Validation of CRISPR activation system in Aedes cells using multicistronic plasmid vectors.

Aedes mosquitoes transmit several pathogens including flaviviruses to humans which result in high morbidity and mortality. Owing to adaptability and climate change, these mosquito vectors are predicted to establish in new geographical areas thus exposing larger populations to the risk of infection. Therefore, control of Aedes vector is necessary to prevent disease transmission. Recently, genetic approaches to vector control have shown promise; however, the tools and methods for manipulating the mosquito genome are rather limited. While CRISPR-Cas9 system has been adapted for gene editing purposes in Aedes mosquito, the dCas9-based transcription control of genes remain unexplored. In this study we report implementation of the CRISPR activation system in Aedes cells. For this we designed, constructed and tested a bi-partite plasmid-based strategy that allows expression of the dCas9-VPR and targeting guide RNA together with a reporter cassette. Quantitative analysis of the fluorescent reporter gene levels showed a robust over-expression validating CRISPR activation in Aedes cells. This strategy and the biological parts will be useful resource for synthetic transcription factor-based robust upregulation of Aedes genes for application of synthetic biology approaches for vector control.

Spatial analysis of chronic obstructive pulmonary disease and its risk factors in an urban area of Trivandrum, Kerala, India.

Background: Chronic obstructive pulmonary disease (COPD) is the second leading cause of death in India. The objective of this study was to map COPD cases and its risk factors and to determine the association between them using geographic information system (GIS) in a semi-urban area of Trivandrum, South India. Materials and Methods: This community-based cross-sectional, descriptive study (n = 494) was conducted in a subcenter area of a primary health center. Location was mapped and COPD population screener questionnaire was administered to all the study subjects enrolled by census method. Lifetime firewood exposure (person-hours) and tobacco smoking were enquired and distance from road was mapped using portable differential global positioning system. The association with COPD was assessed by kriging and hotspot analysis using ArcGIS software. Results: The prevalence of COPD (6.5%) was comparable to national prevalence estimates. Spatial maps showed COPD case clustering in areas with higher firewood exposure, greater smoking exposure, and in households with closer proximity to local roads. A particular high-risk cluster was obtained which had a significant association with all the risk factors. Conclusion: GIS technology is useful in identification of spatial clustering of COPD cases and its environmental risk factors, making it an important tool for targeted interventions for COPD.

Impact of Right Ventricular Function on Left Ventricular Torsion and Ventricular Deformations in Pulmonary Artery Hypertension Patients.

INTRODUCTION: Ventricular interdependence in pulmonary arterial hypertension (PAH) by the use of most recent echocardiographic techniques is still rare. The current case-controlled study aims to assess left ventricular (LV) torsion in patients with PAH. METHODS: The study included 42 cases of moderate to severe PAH and 42 age and gender-matched healthy controls between March 2016 and January 2018. All the patients and controls undergo routine practice echocardiography using the Vivid 7-echocardiography (2.5MHz transducer) system. RESULTS: The LV twisting parameters, peak basal rotation, peak apical rotation, and twist were similar among both cases and controls, however, LV torsion was significantly (p=0.04) impacted. Right ventricular (RV) longitudinal deformation was clinically significant in the cases compared to controls: RV systolic strain imaging (p=0.001, 95% CI-9.75 to -2.65), RV systolic strain rate (p=0.01, 95% CI-0.99 to -0.09), and RV late diastolic strain rate (p=0.01, 95% CI-0.64 to -0.85). Although PAH did not impact longitudinal LV deformations significantly. At basal level circumferential strain and strain rate were significantly impacted (p=0.005, 95% CI-4.38 to -0.70; p=0.004, 95% CI-0.35 to -0.07) in the PAH group, while the radial strain was preserved. All RV echocardiographic parameters and LV end-diastolic dimension, LV end-systolic volume in the PAH were affected significantly (p=0.002, 95% CI-19.91 to -4.46; p=0.01, 95% CI-8.44 to -2.77). However, only a weak correlation (p=0.05, r =-0.20) was found between tricuspid annular plane systolic excursion and LV Tei index. CONCLUSION: RV pressure overload directly affects RV longitudinal systolic deformation further influences the interventricular septal and LV geometry, which impaired LV torsion.

Spatial analysis of hypothyroidism and ground water pH in an Urban Area of Kerala using the geographic information system.

Hypothyroidism is a disease assuming increasing relevance. The causative role of acidic nature of drinking water has not yet been investigated in Kerala. We attempted to determine the spatial association between the occurrence of self-reported hypothyroidism and pH of ground water using the geographic information system. The cross-sectional study was conducted among 1649 individuals residing in the subcenter area in urban Trivandrum. Self-reported hypothyroidism was obtained by the interview. Differential Global Positioning System was used to record the location of each house and its drinking water source. PH of 50 open-well water samples was estimated. The prevalence of self-reported hypothyroidism was 4.24%. Maps depicting pH distribution and occurrence of hypothyroidism were prepared. Most of the areas had acidic ground water. Geo-statistical analysis revealed the occurrence of statistically significant clustering of hypothyroid individuals in areas having acidic ground water. The study brings out possible linkage between hypothyroidism and acidic water intake necessitating detailed epidemiological investigations for drawing more robust associations.

A cross-sectional study to assess prevalence and factors associated with mild cognitive impairment among older adults in an urban area of Kerala, South India.

OBJECTIVES: To assess the prevalence and factors associated with mild cognitive impairment (MCI) among older adults in an urban area of South India. SETTING: The study was conducted in the capital city of Thiruvananthapuram in the South Indian state of Kerala. PARTICIPANTS: The study participants were community-dwelling individuals aged 60 years and above. PRIMARY OUTCOME MEASURE: MCI was the primary outcome measure and was defined using the criteria by European Alzheimer's Disease Consortium. Cognitive assessment was done using the Malayalam version of Addenbrooke's Cognitive Examination tool. Data were also collected on sociodemographic variables, self-reported comorbidities like hypertension and diabetes, lifestyle factors, depression, anxiety and activities of daily living. RESULTS: The prevalence of MCI was found to be 26.06% (95% CI of 22.12 to 30.43). History of imbalance on walking (adjusted OR 2.75; 95 % CI of 1.46 to 5.17), presence of depression (adjusted OR 2.17, 95 % CI of 1.21 to 3.89), anxiety (adjusted OR 2.22; 95 % CI of 1.21 to 4.05) and alcohol use (adjusted OR 1.99; 95 % CI of 1.02 to 3.86) were positively associated with MCI while leisure activities at home (adjusted OR 0.33; 95 % CI of 0.11 to 0.95) were negatively associated. CONCLUSION: The prevalence of MCI is high in Kerala. It is important that the health system and the government take up urgent measures to tackle this emerging public health issue.

Seroprevalence of dengue infection in pregnant women and placental antibody transfer.

BACKGROUND: Pregnant women and infants are vulnerable for developing severe dengue. This study was conducted to determine the seroprevalence of dengue infections among pregnant women, their offsprings and its association with outcomes. METHODS: A cross-sectional study was conducted among pregnant women, admitted for delivery in a tertiary mother and child hospital in Thiruvananthapuram. Blood specimens (2 ml) were collected from the women during hospitalization in the first stage along with blood samples being drawn for other investigations. Umbilical cord blood was collected from the neonates. The samples were tested using IgG enzyme-linked immunosorbent assay (ELISA). Quantitative titres were also obtained, and index ratios were calculated using optical density values. RESULTS: Seroprevalence of dengue in antenatal women was 6.9% (95% confidence interval [CI]: 3.4-12.48). Among cord blood samples, the seropositivity was 10.8% (95% CI: 6.3-16.6). A significant correlation (Spearman rho: 0.653 and p value <0.001) was obtained between maternal and cord sample IgG index ratios. Agreement between maternal and cord blood IgG values was obtained using kappa as 0.742. The mean weight of newborns born to IgG-positive mothers was significantly lower than babies of IgG-negative mothers (2.3 vs 2.8; t = 2.64; p = 0.01). The mean gestational age at delivery was also significantly lower in IgG-positive mothers (36.29 vs 38.04 weeks; t = 2.48; p = 0.01). CONCLUSION: Seroprevalence of dengue in antenatal women and in their offsprings is lower than other areas endemic for dengue. Dengue infection (any time before pregnancy) may result in preterm delivery and low birth weights.

Role of Geographic Information System in Assessing Determinants of Cardiovascular Disease: An Experience From a Low- and Middle-Income Country.

Cardiovascular disease (CVD) is the leading cause of morbidity and mortality in the world. The determinants of CVD in an urban population using conventional and geographic information system techniques were attempted as a community-based census-type cross-sectional study in Kerala, India, among 1649 individuals residing in 452 households. Sociodemographic details, risk factor exposures, and self-reported disease prevalence were determined. Location of houses, wells from which subjects drew drinking water, and distances of the house from the outer road (proxy for air pollution) were mapped using differential global positioning system and pH of water samples determined. Prevalence of CVD was 5.8%. Significant predictors of CVD were male gender, diabetes mellitus, hypertension, and hypothyroidism. Statistically significant spatial association was found between CVD and groundwater pH. Geographic information system technology is useful in identification of spatial clustering and disease hotspots for designing preventive strategies targeting CVD.

Investigation of an outbreak of measles: failure to vaccinate or vaccine failure in a community of predominantly fishermen in Kerala.

Measles outbreaks continue to occur in developing countries. This study attempted to explore the context of an outbreak of measles in a community of predominantly fishermen in Kerala to find out whether the outbreak was the result of a failure to vaccinate or failure of the vaccine itself. A cross sectional study was conducted in Mukkola village of Thiruvananthapuram district, Kerala, India. A total of 215 children of ages between 9 and 35 months were studied. Documented evidence of measles vaccination was available only in 71.6% (65.57-77.62) of the children. The risk factors for not being immunized against measles were being third or higher in birth order and having: a father whose occupation is fishing, low family income, lower parental education, Muslim religion and poor knowledge regarding measles and its vaccine. Of the 215 children studied, 43 had a history of measles. Thirty percent of these 43 children were younger than the age of vaccination. Unvaccinated children, children third or higher in birth order and children of families with more than 5 members had a significantly higher risk of contracting measles. Vaccine effectiveness was 76.6% (95% CI: 75.96-77.99). The prevalence of missed vaccination opportunities was found to be 15.8% (34/215). Even with the relatively low vaccine effectiveness, this outbreak could have been prevented by higher vaccination coverage. Lowering the age at administration of the first dose of measles vaccine needs to be considered. Effective utilization of opportunities for vaccination could enhance coverage and prevent outbreaks in the future.

Detection of numerical chromosomal abnormalities in epithelial ovarian neoplasms by fluorescence in situ hybridization (FISH) and a review of the current literature.

Preliminary retrospective chromosomal analysis was performed using fluorescence in situ hybridization (FISH) with alphoid DNA probes for chromosomes 1, 3, 6, 8, 12, 17, and X. Twenty-four epithelial ovarian tumors were examined in this pilot study, including 8 borderline (LMP) serous tumors, 9 serous carcinoma, and 7 mucinous carcinoma. Hybridization signals were counted to demonstrate the frequency of aneusomy, trace chromosomal progression, and identify the predominance of chromosome copy number abnormalities that are specific to a particular histotype. The preliminary results revealed almost an equal number of mean aneusomies in serous (58.13 +/- 13%) and mucinous (64.33 +/- 10%) carcinoma, both of which were slightly higher than borderline serous tumors (50.57 +/- 17%). Hyposomies 3 and X were significantly higher in mucinous than in serous ovarian carcinomas, and lowest in borderline serous tumors (P<0.05 and P<0.01). Signal losses were a more frequent abnormality in all three histologic subtypes. Mucinous carcinomas showed a loss of chromosomes 8 (45.00 +/- 28%) and 3 (43.14 +/- 16%), in addition to a loss of chromosome X (56.29 +/- 12%). Serous carcinomas showed a gain of chromosome 1 (39.44 +/- 32%), followed by losses of chromosomes 6 (37.00 +/- 20%), 17 (36.44 +/- 19%), and 8 (36.89 +/- 19%). In borderline serous tumors, the most frequent findings were losses of chromosomes 6 (38.00 +/- 17%), 12 (36.88 +/- 17%), and 3 (36.13 +/- 21%). However, further research is necessary to substantiate these preliminary results and elucidate their clinical significance. A brief review of the literature pertaining to interphase cytogenetics in ovarian epithelial tumors is discussed also.