Evaluation of a new flow cytometry based method for detection of BCR-ABL1 fusion protein in chronic myeloid leukemia

BACKGROUND: Philadelphia chromosome, a hallmark of chronic myeloid leukemia (CML), plays a key role in disease pathogenesis. It reflects a balanced reciprocal translocation between long arms of chromosomes 9 and 22 involving BCR and ABL1 genes, respectively. An accurate and reliable detection of BCR-ABL fusion gene is necessary for the diagnosis and monitoring of CML. Previously, many technologies, most of which are laborious and time consuming, have been developed to detect BCR-ABL chimeric gene or chromosome. METHODS: A new flow cytometric immunobead assay was used for detection of BCR-ABL fusion proteins and applicability, sensitivity, reliability, efficacy and rapidity of this method was evaluated. RESULTS: From February 2009 to January 2014, a total 648 CML patients were investigated for the status of BCR-ABL1 protein. Among them, 83 patients were enrolled for comparative study of BCR-ABL1 positivity by three routinely used procedures like karyotyping, and quantitative real time PCR (RT-PCR) as well as immunobead flow cytometry assay. BCR-ABL protein analysis was found consistent, more sensitive (17% greater sensitivity) and reliable than the conventional cytogenetics, as flow cytometry showed 95% concordance rate to RT-PCR. CONCLUSION: BCR-ABL fusion protein assay using a new flow cytometric immunobead might be useful in the diagnosis and monitoring CML patients.

E84Gmutation in dihydrofolate reductase from drug resistant strains of Mycobacterium tuberculosis [Mumbai, India] leads to increased interaction with Trimethoprim 5

Dihydrofolate reductase [DHFR][dfrA gene] is an essential enzyme for cell survival and an unexplored target in Mycobacterium tuberculosis [Mtb]. This study was carried out to analyze mutations in the dfrA gene amongst 20 clinical DNA samples from Mtb isolates obtained from Mumbai, India. Sequencing of the PCR amplified dfrA gene from these DNA isolates revealed a point mutation in one strain, leading to a glutamic acid to glycine change. In silico simulation studies revealed a surface alteration in the enzyme due to this E84G mutation. The amplified mutant gene was cloned and expressed. The mutant protein was assessed against known DHFR inhibitors: Methotrexate and Trimethoprim. An increased affinity for inhibitor Trimethoprim and native substrate dihydrofolate was observed with the mutant. Methotrexate did not vary in its activity with both the enzyme forms. The Glu84Gly point mutation may lead to a variation in the strain which may cause resistance in the future

Measurement and Modeling of Job Stress of Electric Overhead Traveling Crane Operators

BACKGROUND: In this study, the measurement of job stress of electric overhead traveling crane operators and quantification of the effects of operator and workplace characteristics on job stress were assessed. METHODS: Job stress was measured on five subscales: employee empowerment, role overload, role ambiguity, rule violation, and job hazard. The characteristics of the operators that were studied were age, experience, body weight, and body height. The workplace characteristics considered were hours of exposure, cabin type, cabin feature, and crane height. The proposed methodology included administration of a questionnaire survey to 76 electric overhead traveling crane operators followed by analysis using analysis of variance and a classification and regression tree. RESULTS: The key findings were: (1) the five subscales can be used to measure job stress; (2) employee empowerment was the most significant factor followed by the role overload; (3) workplace characteristics contributed more towards job stress than operator's characteristics; and (4) of the workplace characteristics, crane height was the major contributor CONCLUSION: The issues related to crane height and cabin feature can be fixed by providing engineering or foolproof solutions than relying on interventions related to the demographic factors.

Nutritional status of school going adolescents in a rural block of darjeeling, West Bengal, India

Background: Poor nutritional status during adolescence is an important determinant of health outcome. The adolescents have different needs and have diverse problems. This study was conducted to assess the nutritional status and find out the morbidity pattern of the school going adolescents. Methods: A cross-sectional study was conducted among school going adolescents, 11-19 years studying in various schools of Naxalbari block of Darjeeling district in 2007. A two stage sampling design was used to draw representative sample. Participants were clinically examined, anthropometric measurements were made and pre-tested questionnaire was dispensed. BMI-forage and height-for-age were considered as the indicators. Results: The study population comprise of 420 participants [39% girls]. 40% of the adolescents were under nourished. Severe malnutrition was found in 27% of male and 29% of female population. 44% of the study participants were stunted and 14% were severely stunted. Pallor was present in 40% participants. One fourth of the adolescents complained of psychological problems. Conclusion: There is an urgent need to intensify efforts to improve the nutritional profile of children to optimize human resource development