Development of a novel chelation-based recycling strategy for the efficient decontamination of hazardous petroleum refinery spent catalysts.

The conventional hydrometallurgical methods for recycling refinery spent hydroprocessing catalysts are ineffective in simultaneously removing all metals (Ni, V, and Mo) in a single-stage operation. In this study, a novel octadentate chelating agent, diethylenetriaminepentaacetic acid (DTPA-C14H23N3O10), has been proposed for the first time to remove toxic metals (Ni, V, and Mo) in a single stage of operation from an industrial spent atmospheric residue desulfurization (ARDS) catalysts. It was discovered that the efficient formation of metal-DTPA complexes was attained under the optimum experimental conditions (60 °C, stirring - 150 rpm, S/L ration (w/v) of 2.5%, 7.5% DTPA, and medium pH-9) that resulted in the high removal of Mo (83.6%), V (81.3%) and Ni (64.1%) from the spent ARDS catalyst. Kinetic studies suggest that the leaching process followed a semi-empirical Avrami equation (R2 > 0.92), which predicted that the diffusion control reaction controlled the leaching. Species distribution and ecological risk analysis of the remaining metals in the insoluble residue (mostly Al2O3) indicated that the potential bioavailability of the remaining metals (except Ni) was significantly decreased, and residue poses a low ecological and contamination risk (individual contamination factor <1). Furthermore, the textural properties of the residue (BET surface area-103 m2/g and pore volume- 0.49 ml/g) were dramatically improved, suggesting that fresh hydroprocessing catalyst support can be synthesized using the leached residue. Compared to the conventional processes, the proposed chelating process is highly selective, closed-loop, and achieved high metal recovery in a single-stage operation while decreasing the environmental risks of the hazardous spent catalysts.

Identification of Quinolinones as Antivirals against Venezuelan Equine Encephalitis Virus.

Venezuelan equine encephalitis virus (VEEV) is a reemerging alphavirus that can cause encephalitis resulting in severe human morbidity and mortality. Using a high-throughput cell-based screen, we identified a quinolinone compound that protected against VEEV-induced cytopathic effects. Analysis of viral replication in cells identified several quinolinone compounds with potent inhibitory activity against vaccine and virulent strains of VEEV. These quinolinones also displayed inhibitory activity against additional alphaviruses, such as Mayaro virus and Ross River virus, although the potency was greatly reduced. Time-of-addition studies indicated that these compounds inhibit the early-to-mid stage of viral replication. Deep sequencing and reverse genetics studies identified two unique resistance mutations in the nsP2 gene (Y102S/C; stalk domain) that conferred VEEV resistance on this chemical series. Moreover, introduction of a K102Y mutation into the nsP2 gene enhanced the sensitivity of chikungunya virus (CHIKV) to this chemical series. Computational modeling of CHIKV and VEEV nsP2 identified a highly probable docking alignment for the quinolinone compounds that require a tyrosine residue at position 102 within the helicase stalk domain. These studies identified a class of compounds with antiviral activity against VEEV and other alphaviruses and provide further evidence that therapeutics targeting nsP2 may be useful against alphavirus infection.

A multicentre point prevalence survey (PPS) of antimicrobial use amongst admitted patients in tertiary care centres in India.

OBJECTIVES: Data from point prevalence surveys (PPSs) in India are scarce. Conducting PPSs is especially challenging in the absence of electronic medical records, a lack of dedicated resources and a high patient load in resource-poor settings. This multicentre survey was conducted to provide background data for planning and strengthening antimicrobial stewardship programmes across the country. METHODS: This inpatient PPS was conducted over 2 weeks in May 2019 simultaneously across five study centres in India. Data about patient characteristics, indications for antimicrobials use and details of each antimicrobial prescribed including supportive investigation reports were collected in predesigned forms. RESULTS: A total of 3473 admitted patients in wards and ICUs were covered across five study centres. Of these, 1747 (50.3%) patients were on antimicrobials, with 46.9% patients being on two or more antimicrobials. Out of the total antimicrobials prescribed, 40.2% of the antimicrobials were prescribed for community-acquired infection requiring hospitalization followed by surgical prophylaxis (32.6%). Third-generation cephalosporins and drugs from the 'Watch' category were prescribed most commonly. Only 22.8% of the antimicrobials were based on microbiology reports. CONCLUSIONS: The survey demonstrated a high use of antimicrobials in admitted patients with a considerable proportion of drugs from the 'Watch' category. The targets for interventions that emerged from the survey were: improving surgical prophylaxis, decreasing double anaerobic cover, initiating culture of sending cultures and de-escalation with targeted therapy.

Effect of school-based educational water, sanitation, and hygiene intervention on student's knowledge in a resource-limited setting.

BACKGROUND: Globally, diarrhea is one of the major causes of under-5 mortality. India accounts for the highest number of childhood deaths from diarrhea globally. Therefore, facilitating the implementation of water, sanitation, and hygiene (WASH)-related interventions in schools and communities is crucial. In the present study, we investigated the effectiveness of a school-based educational WASH intervention in improving students' knowledge on prevention and management of diarrhea in Ujjain district, India. METHODS: The present pre-post intervention study with a two-stage (schools and classrooms) cluster sampling was conducted on 1,781 students studying in grades: 8th-12th; age: 14-19 years) in schools located in Ujjain, Madhya Pradesh, India. The intervention comprised an educational training session using a WASH training module. The means of pre- and post-intervention scores were compared using repeated measure analysis of variance. A multivariate quantile regression model was used to test the correlation between the change in score after intervention and the independent variables. A P value of <0.05 was considered statistically significant. RESULTS: The proportions of students possessing knowledge on the treatment of diarrhea, use of zinc tablets during an episode of diarrhea, and the symptoms and signs of severe pediatric diarrhea were 28%, 27%, and 27%, respectively, before intervention. These proportions increased (P<0.001) after the educational intervention to 72%, 73%, and 74%, respectively. The mean post-intervention knowledge score (34.13) was higher than the mean pre-intervention score (15.17) (F = 16513.36, P< 0.001). Age was associated with the knowledge score at the 25th and higher quantile (q). Gender exhibited a greater effect at q10th. School location was positively associated at q25th and higher. School type was strongly associated at low quantiles (q10th and q25th). School medium exhibited a greater association at low quantiles (≤q25th). CONCLUSION: WASH- and diarrhea-related knowledge among higher secondary school students increased after the educational intervention. Further research is required to evaluate the sociodemographic characteristics associated with change in the knowledge score to better evaluate school-based educational WASH interventions and improve the management and prevention of diarrhea.

Effects of improved information and volunteer support on segregation of solid waste at the household level in urban settings in Madhya Pradesh, India (I-MISS): protocol of a cluster randomized controlled trial.

BACKGROUND: Segregation of household waste at the source is an effective and sustainable strategy for management of municipal waste. However, household segregation levels remain insufficient as waste management approaches are mostly top down and lack local support. The realisation and recognition of effective, improved and adequate waste management may be one of the vital drivers for attaining environmental protection and improved health and well-being. The presence of a local level motivator may promote household waste segregation and ultimately pro-environmental behaviour. The present cluster randomized control trial aims to understand if volunteer based information on waste segregation (I-MISS) can effectively promote increased waste segregation practices at the household level when compared with existing routine waste segregation information in an urban Indian setting. METHODS: This paper describes the protocol of an 18 month two-group parallel,cluster randomised controlled trialin the urban setting of Ujjain, Madhya Pradesh, India. Randomization will be conducted at ward level, which is the last administrative unit of the municipality. The study will recruit 425 households in intervention and control groups. Assessments will be performed at baseline (0 months), midline (6 months), end line (12 months) and post intervention (18 months). The primary outcome will be the comparison of change in proportion of households practicing waste segregation and change in proportion of mis-sorted waste across the study period between the intervention and control groups as assessed by pick analysis. Intention to treat analysis will be conducted. Written informed consent will be obtained from all participants. DISCUSSION: The present study is designed to study whether an external motivator, a volunteer selected from the participating community and empowered with adequate training, could disseminate waste segregation information to their community, thus promoting household waste segregation and ultimately pro-environmental behaviour. The study envisages that the volunteers could link waste management service providers and the community, give a local perspective to waste management, and help to change community habits through information, constant communication and feedback. TRIAL REGISTRATION: The study is registered prospectively with Indian Council of Medical Research- Clinical Trial Registry of India ( CTRI/2020/03/024278 ).

Fungal bioleaching of metals from refinery spent catalysts: A critical review of current research, challenges, and future directions.

Petroleum refining operations such as hydroprocessing and fluid catalytic cracking (FCC) generate huge quantities of spent catalysts containing toxic and valuable metals (Ni, V, Mo, Co, W, Al, etc.), the management of which is a serious environmental issue. Besides environmental concerns, the different metals present in the spent catalysts are also a valuable commodity to modern industries. Therefore, these spent catalysts also provide an opportunity to use it as a source of value to the refiners. In recent years, a biotechnological based leaching process 'bioleaching' has emerged as a promising eco-friendly technique for the extraction of metals from these refinery spent catalysts. Among various bioleaching agents such as archean, bacterial, or fungi, the process mediated by the fungi (Aspergillus niger, Penicillium simplicissimum, and many others) is gaining attention owing to the high metal extraction ability of the various fungal produced metabolites (organic acids) under moderately acidic conditions. Furthermore, the ability of these fungi to withstand wide process conditions (pH, spent catalyst concentration, substrate types, etc.), high metal toxicity and use of low-cost organic substrate make them an ideal candidate for bioleaching. In this review article, we shed light on the role and mechanisms of fungi involved in extracting different metals from spent hydroprocessing and FCC catalysts. Key process parameters that affect the efficiency of fungal based bioleaching are discussed. The techno-economic challenges associated with the process are elaborated, and the needed future research directions to promote its commercial applications are highlighted. Based on our analysis, it can be argued that the fungi bioleaching has potential, however, some challenges (slower kinetics, and health and safety) should be addressed before the process can be scaled up for the commercial application.

Stable high frequencies of sulfadoxine-pyrimethamine resistance associated mutations and absence of K13 mutations in Plasmodium falciparum 3 and 4 years after the introduction of artesunate plus sulfadoxine-pyrimethamine in Ujjain, Madhya Pradesh, India.

BACKGROUND: Artesunate plus sulfadoxine-pyrimethamine (ASP) is first-line treatment for uncomplicated Plasmodium falciparum malaria in most of India, except for six North-eastern provinces where treatment failure rates were high. In Ujjain, central India, the frequency of mutations associated with increased drug tolerance, but not overt resistance to sulfadoxine and pyrimethamine were 9% and > 80%, respectively, in 2009 and 2010, just prior to the introduction of ASP. The frequency of drug resistance associated mutations in Ujjain in 2015-2016 after 3-4 years of ASP use, are reported. METHODS: Blood samples from patients with P. falciparum mono-infection verified by microscopy were collected on filter-paper at all nine major pathology laboratories in Ujjain city. Codons pfdhfr 16-185, pfdhps 436-632 and K13 407-689 were identified by sequencing. Pfcrt K76T and pfmdr1 N86Y were identified by restriction fragment length polymorphism. RESULTS: Sulfadoxine-pyrimethamine resistance-associated pfdhfr 108 N and 59R alleles were found in 100/104 (96%) and 87/91 (96%) samples, respectively. Pfdhps 437G was found in 10/105 (10%) samples. Double mutant pfdhfr 59R + 108 N were found in 75/81 (93%) samples. Triple mutant pfdhfr 59R + 108 N and pfdhps 437G were found in 6/78 (8%) samples. Chloroquine-resistance-associated pfcrt 76T was found in 102/102 (100%). Pfmdr1 N86 and 86Y were identified in 83/115 (72%) and 32/115 (28%) samples, respectively. CONCLUSION: The frequency of P. falciparum with reduced susceptibility to sulfadoxine-pyrimethamine remained high, but did not appear to have increased significantly since the introduction of ASP. No polymorphisms in K13 associated with decreased artemisinin susceptibility were found. ASP probably remained effective, supporting continued ASP use.

Incidence, clinical profile, and risk factors for serious bacterial infections in children hospitalized with fever in Ujjain, India.

BACKGROUND: Fever is a cause for concern for both parents and the treating pediatrician and a common reason for antibiotic overuse. However, the proportion of children hospitalized for fever with serious bacterial infection (SBI) is uncertain. We aimed to evaluate the epidemiological, clinical, hematological, and biochemical risks for SBI among the children admitted with fever. METHOD: This prospective study was conducted in a rural teaching hospital in India on consecutive children, aged 3 months-12 years, presenting with fever 100 °F (37.7 °C) or higher. The presence of SBI was confirmed with one of the following criteria: (a) a positive blood culture; (b) roentgenographically confirmed pneumonia with high titres of C-reactive protein; (c) a culture-confirmed urinary tract infection; (d) enteric fever diagnosed clinically in addition to either a positive blood culture or high Widal titers; and (e) meningitis diagnosed clinically in addition to either a positive blood culture or cerebrospinal fluid culture. A predefined questionnaire was filled. RESULTS: A total of 302 children were included in the study, out of which 47% (95% CI 41.4-52.7%) presented with SBI. The factors associated with confirmed SBI in bivariate analysis were history of previous hospitalization, history of chronic illness, history of medication in the previous 1 week, a partially immunized child, history of common cold, moderate-grade fever, toxic look, significant lymphadenopathy, absence of BCG scar, delayed development, irritability, breathlessness, respiratory distress, poor feeding, significant weight loss, suspected urinary tract infection, hyponatremia, hypokalemia, and abnormal leucocyte count. The final generalized logistic regression model revealed partially immunized child (RR 4.26), breathlessness (RR 1.80), weight loss (RR 2.28), and suspected urinary tract infection (RR 1.95) as risk factors for the increased risk of SBI. CONCLUSION: The study identified multiple risk factors for SBI. Pediatricians can be made aware of these risk factors. Further studies are warranted to identify age-specific risk factors for SBI because most clinicians depend on clinical signs and symptoms to identify SBI.

Falls in older aged adults in 22 European countries: incidence, mortality and burden of disease from 1990 to 2017.

INTRODUCTION: Falls in older aged adults are an important public health problem. Insight into differences in fall-related injury rates between countries can serve as important input for identifying and evaluating prevention strategies. The objectives of this study were to compare Global Burden of Disease (GBD) 2017 estimates on incidence, mortality and disability-adjusted life years (DALYs) due to fall-related injury in older adults across 22 countries in the Western European region and to examine changes over a 28-year period. METHODS: We performed a secondary database descriptive study using the GBD 2017 results on age-standardised fall-related injury in older adults aged 70 years and older in 22 countries from 1990 to 2017. RESULTS: In 2017, in the Western European region, 13 840 per 100 000 (uncertainty interval (UI) 11 837-16 113) older adults sought medical treatment for fall-related injury, ranging from 7594 per 100 000 (UI 6326-9032) in Greece to 19 796 per 100 000 (UI 15 536-24 233) in Norway. Since 1990, fall-related injury DALY rates showed little change for the whole region, but patterns varied widely between countries. Some countries (eg, Belgium and Netherlands) have lost their favourable positions due to an increasing fall-related injury burden of disease since 1990. CONCLUSIONS: From 1990 to 2017, there was considerable variation in fall-related injury incidence, mortality, DALY rates and its composites in the 22 countries in the Western European region. It may be useful to assess which fall prevention measures have been taken in countries that showed continuous low or decreasing incidence, death and DALY rates despite ageing of the population.

Prevalence and perceptions of infant massage in India: study from Maharashtra and Madhya Pradesh states.

BACKGROUND: Knowledge of the prevailing infant care practices and their effects is important to inform practice and public programs. Infant massage is a traditional practice in India but remains less studied. This study was conducted to study the prevalence and perceptions of infant massage practices in two states of India. METHODS: A total of 1497 caretakers of children under 18 months of age were interviewed in a cross-sectional study at immunisation units of medical schools in Maharashtra (MH) and Madhya Pradesh (MP) states and through home visits in villages in MH during March through August 2018. RESULTS: Infant massage was a prevalent practice (93.8% [95%CI: 92.4,94.9]) in both study states - 97.9%[95%CI:96.9,98.8] in MH and 85.3%[95%CI: 81.9,88.3] in MP - and the prevalence did not vary between male (94.5%) and female (93.5%) infants (p = 0.44). Massage was mostly initiated in the first week of life (82%); it is widely viewed as a traditional practice. It was common to massage the baby once daily (77%), before bathing (77%), and after feeding (57%). Massage was mostly conducted using oils (97%). In MH, preferred oils were a sesame oil-based proprietary traditional medicine oil (36%) and coconut oil (18%) while olive (29%) and mustard (20%) oils were most popular in MP. Commonly reported application techniques included gentle massage with minimal pressure, pressing (30%) and manually stretching certain joints (60%). Commonly reported perceived benefits of infant massage included increased bone strength, better sleep and growth, while no harm was perceived (95%). CONCLUSION: Infant oil massage is a highly prevalent traditional practice in MH and MP. Clear guidance on the use of massage, choice of oil, and techniques for application is required to optimize benefits and minimize risks of this popular traditional practice.

Identifying the know-do gap in evidence-based neonatal care practices among informal health care providers-a cross-sectional study from Ujjain, India.

BACKGROUND: More than a quarter of global neonatal deaths are reported from India, and a large proportion of these deaths are preventable. However, in the absence of robust public health care systems in several states in India, informal health care providers (IHCPs) with no formal medical education are the first contact service providers. The aim of this study was to assess the knowledge of IHCPs in basic evidence-based practices in neonatal care in Ujjain district and investigated factors associated with differences in levels of knowledge. METHODS: A cross-sectional survey was conducted using a questionnaire with multiple-choice questions covering the basic elements of neonatal care. The total score of the IHCPs was calculated. Multivariate quantile regression model was used to look for association of IHCPs knowledge score with: the practitioners' age, years of experience, number of patients treated per day, and whether they attended children in their practice. RESULTS: Of the 945 IHCPs approached, 830 (88%) participated in the study. The mean ± SD score achieved was 22.3 ± 7.7, with a median score of 21 out of maximum score of 48. Although IHCPs could identify key tenets of enhancing survival chances of neonates, they scored low on the specifics of cord care, breastfeeding, vitamin K use to prevent neonatal hemorrhage, and identification and care of low-birth-weight babies. The practitioners particularly lacked knowledge about neonatal resuscitation, and only a small proportion reported following up on immunizations. Results of quantile regression analysis showed that more than 5 years of practice experience and treating more than 20 patients per day had a statistically significant positive association with the knowledge score at higher quantiles (q75th and q90th) only. IHCPs treating children had significantly better scores across quantiles accept at the highest quantile (90th). CONCLUSIONS: The present study highlighted that know-do gap exists in evidence-based practices for all key areas of neonatal care tested among the IHCPs. The study provides the evidence that some IHCPs do possess knowledge in basic evidence-based practices in neonatal care, which could be built upon by future educational interventions. Targeting IHCPs can be an innovative way to reach a large rural population in the study setting and to improve neonatal care services.

Column bioleaching of metals from refinery spent catalyst by Acidithiobacillus thiooxidans: Effect of operational modifications on metal extraction, metal precipitation, and bacterial attachment.

The feasibility of column bioleaching in the recovery of valuable metals (Ni, V, Mo, and Al) from an uncrushed petroleum refinery spent hydroprocessing catalyst using Acidithiobacillus thiooxidans has been reported. Different operational strategies such as submerged bioleaching in continuous mode, submerged bioleaching in resting period mode, free flow bioleaching in continuous mode, and free flow bioleaching in resting period mode were tested to find out the optimum bioleaching strategy for the recovery of metals from spent hydroprocessing catalyst. Among various operational modifications, submerged bioleaching in continuous mode was considered as the best strategy in which about 82.9% of Ni, 33.4% of Al, and 22.7% of Mo were leached after 315 h of column operation. The maximum yield of V (53.6%) in this column was achieved in 105 h, after which, a rapid decrease in its yield was observed, possibly due to its precipitation. The field emission scanning electron microscopy (FESEM) analysis revealed the presence of V in precipitates. The modified kinetic models showed that the leaching of Al, V and Mo followed the chemical control model, whereas the dissolution of Ni was controlled by diffusion control reaction. The bacterial attachment study with FESEM indicated that the metal toxicity was induced on bacterial cells attached to the sulfur particles. The results of the current study indicate that column bioleaching of spent hydroprocessing catalyst is effective in leaching of Ni and V, whereas leaching of Al and Mo require further treatments.

Changes in the fractionation profile of Al, Ni, and Mo during bioleaching of spent hydroprocessing catalysts with Acidithiobacillus ferrooxidans.

Spent hydroprocessing catalysts are known to contain a variety of potentially toxic metals and therefore studies on the bioavailability and mobility of these metals are critical for understanding the possible environmental risks of the spent catalysts. This study evaluates the different chemical fractions/forms of aluminium (Al), nickel (Ni), and molybdenum (Mo) in spent hydroprocessing catalyst and the changes they undergo during bioleaching with Acidithiobacillus ferrooxidans. In the spent catalyst (prior to bioleaching), Al was primarily present in its residual form, suggesting its low environmental mobility. However, Ni comprised mainly an exchangeable fraction, indicating its high environmental mobility. Molybdenum was mainly in the oxidizable form (47.1%), which indicated that highly oxidizing conditions were required to liberate it from the spent catalyst. During bioleaching the exchangeable, reducible and oxidizable fractions of all the metals were leached, whereas the residual fractions remained largely unaffected. At the end of bioleaching process, the metals remaining in the bioleached sample were predominantly in the residual fraction (98.3-99.5%). The 'risk assessment code' (RAC) and IR analysis also demonstrated that the environmental risks of the bioleached residue were significantly lower compared to the untreated spent catalyst. The results of this study suggest that bioleaching is an effective method in removing the metals from spent catalysts and the bioleached residue poses little environmental risk.

Does Risperidone Long Acting Injectable Depot (RLAI) Reduce number of Admissions to Hospital.

BACKGROUND: Adherence to treatment is a major issue in relapse prevention in schizophrenia. Injectable depot has been claimed to improve non-adherence. A study in the North of England has shown that risperidone long acting injectable form has reduced number of admissions and number of days stayed in hospital. Our study aimed to replicate the previous study in the South of England. METHODS: A retrospective study was conducted in the South Essex Foundation University NHS trust. We selected every fifth patient on the Hospital Pharmacy list for risperidone long acting injectable (RLAI). We collected information on. Age, sex, diagnosis and medication ,regular follow up, investigation of each patient which included weight, FBS, S. lipid and hormones at the start of treatment, at three months and six months intervals. We recorded reasons for starting RLAI. Number of antipsychotics prescribed before RLAI, illness duration, number of admissions and days stayed in hospital in each episode before and after RLAI. Patients were included if they stayed for one year or on RLAI. RESULTS: We reviewed 65 case notes. The most common reason for starting RLAI was non compliance. 80% of our sample had the illness for more than five years. Half of them had physical morbidity. After RLAI was commenced there was a significant association with number and length of admissions. CONCLUSIONS: We have found an association between prescription of RLAI and reduction in number and length of admission. This needs to be confirmed in a randomised controlled trial.

Unique functional and structural properties of the LRRK2 protein ATP-binding pocket.

Pathogenic mutations in the LRRK2 gene can cause late-onset Parkinson disease. The most common mutation, G2019S, resides in the kinase domain and enhances activity. LRRK2 possesses the unique property of cis-autophosphorylation of its own GTPase domain. Because high-resolution structures of the human LRRK2 kinase domain are not available, we used novel high-throughput assays that measured both cis-autophosphorylation and trans-peptide phosphorylation to probe the ATP-binding pocket. We disclose hundreds of commercially available activity-selective LRRK2 kinase inhibitors. Some compounds inhibit cis-autophosphorylation more strongly than trans-peptide phosphorylation, and other compounds inhibit G2019S-LRRK2 more strongly than WT-LRRK2. Through exploitation of structure-activity relationships revealed through high-throughput analyses, we identified a useful probe inhibitor, SRI-29132 (11). SRI-29132 is exquisitely selective for LRRK2 kinase activity and is effective in attenuating proinflammatory responses in macrophages and rescuing neurite retraction phenotypes in neurons. Furthermore, the compound demonstrates excellent potency, is highly blood-brain barrier-permeant, but suffers from rapid first-pass metabolism. Despite the observed selectivity of SRI-29132, docking models highlighted critical interactions with residues conserved in many protein kinases, implying a unique structural configuration for the LRRK2 ATP-binding pocket. Although the human LRRK2 kinase domain is unstable and insoluble, we demonstrate that the LRRK2 homolog from ameba can be mutated to approximate some aspects of the human LRRK2 ATP-binding pocket. Our results provide a rich resource for LRRK2 small molecule inhibitor development. More broadly, our results provide a precedent for the functional interrogation of ATP-binding pockets when traditional approaches to ascertain structure prove difficult.

De novo assembly and characterization of transcriptomes of early-stage fruit from two genotypes of Annona squamosa L. with contrast in seed number.

BACKGROUND: Annona squamosa L., a popular fruit tree, is the most widely cultivated species of the genus Annona. The lack of transcriptomic and genomic information limits the scope of genome investigations in this important shrub. It bears aggregate fruits with numerous seeds. A few rare accessions with very few seeds have been reported for Annona. A massive pyrosequencing (Roche, 454 GS FLX+) of transcriptome from early stages of fruit development (0, 4, 8 and 12 days after pollination) was performed to produce expression datasets in two genotypes, Sitaphal and NMK-1, that show a contrast in the number of seeds set in fruits. The data reported here is the first source of genome-wide differential transcriptome sequence in two genotypes of A. squamosa, and identifies several candidate genes related to seed development. RESULTS: Approximately 1.9 million high-quality clean reads were obtained in the cDNA library from the developing fruits of both the genotypes, with an average length of about 568 bp. Quality-reads were assembled de novo into 2074 to 11004 contigs in the developing fruit samples at different stages of development. The contig sequence data of all the four stages of each genotype were combined into larger units resulting into 14921 (Sitaphal) and 14178 (NMK-1) unigenes, with a mean size of more than 1 Kb. Assembled unigenes were functionally annotated by querying against the protein sequences of five different public databases (NCBI non redundant, Prunus persica, Vitis vinifera, Fragaria vesca, and Amborella trichopoda), with an E-value cut-off of 10(-5). A total of 4588 (Sitaphal) and 2502 (NMK-1) unigenes did not match any known protein in the NR database. These sequences could be genes specific to Annona sp. or belong to untranslated regions. Several of the unigenes representing pathways related to primary and secondary metabolism, and seed and fruit development expressed at a higher level in Sitaphal, the densely seeded cultivar in comparison to the poorly seeded NMK-1. A total of 2629 (Sitaphal) and 3445 (NMK-1) Simple Sequence Repeat (SSR) motifs were identified respectively in the two genotypes. These could be potential candidates for transcript based microsatellite analysis in A. squamosa. CONCLUSION: The present work provides early-stage fruit specific transcriptome sequence resource for A. squamosa. This repository will serve as a useful resource for investigating the molecular mechanisms of fruit development, and improvement of fruit related traits in A. squamosa and related species.

Impacts of Long-Term Irrigation of Domestic Treated Wastewater on Soil Biogeochemistry and Bacterial Community Structure.

Freshwater scarcity and regulations on wastewater disposal have necessitated the reuse of treated wastewater (TWW) for soil irrigation, which has several environmental and economic benefits. However, TWW irrigation can cause nutrient loading to the receiving environments. We assessed bacterial community structure and associated biogeochemical changes in soil plots irrigated with nitrate-rich TWW (referred to as pivots) for periods ranging from 13 to 30 years. Soil cores (0 to 40 cm) were collected in summer and winter from five irrigated pivots and three adjacently located nonirrigated plots. Total bacterial and denitrifier gene abundances were estimated by quantitative PCR (qPCR), and community structure was assessed by 454 massively parallel tag sequencing (MPTS) of small-subunit (SSU) rRNA genes along with terminal restriction fragment length polymorphism (T-RFLP) analysis of nirK, nirS, and nosZ functional genes responsible for denitrification of the TWW-associated nitrate. Soil physicochemical analyses showed that, regardless of the seasons, pH and moisture contents (MC) were higher in the irrigated (IR) pivots than in the nonirrigated (NIR) plots; organic matter (OM) and microbial biomass carbon (MBC) were higher as a function of season but not of irrigation treatment. MPTS analysis showed that TWW loading resulted in the following: (i) an increase in the relative abundance of Proteobacteria, especially Betaproteobacteria and Gammaproteobacteria; (ii) a decrease in the relative abundance of Actinobacteria; (iii) shifts in the communities of acidobacterial groups, along with a shift in the nirK and nirS denitrifier guilds as shown by T-RFLP analysis. Additionally, bacterial biomass estimated by genus/group-specific real-time qPCR analyses revealed that higher numbers of total bacteria, Acidobacteria, Actinobacteria, Alphaproteobacteria, and the nirS denitrifier guilds were present in the IR pivots than in the NIR plots. Identification of the nirK-containing microbiota as a proxy for the denitrifier community indicated that bacteria belonged to alphaproteobacteria from the Rhizobiaceae family within the agroecosystem studied. Multivariate statistical analyses further confirmed some of the above soil physicochemical and bacterial community structure changes as a function of long-term TWW application within this agroecosystem.

Protocol: a 'One health' two year follow-up, mixed methods study on antibiotic resistance, focusing children under 5 and their environment in rural India.

BACKGROUND: Antibiotic resistance has been referred to as 'the greatest malice of the 21st century' and a global action plan was adopted by the World Health Assembly in 2015. There is a wealth of independent studies regarding antibiotics and resistant bacteria in humans, animals and their environment, however, integrated studies are lacking, particularly ones that simultaneously also take into consideration the health related behaviour of participants and healthcare providers. Such, 'One health' studies are difficult to implement, because of the complex teamwork that they entail. This paper describes the protocol of a study that investigates 'One health' issues regarding antibiotic use and antibiotic resistance in children and their environment in Indian villages. METHODS/DESIGN: Both quantitative and qualitative studies are planned for a cohort of children, from 6 villages, and their surrounding environment. Repeated or continues data collection is planned over 2 years for quantitative studies. Qualitative studies will be conducted once. Studies include parents' health seeking behavior for their children (1-3 years of age at the onset), prescribing pattern of formal and informal healthcare providers, analysis of phenotypic antibiotic resistance of Escherichia coli from samples of stool from children and village animals, household drinking water, village source water and waste water, and investigation on molecular mechanisms governing resistance. Analysis of interrelationship of these with each other will also be done as basis for future interventions. Ethics approval has been obtained from the Institutional Ethics Committee R.D. Gardi Medical College, Ujjain, India (No: 2013/07/17-311). DISCUSSION: The findings of the study presented in this protocol will add to our knowledge about the multi-factorial nature of causes governing antibiotic use and antibiotic resistance from a 'One health' perspective. Our study will be the first of its kind addressing antibiotic use and resistance issues related to children in a One-health approach, particularly for rural India.

Feasibility of a Mobile Phone-Based Surveillance for Surgical Site Infections in Rural India.

OBJECTIVES: To assess the feasibility of using mobile communication technology in completing a 30-day follow-up of surgical site infection (SSI). SUBJECTS AND METHODS: SSIs are infections occurring up to 30 days after an operative procedure. This prospective exploratory study was conducted in a cohort of patients who were admitted and operated on in the general surgery wards of a rural hospital in India from October 2010 to June 2011. At the time of discharge, all patients were requested to follow-up in the surgical outpatient clinic at 30 days after surgery. If this was not done, a mobile phone-based surveillance was done to complete the follow-up. RESULTS: The mean age of the 536 operated-on patients was 40 years (95% confidence interval [CI], 38-41 years). The mean duration of hospital stay was 10.7 days (95% CI, 9.9-11.6 days). Most (81%) operated-on patients were from rural areas, and 397 (75%) were male. Among the operated-on patients the ownership of mobile phones was 75% (95% CI, 73-78%). The remaining 25% of patients (n=133) used a shared mobile phone. For 380 patients (74.5%) the follow-up was completed by mobile phones. The SSI rate at follow-up was 6.3% (n=34). In 10 patients, an SSI was detected over the mobile phone. CONCLUSIONS: Mobile communication technology is feasible to be used in rural settings to complete case follow-up for SSIs.

Clinical profile and outcome of patients with acromegaly according to the 2014 consensus guidelines: Impact of a multi-disciplinary team.

AIM: The diagnosis and treatment of acromegaly, a rare and possibly curable disease, has undergone a paradigm shift in the past few decades. Our aim was to study the changing trends in clinical presentation, management and outcome of the disease in the last fifteen years. METHODOLOGY: 271 consecutive patients with acromegaly treated at the Departments of Endocrinology and Neurosurgery, PGIMER, Chandigarh, between 2000 and 2014, were included in the study. Clinical and hormonal profiles, comorbidities, treatment modalities, outcome and mortality data were evaluated. The cure rate was assessed according to the present consensus criteria. RESULTS: The gender distribution was equal with the mean age (±SD) of 37.1 ± 12.3 years at diagnosis. The average lag period to diagnosis was 4.7 ± 4.2 years. The most common presenting manifestations were acral enlargement and headache followed by visual deficits. The overall mortality rate was 5%, with the perioperative mortality being 1.5%. The most prevalent comorbidities in our series were hypertension (17.7%), diabetes mellitus (16.2%), arthropathy (11.8%) and obstructive sleep apnea (10.3%). Overall, 2 patients in our series suffered from extra-pituitary neoplasms and 12 patients had apoplexy as the presenting manifestation. As per the present consensus criteria, cure rate in our series was 28.5%. The cure rate was only 7.9% when many surgeons were operating. It increased to 25.5% when surgeries were being performed by one surgeon exclusively; and, when a sub-specialty clinic exclusively for pituitary diseases was set up, the cure rates improved upto 56%. CONCLUSION: Acromegaly has wide-ranging manifestations from acral enlargement to altered sensorium; incidental diagnosis was not prevalent in our series. Majority of the cases were due to the presence of a pituitary macroadenoma. Better cure rate can be achieved only when a dedicated group of multi-disciplinary team is involved.