👤 Manish Shankar

Multidrug-resistant tuberculosis continues to be a major threat to mankind and is a major social and economic burden to society. Line probe assay (LPA) is a method for detecting Mycobacterium tuberculosis (MTb) in combination with resistance to rifampicin and isoniazid by first-line LPA, resistance to fluoroquinolones (FQ) and second-line injectable drugs (SLID) by second-line LPA in sputum smear-positive specimens (direct testing) and cultured isolates (indirect testing). LPA uses a DNA-based reverse hybridization method, which determines the drug resistance profile through the pattern of binding of DNA amplicons to probes that target specific areas of the MTb genome for MTb detection and to most common mutations conferring resistance to various drugs and/or the corresponding wild-type DNA sequence. LPA is endorsed by the National Tuberculosis Elimination Programme (NTEP), but very limited data are available, especially from high-burden areas such as Bihar, where a rapid, accurate, cost-effective technique like LPA can play a crucial role in early diagnosis and initiation of treatment and ultimately contribute to the effective elimination of the disease. This cross-sectional study was performed at Indira Gandhi Institute of Medical Sciences, Patna, a tertiary care centre, and aimed at the molecular characterization of Rifampicin resistant tuberculosis (RR-TB) isolates using the line probe assay method. The study was conducted between November 2022 and May 2024 with 116 samples obtained from both pulmonary tuberculosis(PTB) and extrapulmonary tuberculosis(EPTB) cases that were found to be rifampicin-resistant on CBNAAT (Cartridge based nucleic acid amplification test-Gene Xpert Mtb/RIF). In this study, 116 patients were enrolled, of whom 80(68.9 %) were pre-XDR-TB cases and 36(31 %) were MDR-TB cases. The most common pattern of mutation associated with rifampicin rpoB gene was WT8 MUT3 S531L and, Isoniazid inhA gene was WT1 MUT1 c-15 t. High-level isoniazid resistance involving KatG mutation was present in 111 (95.7 %) cases and the most common mutation associated was MUT1 -S315T1. Overall prevalence of fluoroquinolone resistance in this study was 68.9 %. There is a wide prevalence of high-level isoniazid resistance and fluoroquinolone resistance among RR-TB patients, indicating the rapid emergence and transmission of resistant strains in the community. This underscores the need for enforced interventions, such as screening for MDR-TB before starting therapy and surveillance of fluoroquinolone susceptibility. Molecular characterization of RR-TB strains by Line probe assay method can play a critical role in the rapid determination of pattern of resistance in the circulating strains and hence guide tailored therapy at the earliest opportunity especially in high burden setting with limited infrastructure. Show less

Pre-pregnancy overweight and obesity promote deleterious health impacts on both mothers during pregnancy and the offspring. Significant changes in the maternal peripheral blood mononuclear cells (PBMCs) gene expression due to obesity are well-known. However, the impact of pre-pregnancy overweight on immune cell gene expression during pregnancy and its association with maternal and infant outcomes is not well explored. Blood samples were collected from healthy normal weight (NW, pre-pregnancy BMI 18.5-24.9) or overweight (OW, pre-pregnancy BMI 25-29.9) 2nd parity pregnant women at 12, 24 and 36 weeks of pregnancy. PBMCs were isolated from the blood and subjected to mRNA sequencing. Maternal and infant microbiota were analyzed by 16S rRNA gene sequencing. Integrative multi-omics data analysis was performed to evaluate the association of gene expression with maternal diet, gut microbiota, milk composition, and infant gut microbiota. Gene expression analysis revealed that 453 genes were differentially expressed in the OW women compared to NW women at 12 weeks of pregnancy, out of which 354 were upregulated and 99 were downregulated. Several up-regulated genes in the OW group were enriched in inflammatory, chemokine-mediated signaling and regulation of interleukin-8 production-related pathways. At 36 weeks of pregnancy healthy eating index score was positively associated with several genes that include, DTD1, ELOC, GALNT8, ITGA6-AS1, KRT17P2, NPW, POT1-AS1 and RPL26. In addition, at 36 weeks of pregnancy, genes involved in adipocyte functions, such as NG2 and SMTNL1, were negatively correlated to human milk 2'FL and total fucosylated oligosaccharides content collected at 1 month postnatally. Furthermore, infant Akkermansia was positively associated with maternal PBMC anti-inflammatory genes that include CPS1 and RAB7B, at 12 and 36 weeks of pregnancy. These findings suggest that prepregnancy overweight impacts the immune cell gene expression profile, particularly at 12 weeks of pregnancy. Furthermore, deciphering the complex association of PBMC's gene expression levels with maternal gut microbiome and milk composition and infant gut microbiome may aid in developing strategies to mitigate obesity-mediated effects. Show less

Introduction: Despite significant advances in pediatric burn care, bloodstream infections (BSIs) remain a compelling challenge during recovery. A personalized medicine approach for accurate prediction of BSIs before they occur would contribute to prevention efforts and improve patient outcomes. Methods: We analyzed the blood transcriptome of severely burned (total burn surface area [TBSA] ≥20%) patients in the multicenter Inflammation and Host Response to Injury ("Glue Grant") cohort. Our study included 82 pediatric (aged <16 years) patients, with blood samples at least 3 days before the observed BSI episode. We applied the least absolute shrinkage and selection operator (LASSO) machine-learning algorithm to select a panel of biomarkers predictive of BSI outcome. Results: We developed a panel of 10 probe sets corresponding to six annotated genes ( ARG2 [ arginase 2 ], CPT1A [ carnitine palmitoyltransferase 1A ], FYB [ FYN binding protein ], ITCH [ itchy E3 ubiquitin protein ligase ], MACF1 [ microtubule actin crosslinking factor 1 ], and SSH2 [ slingshot protein phosphatase 2 ]), two uncharacterized ( LOC101928635 , LOC101929599 ), and two unannotated regions. Our multibiomarker panel model yielded highly accurate prediction (area under the receiver operating characteristic curve, 0.938; 95% confidence interval [CI], 0.881-0.981) compared with models with TBSA (0.708; 95% CI, 0.588-0.824) or TBSA and inhalation injury status (0.792; 95% CI, 0.676-0.892). A model combining the multibiomarker panel with TBSA and inhalation injury status further improved prediction (0.978; 95% CI, 0.941-1.000). Conclusions: The multibiomarker panel model yielded a highly accurate prediction of BSIs before their onset. Knowing patients' risk profile early will guide clinicians to take rapid preventive measures for limiting infections, promote antibiotic stewardship that may aid in alleviating the current antibiotic resistance crisis, shorten hospital length of stay and burden on health care resources, reduce health care costs, and significantly improve patients' outcomes. In addition, the biomarkers' identity and molecular functions may contribute to developing novel preventive interventions. Show less

Viruses are masters at using cellular pathways to aid their replication. Cryo-electron tomography of poliovirus-infected cells revealed how it utilizes macroautophagy to its advantage. Assembly of these non-enveloped virions takes place directly on membranes and requires PIK3C3/VPS34 activity to be completed, whereas the canonical autophagy inducer ULK1 restricts virus assembly. The tomograms further revealed that enterovirus-induced autophagy is selective for RNA-loaded virions, which may help ensure maximum infectivity of the virus-laden vesicles released through secretory autophagy. Show less

LY2409021 is a glucagon receptor antagonist that was associated with hepatic steatosis and elevated aminotransferases in phase 2 diabetes studies. We investigated the relationship between selected genetic variants and hepatic steatosis and elevated alanine aminotransferases (ALTs) associated with LY2409021. Patients participated in a 6-week placebo-controlled trial (I1R-MC-GLDI [GLDI], n = 246) and a 52-week placebo- and active comparator-controlled trial (I1R-MC-GLDJ [GLDJ], n = 158). GLDJ had endpoints at 6 months, including measures of hepatic fat fraction (HFF) by magnetic resonance imaging. The five genes tested were patatin-like phospholipase domain containing 3 ( Show less

A diagnosis of granulocytic sarcoma was made in a 2-year-old child based on the detection of myelomonocytic blasts in tissue obtained from a subcutaneous nodule with no evidence of concomitant disease in the bone marrow. The child responded to systemic chemotherapy and is in remission 3 years later. An identical clone with an in frame fusion of the MLL and AF10 genes was identified from both tissue and bone marrow samples. The generation of an in frame MLL-AF10 fusion requires complex intra- and interchromosomal exchanges between chromosomes 10 and 11. In this case, an intrachromosomal rearrangement of chromosome 5 was also observed. This case illustrates the presence of systemic disease in extramedullary leukaemia, its response to systemic rather than topical therapy and suggests that the events leading to chromosomal translocations in leukaemia may be part of a generalized intracellular event. Show less