Photosynthetic efficiency and transcriptome analysis of Dunaliella salina under hypersaline: a retrograde signaling mechanism in the chloroplast.

Understanding the molecular mechanisms of environmental salinity stress tolerance and acclimation strategies by photosynthetic organisms facilitates accelerating the genetic improvement of tolerant economically important crops. In this study, we have chosen the marine algae Dunaliella (D.) salina, a high-potential and unique organism that shows superior tolerance against abiotic stresses, especially hypersaline conditions. We have grown the cells in three different salt concentrations 1.5M NaCl (control), 2M NaCl, and 3M NaCl (hypersaline). Fast chlorophyll fluorescence analysis showed increased initial fluorescence (Fo) and decreased photosynthetic efficiency, indicating hampered photosystem II utilization capacity under hypersaline conditions. Also, the reactive oxygen species (ROS) localization studies and quantification revealed elevated accumulation of ROS was observed in the chloroplast in the 3M condition. Pigment analysis shows a deficit in chlorophyll content and increased carotenoid accumulation, especially lutein and zeaxanthin content. This study majorly explored the chloroplast transcripts of the D. salina cell as it is the major environmental sensor. Even though most of the photosystem transcripts showed moderate upregulation in hypersaline conditions in the transcriptome study, the western blot analysis showed degradation of the core as well as antenna proteins of both the photosystems. Among the upregulated chloroplast transcripts, chloroplast Tidi, flavodoxin IsiB, and carotenoid biosynthesis-related protein transcripts strongly proposed photosynthetic apparatus remodeling. Also, the transcriptomic study revealed the upregulation of the tetrapyrrole biosynthesis pathway (TPB) and identified the presence of a negative regulator of this pathway, called the s-FLP splicing variant. These observations point towards the accumulation of TPB pathway intermediates PROTO-IX, Mg-PROTO-IX, and P-Chlide, those earlier reported as retrograde signaling molecules. Our comparative transcriptomic approach along with biophysical and biochemical studies in D. salina grown under control (1.5 M NaCl) and hypersaline (3M NaCl) conditions, unveil an efficient retrograde signaling mechanism mediated remodeling of photosynthetic apparatus.

Prevalence of latent structural heart disease in Nepali schoolchildren.

BACKGROUND: The present study aimed to quantify the burden of structural heart disease in Nepali children. METHODS: We performed a school-based cross-sectional echocardiographic screening study with cluster random sampling among children 5-16 years of age. RESULTS: Between December 2012 and January 2019, 6573 children (mean age 10.6 ± 2.9 years) from 41 randomly selected schools underwent echocardiographic screening. Structural heart disease was detected in 14.0 per 1000 children (95% CI 11.3-17.1) and was congenital in 3.3 per 1000 (95% CI 2.1-5.1) and rheumatic in 10.6 per 1000 (95% CI 8.3-13.4). Rates of rheumatic heart disease were higher among children attending public as compared to private schools (OR 2.8, 95% CI 1.6-5.2, p = 0.0001). CONCLUSION: Rheumatic heart disease accounted for three out of four cases of structural heart disease and was more common among children attending public as compared to private schools.

Livestock vaccination programme participation among smallholder farmers on the outskirts of National Parks and Tiger Reserves in the Indian states of Madhya Pradesh and Assam.

Effective livestock vaccination has the potential to raise prosperity and food security for the rural poor in low and middle income countries. To understand factors affecting access to vaccination services, and guide future policy, smallholder farmers in three locations in India were questioned about vaccination of their cattle and buffalo, with particular reference to foot and mouth disease (FMD), haemorrhagic septicaemia (HS) and blackquarter (BQ). In the three regions 51%, 50%, and 31% of respondents reported vaccinating their livestock; well below any threshold for effective population level disease control. However, within the third region, 65% of respondents in villages immediately surrounding the Kaziranga National Park reported vaccinating their cattle. The majority of respondents in all three regions were aware of FMD and HS, awareness of BQ was high in the Kanha and Bandhavgarh regions, but much lower in the Kaziranga region. The majority of respondents had positive attitudes to vaccination; understood vaccination protected their animals from specific diseases; and wished to immunise their livestock. There was no significant association between the age or gender of respondent and the immunisation of their livestock. Common barriers to immunisation were: negative attitudes to vaccination; lack of awareness of date and time of vaccination events; and difficulty presenting animals. Poor access to vaccination services was significantly associated with not vaccinating livestock. Fear of adverse reactions to vaccines was not significantly associated with not vaccinating livestock. Respondents who reported that vets or animal health workers (AHWs) were their main source of animal health knowledge were significantly more likely to have immunised their livestock in the last twelve months. Participants cited poor communication from vaccinators as problematic, both in publicising immunisation programmes, and explaining the purpose of vaccination. Where vaccinations were provided free of charge, farmers commonly displayed passive attitudes to accessing vaccination services, awaiting organised "immunisation drives" rather than seeking vaccination themselves. Based on these findings the following recommendations are made to improve participation and effectiveness of immunisation programmes. Programmes should be planned to integrate with annual cycles of: disease risk, agricultural activity, seasonal climate, social calendar of villages; and maximise efficiency for vaccinators. Dates and times of immunisation in each village must be well publicised, as respondents frequently reported missing the vaccinators. Relevant farmer education should precede immunisation programmes to mitigate against poor knowledge or negative attitudes. Immunisation drives must properly engage beneficiaries, particularly ensuring that services are accessible to female livestock keepers, and sharing some responsibilities with local farmers. Payment of a small monetary contribution by animal keepers could be considered to encourage responsibility for disease prevention, making vaccination an active process by farmers.

Effectiveness of Systematic Echocardiographic Screening for Rheumatic Heart Disease in Nepalese Schoolchildren: A Cluster Randomized Clinical Trial.

Importance: Echocardiographic screening allows for early detection of subclinical stages of rheumatic heart disease among children in endemic regions. Objective: To investigate the effectiveness of systematic echocardiographic screening in combination with secondary antibiotic prophylaxis on the prevalence of rheumatic heart disease. Design, Setting, and Participants: This cluster randomized clinical trial included students 9 to 16 years of age attending public and private schools in urban and rural areas of the Sunsari district in Nepal that had been randomly selected on November 17, 2012. Echocardiographic follow-up was performed between January 7, 2016, and January 3, 2019. Interventions: In the experimental group, children underwent systematic echocardiographic screening followed by secondary antibiotic prophylaxis in case they had echocardiographic evidence of latent rheumatic heart disease. In the control group, children underwent no echocardiographic screening. Main Outcomes and Measures: Prevalence of the composite of definite or borderline rheumatic heart disease according to the World Heart Federation criteria in experimental and control schools as assessed 4 years after intervention. Results: A total of 35 schools were randomized to the experimental group (n = 19) or the control group (n = 16). After a median of 4.3 years (interquartile range [IQR], 4.0-4.5 years), 17 of 19 schools in the experimental group (2648 children; median age at follow-up, 12.1 years; IQR, 10.3-12.5 years; 1308 [49.4%] male) and 15 of 16 schools in the control group (1325 children; median age at follow-up, 10.6 years; IQR, 10.0-12.5 years; 682 [51.5%] male) underwent echocardiographic follow-up. The prevalence of definite or borderline rheumatic heart disease was 10.8 per 1000 children (95% CI, 4.7-24.7) in the control group and 3.8 per 1000 children (95% CI, 1.5-9.8) in the experimental group (odds ratio, 0.34; 95% CI, 0.11-1.07; P = .06). The prevalence in the experimental group at baseline had been 12.9 per 1000 children (95% CI, 9.2-18.1). In the experimental group, the odds ratio of definite or borderline rheumatic heart disease at follow-up vs baseline was 0.29 (95% CI, 0.13-0.65; P = .008). Conclusions and Relevance: School-based echocardiographic screening in combination with secondary antibiotic prophylaxis in children with evidence of latent rheumatic heart disease may be an effective strategy to reduce the prevalence of definite or borderline rheumatic heart disease in endemic regions. Trial Registration: ClinicalTrials.gov Identifier: NCT01550068.

Anaemia among Patients of Heart Failure in a Tertiary Care Centre of Nepal: A Descriptive Cross-sectional Study.

INTRODUCTION: Anaemia is an important comorbidity common in patients with heart failure and is associated with poor clinical status and worse outcomes. In Nepal few studies have evaluated anaemia amongst patients suffering from heart failure. We intended to find out the prevalence of anaemia in patients with heart failure in a tertiary care centre. METHODS: This is a descriptive cross-sectional study conducted among patients of heart failure presenting to tertiary care hospital in eastern Nepal from April 2017 to January 2018. Ethical approval was taken from the Institutional Review Committee of a tertiary care centre (reference number: IRC/0842/016). Using the convenience sampling method, 100 patients were enrolled in the study. Blood samples from the patients were taken for haemoglobin and serum iron studies. Data was analysed using Statistical Package for Social Sciences version 11. Point estimate at 95% Confidence Interval was calculated, with frequency and percentage. RESULTS: Among 100 patients with heart failure, 82 (82%) (74.47-89.53 at 95% Confidence Interval) had anaemia. Mean haemoglobin level of the study population was 10.40±2.73 g/dl. Fifty four (54%) of patients had iron deficiency status irrespective of presence or absence of anaemia. CONCLUSIONS: Prevalence of anaemia among patients of heart failure in our study was found to be higher than various other homologous international studies.

Animal health perceptions and challenges among smallholder farmers around Kaziranga National Park, Assam, India: A study using participatory epidemiological techniques.

Improvements to smallholder farming are essential to improvements in rural prosperity. Small farmers in the Kaziranga region of Assam operate mixed farming enterprises in a resource limited environment, which is subject to seasonal flooding. Participatory techniques, were used to elucidate the animal health challenges experienced in this landscape in order to inform and guide future animal health education and interventions. The flooding is essential for agricultural activities, but is a source of major losses and disruption. Farmers experience significant losses to their crops due to raiding by wild species such as elephants; predation of livestock by wild carnivores is also of concern. Access to veterinary services and medicines is limited by both financial and geographic constraints. Interviewees discussed nutritional and management issues such as poor availability of fodder and grazing land, while meeting attendees preferred to concentrate discussions on animal health issues. Livestock keepers were adept and consistent at describing disease syndromes. The key challenges identified by farmers were: foot-and-mouth disease; Newcastle disease; haemorrhagic septicaemia; chronic fasciolosis; diarrhoea; bloating diseases; goat pox; and sarcoptic mange. Improvements in the efficiency of farming in this region is a prerequisite for the local achievement of United Nations Sustainable development goals. There exist clear opportunities to increase productivity and prosperity among farmers in this region through a combination of vaccination programmes and planned animal management schemes, driven by a programme of participatory farmer education.

Colostrum provision and care of calves among smallholder farmers in the Kaziranga region of Assam, India.

Smallholder cattle farming in Assamese villages is sub-optimal in terms of calf survivability, growth, age at first service, and milk yield. Proper understanding of the local situation is essential to formulate appropriate, locally driven, livestock keeper education to sustainably improve animal health, welfare and productivity. In-depth interviewing and direct observation were used to understand the farming strategies, husbandry practices and challenges to health and productivity in a cluster of typical villages in the Kaziranga region of Assam, India, where resource use is balanced between the needs of humans and livestock, with competition from wild species. Knowledge of the importance of colostrum consumption by calves is poor. Timely consumption of sufficient colostrum (locally called "phehu") by calves was clearly sub-optimal in the majority of households. The reasons behind this are nuanced, but the practice of collecting colostrum from newly calved cows to make confectionery for human consumption is an important contributory factor. Care of the umbilicus of the newborn is not routine practice in the locality. Local women are the key group assisting with young and sick animals, including cases of simple dystocia and retained foetal membranes. Cows are usually milked once daily, to attempt to balance the needs for milk of household with those of the calf, which can result in suboptimal nutrition for calves. There are clear opportunities to improve animal health and productivity through locally provided farmer education, particularly with reference to colostrum provision, and the engagement of women farmers in any such programme is key to success.

Preparing Intensive Care Unit in Resource-Constraint Setting Amid COVID-19 Pandemic: Our Experience and Review.

COVID-19 pandemic is an emerging, rapidly evolving public health emergency where a nation's health-care system can face a marked surge in demand for intensive care unit (ICU) beds and organ support. In regions with insufficient medical resources, it may further aggravate the existing shortage, limiting an ICU's ability to provide the normal standard of care. It can present ethically or legally demanding questions about how to prioritize the allocation of life-saving medical resources. In developing countries like India, still many hospitals are challenged by competing priorities and remain underprepared. In the wake of COVID-19 pandemic, to guide the intensive care disaster planners in regions with low resources and to ensure ICU readiness, this review shares our experience and strategies for preparing ICU with existing and alternative resources, focusing on space, equipment, and health-care workers' safety and training.

Spread and impact of goat pox ("sagolay bohonta") in a village smallholder community around Kaziranga National Park, Assam, India.

During September and October 2017, a highly fatal outbreak of a disease clinically indistinguishable from goat pox occurred in the villages around the Kaziranga National Park, Assam, India. This was investigated through clinical examination of affected animals, individual interviews with goat keepers and participatory village meetings. Laboratory confirmation was impractical due to the isolation and poverty of the affected community and unnecessary due to the specific nature of the clinical signs. Respondents reported not having encountered the disease previously, and it would appear that a naïve local population developed within an endemically affected region because of a trend to avoid purchasing animals from outside the village. Local grazing practices appear to have had a role in both the spread and control of the outbreak. Goats are an important form of savings and cash income to people in the locality, and the outbreak may result in considerable financial hardship for affected goat keepers. We provide a detailed description of the clinical disease and the spread of the outbreak in the locality. Awareness of the disease with reference to farming practices will provide opportunities for future disease control to enhance animal welfare and rural prosperity.

Correction: Delivery and evaluation of participatory education for animal keepers led by veterinarians and para-veterinarians around the Kanha Tiger Reserve, Madhya Pradesh, India.

[This corrects the article DOI: 10.1371/journal.pone.0200999.].

Delivery and evaluation of participatory education for animal keepers led by veterinarians and para-veterinarians around the Kanha Tiger Reserve, Madhya Pradesh, India.

AIM: We aim to investigate local perceptions of animal health challenges; current animal health knowledge; and methods to provide effective, relevant education to animal keepers in the Kanha Tiger Reserve area. MATERIALS AND METHODS: A farmer education programme was undertaken in the Kanha Tiger Reserve area. Local animal health priorities were investigated through participatory village meetings (n = 38), individual animal keeper questionnaires (n = 100) and a written survey of local paravets (n = 16). Educational interventions were: veterinary surgeon led education meeting (VE); paravet led education meeting (PVE); distribution of printed materials (PM). 230 village meetings were carried out across 181 villages, contacting 3791 animal keepers. 20 villages received printed materials. Information was gathered on perceptions of local animal health challenges and current remedies. Efficacy of knowledge transfer was assessed four to five months later using a purposeful sample of 38 villages. RESULTS: Group meetings identified ticks (35/38), foot and mouth disease (FMD) (31/38) and diarrhoea (30/38) as the greatest animal health challenges. Individual interviews identified haemorrhagic septicaemia (HS) (87/100), blackquarter (BQ) (66/100) and plastic ingestion (31/100). Paravets identified FMD (7/16), BQ (6/16) and HS (6/16), and also indicated that animal husbandry and socio-economic factors were important. Current treatments were primarily home remedies and herbalism, but also included contacting a paravet, use of pharmaceuticals and faith healing. Animal treatment knowledge prior to intervention was not significantly different between groups (P = 0.868). Following intervention animal health knowledge was assessed: PVE performed better than controls (P = 0.001) and PM (P = 0.003); VE performed better than controls (P = 0.009). There was no significant difference between VE and PVE (P = 0.666) nor PM and controls (P = 0.060). CONCLUSIONS AND RECOMMENDATIONS: Open access participatory village meetings are an effective way to provide animal health education. In this region distribution of posters and leaflets did not appear to be an effective way to contact animal keepers. Meetings led by paravets can be as effective as those led by veterinarians and paravets can rapidly and sustainably contact large numbers of animal keepers. Investigation of the local animal health situation is essential to ensure education is relevant and accessible to intended recipients. Interventions must be carefully planned to maximise engagement of all sections of the community, particularly women.

Prevalence of Subclinical Rheumatic Heart Disease in Eastern Nepal: A School-Based Cross-sectional Study.

IMPORTANCE: Although rheumatic heart disease has been nearly eradicated in high-income countries, 3 in 4 children grow up in parts of the world where it is still endemic. OBJECTIVES: To determine the prevalence of clinically silent and manifest rheumatic heart disease as a function of age, sex, and socioeconomic status and to estimate age-specific incidence. DESIGN, SETTING, AND PARTICIPANTS: In this school-based cross-sectional study with cluster sampling, 26 schools in the Sunsari district in Eastern Nepal with 5467 eligible children 5 to 15 years of age were randomly selected from 595 registered schools. After exclusion of 289 children, 5178 children were enrolled in the present study from December 12, 2012, through September 12, 2014. Data analysis was performed from October 1, 2014, to April 15, 2015. EXPOSURES: Demographic and socioeconomic characteristics were acquired in a standardized interview by means of a questionnaire customized to the age of the children. A focused medical history was followed by a brief physical examination. Cardiac auscultation and transthoracic echocardiography were performed by 2 independent physicians. MAIN OUTCOMES AND MEASURES: Rheumatic heart disease according to the World Heart Federation criteria. RESULTS: The median age of the 5178 children enrolled in the study was 10 years (interquartile range, 8-13 years), and 2503 (48.3%) were female. The prevalence of borderline or definite rheumatic heart disease was 10.2 (95% CI, 7.5-13.0) per 1000 children and increased with advancing age from 5.5 (95% CI, 3.5-7.5) per 1000 children 5 years of age to 16.0 (95% CI, 14.9-17.0) in children 15 years of age, whereas the mean incidence remained stable at 1.1 per 1000 children per year. Children with rheumatic heart disease were older than children without rheumatic heart disease (median age [interquartile range], 11 [9-14] years vs 10 [8-13] years; P = .03), more commonly female (34 [64.2%] vs 2469 [48.2%]; P = .02), and more frequently went to governmental schools (40 [75.5%] vs 2792 [54.5%]; P = .002). Silent disease (n = 44) was 5 times more common than manifest disease (n = 9). CONCLUSIONS AND RELEVANCE: Rheumatic heart disease affects 1 in 100 schoolchildren in Eastern Nepal, is primarily clinically silent, and may be more common among girls. The overall prevalence and the ratio of manifest to subclinical disease increase with advancing age, whereas the incidence remains stable at 1.1 per 1000 children per year. Early detection of silent disease may help prevent progression to severe valvular damage.