 88.6% sensitivity and 96.4% specificity. Among all sample types included in our meta-analysis, urine showed the best performance for EPTB diagnosis. This pilot study supports the use of stool as an alternative non-invasive sample on Xpert MTB/RIF Ultra for rapid testing, suitable for both PTB and EPTB diagnosis.

Latent Tuberculosis Infection Diagnosis among Household Contacts in a High Tuberculosis-Burden Area: a Comparison between Transcript Signature and Interferon Gamma Release Assay.

Diagnosis of latent tuberculosis infection (LTBI) using biomarkers in order to identify the risk of progressing to active TB and therefore predicting a preventive therapy has been the main bottleneck in eradication of tuberculosis. We compared two assays for the diagnosis of LTBI: transcript signatures and interferon gamma release assay (IGRA), among household contacts (HHCs) in a high tuberculosis-burden population. HHCs of active TB cases were recruited for our study; these were confirmed to be clinically negative for active TB disease. Eighty HHCs were screened by IGRA using QuantiFERON-TB Gold Plus (QFT-Plus) to identify LTBI and uninfected cohorts; further, quantitative levels of transcript for selected six genes (TNFRSF10C, ASUN, NEMF, FCGR1B, GBP1, and GBP5) were determined. Machine learning (ML) was used to construct models of different gene combinations, with a view to identify hidden but significant underlying patterns of their transcript levels. Forty-three HHCs were found to be IGRA positive (LTBI) and thirty-seven were IGRA negative (uninfected). FCGR1B, GBP1, and GBP5 transcripts differentiated LTBI from uninfected among HHCs using Livak method. ML and ROC (Receiver Operator Characteristic) analysis validated this transcript signature to have a specificity of 72.7%. In this study, we compared a quantitative transcript signature with IGRA to assess the diagnostic ability of the two, for detection of LTBI cases among HHCs of a high-TB burden population; we concluded that a three gene (FCGR1B, GBP1, and GBP5) transcript signature can be used as a biomarker for rapid screening. IMPORTANCE The study compares potential of transcript signature and IGRA to diagnose LTBI. It is first of its kind study to screen household contacts (HHCs) in high TB burden area of India. A transcript signature (FCGR1B, GBP1, & GBP5) is identified as potential biomarker for LTBI. These results can lead to development of point-of-care (POC) like device for LTBI screening in a high TB burdened area.

Early implementation challenges in electronic referral and feedback mechanism for patients with tuberculosis using Nikshay - A mixed-methods study from a medical college TB referral unit of Delhi, India.

BACKGROUND: Diagnosis, notification and timely initiation of treatment is an important cornerstone for the elimination of tuberculosis (TB). The referral and feedback mechanism under National Tuberculosis Programme of India has been changed from paper-based to web-based electronic system (Nikshay) since 2018. The current study was carried out to assess the effect of Nikshay in referral and receipt of feedback on treatment initiation and to understand the early implementation challenges. METHODS: A mixed-methods study was conducted in a medical college referral unit (MCRU) of Delhi, India. The electronic TB notification data for July 2018-March 2019 were abstracted from Nikshay portal and analysed. Unadjusted and adjusted relative risk (aRR) was calculated to assess the factors associated with the receipt of feedback. Themes and subthemes were generated from qualitative data obtained through key-informant interviews of healthcare providers. RESULTS: Of the total 4395 patients handled by MCRU during the study period, 3315 (75.4%) were referred out within and outside Delhi for treatment. Feedback was received among 797 (24.0%) of the patients who were referred out. Patients with extrapulmonary TB (aRR: 1.3, confidence interval (CI): 1.1-1.8), previously treated (aRR: 1.2, CI: 1.2-1.3) and registered for drug-resistant TB care (aRR: 1.4, CI: 1.1-1.8), had high chance of receiving feedback. Four broad themes emerged, namely, (a) awareness of programme and Nikshay; (b) tracking of patients; (c) user-friendly portal and (d) workload. CONCLUSION: The low feedback on treatment initiation of patients with TB needs further research after health system-level quality improvement interventions. Real-time tracking of patients is the need of the hour towards the path for TB elimination.

A comparison of patient treatment pathways among multidrug-resistant and drug-sensitive TB cases in Delhi, India: A cross-sectional study.

BACKGROUND: The delay in the diagnosis and treatment initiation of patients with MDR-TB worsens individual prognosis and increases the risk of disease transmission in the community. These delays have been attributed to delay in treatment-seeking by the patient and shifting to multiple healthcare facilities before being tested and diagnosed through India's National Tuberculosis Elimination Program (NTEP). OBJECTIVE: to identify treatment pathways in patients with MDR-TB from the time of onset of symptoms and treatment seeking until diagnosis at a PMDT site and subsequent treatment initiation. We also compared these characteristics with those of patients with DS-TB. METHODS: We recruited a total of 168 patients with MDR-TB and DS-TB each, in Delhi. Data were analyzed using IBM SPSS Version 25. RESULTS: The mean (SD) patient delay for initial treatment-seeking was 20.9 (15.9) days in patients with MDR-TB, and 16.1 (17.1) days in patients with DS-TB (p < 0.001). The median time from visit to the first healthcare facility (HCF) until confirmation of MDR-TB diagnosis was 78.5 days, and until treatment initiation was 102.5 days. Among patients with DS-TB, the time interval from a visit to the first HCF until the initiation of ATT-DOTS was 61.5 days.. Patients diagnosed with DS-TB, whose first source of treatment was a private facility (n = 49), reported a significant delay in the initiation of ATT-DOTS (p < 0.001). CONCLUSIONS: Despite the introduction of universal drug sensitivity testing in individuals having presumptive MDR-TB, a significant delay in the diagnosis and initiation of effective MDR-TB treatment persists as a major public health challenge in India.

Does Drug-Resistant Extrapulmonary Tuberculosis Hinder TB Elimination Plans? A Case from Delhi, India.

Extrapulmonary drug-resistant tuberculosis (DR-EPTB) poses a formidable diagnostic and therapeutic challenge.Besides associated with high morbidity, it is a major financial burden for the patient and the health system. In spite of this, it has often been neglected as it does not "pose" a visible public health threat. We study clinical profiles, treatment outcomes, and factors associated with unfavourable outcomes among DR-EPTB patients under programmatic settings in New Delhi, India, and evaluate how this could impact TB elimination. A retrospective analysis of all DR-EPTB patients registered at three nodal DR-TB centres in Delhi in 2016 was carried out. Of the 1261 DR-TB patients registered, 203 (16%) were DR-EPTB, with lymph nodes (118, 58%) being the most common site, followed by bone (69, 34%). Nearly 29% (n = 58) experienced adverse drug reactions with severe vomiting (26, 13 %), joint pain (21, 10%) and behavioral disorder (15, 7%). History of previous TB treatment was observed in a majority of the cases (87.7%). Nearly one-third of DR-EPTB cases (33%) had unfavourable treatment outcomes, with loss-to-follow-up (n = 40, 58%) or death (n = 14, 20%) being the most common unfavourable outcomes. In the adjusted analysis, weight band 31-50 kilograms (aRR = 1.8, 1.2-3.4) and h/o previous TB (aRR = 2.1, 1.1-4.8) were mainly associated with unfavourable outcomes. TB elimination efforts need to focus on all forms of TB, including DR-EPTB, leaving no one behind, in order to realise the dream of ending TB.

Diagnostic performance of non-invasive, stool-based molecular assays in patients with paucibacillary tuberculosis.

Timely diagnosis of paucibacillary tuberculosis (TB) which includes smear-negative pulmonary TB (PTB) and extra-pulmonary TB (EPTB) remains a challenge. This study was performed to assess the diagnostic utility of stool as a specimen of choice for detection of mycobacterial DNA in paucibacillary TB patients in a TB-endemic setting. Stool samples were collected from 246 subjects including 129 TB patients (62 PTB and 67 EPTB) recruited at TB hospital in Delhi, India. Diagnostic efficacy of stool IS6110 PCR (n = 228) was measured, using microbiologically/clinically confirmed TB as the reference standard. The clinical sensitivity of stool PCR was 97.22% (95% confidence interval (CI), 85.47-99.93) for detection of Mycobacterium tuberculosis in stool samples of smear-positive PTB patients and 76.92% (CI, 56.35-91.03) in samples from smear-negative PTB patients. Overall sensitivity of PCR for EPTB was 68.66% (CI, 56.16-79.44), with the highest sensitivity for stool samples from patients with lymph node TB (73.5%), followed by abdominal TB (66.7%) and pleural effusion (56.3%). Stool PCR presented a specificity of 95.12%. The receiver operating characteristic curve also indicated the diagnostic utility of stool PCR in TB detection (AUC: 0.882). The performance characteristic of the molecular assay suggests that stool DNA testing has clinical value in detection of TB.

Nonmelanoma skin cancer treatment training varies across different medical specialists.

BACKGROUND: Physicians from various specialties treat patients with nonmelanoma skin cancer (NMSC). The isolation of specialties from each other may result in different approaches to skin cancer training. PURPOSE: Our purpose was to determine the type and amount of NMSC surgical training that is received during dermatology, general surgery, internal medicine, otolaryngology, and plastic surgery residencies. METHODS: E-mail contact information for residency program directors of all accredited programs in each specialty was compiled through the American Medical Association's online residency database. A total of 920 residency program directors were emailed surveys concerning the training of residents in the treatment of NMSC. RESULTS: Forty-two of 920 surveys were returned. All surveyed specialty groups, except internal medicine, had training in NMSC treatment including simple excision, split thickness skin grafts, and tissue rearrangement. A majority of the dermatology and plastic surgery programs instruct their residents in Mohs micrographic surgery and full thickness skin grafts. Electrodessication and curettage was most often instructed in dermatology, general surgery, and plastic surgery programs. CONCLUSION: Greater consistency in NMSC treatment training may be beneficial. Because different approaches may be best suited to particular clinical situations, NMSC treatment training should include adequate exposure to all NMSC treatment techniques.

US dermatopathology fellows career survey: 2004-2005.

BACKGROUND: Graduates of a dermatopathology fellowship may choose an academic career or a career in private practice. OBJECTIVE: To assess career plans of 2004-2005 dermatopathology fellows and to correlate an academic career choice with factors identified in a national survey of US dermatopathology fellowship programs. METHODS: Surveys were mailed to 60 trainees at 45 dermatopathology fellowship programs across the United States. Pearson correlation analysis was used to interpret the data. RESULTS: Thirty-five surveys (58% response rate) were returned. Top five factors that correlated positively with an academic career choice were graduating from a non-US medical school, performing research during fellowship, importance of research in a career decision, completing a dermatology residency and publication requirement in fellowship. Top five factors that correlated positively with choosing a career in private practice were loan debt, importance of salary/earning potential, importance of job availability, being married and having an employed spouse. LIMITATIONS: Study limitations are a small sample size and potential response bias. CONCLUSION: Supporting research during fellowship, supporting applicants who completed a dermatology residency or graduated from a foreign medical school, providing loan forgiveness/repayment and increasing earning/salary potential in academic practice may encourage more young physicians to join the academic workforce.

Advanced squamous cell carcinoma of lower gingivobuccal complex: patterns of spread and failure.

BACKGROUND: Carcinoma of the gingivobuccal complex is commonly associated with the use of smokeless tobacco known as "quid." METHODS: We conducted a retrospective chart review of 511 patients with advanced cancer of gingivobuccal complex surgically treated during 1994 to 1995. We evaluated patterns of disease failure in these patients and correlated disease-free survival with various prognostic factors. RESULTS: During a median follow-up of 46 months, 159 locoregional recurrences and 11 distant metastases were detected in 148 patients. Seventy-nine percent of the recurrences appeared within 18 months of surgery, and the median survival for patients with recurrent disease was less than 4 months. Two-year and 5-year disease-free survival rates were 64% and 57%, respectively. On multivariate analysis, disease-free survival showed significant correlation with skin involvement and extracapsular spread. CONCLUSIONS: Gingivobuccal cancers usually fail locoregionally. Soft tissue infiltration and extracapsular spread of nodal disease influence disease-free survival.