Asunto(s)
Protrombina , Sangrado por Deficiencia de Vitamina K , Humanos , Recién Nacido , Sangrado por Deficiencia de Vitamina K/diagnóstico , Protrombina/genética , Biomarcadores/sangre , Masculino , Deficiencia de Vitamina K/complicaciones , Femenino , Vitamina K/uso terapéutico , Precursores de ProteínasRESUMEN
New or mild heart failure (HF) is mainly caused by left ventricular dysfunction. We hypothesised that gene expression differ between the left (LV) and right ventricle (RV) and secondly by type of LV dysfunction. We compared gene expression through myocardial biopsies from LV and RV of patients undergoing elective coronary bypass surgery (CABG). Patients were categorised based on LV ejection fraction (EF), diastolic function and NT-proBNP into pEF (preserved; LVEF ≥ 45%), rEF (reduced; LVEF < 45%) or normal LV function. Principal component analysis of gene expression displayed two clusters corresponding to LV and RV. Up-regulated genes in LV included natriuretic peptides NPPA and NPPB, transcription factors/coactivators STAT4 and VGLL2, ion channel related HCN2 and LRRC38 associated with cardiac muscle contraction, cytoskeleton, and cellular component movement. Patients with pEF phenotype versus normal differed in gene expression predominantly in LV, supporting that diastolic dysfunction and structural changes reflect early LV disease in pEF. DKK2 was overexpressed in LV of HFpEF phenotype, potentially leading to lower expression levels of ß-catenin, α-SMA (smooth muscle actin), and enhanced apoptosis, and could be a possible factor in the development of HFpEF. CXCL14 was down-regulated in both pEF and rEF, and may play a role to promote development of HF.
Asunto(s)
Insuficiencia Cardíaca , Disfunción Ventricular Izquierda , Humanos , Insuficiencia Cardíaca/diagnóstico , Insuficiencia Cardíaca/genética , Ventrículos Cardíacos , Volumen Sistólico/fisiología , Ecocardiografía , Perfilación de la Expresión Génica , Biopsia , Función Ventricular IzquierdaRESUMEN
Inhibition of Reactive Oxygen Species (ROS) is one of the strategies that Mycobacterium tuberculosis (Mtb) employs as its defence mechanism. In this study, the role of PPE15 (Rv1039c), a late-stage protein, has been investigated in modulating the cellular ROS. We discovered PPE15 to be a secretory protein that downregulates ROS generation in THP1 macrophages. Our in-silico analysis revealed the presence of a eukaryote-like SH3 (SH3e) domain in PPE15. The predicted SH3e-domain of PPE15 was found to interact with cytosolic components of NADPH Oxidase (NOX), p67phox and p47phox through molecular docking. In-vitro experiments using THP1 macrophages showed a diminished NADP/NADPH ratio, indicating reduced NOX activity. We also observed increased levels of p67phox and p47phox in the cytoplasmic fraction of PPE15 treated macrophages as compared to the plasma membrane fraction. To understand the role of the SH3e-domain in ROS modulation, this domain was deleted from the full-length PPE15 (PPE15-/-SH3). We observed an increase in cellular ROS and NADP/NADPH ratio in response to PPE15-/-SH3 protein. The interaction of PPE15-/-SH3 with p67phox or p47phox was also reduced in the cytoplasm, indicating migration of NOX subunits to the plasma membrane. Additionally, M. smegmatis expressing PPE15 was observed to be resistant to oxidative stress with significant intracellular survival in THP1 macrophages as compared to M. smegmatis expressing PPE15-/-SH3. These observations suggest that the SH3e-domain of PPE15 interferes with ROS generation by sequestering NOX components that inhibit NOX assembly at the cell membrane. Therefore, PPE15 acts like a molecular mimic of SH3-domain carrying eukaryotic proteins that can be employed by Mtb at late stages of infection for its survival. These findings give us new insights about the pathogen evading strategy of Mtb which may help in improving the therapeutics for TB treatment.
Asunto(s)
Mycobacterium tuberculosis , Especies Reactivas de Oxígeno/metabolismo , NADP/metabolismo , Dominios Homologos src , Simulación del Acoplamiento Molecular , NADPH Oxidasas/genética , NADPH Oxidasas/metabolismo , EucariontesRESUMEN
Mammalian cell entry (mce) operons play a vital role in cell invasion and survival of M. tuberculosis. Of the mce genes, the function of Rv0590A is still unknown. The present study was performed to investigate the function and immunogenic properties of the protein Rv0590A. Human leukemia monocytic cell line (THP-1) derived macrophages were infected with M. tuberculosis H37Rv at 3, 6, and 24 h of infection. The maximum colony forming units (CFU) were observed at 6 h (p < 0.005), followed by 3 h after infection. M. tuberculosis H37Rv and clinical isolates representative of Delhi/CAS, EAI, Beijing, Haarlem and Euro-American-superlineage were included in the study for expression analysis of mce1A, mce2A, mce3A, mce4A, and Rv0590A genes. Maximum upregulation of all mce genes was observed at 3 h of infection. All the five clinical isolates and H37Rv upregulated Rv0590A at various time points. Macrophage infection with M. tuberculosis H37Rv-overexpressing Rv0590A gene showed higher intracellular CFU as compared to that of wild-type H37Rv. Further, purified Rv0590A protein stimulated the production of TNFα, IFNγ, and IL-10 in macrophages. Thus, Rv0590A was found to be involved in cell invasion and showed good immunological response.
Asunto(s)
Mycobacterium tuberculosis , Tuberculosis , Animales , Humanos , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Internalización del Virus , Mycobacterium tuberculosis/genética , Antígenos Bacterianos/genética , MamíferosRESUMEN
Background: Understanding the protein's subcellular localization and secretory nature can greatly improve the target identification for diagnostic assays and drug discovery, although their identification in laboratory experiments is a time-consuming and labor-intensive process. In order to identify proteins that could be targeted for therapeutic intervention or the development of diagnostic assays, we used a variety of computational tools to predict the subcellular localization or secretory nature of mycobacterial proline-glutamate/proline-proline-glutamate (PE/PPE) proteins. Methods: PSORTb version 3.0.3, TBpred, and Gpos-mPLoc analyses were performed on 30 selected PE/PPE protein sequences, while, SignalP 6.0, SignalP 5.0, Phobius, PSORTb version 3.0.3 and TBpred were used for signal sequence predictions. Results: Gpos-mPLoc and TBpred had the highest concordance for extracellular prediction, while PSORTb and TBpred had the highest concordance for prediction of membrane localization. The tools for predicting the secretory nature of proteins had little agreement. Conclusion: Multiple computational tools must be considered to provide an indication of the subcellular localization of PE/PPE proteins. Laboratory experiments should be used to confirm the findings of the tools.
Asunto(s)
Mycobacterium tuberculosis , Humanos , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/metabolismo , Ácido Glutámico/metabolismo , Algoritmos , Internet , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismoRESUMEN
T cells are an important component of adaptive immunity and T-cell-derived lymphomas are very complex due to many functional sub-types and functional elasticity of T-cells. As with other tumors, tissues specific factors are crucial in the development of T-cell lymphomas. In addition to neoplastic cells, T- cell lymphomas consist of a tumor micro-environment composed of normal cells and stroma. Numerous studies established the qualitative and quantitative differences between the tumor microenvironment and normal cell surroundings. Interaction between the various component of the tumor microenvironment is crucial since tumor cells can change the microenvironment and vice versa. In normal T-cell development, T-cells must respond to various stimulants deferentially and during these courses of adaptation. T-cells undergo various metabolic alterations. From the stage of quiescence to attention of fully active form T-cells undergoes various stage in terms of metabolic activity. Predominantly quiescent T-cells have ATP-generating metabolism while during the proliferative stage, their metabolism tilted towards the growth-promoting pathways. In addition to this, a functionally different subset of T-cells requires to activate the different metabolic pathways, and consequently, this regulation of the metabolic pathway control activation and function of T-cells. So, it is obvious that dynamic, and well-regulated metabolic pathways are important for the normal functioning of T-cells and their interaction with the microenvironment. There are various cell signaling mechanisms of metabolism are involved in this regulation and more and more studies have suggested the involvement of additional signaling in the development of the overall metabolic phenotype of T cells. These important signaling mediators include cytokines and hormones. The impact and role of these mediators especially the cytokines on the interplay between T-cell metabolism and the interaction of T-cells with their micro-environments in the context of T-cells lymphomas are discussed in this review article.
RESUMEN
OBJECTIVE: To evaluate whether magnesium sulfate and therapeutic hypothermia in combination decreases mortality and/or major neurodevelopmental disability at 1 y of age among term neonates with hypoxic-ischemic encephalopathy. METHODS: A total of 134 term neonates were randomized to receive intravenous magnesium sulfate at a dose of 250 mg/kg (at 8 mg/kg/min) once daily for 3 d starting within 6 h after birth along with therapeutic hypothermia in the intervention group and therapeutic hypothermia alone in the comparator group. The primary outcome was the composite outcome of mortality and/or major neurodevelopmental disability (Developmental Assessment Scale for Indian Infants score < 70) at 1 y of age. RESULTS: A total of 115 infants were included in the primary analysis. The composite primary outcome occurred in 14 (24%) infants in the intervention group and 19 (33%) infants in the comparator group, and the difference was not statistically significant (p = 0.30; relative risk 0.72; 95% confidence interval 0.40-1.30). The secondary outcomes including neonatal mortality, major neurodevelopmental disability at 1 y of age, neurological status at discharge, level of oxidative stress markers, and adverse effects including hypotension and respiratory depression requiring support were also comparable between the groups. CONCLUSIONS: The combination of magnesium sulfate and therapeutic hypothermia did not improve the composite outcome of neonatal mortality and/or major neurodevelopmental disability at 1 y of age. TRAIL REGISTRATION: Clinical Trials Registry of India (CTRI/2018/06/014594), prospectively registered.
Asunto(s)
Hipotermia Inducida , Hipoxia-Isquemia Encefálica , Recién Nacido , Lactante , Humanos , Sulfato de Magnesio/uso terapéutico , Sulfato de Magnesio/efectos adversos , Hipoxia-Isquemia Encefálica/terapia , Mortalidad Infantil , Hipotermia Inducida/efectos adversos , Administración IntravenosaRESUMEN
Kidney disease is a complex disease with several different etiologies and underlying associated pathophysiology. This is reflected by the lack of effective treatment therapies in chronic kidney disease (CKD) that stop disease progression. However, novel strategies, recent scientific breakthroughs, and technological advances have revealed new possibilities for finding novel disease drivers in CKD. This review describes some of the latest advances in the field and brings them together in a more holistic framework as applied to identification and validation of disease drivers in CKD. It uses high-resolution 'patient-centric' omics data sets, advanced in silico tools (systems biology, connectivity mapping, and machine learning) and 'state-of-the-art' experimental systems (complex 3D systems in vitro, CRISPR gene editing, and various model biological systems in vivo). Application of such a framework is expected to increase the likelihood of successful identification of novel drug candidates based on strong human target validation and a better scientific understanding of underlying mechanisms.
RESUMEN
Environmental and genetic factors cause defects in pancreatic islets driving type 2 diabetes (T2D) together with the progression of multi-tissue insulin resistance. Mass spectrometry proteomics on samples from five key metabolic tissues of a cross-sectional cohort of 43 multi-organ donors provides deep coverage of their proteomes. Enrichment analysis of Gene Ontology terms provides a tissue-specific map of altered biological processes across healthy, prediabetes (PD), and T2D subjects. We find widespread alterations in several relevant biological pathways, including increase in hemostasis in pancreatic islets of PD, increase in the complement cascade in liver and pancreatic islets of PD, and elevation in cholesterol biosynthesis in liver of T2D. Our findings point to inflammatory, immune, and vascular alterations in pancreatic islets in PD that are hypotheses to be tested for potential contributions to hormonal perturbations such as impaired insulin and increased glucagon production. This multi-tissue proteomic map suggests tissue-specific metabolic dysregulations in T2D.
Asunto(s)
Diabetes Mellitus Tipo 2 , Estado Prediabético , Humanos , Diabetes Mellitus Tipo 2/diagnóstico , Estado Prediabético/diagnóstico , Proteómica , Glucagón/metabolismo , Proteoma/metabolismo , Estudios Transversales , Insulina/genética , Redes y Vías Metabólicas/genética , ColesterolRESUMEN
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.
Asunto(s)
Antibióticos Antituberculosos , Mycobacterium tuberculosis , Tuberculosis , Antibióticos Antituberculosos/farmacología , Farmacorresistencia Bacteriana , Humanos , Mycobacterium tuberculosis/genética , Proyectos Piloto , Rifampin , Sensibilidad y Especificidad , Esputo/microbiología , Tuberculosis/diagnósticoRESUMEN
In an attempt to uncover genotypic indicators for isoniazid (INH) resistance in M. tuberculosis, in addition to the canonical mutations in genes associated with INH resistance, including katG, inhA and fabG promoter; we analyzed, two INH monoresistant isolates, ASTS24/13 (INHR1) and SHR1/14 (INHR2). Targeted Sanger sequencing detected a canonical mutation at katG315 only in INHR2. Infection of THP-1 cells and exposure to antituberculosis drugs led to two-fold increase in the minimum inhibitory concentration of INH in INHR2. Whole genome sequences revealed that INHR1 and INHR2 belonged to Delhi Central Asian Strain and East African Indian lineages, respectively. The sequences were compared with INH susceptible isolates with the same lineage as the INH monoresistant strains. INHR1 had a novel unique mutation STOP420Trp in the efflux pump gene Rv0849, while INHR2 had a novel mutation Arg579Ser in efflux pump gene mmpL5. Comparison of lipid associated genes showed novel mutations in INHR1 in fadE16, fadD3 and fbpD; while INHR2 had mutations in fadE1, Rv0145, Rv1425, fadD9 and mmaA3. Both isolates also demonstrated novel mutations in cell wall associated genes. Our study highlights the importance of searching for alternate mechanisms of INH resistance that may contribute to the development of more comprehensive diagnostic tools.
Asunto(s)
Isoniazida , Mycobacterium tuberculosis , Antituberculosos/farmacología , Antituberculosos/uso terapéutico , Proteínas Bacterianas/genética , Catalasa/genética , Farmacorresistencia Bacteriana/genética , Isoniazida/farmacología , Pruebas de Sensibilidad Microbiana , Mutación , Polimorfismo Genético , Secuenciación Completa del GenomaRESUMEN
BACKGROUND: Among term and late preterm infants, hypoxic ischemic encephalopathy (HIE) is an important cause of mortality, and neurologic morbidity among survivors. OBJECTIVE: The primary objective was to study the incidence of survival to discharge among late preterm and term infants with moderate or severe HIE. Secondary objectives were to explore variation in the management of HIE across participating sites and to identify the predictors of survival. SETTING: Indian Neonatal Collaborative (INNC), a network of 28 neonatal units in India. STUDY DESIGN: Retrospective cohort. PARTICIPANTS: Late preterm (34-36 weeks) and term (37-42 weeks) infants with moderate to severe HIE from 2018-2019. OUTCOME: The primary outcome was survival to discharge (including discharged home and transfer to other hospital). A multivariate logistic regression model was constructed to identify the predictors of survival. RESULTS: Of 352 infants with moderate or severe HIE, 59% received therapeutic hypothermia. Survival to discharge among infants with moderate or severe HIE was 82%. Severe HIE (aOR 0.04; 95% CI 0.02-0.10), persistent pulmonary hypertension (PPHN) (aOR 0.22; 95% CI 0.08-0.61) and requirement of epinephrine during resuscitation (aOR 0.21; 95% CI 0.05-0.84) were independently associated with decreased odds of survival to discharge. CONCLUSION: Survival to discharge among infants with moderate or severe HIE was 82%. Severe HIE, requirement of epinephrine during resuscitation and PPHN decreased the odds of survival.
Asunto(s)
Hipotermia Inducida , Hipoxia-Isquemia Encefálica , Estudios de Cohortes , Humanos , Hipoxia-Isquemia Encefálica/epidemiología , Hipoxia-Isquemia Encefálica/terapia , Lactante , Recién Nacido , Recien Nacido Prematuro , Estudios RetrospectivosRESUMEN
Background: Rapidly growing mycobacteria (RGM) are increasingly being recognized as potential pathogens. RGM, particularly Mycobacterium abscessus, Mycobacterium fortuitum, and Mycobacterium chelonae, have been observed in both pulmonary and extrapulmonary infections including cutaneous, soft-tissue, and wound infections. However, there are limited reports of these potential pathogens from skin and soft-tissue infections. Moreover, the drug susceptibility profile of RGM is largely unknown in several regions of the world. Methods: We analyzed reports on RGM isolated from skin and soft-tissue infections globally for details of RGM species and drug susceptibility profile. We also analyzed the drug susceptibility profile of four RGM isolates, obtained from skin and soft-tissue infections in our laboratory, by broth microdilution method. Results: In the reports reviewed, the most common RGM isolated from skin and soft-tissue infections were M. abscessus (184/475, 38.7%), M. fortuitum (150/475, 31.5%), M. chelonae (72/475, 15%), and M. chelonae-M. abscessus complex (46/475, 9.6%). However, drug susceptibility was tested only in 26/39 (66.6%) reports. In our own laboratory, we obtained three isolates of M. abscessus and one isolate of M. fortuitum from one case of breast abscess and three cases of postsurgical wound infections. Maximum susceptibility of M. abscessus was observed to clarithromycin, amikacin, and linezolid. The M. fortuitum isolate was susceptible to clarithromycin, amikacin, clofazimine, and linezolid. Conclusion: Paucity of information available on RGM isolated from skin and soft-tissue infections highlights the need to be aware of the pathogenic potential and the drug susceptibility profile of these organisms.
Asunto(s)
Infecciones por Mycobacterium no Tuberculosas , Mycobacterium , Amicacina , Antibacterianos/farmacología , Claritromicina , Humanos , Pruebas de Sensibilidad Microbiana , Micobacterias no TuberculosasRESUMEN
There is an avalanche of biomedical data generation and a parallel expansion in computational capabilities to analyze and make sense of these data. Starting with genome sequencing and widely employed deep sequencing technologies, these trends have now taken hold in all omics disciplines and increasingly call for multi-omics integration as well as data interpretation by artificial intelligence technologies. Here, we focus on mass spectrometry (MS)-based proteomics and describe how machine learning and, in particular, deep learning now predicts experimental peptide measurements from amino acid sequences alone. This will dramatically improve the quality and reliability of analytical workflows because experimental results should agree with predictions in a multi-dimensional data landscape. Machine learning has also become central to biomarker discovery from proteomics data, which now starts to outperform existing best-in-class assays. Finally, we discuss model transparency and explainability and data privacy that are required to deploy MS-based biomarkers in clinical settings.
Asunto(s)
Inteligencia Artificial , Proteómica , Biomarcadores/análisis , Espectrometría de Masas/métodos , Proteómica/métodos , Reproducibilidad de los ResultadosAsunto(s)
Ictericia Obstructiva , Síndrome de Respuesta Inflamatoria Sistémica , Preescolar , Humanos , Ictericia Obstructiva/diagnóstico , Ictericia Obstructiva/etiología , Masculino , Síndrome de Respuesta Inflamatoria Sistémica/complicaciones , Síndrome de Respuesta Inflamatoria Sistémica/diagnósticoAsunto(s)
Infecciones por Flavobacteriaceae , Flavobacteriaceae , Meningitis Bacterianas , Meningitis , Sepsis Neonatal , Sepsis , Infecciones por Flavobacteriaceae/diagnóstico , Infecciones por Flavobacteriaceae/tratamiento farmacológico , Humanos , Recién Nacido , Meningitis/diagnóstico , Meningitis/etiología , Meningitis Bacterianas/diagnóstico , Sepsis Neonatal/diagnóstico , Sepsis/diagnósticoRESUMEN
The study evaluated the short-term outcome of children diagnosed with conversion disorder and compared their pre- and post-intervention psychological functioning. Fifty children consecutively diagnosed with conversion disorder over a period of one year were recruited from the pediatrics department of a tertiary care teaching hospital in North India. The adverse life events were assessed by the Life Events Scale for Indian Children, emotional and behavioral difficulties by the Childhood Psychopathology Measurement Schedule (CPMS), and adjustment by the Pre-Adolescent Adjustment Scale (PAAS). Majority of the children improved at follow up at 3 mo after initiation of treatment. In addition, total scores on the CPMS significantly declined (t = 5.12, P = 0.0001) and self-reported adjustment improved on the PAAS (t = 5.81, P = 0.0001) as compared to functioning before the initiation of therapy. Timely recognition and multi-disciplinary management can lead to successful outcome and improved functioning in most children diagnosed with conversion disorder.
Asunto(s)
Cognición , Emociones , Adolescente , Niño , Humanos , IndiaRESUMEN
OBJECTIVE: The present study aims to implement family-centered care (FCC) in neonatal intensive care unit (NICU). FCC facilitates mother-infant bonding with benefits for both families and health system. The authors used quality-improvement (QI) methods to implement FCC in level-2 NICU from an existing baseline of 30% to 80% over a period of 6 mo. METHODS: Using the Institute of Healthcare model for improvement, the authors implemented strategies for family participation in caregiving activities, oral feeding and kangaroo care for neonates admitted in level-2 NICU. Eligibility criteria included the availability of at least one family member, preferably the mother for at least 6 h/d and a stable neonate based on physiological criteria irrespective of gestational age. The key interventions were: (1) adoption of a unit protocol for FCC with expanded visitation hours; (2) parental education through audio-visual aids, and (3) capacity building through training and peer support. RESULT: Between August 2019 and January 2020, 1587 neonates were admitted to the NICU and 505 admitted in level-2 were enrolled. The proportion of eligible mother-infant dyads participating in FCC increased from a baseline of 32% to 44% during intervention and to 66% in the postintervention phase. The number of days per month FCC was tracked increased from 67% in the baseline to 82% in postintervention phase. There was no increase in the incidence of sepsis after implementation of FCC. CONCLUSION: Orientation of parents to FCC using audio-visual aids, provision of hands-on training and peer-support facilitated them to become active participants in their neonates' care.