Antimicrobial resistance heterogeneity among multidrug-resistant Gram-negative pathogens: Phenotypic, genotypic, and proteomic analysis.

Microbes evolve rapidly by modifying their genomes through mutations or through the horizontal acquisition of mobile genetic elements (MGEs) linked with fitness traits such as antimicrobial resistance (AMR), virulence, and metabolic functions. We conducted a multicentric study in India and collected different clinical samples for decoding the genome sequences of bacterial pathogens associated with sepsis, urinary tract infections, and respiratory infections to understand the functional potency associated with AMR and its dynamics. Genomic analysis identified several acquired AMR genes (ARGs) that have a pathogen-specific signature. We observed that blaCTX-M-15, blaCMY-42, blaNDM-5, and aadA(2) were prevalent in Escherichia coli, and blaTEM-1B, blaOXA-232, blaNDM-1, rmtB, and rmtC were dominant in Klebsiella pneumoniae. In contrast, Pseudomonas aeruginosa and Acinetobacter baumannii harbored blaVEB, blaVIM-2, aph(3'), strA/B, blaOXA-23, aph(3') variants, and amrA, respectively. Regardless of the type of ARG, the MGEs linked with ARGs were also pathogen-specific. The sequence type of these pathogens was identified as high-risk international clones, with only a few lineages being predominant and region-specific. Whole-cell proteome analysis of extensively drug-resistant K. pneumoniae, A. baumannii, E. coli, and P. aeruginosa strains revealed differential abundances of resistance-associated proteins in the presence and absence of different classes of antibiotics. The pathogen-specific resistance signatures and differential abundance of AMR-associated proteins identified in this study should add value to AMR diagnostics and the choice of appropriate drug combinations for successful antimicrobial therapy.

Immediate "Kangaroo Mother Care" and Survival of Infants with Low Birth Weight.

BACKGROUND: "Kangaroo mother care," a type of newborn care involving skin-to-skin contact with the mother or other caregiver, reduces mortality in infants with low birth weight (<2.0 kg) when initiated after stabilization, but the majority of deaths occur before stabilization. The safety and efficacy of kangaroo mother care initiated soon after birth among infants with low birth weight are uncertain. METHODS: We conducted a randomized, controlled trial in five hospitals in Ghana, India, Malawi, Nigeria, and Tanzania involving infants with a birth weight between 1.0 and 1.799 kg who were assigned to receive immediate kangaroo mother care (intervention) or conventional care in an incubator or a radiant warmer until their condition stabilized and kangaroo mother care thereafter (control). The primary outcomes were death in the neonatal period (the first 28 days of life) and in the first 72 hours of life. RESULTS: A total of 3211 infants and their mothers were randomly assigned to the intervention group (1609 infants with their mothers) or the control group (1602 infants with their mothers). The median daily duration of skin-to-skin contact in the neonatal intensive care unit was 16.9 hours (interquartile range, 13.0 to 19.7) in the intervention group and 1.5 hours (interquartile range, 0.3 to 3.3) in the control group. Neonatal death occurred in the first 28 days in 191 infants in the intervention group (12.0%) and in 249 infants in the control group (15.7%) (relative risk of death, 0.75; 95% confidence interval [CI], 0.64 to 0.89; P = 0.001); neonatal death in the first 72 hours of life occurred in 74 infants in the intervention group (4.6%) and in 92 infants in the control group (5.8%) (relative risk of death, 0.77; 95% CI, 0.58 to 1.04; P = 0.09). The trial was stopped early on the recommendation of the data and safety monitoring board owing to the finding of reduced mortality among infants receiving immediate kangaroo mother care. CONCLUSIONS: Among infants with a birth weight between 1.0 and 1.799 kg, those who received immediate kangaroo mother care had lower mortality at 28 days than those who received conventional care with kangaroo mother care initiated after stabilization; the between-group difference favoring immediate kangaroo mother care at 72 hours was not significant. (Funded by the Bill and Melinda Gates Foundation; Australian New Zealand Clinical Trials Registry number, ACTRN12618001880235; Clinical Trials Registry-India number, CTRI/2018/08/015369.).

Effect of ivermectin, amitraz and fipronil on midgut epithelium and digestive enzyme profile in Rhipicephalus microplus ticks (Acari: Ixodidae).

Blood feeding and digestion are vital physiological activities essential for the survival and reproduction of ticks. Chemical acaricides viz., ivermectin, amitraz and fipronil, are known to act on the central nervous system, resulting in the mortality of ticks. The present study is focused on the effect of these acaricides on the midgut and gut enzymes of Rhipicephalus microplus. The ultra-thin sections of midgut of ivermectin-treated ticks showed irregular basal membrane and ruptured digestive vesicles. Amitraz treatment resulted in a notable decrease in digestive cells with pleats in the basal membrane, while fipronil-exposed ticks exhibited reduced digestive cells, loss of cellular integrity, and disintegration of the basal membrane and muscle layer. The gut tissue homogenate of ivermectin and fipronil treated ticks showed a significant reduction of cathepsin D level, 76.54 ± 3.20 µg/mL and 92.67 ± 3.72 µg/mL, respectively, as compared to the control group (150.0 ± 3.80 µg/mL). The leucine aminopeptidase level (4.27 ± 0.08 units/mL) was significantly decreased in the ivermectin treated ticks compared to other treatment groups. The acid phosphatase activity (29.16 ± 0.67 µmole/min/L) was reduced in the ivermectin treated group whereas, increased activity was observed in the fipronil and amitraz treated groups. All the treatment groups revealed increased alkaline phosphatase levels (17.47-26.72 µmole/min/L). The present finding suggests that in addition to the established mechanism of action of the tested acaricides on the nervous system, the alterations in the cellular profile of digestive cells and enzymes possibly affect the blood digestion process and thus the synthesis of vital proteins which are essential for vitellogenesis, and egg production in ticks.

Liposomal formulation of model pentathiepin improves solubility and stability toward glutathione while preserving anticancer activity.

The biological properties of pentathiepins have been attracting increased attention in recent years. Experiments have shown a wide range of effects of pentathiepins in vitro, such as induction of apoptosis and alteration of mitochondrial membrane potential in cancer cells, and inhibition of antioxidant enzymes, for example, glutathione peroxidase 1 (GPx1). Biological evaluation is sometimes limited due to low aqueous solubility, high lipophilicity, and poor stability toward thiols, for example, glutathione (GSH). To assess whether liposomes are suitable as drug carriers to overcome these drawbacks, a model pentathiepin was formulated in a liposomal preparation. The success of loading liposomes with pentathiepins was evaluated by using ultraviolet-visible light (UV-Vis) spectroscopy, dynamic light scattering (DLS), and high-performance liquid chromatography (HPLC). Through inclusion into 100-nm-sized 1,2-dioleoyl-sn-glycero-3-phosphocholine liposomes, the aqueous solubility of a representative pentathiepin could be increased by several orders of magnitude to ca. 400 µM. The stability of the pentathiepin in the presence of GSH was increased fourfold as determined by UV-Vis spectroscopy. In antiproliferation experiments with two human cancer cell lines, no decrease in potency in the liposomal loaded pentathiepin compared to the free pentathiepin was found. In conclusion, liposomes are a suitable carrier for pentathiepins and improve both solubility and stability in the presence of thiols without compromising anticancer activity.

First report of molecular confirmation and phylogenetic analysis of ocular seteriasis in buffalo in India using 12S rRNA.

An adult Indian buffalo (Bubalus bubalis) presented with corneal opacity, irritation, and excessive lacrimation from the left eye in the Referral Veterinary Polyclinic-Teaching Veterinary Clinical Complex (RVC-TVCC), Indian Veterinary Research Institute, Izatnagar. Clinical examination revealed a whitish thread-like worm in the left eye's anterior chamber. The worm was surgically removed from the eye with supportive nerve blocks. Light microscopy was used for parasite morphological identification, which provided insight into the worm as female Setaria sp. Genomic DNA was isolated, and polymerase chain reaction amplification of 12S rRNA was conducted for molecular confirmation of the parasite. The amplicon was sequenced and analysed by bioinformatics software. Sequence data showed an amplicon size of 243 bp. Phylogenetic analysis with reference data from the NCBI Genbank database revealed the worm was S. digitata, with a similarity of 99.17%. The common predilection site of S. digitata is in the peritoneal cavity of natural hosts like cattle and buffalo and is mostly non-pathogenic. The aberrant migration of the parasite larva to the brain and eye commonly occurs in goats, sheep, and horses, causing clinical conditions like cerebrospinal nematodiasis (lumbar paralysis) and ocular setariasis, respectively. Nevertheless, until now, there have been no reports of ocular setariasis in buffalo. This report is the first unusual occurrence of ocular setariasis in buffalo and its molecular confirmation and phylogenetic analysis using 12S rRNA.

Economic impact of predominant ticks and tick-borne diseases on Indian dairy production systems.

As per 20th livestock census, India possessing 193.46 million cattle and 109.85 million buffaloes in organized and unorganized sectors. These animals are suffering from tick infestation almost throughout the year causing both direct and indirect losses. Repeated use of chemical acaricides for tick management resulted in the establishment of acaricide resistant tick populations, insecticide residues in livestock products and environmental pollution. In the present study, analysis of the data generated during 1990-2021 revealed pooled prevalence of infestations in cattle and buffaloes was 53% [95% CI = 47-60%] and 38% [95% CI = 28-49%], respectively. Based on peripheral blood smear examination the prevalence of theileriosis, babesiosis and anaplasmosis in cattle is estimated as 17% [95% CI = 12-24%], 4% [95% CI = 3-6%] and 5% [95% CI = 3-7%], respectively, while in buffalo the prevalence of infection is estimated as 7% [95% CI = 2-21%], 2% [95% CI = 0-5%] and 8% [95% CI = 2-36%],respectively.To estimate economic impact, both direct (reduction in milk production, cost of treatment, leather depreciation) and indirect (milk loss and treatment cost) losses were taken into consideration. Loss of milk production was predicted as 13.91, 56.91 and 85.34L/cross-bred cow/lactation in low, moderate and high tick infestation conditions, respectively. Whereas, 20.10, 7.01L milk/buffalo/lactation in Hyalomma spp. and Rhipicephalus spp. infestation was estimated. Similarly,the estimated loss of milk production due to clinical theileriosis, babesiosis and anaplasmosis was 57.96, 30.96 and 59.22L, respectively. The cumulative (milk loss, treatment cost and leather loss) loss due to tick infestation was calculated as 46199.31 million INR (USD595.07 million) while due to TBDs 14877.15 million INR (USD191.15 million) = 61076.46 million INR (USD787.63 million). The data provided base line information for the policy maker to develop strategies at government level so that the significantly high cumulative loss of 787.63 million USD due to ticks and tick borne diseases (TTBDs) can be minimized.

Seasonal changes of trace elements, nutrients, dissolved organic matter, and coastal acidification over the largest oyster reef in the Western Mississippi Sound, USA.

Seasonal changes of trace elements, nutrients, dissolved organic matter (DOM), and carbonate system parameters were evaluated over the largest deteriorating oyster reef in the Western Mississippi Sound using data collected during spring, summer, and winter of 2018, and summer of 2019. Higher concentrations of Pb (224%), Cu (211%), Zn (2400%), and Ca (240%) were observed during winter of 2018 compared to summer 2019. Phosphate and ammonia concentrations were higher (> 800%) during both summers of 2018 and 2019 than winter of 2018. Among the three distinct DOM components identified, two terrestrial humic-like components were more abundant during both spring (12% and 36%) and summer (11% and 33%) of 2018 than winter of 2018, implying a relatively lesser supply of humic-like components from terrestrial sources during winter. On the other hand, the protein-like component was more abundant during summer of 2019 compared to rest of the study period, suggesting a higher rate of autochthonous production during summer 2019. In addition, to their significant depth-wise variation, ocean acidification parameters including pH, pCO2, CO32-, and carbonate saturation states were all higher during both summers of 2018 and 2019. The measured variables such as trace elements, organic carbon, suspended particulates, and acidification parameters exhibited conservative mixing behavior against salinity. These observations have strong implications for the health of the oyster reefs, which provides ecologically important habitats and supports the economy of the Gulf Coast.

Delayed Administration of Antibiotics Beyond the First Hour of Recognition Is Associated with Increased Mortality Rates in Children with Sepsis/Severe Sepsis and Septic Shock.

OBJECTIVE: To compare the risk of mortality and other clinical outcomes in children with sepsis, severe sepsis, or septic shock who received antibiotics within the first hour of recognition (early antibiotics group) with those who received antibiotics after the first hour (delayed antibiotics group). STUDY DESIGN: In this prospective cohort study, we enrolled children <17 years of age presenting to the pediatric emergency and diagnosed with sepsis or septic shock without prior antibiotic therapy. Primary outcome was mortality and the secondary outcomes were day 1 Pediatric Logistic Organ Dysfunction score, ventilator-free days, and hospital-free days. These outcomes were compared between the early and the delayed antibiotic groups. The reference point for defining early and delayed antibiotic groups was time 0, which was measured from the time the patient was diagnosed to have sepsis, severe sepsis, or septic shock to the time of administration of the first dose of antibiotics. RESULTS: About three-fourths (77%) of the 441 children enrolled had septic shock. A total of 241 (55%) and 200 (45%) children were in the delayed and early antibiotic groups, respectively. Children in the delayed group had significantly higher odds of mortality than those in the early group (29% vs 20%; aOR 1.83; 95% CI, 1.14-2.92; P = .01). The time to shock reversal was significantly shorter, and the ventilator-free days and hospital-free days were significantly greater, in the early antibiotic group. There was no difference between the groups with regard to any of the other clinical outcomes. CONCLUSIONS: Delayed administration of antibiotics beyond 1 hour of recognition was associated with higher mortality rates in children with sepsis, severe sepsis, and septic shock. Antibiotics should be administered within the first hour, along with other resuscitative measures, in these children.

Functional Outcomes at 1 Year After PICU Discharge in Critically Ill Children With Severe Sepsis.

OBJECTIVES: To evaluate the functional outcomes in critically ill children with severe sepsis using the Pediatric Overall Performance Category scale and Pediatric Cerebral Performance Category scale and to evaluate the risk factors for "worse outcomes." DESIGN: Prospective observational cohort study. SETTING: Tertiary care PICU from September 2017 to October 2019. PATIENTS: One hundred twenty-one children with severe sepsis, 2 months to 17 years old, admitted to PICU. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Pediatric Overall Performance Category and Pediatric Cerebral Performance Category scores were recorded at admission, PICU discharge, at 3 months and 1 year after discharge. "New disability" was defined as Pediatric Overall Performance Category and Pediatric Cerebral Performance Category score change "from baseline score" by greater than or equal to 1 category. Risk factors for "worse outcomes" (defined as "death" or "new disability") were evaluated by univariate and multivariate analysis. At admission, 33% (n = 39) had mild to moderate "overall disability" (Pediatric Overall Performance Category) and 26% (n = 32) had mild to moderate "cognitive disability" (Pediatric Cerebral Performance Category). At PICU discharge (n = 89 children), 50.5% (n = 45) had "new disability" in overall function (Pediatric Overall Performance Category scores) and 28% (n = 25) had "new disability" in cognitive function (Pediatric Cerebral Performance Category scores). At 3 months follow-up (n = 85 children), "new disability" at PICU discharge improved in 65% (n = 28/43) and 50% (n = 12/24) of those with "overall disability" (Pediatric Overall Performance Category) and "cognitive disability" (Pediatric Cerebral Performance Category), respectively. At 1-year follow-up (n = 84 children), only 5% (n = 2/43) had residual "new disability" in overall function (Pediatric Overall Performance Category) and 14% (n = 3/21) had residual "new disability" in cognitive function. PICU mortality was 26% (n = 32). The proportion with "worse outcomes" was 64% (n = 77). Risk factors for worse outcomes on univariate analysis included higher Pediatric Index of Mortality-3 scores, day 1 pediatric Sequential Organ Failure Assessment score, receiving cardiopulmonary resuscitation during the ICU stay, and treatments received such as ventilation and Vasoactive-Inotrope Score. On multivariate analysis, only day 1 pediatric Sequential Organ Failure Assessment score and receiving cardiopulmonary resuscitation during the ICU stay were found to be statistically significant. CONCLUSIONS: Children with severe sepsis had significant "new onset" mild to moderate functional disability at PICU discharge, and most of these children recovered within 1 year after PICU discharge. Day 1 pediatric Sequential Organ Failure Assessment score and patient receiving cardiopulmonary resuscitation during the ICU stay were found to be the significant risk factors of "worse outcomes."

Molecular characterization, computational analysis and expression profiling of Dmrt1 gene in Indian major carp, Labeo rohita (Hamilton 1822).

Sexual dimorphism of fish morphology, physiology and behavior is diverse and complex in nature. Doublesex and mab-3 related transcription factor (Dmrt) is a large protein family whose function is sexual development and differentiation in vertebrates. Here, we report a full-length cDNA sequence of Labeo rohita (rohu) Dmrt1 of 907 bp length having 798 bp of open reading frame encoding 265 amino acids. The molecular weight of rohu DMRT1 protein was found to be 28.74 KDa and isoelectric point was 7.53. DMRT1 protein contains 23 positively and 24 negatively charged amino acids with a GRAVY score of -0.618. A characteristic DM domain was found in DMRT1 protein, which is a novel DNA-binding domain. Phylogenetic analysis showed maximum similarity with Cyprinus carpio when compared with DMRT1 of other vertebrates. Molecular docking study identified active sites to be targeted for drug designing. Rohu DMRT1 was observed to interact with other proteins such as FOXL2, CYP19a1a, AMH and SOX9a. Differential expression study revealed higher expression in testis tissue implying its role in male sex differentiation and testicular development. The information generated in the present work could facilitate further research to resolve the issues related to gonadal maturation and reproduction of commercially important aquaculture species.

Low dose fentanyl infusion versus 24% oral sucrose for pain management during laser treatment for retinopathy of prematurity-an open label randomized clinical trial.

To compare the efficacy of low dose fentanyl infusion and 24% oral sucrose in providing optimal pain relief during laser for retinopathy of prematurity (ROP), we enrolled fifty-eight spontaneously breathing preterm infants undergoing laser. The preterm infants were randomized to either fentanyl infusion (1 mcg/kg/hr) or 24% oral sucrose (2 ml). We evaluated and compared the proportion of time spent crying during the procedure, salivary cortisol before and after the procedure, premature infant pain profile- revised (PIPP-R) scores during the procedure, apnoea during and after the procedure, need for mechanical ventilation, and feed intolerance and urinary retention 24 h after the procedure between the two groups. We found that the proportion of time spent crying during the procedure was significantly less in the fentanyl group [62.5% (50.7-74.2) vs 73.8% (55.6-83.4); P = 0.02]. Average PIPP-R score during the procedure was significantly less in the fentanyl group [7.2 vs 9.0; (mean difference of - 1.8; P = 0.01)]. There was no difference in other outcomes between the two groups.Conclusion: During laser for ROP, low dose fentanyl infusion was found to be efficacious in reducing pain as compared with 24% sucrose.What is Known:• Preterm infants undergoing laser photocoagulation for ROP suffer significant amount of pain.• Standard of care for pain relief in infants undergoing laser therapy in developed countries is general anesthesia (GA) or combination of sedation, analgesia, and paralysis (SAP).What is New:• During laser photocoagulation for ROP, fentanyl infusion at low dose (1 mcg/kg/hr) is efficacious in reducing pain as compared to 24% oral sucrose.

Delayed cord clamping in Rh-alloimmunised infants: a randomised controlled trial.

Despite advancement in medical care, Rh alloimmunisation remains a major cause of neonatal hyperbilirubinaemia, neuro-morbidity, and late-onset anaemia. Delayed cord clamping (DCC), a standard care now-a-days, is yet not performed in Rh-alloimmunised infants due to paucity of evidence. Hence, we randomised these infants of 28- to 41-week gestation to delayed cord clamping (N = 36) or early cord clamping (N = 34) groups. The primary outcome variable was venous packed cell volume (PCV) at 2 h of birth. The secondary outcomes were incidence of double volume exchange transfusion (DVET) and partial exchange transfusion (PET), duration of phototherapy (PT), functional echocardiography (parameters measured: superior vena cava flow, M-mode fractional shortening, left ventricular output, myocardial perfusion index, and inferior vena cava collapsibility) during hospital stay, and blood transfusion (BT) until 14 weeks of life. Neonates were managed as per unit protocol. The baseline characteristics of enrolled infants were comparable between the groups. The median (IQR) gestation and mean (SD) birth weight of enrolled infants were 35 (33-37) weeks and 2440 (542) g, respectively. The DCC group had a higher mean PCV at 2 h of life (48.4 ± 9.2 vs. 43.5 ± 8.7, mean difference 4.9% (95% CI 0.6-9.1), p = 0.03). However, incidence of DVET and PET, duration of PT, echocardiography parameters, and BT until 14 weeks of postnatal age were similar between the groups.Conclusion: DCC in Rh-alloimmunised infants improved PCV at 2 h of age without significant adverse effects.Trial registration: Clinical Trial Registry of India (CTRI), Ref/2016/11/012572 http://ctri.nic.in/Clinicaltrials, date of trial registration 19.12.2016, date of first patient enrolment 1 January 2017.What is Known:•Delayed cord clamping improves haematocrit, results in better haemodynamic stability, and decreases the need of transfusion in early infancy.•However, due to lack of evidence, potential risk of hyperbilirubinaemia, and exacerbation of anaemia (following delayed cord clamping), early cord clamping is the usual norm in Rh-alloimmunised infantsinfants.What is New:•Delayed cord clamping in Rh-alloimmunised infants improves haematocrit at 2 h of life without any increase in incidence of serious adverse effects.

Concordance of glycaemic and cardiometabolic traits between Indian women with history of gestational diabetes mellitus and their spouses: an opportunity to target the household.

AIMS/HYPOTHESIS: The aim of this study was to investigate the concordance of dysglycaemia (prediabetes or diabetes) and cardiometabolic traits between women with a history of gestational diabetes mellitus (GDM) and their spouses. METHODS: Using hospital medical records, women with GDM (diagnosed between 2012 and 2016) and their spouses were invited to participate in the study and to attend a scheduled hospital visit in a fasting state. Sociodemographic, anthropometric and medical data were collected, and a 75 g OGTT with serum insulin estimation, HbA1c measurement and fasting lipid profile were performed at the visit. Prediabetes and diabetes were defined using ADA criteria and the metabolic syndrome was defined using IDF criteria. RESULTS: A total of 214 couples participated in the study. Women were tested at a mean ± SD age of 32.4 ± 4.6 years and median (quartile [q]25-q75) of 19.5 (11-44) months following the index delivery, while men were tested at a mean ± SD age of 36.4 ± 5.4 years. A total of 72 (33.6%) couples showed concordance for dysglycaemia, while 99 (46.3%) and 51 (23.8%) couples were concordant for overweight/obesity and the metabolic syndrome, respectively. A total of 146 (68.2%) couples showed concordance for any of the above three factors. The presence of dysglycaemia in one partner was associated with an increased risk of dysglycaemia in the other partner (OR 1.80 [95% CI 1.04, 3.11]). Similarly, being overweight/obese (OR 2.19 [95% CI 1.22, 3.93]) and presence of the metabolic syndrome (OR 2.01 [95% CI 1.16, 3.50]) in one partner was associated with an increased risk of these conditions in the other partner. Both women and men were more likely to have dysglycaemia if they had a partner with dysglycaemia. Women with a partner with dysglycaemia had a significantly higher BMI, waist circumference and diastolic BP, and a significantly higher probability of low HDL-cholesterol (<1.29 mmol/l) and the metabolic syndrome compared with women with a normoglycaemic partner. No such differences were observed for men with or without a partner with dysglycaemia. CONCLUSIONS/INTERPRETATION: The high degree of spousal concordance found in this study suggests social clustering of glycaemic and cardiometabolic traits among biologically unrelated individuals. This provides us with an opportunity to target the behavioural interventions at the level of the 'married couple', which may be a novel and cost-effective method of combating the current diabetes epidemic.

Prokaryotic expression, purification and evaluation of goatpox virus ORF117 protein as a diagnostic antigen in indirect ELISA to detect goatpox.

Goatpox is an economically significant transboundary viral disease of goats that is caused by goatpox virus (GTPV). This study describes the prokaryotic expression of the GTPV ORF117 protein, a homologue of vaccinia virus A27L, and evaluation of its diagnostic potential in ELISA. The GTPV ORF117 gene was cloned into the pET32a vector to express recombinant ORF117 protein (rA27L) in E. coli BL21-CodonPlus (DE3)-RIPL. The bacterial expression of the protein was confirmed by western blot analysis using anti-GTPV polyclonal antibodies that detected rA27L, which is ~ 35 kDa in size. rA27L was affinity purified under native conditions and used to assess the antibody response in an optimized indirect ELISA. The purified antigen specifically reacted with anti-GTPV and anti-SPPV serum in ELISA. A preliminary screening of random and purposive serum samples (n = 520) from sheep and goats using this optimized ELISA gave a positivity rate of 19.4 % with a diagnostic specificity of 88.7% and diagnostic sensitivity of 98.5% when compared to the gold standard serum neutralization test. Our results suggest that the indirect ELISA based on the rA27L protein has potential for serosurveillance and seromonitoring of GTPV in goats.

Effect of Severe Vitamin D Deficiency at Admission on Shock Reversal in Children With Septic Shock: A Prospective Observational Study.

OBJECTIVES:: To evaluate the association of severe vitamin D deficiency with clinically important outcomes in children with septic shock. METHODS:: We enrolled children ≤17 years with septic shock prospectively over a period of 6 months. We estimated 25-hydroxyvitamin D [25 (OH) D] levels at admission and 72 hours. Severe deficiency was defined as serum 25 (OH) <10 ng/mL. We performed univariate and multivariate analysis to evaluate association with clinically important outcomes. RESULTS:: Forty-three children were enrolled in the study. The prevalence of severe vitamin D deficiency was 72% and 69% at admission and 72 hours, respectively. On univariate analysis, severe vitamin D deficiency at admission was associated with lower rates of shock reversal, 74% (23) versus 25% (3); relative risk (95% confidence interval [CI]): 2.9 (1.09-8.08), at 24 hours and greater need for fluid boluses (75 vs 59 mL/kg). On multivariate analysis, nonresolution of shock at 24 hours was significantly associated with severe vitamin D deficiency after adjusting for other key baseline and clinical variables, adjusted odds ratio (95% CI): 12 (2.01-87.01); 0.01. CONCLUSION:: The prevalence of severe vitamin D deficiency is high in children with septic shock admitted to pediatric intensive care unit. Severe vitamin D deficiency at admission seems to be associated with lower rates of shock reversal at 24 hours of ICU stay. Our study provides preliminary data for planning interventional studies in children with septic shock and severe vitamin D deficiency.

Comparison of International Pediatric Sepsis Consensus Conference Versus Sepsis-3 Definitions for Children Presenting With Septic Shock to a Tertiary Care Center in India: A Retrospective Study.

OBJECTIVES: To evaluate the proportion of children fulfilling "Sepsis-3" definition and International Pediatric Sepsis Consensus Conference definition among children diagnosed to have septic shock and compare the mortality risk between the two groups. DESIGN: Retrospective chart review. SETTING: PICU of a tertiary care teaching hospital from 2014 to 2017. PATIENTS: Children (≤ 17 yr old) with a diagnosis of septic shock at admission or during PICU stay. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We applied both International Pediatric Sepsis Consensus Conference and the new "Sepsis-3" definition (sepsis with hypotension requiring vasopressors and a lactate value of ≥ 2 mmol/L) to identify cases of septic shock by these definitions. Key outcomes such as mortality, proportion attaining shock reversal at 24 hours and organ dysfunction were compared between those fulfilling "Sepsis-3" definitions ("Sepsis-3" group) and those fulfilling "International Pediatric Sepsis Consensus Conference" definition ("International Pediatric Sepsis Consensus Conference" group). A total of 216 patients fulfilled International Pediatric Sepsis Consensus Conference definitions of septic shock. Of these, only 104 (48%; 95% CI, 42-55) fulfilled "Sepsis-3" definition. Children fulfilling "Sepsis-3 plus International Pediatric Sepsis Consensus Conference definitions" ("Sepsis-3 and International Pediatric Sepsis Consensus Conference" group) had lower proportion with shock resolution (61% vs 82%; relative risk, 0.73; 95% CI, 0.62-0.88) and higher risk of multiple organ dysfunction (85% vs 68%; 1.24; 1.07-1.45) at 24 hours. The mortality was 48.5% in "Sepsis-3 and International Pediatric Sepsis Consensus Conference" group as compared with 37.5% in the "International Pediatric Sepsis Consensus Conference only" group (relative risk, 1.3; 95% CI, 0.94-1.75). CONCLUSIONS: Less than half of children with septic shock identified by International Pediatric Sepsis Consensus Conference definitions were observed to fulfill the criteria for shock as per "Sepsis-3" definitions. Lack of difference in the risk of mortality between children who fulfilled "Sepsis-3" definition and those who did not fulfill the definition raises questions on the appropriateness of using this definition for diagnosis of septic shock in children.

Effect of a Training Strategy in Improving Medication Fallacies During Pediatric Cardiopulmonary Resuscitation: A Before-and-After Study From a Developing Country.

OBJECTIVE: This study aims to evaluate the effect of structured training on resident performance in improving medication fallacies during pediatric cardiopulmonary resuscitation (CPR). METHODS: This before-and-after study was conducted in the pediatric acute care areas of tertiary care teaching hospitals of a developing country from August to December 2015. Case records of children younger than 18 years who underwent CPR were reviewed. Senior residents rotating through pediatric emergency department and pediatric intensive care unit were evaluated for their knowledge. Incidence of medication fallacies in pediatric CPR and change in the knowledge scores of residents posted in these areas were the main outcome measures. RESULTS: One-hundred records were evaluated (pre-intervention, 54; post-intervention, 46). In the pre-intervention period, 25 had medication fallacies (documentation, 16; dosing, 9). In the post-intervention period, 7 fallacies pertaining to documentation (not dosing) were found. The incidence of severe fallacies decreased from 20% pretraining to 0% posttraining. The mean (SD) knowledge scores of residents increased from 7.9 (2.9) pretraining to 13 (1.4) posttraining. On univariate analysis, fallacies were found to be less if the resident was formally trained (pediatric advanced life support certified), if the patient was older, and during morning and night shifts as compared with evening shift. On multivariate analysis, however, only status of training (posttraining) (adjusted odds ratio, 0.12; 95% confidence interval, 0.02-0.68) and the morning shift (adjusted odds ratio, 0.03; 95% confidence interval, 0.001-0.72) remained significant with lower incidence of fallacies associated with these variables. CONCLUSIONS: Rates of medication fallacies in pediatric CPR declined with structured training. Documentation fallacies may not be eliminated completely with only 1-time training.

Electrocoagulation using commercial grade aluminium electrode for the removal of crystal violet from aqueous solution.

The current research work studies the removal of crystal violet (also known as gentian violet), a kind of dye, from simulated wastewater by electrocoagulation using scrap aluminium roofing sheet as electrode in a batch electrochemical cell. Optimization of different operational parameters - pH, current density, time, initial concentration - was carried out experimentally. The equilibrium was attained at 1 hour and at pH 11. After suitable optimization, a removal of above 90% was achieved at an optimum current density of 20 mA/m2. Also, the adsorption behaviour of crystal violet in electrocoagulation was also studied and the isothermal and kinetic models were proposed to be the Dubinin-Radushkevich model and pseudo-first order model. The mechanism involved during the process was suggested as chemisorption. The adsorption thermodynamic studies were a clear indication that the process is spontaneous and endothermic as well as thermodynamically favourable too. Both chemical and physical characterization of the flocs generated during the electrolysis was explained by Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) respectively.

Effect of photo-biodegradation and biodegradation on the biogeochemical cycling of dissolved organic matter across diverse surface water bodies.

The objective of this research was to quantify the temporal variation of dissolved organic matter (DOM) in five distinct waterbodies in watersheds with diverse types of land use and land cover in the presence and absence of sunlight. The water bodies were an agricultural pond, a lake in a forested watershed, a man-made reservoir, an estuary, and a bay. Two sets of samples were prepared by dispensing unfiltered samples into filtered samples in 1:10 ratio (V/V). The first set was exposed to sunlight (10 hr per day for 30 days) for examining the combined effect of photo-biodegradation, while the second set was stored in dark for examining biodegradation alone. Spectroscopic measurements in tandem with multivariate statistics were used to interpret DOM lability and composition. The results suggest that the agricultural pond behaved differently compared to other study locations during degradation experiments due to the presence of higher amount of microbial humic-like and protein-like components derived from microbial/anthropogenic sources. For all samples, a larger decrease in dissolved organic carbon (DOC) concentration (10.12% ±â€¯9.81% for photo-biodegradation and 6.65% ±â€¯2.83% for biodegradation) and rapid transformation of DOM components (i.e., terrestrial humic-like components into microbial humic and protein-like components) were observed during photo-biodegradation experiments. Results suggest that sunlight facilitated DOM biodegradation, resulting in simpler recalcitrant molecules regardless of original composition. Overall, it was found that combined effects of light and bacteria are more efficient than bacterial effects alone in remineralizing and altering DOM, which highlights the crucial importance of sunlight in transforming aquatic DOM.

Prevalence and Outcome of Sepsis-induced Myocardial Dysfunction in Children with 'Sepsis' 'With' and 'Without Shock'-A Prospective Observational Study.

OBJECTIVE: To estimate the prevalence and effects of sepsis-induced myocardial dysfunction (SIMD) in children with septic shock. METHODOLOGY: Enrolled children with septic shock (n = 31) and sepsis (n = 30) underwent echocardiography and cardiac troponin-I (cTnI) estimation within first 3 h. SIMD was defined as presence of systolic/diastolic dysfunction by echocardiography. RESULTS: The prevalence of SIMD was 71% in 'septic shock' and 23% in 'sepsis'. Diastolic dysfunction (45.2%) was more prevalent than systolic dysfunction (32.3%). Children with SIMD had higher requirement of inotropes [81 vs. 44%; adjusted odds ratio: 1.41 (1.04-1.92)] in first 48 h. cTnI had low sensitivity (62.5%) and specificity (55.1%) for detecting SIMD. On follow-up at 3 months, there was no residual dysfunction in the majority (71.3%). CONCLUSION: SIMD, especially diastolic dysfunction, is common in septic shock and may increase inotrope requirement. It is reversible in majority. Sepsis patients may have asymptomatic underlying SIMD. cTnI does not correlate with the degree of SIMD.