Diet, cardiovascular disease, and mortality in 80 countries.

AIMS: To develop a healthy diet score that is associated with health outcomes and is globally applicable using data from the Prospective Urban Rural Epidemiology (PURE) study and replicate it in five independent studies on a total of 245 000 people from 80 countries. METHODS AND RESULTS: A healthy diet score was developed in 147 642 people from the general population, from 21 countries in the PURE study, and the consistency of the associations of the score with events was examined in five large independent studies from 70 countries. The healthy diet score was developed based on six foods each of which has been associated with a significantly lower risk of mortality [i.e. fruit, vegetables, nuts, legumes, fish, and dairy (mainly whole-fat); range of scores, 0-6]. The main outcome measures were all-cause mortality and major cardiovascular events [cardiovascular disease (CVD)]. During a median follow-up of 9.3 years in PURE, compared with a diet score of ≤1 points, a diet score of ≥5 points was associated with a lower risk of mortality [hazard ratio (HR) 0.70; 95% confidence interval (CI) 0.63-0.77)], CVD (HR 0.82; 0.75-0.91), myocardial infarction (HR 0.86; 0.75-0.99), and stroke (HR 0.81; 0.71-0.93). In three independent studies in vascular patients, similar results were found, with a higher diet score being associated with lower mortality (HR 0.73; 0.66-0.81), CVD (HR 0.79; 0.72-0.87), myocardial infarction (HR 0.85; 0.71-0.99), and a non-statistically significant lower risk of stroke (HR 0.87; 0.73-1.03). Additionally, in two case-control studies, a higher diet score was associated with lower first myocardial infarction [odds ratio (OR) 0.72; 0.65-0.80] and stroke (OR 0.57; 0.50-0.65). A higher diet score was associated with a significantly lower risk of death or CVD in regions with lower than with higher gross national incomes (P for heterogeneity <0.0001). The PURE score showed slightly stronger associations with death or CVD than several other common diet scores (P < 0.001 for each comparison). CONCLUSION: A diet comprised of higher amounts of fruit, vegetables, nuts, legumes, fish, and whole-fat dairy is associated with lower CVD and mortality in all world regions, especially in countries with lower income where consumption of these foods is low.

Acute toxicity studies and protective effects of Cinnamon cassia bark extract in streptozotocin-induced diabetic rats.

The medicinal properties of Cinnamon cassia (C. cassia) bark have been reported for their clinical importance for many diseases including diabetes. However, there is no clear evidence so far regarding dose selection for its hepato- and nephroprotective effect in diabetic condition. Hence, the present study aims at evaluating in vitro antioxidant activity, the acute toxicity, and dose fixation of C. cassia bark for their effective medicinal values in streptozotocin (STZ)-induced rats. All the extracts exhibited potential in vitro antioxidant activity and showed a dose-dependent (1000, 2000, 3000, 4000, and 5000 mg/kg BW) acute toxicity by in vivo model. The levels of aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), urea, and creatinine showed a significant elevation in animals treated with the highest dose. In further studies along with histopathological studies, animals treated with STZ (60 mg/kg BW) followed by a different dose (300, 400, and 500 mg/kg BW) of ethanolic extract of the C. cassia bark and glibenclamide (3 mg/kg BW) revealed that the altered level of mitochondrial enzymes, hepatic, and renal marker in STZ-induced animals were restored in C. cassia bark extract-treated group as of control. These results could be of scientific support for the use of the ethanolic extract of the C. cassia bark in folk medicine for the management of diabetes and its associated complications.

Automatic detection of breast cancer in ultrasound images using Mayfly algorithm optimized handcrafted features.

BACKGROUND: The incidence rates of breast cancer in women community is progressively raising and the premature diagnosis is necessary to detect and cure the disease. OBJECTIVE: To develop a novel automated disuse detection framework to examine the Breast-Ultrasound-Images (BUI). METHODS: This scheme includes the following stages; (i) Image acquisition and resizing, (ii) Gaussian filter-based pre-processing, (iii) Handcrafted features extraction, (iv) Optimal feature selection with Mayfly Algorithm (MA), (v) Binary classification and validation. The dataset includes BUI extracted from 133 normal, 445 benign and 210 malignant cases. Each BUI is resized to 256×256×1 pixels and the resized BUIs are used to develop and test the new scheme. Handcrafted feature-based cancer detection is employed and the parameters, such as Entropies, Local-Binary-Pattern (LBP) and Hu moments are considered. To avoid the over-fitting problem, a feature reduction procedure is also implemented with MA and the reduced feature sub-set is used to train and validate the classifiers developed in this research. RESULTS: The experiments were performed to classify BUIs between (i) normal and benign, (ii) normal and malignant, and (iii) benign and malignant cases. The results show that classification accuracy of > 94%, precision of > 92%, sensitivity of > 92% and specificity of > 90% are achieved applying the developed new schemes or framework. CONCLUSION: In this work, a machine-learning scheme is employed to detect/classify the disease using BUI and achieves promising results. In future, we will test the feasibility of implementing deep-learning method to this framework to further improve detection accuracy.

Glycemic control and its determinants among people with type 2 diabetes mellitus in Ernakulam district, Kerala.

Background: Diabetes is a public health problem of colossal proportions. The National Family Health Survey 5 (2019-2020) has found high blood sugar levels among 13.5% of women and 15.6% of men in India. The high morbidity and mortality in diabetes mellitus are due to uncontrolled hyperglycemia resulting in micro- and macrovascular changes affecting multiple organs in the body. The objectives of this study were to estimate glycemic control and its determinants among type 2 diabetics in the Ernakulam district. Methods: A community-based cross-sectional study was conducted among 364 type 2 diabetics who had the disease for at least 5 years duration in the Ernakulam district of Kerala. Probability proportional to size cluster sampling technique was adopted. A semi-structured questionnaire was used to collect sociodemographic and behavioral profiles. Participants' HbA1c levels were assessed to determine glycemic control. Results: The proportion of people with good glycemic control was 21.4%. Determinants such as female gender adjusted odds ratio (aOR = 2.36, P = 0.005), body mass index >23 kg/m2 (aOR = 2.71, P = 0.002), combined drug treatment with Oral Hypoglycaemic agents (OHA) and insulin (aOR = 3.76, P = 0.004), and poor compliance with medications (aOR = 1.93, P = 0.030) were found to be significantly associated with poor glycemic control. Conclusions: Poor compliance with medications and unhealthy lifestyle choices has resulted in a high proportion of diabetics with poor glycemic control in the district. Women are particularly more vulnerable to uncontrolled hyperglycemia than males. Type 2 diabetes mellitus people should be encouraged to maintain strict glycemic control, which is an important measure for secondary prevention of complications.

Variations in common diseases, hospital admissions, and deaths in middle-aged adults in 21 countries from five continents (PURE): a prospective cohort study.

BACKGROUND: To our knowledge, no previous study has prospectively documented the incidence of common diseases and related mortality in high-income countries (HICs), middle-income countries (MICs), and low-income countries (LICs) with standardised approaches. Such information is key to developing global and context-specific health strategies. In our analysis of the Prospective Urban Rural Epidemiology (PURE) study, we aimed to evaluate differences in the incidence of common diseases, related hospital admissions, and related mortality in a large contemporary cohort of adults from 21 HICs, MICs, and LICs across five continents by use of standardised approaches. METHODS: The PURE study is a prospective, population-based cohort study of individuals aged 35-70 years who have been enrolled from 21 countries across five continents. The key outcomes were the incidence of fatal and non-fatal cardiovascular diseases, cancers, injuries, respiratory diseases, and hospital admissions, and we calculated the age-standardised and sex-standardised incidence of these events per 1000 person-years. FINDINGS: This analysis assesses the incidence of events in 162 534 participants who were enrolled in the first two phases of the PURE core study, between Jan 6, 2005, and Dec 4, 2016, and who were assessed for a median of 9·5 years (IQR 8·5-10·9). During follow-up, 11 307 (7·0%) participants died, 9329 (5·7%) participants had cardiovascular disease, 5151 (3·2%) participants had a cancer, 4386 (2·7%) participants had injuries requiring hospital admission, 2911 (1·8%) participants had pneumonia, and 1830 (1·1%) participants had chronic obstructive pulmonary disease (COPD). Cardiovascular disease occurred more often in LICs (7·1 cases per 1000 person-years) and in MICs (6·8 cases per 1000 person-years) than in HICs (4·3 cases per 1000 person-years). However, incident cancers, injuries, COPD, and pneumonia were most common in HICs and least common in LICs. Overall mortality rates in LICs (13·3 deaths per 1000 person-years) were double those in MICs (6·9 deaths per 1000 person-years) and four times higher than in HICs (3·4 deaths per 1000 person-years). This pattern of the highest mortality in LICs and the lowest in HICs was observed for all causes of death except cancer, where mortality was similar across country income levels. Cardiovascular disease was the most common cause of deaths overall (40%) but accounted for only 23% of deaths in HICs (vs 41% in MICs and 43% in LICs), despite more cardiovascular disease risk factors (as judged by INTERHEART risk scores) in HICs and the fewest such risk factors in LICs. The ratio of deaths from cardiovascular disease to those from cancer was 0·4 in HICs, 1·3 in MICs, and 3·0 in LICs, and four upper-MICs (Argentina, Chile, Turkey, and Poland) showed ratios similar to the HICs. Rates of first hospital admission and cardiovascular disease medication use were lowest in LICs and highest in HICs. INTERPRETATION: Among adults aged 35-70 years, cardiovascular disease is the major cause of mortality globally. However, in HICs and some upper-MICs, deaths from cancer are now more common than those from cardiovascular disease, indicating a transition in the predominant causes of deaths in middle-age. As cardiovascular disease decreases in many countries, mortality from cancer will probably become the leading cause of death. The high mortality in poorer countries is not related to risk factors, but it might be related to poorer access to health care. FUNDING: Full funding sources are listed at the end of the paper (see Acknowledgments).

Cost analysis of dengue from a State in south India.

BACKGROUND & OBJECTIVES: Improved dengue cost estimates offer the potential to provide a baseline measure to determine the cost-effectiveness of interventions. The objective of this study was to estimate the cost of dengue prevention, treatment and fatalities in Kerala, India, over a period of one year. METHODS: The study was done in Kerala, a southern State in India. Costing of treatment was done from a family perspective. It was found by primary data collection in a sample of 83 dengue patients from Thiruvananthapuram district and estimated for Kerala using the reported number of cases in 2016. Costing of prevention was done from the government perspective for the entire State. In-depth interviews with State programme officers and experts in the field were conducted. The present value of lifetime earnings was used to value lives. RESULTS: The cost of treatment of dengue in the State was ₹137 milion (2.16 million US$). The cost of prevention in the State was ₹535 million (8.3 million US$). The cost of fatalities was the highest among costs at ₹1760 million (27.7 million US$). US$ 38 million was the least possible estimate of total cost of dengue. The total out-of-pocket spending (OOPS) of >60 yr was significantly (P<0.05) higher than other age groups. The total OOPS was significantly (P<0.001) higher in private sector compared to public. INTERPRETATION & CONCLUSIONS: Although deaths due to dengue were few, the cost of fatalities was 12 times more than the cost of treatment and three times the cost of prevention. Focusing on mortality reduction and disease prevention in elderly would be beneficial.

Bridging the gap in outreach and compliance with mass drug administration for lymphatic filariasis elimination in an endemic district in Kerala, India: an intervention research approach.

Lymphatic filariasis (LF), a neglected tropical disease is targeted for elimination globally by 2020. National Health Policy of India set the goal by 2017 and annual single-dose mass drug administration (MDA) with anti-filarial drugs is in operation in endemic districts since 2004. Performance and effectiveness of MDA was diverse across the nation and prevalence of infection continues above threshold level in 50% of endemic districts which requires alternative strategies. National programme aims at achieving >65% consumption for transmission control. Post 10 rounds of MDA in an endemic district of Kerala state, a three-arm study identified determinants in gap in outreach and consumption, based on which context-dependent intervention using social group work and networking (SGWN) was implemented and impact assessed. A spill over effect with overall increase in coverage and consumption rates irrespective of arms apparently is due to inclusive improvement in MDA programme whereas, significant relative improvement in intervention arm reflects effectiveness of SGWN strategy. Though reduction in microfilaria prevalence in all arms was observed, it was significant between intervention and comparison arm. This study highlights need for context-dependent communication strategy to improve MDA for accelerating LF elimination by the targeted 2020.

Implementation of Software Agents and Advanced AoA for Disease Data Analysis.

To eliminate the possibilities of getting various contradicting solutions to a single problem during diagnosis, a single regular Agent oriented Approach (AoA) is replaced by Intelligent Artificial Agents that act like human and even dynamically decide in any situations known as Intelligent Searching Approach (ISA) is proposed. These agents are used to analyse the medical forums and results or findings are derived accurately than any manual approach. Multiple Agents have been used to analyse the blogs by dividing the work areas and communicating themselves using Agent Communication Language (ACL) and FIPA. The local solutions thus formed are forwarded to a global agent. This Global Agent controls all operations and makes the decision about the best solution. As the Global Agent controls all other agents, it eradicates unwanted and ineffective communication between the various local agents and hence keeping the time taken for communication at the minimum level. Based on these solutions a prioritization matrix is formed using advanced clustering techniques to create a prioritized content of suggested best solutions. Once the decision is made, the refining process runs several times recursively checking for all possible better solutions solving the input. On completion of this process, the Global Agent returns the exact result of the discussion. This process saves time rather than researching the entire blog for result data. This advanced approach lights a different way of obtaining solution keeping the time taken for discussion and intercommunication between the agents to the minimal level but not compromising on the perfection of the solution at the same time.

Breast Cancer Diagnosis Using Feature Ensemble Learning Based on Stacked Sparse Autoencoders and Softmax Regression.

Nowadays, the most frequent cancer in women is breast cancer (malignant tumor). If breast cancer is detected at the beginning stage, it can often be cured. Many researchers proposed numerous methods for early prediction of this Cancer. In this paper, we proposed feature ensemble learning based on Sparse Autoencoders and Softmax Regression for classification of Breast Cancer into benign (non-cancerous) and malignant (cancerous). We used Breast Cancer Wisconsin (Diagnostic) medical data sets from the UCI machine learning repository. The proposed method is assessed using various performance indices like true classification accuracy, specificity, sensitivity, recall, precision, f measure, and MCC. Simulation and result proved that the proposed approach gives better results in terms of different parameters. The prediction results obtained by the proposed approach were very promising (98.60% true accuracy). In addition, the proposed method outperforms the Stacked Sparse Autoencoders and Softmax Regression based (SSAE-SM) model and other State-of-the-art classifiers in terms of various performance indices. Experimental simulations, empirical results, and statistical analyses are also showing that the proposed model is an efficient and beneficial model for classification of Breast Cancer. It is also comparable with the existing machine learning and soft computing approaches present in the related literature.

Psidium guajava Leaf Extracts and Their Quercetin Protect HepG2 Cell Lines Against CCL4 Induced Cytotoxicity.

The present study was carried out to evaluate the in vitro cytoprotective effects of Psidium guajava and their isolated quercetin fraction to reduce the CCl4 (carbon tetrachloride) induced toxicity in HepG2 cell lines (Hepatocellular carcinoma G2). Silymarin was used as a standard drug to compare the protective effects of plant extracts in infected cell lines. MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) assay, cell viability assay, leakage parameters [Aspartate aminotransferase (AST), Alanine aminotransferase (ALT) and Lactate dehydrogenase (LDH)], lipid peroxidation and reduced glutathione (GSH) levels were used to find out the protection of human derived HepG2 cells against CCl4-induced damage. The levels of cytotoxicity, viability and GSH were reduced. While the activities of AST, ALT, LDH and lipid peroxidation was increased in CCl4-treated groups. The treatment of P. guajava and their isolated quercetin fractions (100, 200, 300 µg/mL) decreased the elevated levels of all these parameters. The results of the present study suggest that the ethanolic extract of P. guajava leaf and their isolated quercetin fractions can able to reduce the CCl4-induced cytotoxicity in HepG2 cell lines.

Burden of dengue in Kerala using disability-adjusted life years from 2006 to 2016.

BACKGROUND: State-specific disability-adjusted life years (DALYs) are seldom calculated. Understanding the health and disease trends in groups of states is useful for finding the heterogeneity of disease epidemiology in the country. OBJECTIVE: The objective of the study was to assess dengue burden in Kerala state, using DALY. METHODS: DALY was computed using the DALY package of R. Incidence was derived using reported and estimated dengue cases for 2006-2016. Mortality was calculated using reported deaths. We computed DALYs using the mortality estimates for the South-East Asia region (SEAR) also. Sensitivity and scenario analyses were done. RESULTS: The highest estimated relative DALY for dengue is 7.22 (95% confidence interval [CI]: 6.66-7.72) per lakh population. The DALY obtained using the mortality rates of SEAR ranged from 19.89 (95% CI: 8.44-29.45) per lakh population to 28.56 (95% CI: 17.04-38.05). We observed a cyclical pattern of increase in DALY every 2-3 years. The DALY in lower age groups is lesser. DALY in females was higher than males. CONCLUSIONS: The dengue-related DALY for the state of Kerala is lower than that of the global burden of disease due to lower mortality rates. Mortality reduction becomes the key to reducing burden, especially in areas of low incidence. The study also forms the benchmark for evaluating and implementing cost-effective measures for dengue control in the state.

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.

Novel 9 amino acid in-frame deletion in the NTRK1 tyrosine kinase domain in a patient with congenital insensitivity to pain with anhydrosis.

Schematic presentation of NTRK1 protein structure. Variants identified in this study are shown in red and previously reported variants associated with CIPA are shown in black (LRM, leucine rich motif; Ig, immunoglobulin-like domain; TM, transmembrane domain; TK, tyrosine kinase domain).

An Efficient Framework for Large Scale Multimedia Content Distribution in P2P Network: I2NC.

Network coding (NC) makes content distribution more effective and easier in P2P content distribution network and reduces the burden of the original seeder. It generalizes traditional network routing by allowing the intermediate nodes to generate new coded packet by combining the received packets. The randomization introduced by network coding makes all packets equally important and resolves the problem of locating the rarest block. Further, it reduces traffic in the network. In this paper, we analyze the performance of traditional network coding in P2P content distribution network by using a mathematical model and it is proved that traffic reduction has not been fully achieved in P2P network using traditional network coding. It happens due to the redundant transmission of noninnovative information block among the peers in the network. Hence, we propose a new framework, called I2NC (intelligent-peer selection and incremental-network coding), to eliminate the unnecessary flooding of noninnovative coded packets and thereby to improve the performance of network coding in P2P content distribution further. A comparative study and analysis of the proposed system is made through various related implementations and the results show that 10-15% of traffic reduced and improved the average and maximum download time by reducing original seeder's workload.

Mortality risk among tobacco users in a rural population of southern India: evidence from a cohort study.

BACKGROUND: Tobacco has long been identified as one of the most hazardous risk factors detrimental to health. To plan and implement anti-tobacco activities in any community, it is necessary to understand the risk it poses in that setting. We assessed the risk of mortality associated with tobacco use in a rural community of Kerala. METHODS: This cohort study (PROLIFE) was done in Varkkala, a rural development block of Thiruvananthapuram district of Kerala. Adults aged 20 years and above were included. Age-adjusted mortality rates were computed for both users and non-users of tobacco. The risk of mortality was plotted using Kaplan-Meier curves. Cox regression was used to compute the age-adjusted hazard ratio of mortality among tobacco users. RESULTS: More than one-fourth of the study population used tobacco. The age-adjusted mortality rates were higher among tobacco users. The major causes of death among both users and non-users of tobacco were similar. The incidence proportion of death among all causes of death was higher for tobacco users. The hazard of mortality was significantly more among tobacco users, with the age-adjusted hazard ratio being 1.225 (1.140-1.315). CONCLUSION: The mortality risk due to tobacco use is high irrespective of the cause of death.

A study on container breeding mosquitoes with special reference to Aedes (Stegomyia) aegypti and Aedes albopictus in Thiruvananthapuram district, India.

BACKGROUND & OBJECTIVES: The district of Thiruvananthapuram reports the maximum number of cases of dengue in the state of Kerala. To determine the larval diversity, density and breeding site preferences of Aedes mosquitoes, during pre-monsoon and monsoon periods in urban and rural areas of Thiruvananthapuram district. METHODS: Based on the daily reports of dengue cases, 70 clusters were identified in Thiruvananthapuram district. A cross-sectional larval survey was done in the domestic and peri-domestic areas of 1750 houses, using the WHO standard techniques. The larval indices were calculated, and the larvae were identified by using taxonomic keys. Urban and rural differences and the variations during pre-monsoon and monsoon seasons were also studied. RESULTS: In the surveyed houses, 15% had mosquito breeding, with 88% having Aedes larvae. The house index, container index and the breteau index were 13.08, 13.28 and 16.57%, respectively. About 86% of the clusters were found positive for Aedes albopictus and 11% for Ae. aegypti. Aedes albopictus was distributed almost equally in rural and urban clusters, whereas the distribution of Ae. aegypti was significantly higher in urban areas (p = 0.03). The most common water holding containers found (outdoor) were of plastic, followed by coconut shells. The breeding preference ratio was highest for tyres. Significantly lesser positivity was found for containers during monsoon period when compared to pre-monsoon period. CONCLUSION: The geographical distribution of Ae. albopictus is significantly high in peri-domestic areas and, therefore, its epidemiological role in the widespread disease occurrence needs to be studied. The discarded tyres being the most preferred breeding sites, where IEC activities will help in source reduction.

Three-year black carbon aerosol synthesis over a pristine location surrounded by hillocks in Haryana state, India.

The three-year Black Carbon (BC) aerosol measurements made during 2020, 2021, and 2022 over a rural location, namely, Panchgaon, surrounded by Aravali hillocks (elevation of about 400-600 m) have been analyzed with an aim to determine their optical and radiative characteristics, seasonal and long-term variations in mass concentration. The affinity between these parameters and associated pollutants and planetary boundary layer height (PBLH), affected by the orography, to delineate their role in mass concentration changes with time have been investigated. The coincident OPAC (Optical Properties of Aerosols and Clouds) Model-derived aerosol optical depth (AOD), and single scattering albedo (SSA) have been compared with the observed BC mass concentration, and also with synchronous satellite measurements. The year-to-year variability analysis of the data reveals that the rate of increase of BC concentration is high. The variability was low due to the reasons explained. It implies that the year-to-year variability in BC concentration at the study site depends on the source strength modulated by the valley-driven meteorology. Added, the percentage departures of BC concentration show positive values (higher concentration) during morning and evening hours, which could be due to more anthropogenic activities while it shows negative values during afternoon hours and lower boundary layer heights. The force exerted by the radiation due to BC aerosols at the bottom of the atmosphere (BOA), and in the atmosphere (ATM) are almost equal in magnitude and negative, while that at the top-of-the-atmosphere (TOA) is smaller and positive, indicating that BC aerosols in the study region cools the atmosphere at the BOA and warms the ATM and TOA, which indirectly reveals the dominant role of long-range transport phenomenon at higher levels as compared to the surface level.

A novel loop-mediated isothermal amplification (LAMP) assay to diagnose feline panleukopenia.

Protoparvovirus carnivoran1, known commonly as feline panleukopenia virus (FPV) is a highly contagious and environmentally stable parvovirus of domestic as well as wild felids. A rapid and robust diagnostic tool will aid in implementing prompt treatment and control measures. A loop-mediated isothermal amplification (LAMP) as a point-of-care diagnostic tool for diagnosing feline panleukopenia was standardised using faecal samples of cats. The assay will reduce the cost and time required to diagnose feline panleukopenia. A set of two outer primers (F3 and B3) and two inner primers (FIP and BIP) were designed to target the viral polypeptide (VP2) gene of FPV. Optimisation of the LAMP reaction was done at 60 °C for one hour after an initial denaturation at 95 °C for five minutes. Visualisation of the result based on the addition of SYBR Green 1 dye offered an easy and reliable diagnosis. The detection limit of the standardised LAMP assay was as low as 1.25 ng/µl of the target DNA. Species specificity of the LAMP primers revealed no amplification of the non-target DNA of any other species except that of the canine parvovirus DNA template. DNA extracted from 100 PCR-positive and 20 PCR-negative faecal samples were subjected to the standardised assay and compared with PCR. Analysis of the results revealed that the LAMP assay was 100% sensitive and 90% specific compared to PCR. The LAMP assay could be a reliable tool for the point-of-care diagnosis of feline panleukopenia in limited resource settings.

Rice (Oryza sativa) lipase: molecular cloning, functional expression and substrate specificity.

Lipases are important biocatalysts showing many interesting properties with industrial applications. Previously, different isoforms of lipases, Lipase-I and Lipase-II from rice (Oryza sativa) have been purified and characterized. Lipase-II identified as the major lipase in rice bran is designated as rice bran lipase (RBL). In this study, we report the cloning and expression of the RBL in Escherichia coli and Pichia pastoris. An exploration of expression in four different E. coli expression systems analyzed: BL21(DE3)pLysS, RIL(DE3)pLysS, Rosetta(DE3)pLysS and Origami(DE3)pLysS indicated that E. coli was not a suitable host. Expression with supplement of rare codons in Rosetta (DE3)pLysS and RIL(DE3)pLysS resulted in highest expression as insoluble inclusion bodies. The hurdles of expression in E. coli were overcome by expression as a secretory protein in P. pastoris X-33. The expression of lipase in shake flasks was optimized to achieve the maximum recombinant lipase activity of 152.6 U/mL. The purified recombinant lipase had a specific activity of 998 U/mg toward triacetin. The pH and temperature optimum of native and recombinant enzymes were pH 7.4 and 25 ± 2 °C, respectively. Both the lipases showed higher activity toward short chain triacylglycerol and unsaturated fatty acid enriched oils. Computational modeling and molecular docking studies reveal that the catalytic efficiency of the lipase correlates with the distance of the nucleophilic Ser(175)-OH and the scissile ester bond. The shorter the distance, the greater is the turnover of the substrate.

Economic impact of chikungunya epidemic: out-of-pocket health expenditures during the 2007 outbreak in Kerala, India.

The southern state of Kerala, India was seriously affected by a chikungunya epidemic in 2007. As this outbreak was the first of its kind, the morbidity incurred by the epidemic was a challenge to the state's public health system. A cross sectional survey was conducted in five districts of Kerala that were seriously affected by the epidemic, using a two-stage cluster sampling technique to select households, and the patients were identified using a syndromic case definition. We calculated the direct health expenditure of families and checked whether it exceed the margins of catastrophic health expenditure (CHE). The median (IQR) total out-of-pocket (OOP) health expenditure in the study population was USD7.4 (16.7). The OOP health expenditure did not show any significant association with increasing per-capita monthly income.The major share (47.4%) of the costs was utilized for buying medicines, but costs for transportation (17.2%), consultations (16.6%), and diagnoses (9.9%) also contributed significantly to the total OOP health expenditure. The OOP health expenditure was high in private sector facilities, especially in tertiary care hospitals. For more than 15% of the respondents, the OOP was more than double their average monthly family income. The chikungunya outbreak of 2007 had significantly contributed to the OOP expenditure of the affected community in Kerala.The OOP health expenditure incurred was high, irrespective of the level of income. Governments should attempt to ensure comprehensive financial protection by covering the costs of care, along with loss of productivity.