Women's Empowerment in the Context of Rural West Bengal: An Index Construction with Exploratory Factor Analysis.

Background: Women's empowerment, a precondition of sustainable development, is a multidimensional and complex concept, often described with three interrelated components: resources, agency, and achievement. There is no universal construct for women's empowerment; rather, it has been assessed based on the context. It had been hardly explored in rural West Bengal. Objectives: This study was formulated to construct a women's empowerment index (WEI), in rural West Bengal, and assess the reliability of the index. Methods: A community-based cross-sectional study focusing on Women's Empowerment, Child Health and Nutrition (WE-CHANT) was conducted in a community development block in West Bengal. Mother (of reproductive age)-child (6-59 months) pairs were recruited from 20 villages by two-stage sampling (n = 268). Mothers were interviewed. Exploratory factor analysis (EFA) with oblique rotation was conducted with 25 measurement variables to construct the WEI. The internal consistency was assessed with Cronbach's alpha, item-rest, average inter-item, inter-domain, and domain-to-index correlation. Results: A 12-item (factor loading ≥0.40) WEI comprising three domains-decision-making power, attitude toward gender-based violence (GBV), and social independence-was constructed with acceptable internal consistency (Cronbach's alpha = 0.747). The multidimensionality of the index was also observed. Conclusion: A concise agency-based WEI was constructed, where decision-making power was explored as the major domain. This index in the context of rural West Bengal could be further utilized to assess women's empowerment and elicit its association with resources and achievements.

Malaria infection and associated household level risks in an urban and rural areas of West Bengal.

BACKGROUND OBJECTIVES: Malaria is one of the most critical public health problem globally as well as nationally. Recent times have seen an increase in the importance of the possible effects of environmental and demographic factors on the local transmission of malaria. We conducted a community-level study between urban and rural areas to find out the factors associated with malaria infection. METHODS: A community based observational cross-sectional study was conducted between October 2020 to September 2022 among 330 households from urban area of Chetla, Kolkata and rural areas of Singur district of West Bengal. Households were selected by two stage random sampling method. Study done by face-to-face interview, review of medical records and observation of the households. SPSS software was used for data analysis. Multivariable logistic regression (hierarchical model) done. (p-value equal to or less than 0.05 was taken as significant). RESULTS: The prevalence of malaria infection among participants and household level risks of malaria infection were found 6.1%, 1.8% and 18.8%, 6.7% respectively in the urban and rural areas. Significant predictors of malaria infection were household with ≥5 family members [AOR=2.91{CI (1.19-7.15)}], households not used any mosquito protective measure [AOR=3.71{CI (1.04-13.13)}], presence of mosquito breeding place [AOR=4.93{CI (1.53-15.93)}], absence of effective window screen [AOR=3.19{CI (1.16-8.71)}], absence of separate kitchen [AOR=2.38{CI (1.06-5.34)}], and inadequate lighting condition of house [AOR=4.35{CI (1.78-10.64)}]. INTERPRETATION CONCLUSION: This study provides important insights into the epidemiology of malaria in a high and also low-transmission setting in West Bengal. Intensified efforts to be strengthen towards community mobilization for preventing breeding places for mosquito. Policy should strengthen investment in housing programme that may complement reduction of malaria transmission. Disease reduction effects of Government and local NGOs should be directed towards those areas in order to decrease disease load.

A randomised phase 2 study of intermittent versus continuous dosing of dabrafenib plus trametinib in patients with BRAFV600 mutant advanced melanoma (INTERIM).

BACKGROUND: BRAF+MEK inhibitors extend life expectancy of patients with BRAFV600 mutant advanced melanoma. Acquired resistance limits duration of benefit, but preclinical and case studies suggest intermittent dosing could overcome this limitation. INTERIM was a phase 2 trial evaluating an intermittent dosing regimen. METHODS: Patients with BRAFV600 mutant advanced melanoma due to start dabrafenib+trametinib were randomised to receive either continuous (CONT), or intermittent (INT; dabrafenib d1-21, trametinib d1-14 every 28 days) dosing. A composite primary endpoint included progression-free survival (PFS) and quality of life (QoL). Secondary endpoints included response rate (ORR), overall survival (OS) and adverse events (AEs). Mutant BRAFV600E ctDNA was measured by droplet digital PCR (ddPCR), using mutant allele frequency of > 1 % as the detection threshold. RESULTS: 79 patients (39 INT, 40 CONT) were recruited; median age 67 years, 65 % AJCC (7th ed) stage IV M1c, 29 % had brain metastases. With 19 months median follow-up, INT was inferior in all efficacy measures: median PFS 8.5 vs 10.7mo (HR 1.39, 95 %CI 0.79-2.45, p = 0.255); median OS 18.1mo vs not reached (HR 1.69, 95 %CI 0.87-3.28, p = 0.121), ORR 57 % vs 77 %. INT patients experienced fewer treatment-related AEs (76 % vs 88 %), but more grade > 3 AEs (53 % vs 42 %). QoL favoured CONT. Detection of BRAFV600E ctDNA prior to treatment correlated with worse OS (HR 2.55, 95 %CI 1.25-5.21, p = 0.01) in both arms. A change to undetected during treatment did not significantly predict better OS. CONCLUSION: INTERIM findings are consistent with other recent clinical trials reporting that intermittent dosing does not improve efficacy of BRAF+MEK inhibitors.

Benchmarking MicrobIEM - a user-friendly tool for decontamination of microbiome sequencing data.

BACKGROUND: Microbiome analysis is becoming a standard component in many scientific studies, but also requires extensive quality control of the 16S rRNA gene sequencing data prior to analysis. In particular, when investigating low-biomass microbial environments such as human skin, contaminants distort the true microbiome sample composition and need to be removed bioinformatically. We introduce MicrobIEM, a novel tool to bioinformatically remove contaminants using negative controls. RESULTS: We benchmarked MicrobIEM against five established decontamination approaches in four 16S rRNA amplicon sequencing datasets: three serially diluted mock communities (108-103 cells, 0.4-80% contamination) with even or staggered taxon compositions and a skin microbiome dataset. Results depended strongly on user-selected algorithm parameters. Overall, sample-based algorithms separated mock and contaminant sequences best in the even mock, whereas control-based algorithms performed better in the two staggered mocks, particularly in low-biomass samples (≤ 106 cells). We show that a correct decontamination benchmarking requires realistic staggered mock communities and unbiased evaluation measures such as Youden's index. In the skin dataset, the Decontam prevalence filter and MicrobIEM's ratio filter effectively reduced common contaminants while keeping skin-associated genera. CONCLUSIONS: MicrobIEM's ratio filter for decontamination performs better or as good as established bioinformatic decontamination tools. In contrast to established tools, MicrobIEM additionally provides interactive plots and supports selecting appropriate filtering parameters via a user-friendly graphical user interface. Therefore, MicrobIEM is the first quality control tool for microbiome experts without coding experience.

The airway microbiota of neonates colonized with asthma-associated pathogenic bacteria.

Culture techniques have associated colonization with pathogenic bacteria in the airways of neonates with later risk of childhood asthma, whereas more recent studies utilizing sequencing techniques have shown the same phenomenon with specific anaerobic taxa. Here, we analyze nasopharyngeal swabs from 1 month neonates in the COPSAC2000 prospective birth cohort by 16S rRNA gene sequencing of the V3-V4 region in relation to asthma risk throughout childhood. Results are compared with previous culture results from hypopharyngeal aspirates from the same cohort and with hypopharyngeal sequencing data from the later COPSAC2010 cohort. Nasopharyngeal relative abundance values of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis are associated with the same species in the hypopharyngeal cultures. A combined pathogen score of these bacteria's abundance values is associated with persistent wheeze/asthma by age 7. No other taxa are associated. Compared to the hypopharyngeal aspirates from the COPSAC2010 cohort, the anaerobes Veillonella and Prevotella, which have previously been implicated in asthma development, are less commonly detected in the COPSAC2000 nasopharyngeal samples, but correlate with the pathogen score, hinting at latent community structures that bridge current and previous results. These findings have implications for future asthma prevention efforts.

Utilisation of adolescent reproductive and sexual health services in a rural area of West Bengal: A mixed-method study.

Introduction: Despite policy actions and strategic efforts for improving the reproductive and sexual health of adolescents by promoting the uptake of adolescent reproductive and sexual health (ARSH) services, the utilisation rate remains significantly low, especially in rural areas of India. This study aimed to assess the utilisation of these services by adolescents in rural West Bengal and its associated determinants. Method: This mixed-method study was conducted from May to September 2021 in the Gosaba rural block of South 24 Parganas, West Bengal. Quantitative data were collected from 326 adolescents using a pre-tested structured questionnaire. Qualitative data were collected via four focus group discussions among 30 adolescents and key-informant interviews among six healthcare workers. Quantitative data were analysed using SPSS, while qualitative data were analysed thematically. Results: Ninety-six (29.4%) adolescents had utilised ARSH services at least once during adolescence. The factors associated with non-utilisation of ARSH services were younger age, female sex, increasing reproductive health stigma and decreasing parent-adolescent communication related to sexual health. Qualitative exploration revealed that unawareness regarding services, perceived lack of privacy and confidentiality at healthcare facilities and disruption of services post-emergence of the COVID-19 pandemic were some major barriers to ARSH service utilisation. Conclusion: A multi-component strategy, including promotion of adolescent-friendly health clinics, community support interventions associated with motivation and counselling of parents regarding the importance of adolescent reproductive health, is needed to improve the utilisation of ARSH services. Necessary steps to correct the deficiencies at the facility level should also be prioritised.

Skin microbiome and its association with host cofactors in determining atopic dermatitis severity.

BACKGROUND: Atopic dermatitis (AD) is a heterogeneous, chronic inflammatory skin disease linked to skin microbiome dysbiosis with reduced bacterial diversity and elevated relative abundance of Staphylococcus aureus (S. aureus). OBJECTIVES: We aimed to characterize the yet incompletely understood association between the skin microbiome and patients' demographic and clinical cofactors in relation to AD severity. METHODS: The skin microbiome in 48 adult moderate-to-severe AD patients was investigated using next-generation deep sequencing (16S rRNA gene, V1-V3 region) followed by denoising (DADA2) to obtain amplicon sequence variant (ASV) composition. RESULTS: In lesional skin, AD severity was associated with S. aureus relative abundance (rS  = 0.53, p < 0.001) and slightly better with the microbiome diversity measure Evenness (rS  = -0.58, p < 0.001), but not with Richness. Multiple regression confirmed the association of AD severity with microbiome diversity, including Shannon (in lesional skin, p < 0.001), Evenness (in non-lesional skin, p = 0.015) or S. aureus relative abundance (p < 0.012), and with patient's IgE levels (p < 0.001), race (p < 0.032), age (p < 0.034) and sex (p = 0.012). The lesional model explained 62% of the variation in AD severity, and the non-lesional model 50% of the variation. CONCLUSIONS: Our results specify the frequently reported "reduced diversity" of the AD-related skin microbiome to reduced Evenness, which was in turn mainly driven by S. aureus relative abundance, rather than to a reduced microbiome Richness. Finding associations between AD severity, the skin microbiome and patient's cofactors is a key aspect in developing new personalized AD treatments, particularly those targeting the AD microbiome.

Efflux-linked accelerated evolution of antibiotic resistance at a population edge.

Efflux is a common mechanism of resistance to antibiotics. We show that efflux itself promotes accumulation of antibiotic-resistance mutations (ARMs). This phenomenon was initially discovered in a bacterial swarm where the linked phenotypes of high efflux and high mutation frequencies spatially segregated to the edge, driven there by motility. We have uncovered and validated a global regulatory network connecting high efflux to downregulation of specific DNA-repair pathways even in non-swarming states. The efflux-DNA repair link was corroborated in a clinical "resistome" database: genomes with mutations that increase efflux exhibit a significant increase in ARMs. Accordingly, efflux inhibitors decreased evolvability to antibiotic resistance. Swarms also revealed how bacterial populations serve as a reservoir of ARMs even in the absence of antibiotic selection pressure. High efflux at the edge births mutants that, despite compromised fitness, survive there because of reduced competition. This finding is relevant to biofilms where efflux activity is high.

Nutritional Status and Intra-household Food Distribution Among Reproductive-Age-Group Women in a Slum Area of Hooghly District, West Bengal: A Mixed-Methods Approach.

Introduction Malnutrition among women of reproductive age (WRA), especially those living in slum areas, is one of the most concerning nutritional issues because of the extreme nutritional stress they face in the form of inequitable intra-household food distribution (IHFD). This study aimed to assess the nutritional status (NS) and its association with IHFD among reproductive-age-group women along with exploring the perspectives of the stakeholders regarding inequitable food distribution. Materials and methods The quantitative part of the convergent parallel mixed-methods design study was conducted among 150 WRA, selected by cluster random sampling from 15 slum areas of Hooghly District, between December 2020 and May 2021. Data were collected using a predesigned pretested schedule with anthropometric measurements. IHFD was quantified by the relative dietary energy adequacy ratio (RDEAR). Ordinal logistic regression was performed to obtain adjusted-proportional odds ratios (aPOR) for higher categories of NS (underweight: reference category). Stratified subgroup analysis was done to assess the influencers of IHFD. For the qualitative part, in-depth interviews were conducted with eight purposively selected in-laws of study participants, and the data were interpreted by thematic analysis. Results The mean age of the study participants was 28.6±6.3 years. The proportion of malnutrition and inequitable IHFD (RDEAR<1) among them was 50% and 46%, respectively. Higher categories of NS were found to be significantly associated with an increase in RDEAR (aPOR=22.6, 95% CI: 2.75-185.45, p-value=0.004). Among underweight and normal NS women, those who were earning members and directly involved in food preparation/production had a greater allocation of food within their households. Physiological intolerance, incapacity of earning, and traditional customs were the most recurring themes transcribed as the barriers to equitable food distribution. Conclusion A high magnitude of malnutrition and its association with inequitable IHFD among WRA warrant policy-level support to increase women's employment opportunities and address gender-based inequities through comprehensive information education communication (IEC) techniques as well.

COVID-appropriate facemask use: A study among patients attending outpatient Departments of a Public Health Institute in West Bengal.

BACKGROUND: In the ongoing COVID-19 pandemic, facemask use has been the most feasible public health measure in preventing transmission of SARS-CoV2 with the dual benefit of controlling the source of infection and decreasing the risk of acquiring infection from infected people. Therefore, COVID-appropriate behavior regarding facemask use is imperative for controlling the pandemic. This study aimed to assess COVID-appropriate mask behavior and factors associated with it among the patients attending outpatient departments of a public health institute in West Bengal. MATERIALS AND METHODS: This cross-sectional study was done among 298 patients attending outpatient departments of Rural and Urban Health Units of a public health institute of Kolkata, during October 2020-February 2021. A pretested questionnaire and an observational checklist based on the World Health Organization guidelines were used to collect data. COVID-appropriate mask behavior was assessed by a five-point domain-wise scale, where good practice regarding facemask usage was considered when the score was three or more. Statistical analyses were done in SPSS 16.0. RESULTS: Good practices regarding COVID-19 appropriate mask-related activities were recorded among 30.2% of the study participants. No difficulties faced during or on wearing a mask (adjusted odds ratio [AOR] = 1.86, 95% confidence interval CI: 1.1-3.33) and higher socioeconomic status (AOR = 2.25, 95% CI: 1.22-4.15) were significantly associated with good practice regarding COVID-19 appropriate facemask usage. CONCLUSION: The magnitude of poor practice regarding COVID-appropriate mask-related behaviors is quite high among the study participants. Awareness regarding correct mask handling, storage, hygiene, disposal, and place of wearing from a reliable source would go a long way to curb the myths and misinformation surrounding mask usage and encourage COVID-appropriate mask behaviors among the general population.

Differential effects of lung inflammation on insulin resistance in humans and mice.

BACKGROUND: The rates of obesity, its associated diseases, and allergies are raising at alarming rates in most countries. House dust mites (HDM) are highly allergenic and exposure often associates with an urban sedentary indoor lifestyle, also resulting in obesity. The aim of this study was to investigate the epidemiological association and physiological impact of lung inflammation on obesity and glucose homeostasis. METHODS: Epidemiological data from 2207 adults of the population-based KORA FF4 cohort were used to test associations between asthma and rhinitis with metrics of body weight and insulin sensitivity. To obtain functional insights, C57BL/6J mice were intranasally sensitized and challenged with HDM and simultaneously fed with either low-fat or high-fat diet for 12 weeks followed by a detailed metabolic and biochemical phenotyping of the lung, liver, and adipose tissues. RESULTS: We found a direct association of asthma with insulin resistance but not body weight in humans. In mice, co-development of obesity and HDM-induced lung inflammation attenuated inflammation in lung and perigonadal fat, with little impact on body weight, but small shifts in the composition of gut microbiota. Exposure to HDM improved glucose tolerance, reduced hepatosteatosis, and increased energy expenditure and basal metabolic rate. These effects associate with increased activity of thermogenic adipose tissues independent of uncoupling protein 1. CONCLUSIONS: Asthma associates with insulin resistance in humans, but HDM challenge results in opposing effects on glucose homeostasis in mice due to increased energy expenditure, reduced adipose inflammation, and hepatosteatosis.

Burden of malnutrition among school-going children in a slum area of Kolkata: A matter of concern.

BACKGROUND: Malnutrition is still a great barrier to the growth and development of children, especially in India. Children from the underprivileged sections of the society like slum children are more vulnerable to both undernutrition and overnutrition. This study aimed to estimate the magnitude of malnutrition among school-going children in a slum area of Kolkata and to elicit the association of nutritional status with parental education. METHODS: A cross-sectional observational study was conducted from November 2017 to May 2018 among 508 school-going children of 6 years to 19 years of age in slums of Chetla, Kolkata. Out of three coeducation high schools catering to those children, two schools were randomly selected. 254 participants from each school were selected using systematic random sampling. Students, unwilling to participate and/or whose parents denied providing informed written consent were excluded. After the interview by a predesigned, pretested schedule, their anthropometric examination was conducted. RESULTS: Prevalence of stunting and severe stunting was present in 9.4% and 1.2% of subjects whereas the prevalence of thinness was 23.6% among them. The prevalence of overnutrition was 11%. Boys are more stunted (OR = 2.13, P = 0.013) and thin (OR = 1.73, P = 0.011) than girls. Poor parental education was significantly associated with both undernourishment (stunting and thinness) as well as overweight and obesity. CONCLUSION: Substantial burden of double malnutrition was observed among school-going slum children. Parental education had an important role to prevent malnutrition of their children. Thus, periodic growth monitoring with the strengthening of the Mid-Day Meal program and awareness generation campaigns regarding malnutrition among parents of slum children are highly needed.

Determinants of Treatment Adherence Among Patients Living With Noncommunicable Diseases: A Mixed-Method Study in a Rural Area of West Bengal.

The multisectoral impact of the COVID-19 pandemic can impair treatment adherence of patients with noncommunicable diseases (NCDs). This mixed-method study, conducted from November 2020 to January 2021, assessed the quantum of their treatment adherence and its determinants in rural West Bengal. Quantitative data were collected from 213 NCD patients while qualitative exploration for barriers of treatment adherence was conducted as 6 in-depth Interviews. Treatment adherence was assessed by "Medication Compliance Questionnaire" and "Adherence to Healthy Lifestyle and Follow-up Advice" Questionnaire. A total of 39.4% were nonadherent to medications while 67.1% had nonadherence to healthy lifestyle and follow-up advice. Significant predictors associated with nonadherence were increasing age, female gender, lower socioeconomic status, decreasing patient empowerment, and decreasing trust in the medical profession. Economic crisis, fear of contagion, and nonavailability of investigation facilities were some new emerging barriers in addition to preexisting barriers of treatment adherence. Therefore, measures for improving patient empowerment and patient-provider relationship by motivation and counseling, taking proper care of vulnerable patients affected by the pandemic, and correcting deficiencies at the health-system level should be given utmost priority.

Hazardous practices related to blood borne viral infection transmission among male barbers: A study in a rural area of West Bengal.

BACKGROUND: Barbers are occupationally predisposed to blood borne viral infection (BBVI) such as human immunodeficiency virus, hepatitis B and C. Unhygienic workplaces, improper disinfection and inadequate wound-care can increase the susceptibility of both clients and barbers to BBVI. There is paucity of studies on practices regarding BBVI among barbers, especially in India. OBJECTIVES: To identify the pattern and predictors of practices related to BBVI transmission among male barbers in a rural area of West Bengal. METHODS: A cross-sectional study was conducted from June 2017 to August 2019 in Indas block of Bankura district among 138 male barbers, who were permanent residents and worked for >6 months. Barbers, randomly selected from 10 Gram Panchayats according to probability proportionate to size, were interviewed. Two barbering sessions were observed. Data were analyzed using SPSS version 16 software. Univariate and multiple linear regression was carried out to identify predictors of better practices. P < 0.05 was considered for statistical significance. RESULTS: Dangerous practices such as reuse of blades (8, 5.8%) and face-towel (77, 55.8%), sale/distribution of used blades (77, 55.8%), improper disinfection of sharps (79, 57.2%), reuse of alum without disinfection (129, 93.5%), and improper wound care (71, 51.4%) were observed. Predictors of better practice regarding BBVI were better attitude regarding BBVI (B = 0.172, standard error [SE] = 0.046, P = 0.000) and increased years of schooling (B = 0.054, SE = 0.021, P = 0.012). CONCLUSION: Inimical infection control practices were noticed. They are serving the general people and therefore they should be imbibed with correct BBVI knowledge and must do away with all hazardous practice during their barbering activities.

Why are our elderly distressed? A cross-sectional study in a rural community of West Bengal.

CONTEXT: Population across the globe are aging rapidly due to demographic transition. More than 50% of the elderly in India suffer from one or more chronic diseases and psychological distress is one of the most common morbidities among them. AIMS AND OBJECTIVE: This study was conducted with the objective to assess the status of psychological distress among the elderly. SETTINGS AND DESIGN: A community based, cross-sectional study among 347 elderly aged 60 years and above. METHODS AND MATERIALS: This study was done in 30 villages (clusters) in Singur block of West Bengal. A predesigned and pretested schedule was used to collect data. STATISTICAL ANALYSIS: Data were analysed by univariate and multivariate analyses. RESULTS: The mean (± SD) age of the elderly was 67.67 (± 7.15) years, majority (67.4%) were in the 60-69 years age group. Most (62.8%) of the elderly were suffering from psychological distress which is significantly associated with below primary education level [AOR (95% CI)] = [25.09 (11.88-52.96)], living without spouse and child [AOR (95% CI) =7.88 (3.90-15.89)], financial dependence [AOR (95% CI) =20.23 (7.58-54.00)], dependent functional activity (assessed by ADL) [AOR (95% CI) =3.84 (1.25-11.76)], and decision for healthcare taken by others [AOR (95% CI)] = [3.84 (1.25-11.76)]. CONCLUSIONS: Alarmingly, the proportion of psychological distress was found to be high among the elderly of this rural area. Therefore, all steps must be taken with special focus on the mental health of the old people so that they may continue to contribute to the upliftment of the society.

Skin pH-dependent Staphylococcus aureus abundance as predictor for increasing atopic dermatitis severity.

BACKGROUND: Atopic eczema (atopic dermatitis, AD) is characterized by disrupted skin barrier associated with elevated skin pH and skin microbiome dysbiosis, due to high Staphylococcus aureus loads, especially during flares. Since S aureus shows optimal growth at neutral pH, we investigated the longitudinal interplay between these factors and AD severity in a pilot study. METHOD: Emollient (with either basic pH 8.5 or pH 5.5) was applied double-blinded twice daily to 6 AD patients and 6 healthy (HE) controls for 8 weeks. Weekly, skin swabs for microbiome analysis (deep sequencing) were taken, AD severity was assessed, and skin physiology (pH, hydration, transepidermal water loss) was measured. RESULTS: Physiological, microbiome, and clinical results were not robustly related to the pH of applied emollient. In contrast to longitudinally stable microbiome in HE, S aureus frequency significantly increased in AD over 8 weeks. High S aureus abundance was associated with skin pH 5.7-6.2. High baseline S aureus frequency predicted both increase in S aureus and in AD severity (EASI and local SCORAD) after 8 weeks. CONCLUSION: Skin pH is tightly regulated by intrinsic factors and limits the abundance of S aureus. High baseline S aureus abundance in turn predicts an increase in AD severity over the study period. This underlines the importance and potential of sustained intervention regarding the skin pH and urges for larger studies linking skin pH and skin S aureus abundance to understand driving factors of disease progression.

Coevolution of the translational machinery optimizes initiation with unusual initiator tRNAs and initiation codons in mycoplasmas.

Initiator tRNAs (i-tRNAs) are characterized by the presence of three consecutive GC base pairs (GC/GC/GC) in their anticodon stems in all domains of life. However, many mycoplasmas possess unconventional i-tRNAs wherein the highly conserved sequence of GC/GC/GC is represented by AU/GC/GC, GC/GC/GU or AU/GC/GU. These mycoplasmas also tend to preferentially utilize non-AUG initiation codons. To investigate if initiation with the unconventional i-tRNAs and non-AUG codons in mycoplasmas correlated with the changes in the other components of the translation machinery, we carried out multiple sequence alignments of genes encoding initiation factors (IF), 16S rRNAs, and the ribosomal proteins such as uS9, uS12 and uS13. In addition, the occurrence of Shine-Dalgarno sequences in mRNAs was analyzed. We observed that in the mycoplasmas harboring AU/GC/GU i-tRNAs, a highly conserved position of R131 in IF3, is represented by P, F or Y and, the conserved C-terminal tail (SKR) of uS9 is represented by the TKR sequence. Using the Escherichia coli model, we show that the change of R131 in IF3 optimizes initiation with the AU/GC/GU i-tRNAs. Also, the SKR to TKR change in uS9 was compatible with the R131P variation in IF3 for initiation with the AU/GC/GU i-tRNA variant. Interestingly, the mycoplasmas harboring AU/GC/GU i-tRNAs are also human pathogens. We propose that these mycoplasmas might have evolved a relaxed translational apparatus to adapt to the environment they encounter in the host.

Ubiquitin Ligase COP1 Controls Hepatic Fat Metabolism by Targeting ATGL for Degradation.

Optimal control of hepatic lipid metabolism is critical for organismal metabolic fitness. In liver, adipose triglyceride lipase (ATGL) serves as a major triacylglycerol (TAG) lipase and controls the bulk of intracellular lipid turnover. However, regulation of ATGL expression and its functional implications in hepatic lipid metabolism, particularly in the context of fatty liver disease, is unclear. We show that E3 ubiquitin ligase COP1 (also known as RFWD2) binds to the consensus VP motif of ATGL and targets it for proteasomal degradation by K-48 linked polyubiquitination, predominantly at the lysine 100 residue. COP1 thus serves as a critical regulator of hepatocyte TAG content, fatty acid mobilization, and oxidation. Moreover, COP1-mediated regulation of hepatic lipid metabolism requires optimum ATGL expression for its metabolic outcome. In vivo, adenovirus-mediated depletion of COP1 ameliorates high-fat diet-induced steatosis in mouse liver and improves liver function. Our study thus provides new insights into the regulation of hepatic lipid metabolism by the ubiquitin-proteasome system and suggests COP1 as a potential therapeutic target for nonalcoholic fatty liver disease.

Agreement between MRI and pathologic breast tumor size after neoadjuvant chemotherapy, and comparison with alternative tests: individual patient data meta-analysis.

BACKGROUND: Magnetic resonance imaging (MRI) may guide breast cancer surgery by measuring residual tumor size post-neoadjuvant chemotherapy (NAC). Accurate measurement may avoid overly radical surgery or reduce the need for repeat surgery. This individual patient data (IPD) meta-analysis examines MRI's agreement with pathology in measuring the longest tumor diameter and compares MRI with alternative tests. METHODS: A systematic review of MEDLINE, EMBASE, PREMEDLINE, Database of Abstracts of Reviews of Effects, Heath Technology Assessment, and Cochrane databases identified eligible studies. Primary study authors supplied IPD in a template format constructed a priori. Mean differences (MDs) between tests and pathology (i.e. systematic bias) were calculated and pooled by the inverse variance method; limits of agreement (LOA) were estimated. Test measurements of 0.0 cm in the presence of pathologic residual tumor, and measurements >0.0 cm despite pathologic complete response (pCR) were described for MRI and alternative tests. RESULTS: Eight studies contributed IPD (N = 300). The pooled MD for MRI was 0.0 cm (LOA: +/-3.8 cm). Ultrasound underestimated pathologic size (MD: -0.3 cm) relative to MRI (MD: 0.1 cm), with comparable LOA. MDs were similar for MRI (0.1 cm) and mammography (0.0 cm), with wider LOA for mammography. Clinical examination underestimated size (MD: -0.8 cm) relative to MRI (MD: 0.0 cm), with wider LOA. Tumors "missed" by MRI typically measured 2.0 cm or less at pathology; tumors >2.0 cm were more commonly "missed" by clinical examination (9.3 %). MRI measurements >5.0 cm occurred in 5.3 % of patients with pCR, but were more frequent for mammography (46.2 %). CONCLUSIONS: There was no systematic bias in MRI tumor measurement, but LOA are large enough to be clinically important. MRI's performance was generally superior to ultrasound, mammography, and clinical examination, and it may be considered the most appropriate test in this setting. Test combinations should be explored in future studies.

Sulfonoquinovosyl diacylglyceride selectively targets acute lymphoblastic leukemia cells and exerts potent anti-leukemic effects in vivo.

DNA topoisomerase II inhibitors e.g. doxorubicin and etoposide are currently used in the chemotherapy for acute lymphoblastic leukemia (ALL). These inhibitors have serious side effects during the chemotherapy e.g. cardiotoxicity and secondary malignancies. In this study we show that sulfonoquinovosyl diacylglyceride (SQDG) isolated from Azadirachta indica exerts potent anti-ALL activity both in vitro and in vivo in nude mice and it synergizes with doxorubicin and etoposide. SQDG selectively targets ALL MOLT-4 cells by inhibiting catalytic activity of topoisomerase I enzyme and inducing p53 dependent apoptotic pathway. SQDG treatment induces recruitment of ATR at chromatin and arrests the cells in S-phase. Down-regulation of topoisomerase I or p53 renders the cells less sensitive for SQDG, while ectopic expression of wild type p53 protein in p53 deficient K562 cells results in chemosensitization of the cells for SQDG. We also show that constant ratio combinations of SQDG and etoposide or SDQG and doxorubicin exert synergistic effects on MOLT-4 cell killing. This study suggests that doses of etoposide/doxorubicin can be substantially reduced by combining SQDG with these agents during ALL chemotherapy and side effects caused can be minimized. Thus dual targeting of topoisomerase I and II enzymes is a promising strategy for improving ALL chemotherapy.