Complete Genome Sequencing Reveals Unusual Equine Rotavirus A of Bat Origin from India.

Rotaviruses are the most common viral agents associated with foal diarrhea. Between 2014 and 2017, the annual prevalence of rotavirus in diarrheic foals ranged between 18 and 28% in Haryana (India). Whole-genome sequencing of two equine rotavirus A (ERVA) isolates (RVA/Horse-wt/IND/ERV4/2017 and RVA/Horse-wt/IND/ERV6/2017) was carried out to determine the genotypic constellations (GCs) of ERVAs. The GCs of both the isolates were G3-P[3]-I8-R3-C3-M3-A9-N3-T3-E3-H6, a unique combination reported for ERVAs so far. Both the isolates carried VP6 of genotype I8, previously unreported from equines. Upon comparison with RVAs of other species, the GC of both isolates was identical to that of a bat rotavirus strain, MSLH14, isolated from China in 2012. The nucleotide sequences of the genes encoding VP3, NSP2, and NSP3 shared >95.81% identity with bat RVA strains isolated from Africa (Gabon). The genes encoding VP1, VP2, VP7, NSP1, and NSP4 shared 94.82% to 97.12% nucleotide identities with the human strains which have zoonotic links to bats (RCH272 and MS2015-1-0001). The VP6 genes of both strains were distinct and had the highest similarity of only 87.08% with that of CMH222, a human strain of bat origin. The phylogenetic analysis and lineage studies revealed that VP7 of both isolates clustered in a new lineage (lineage X) of the G3 genotype with bat, human, and alpaca strains. Similarly, VP4 clustered in a distinct P[3] lineage. These unusual findings highlight the terra incognita of the genomic diversity of equine rotaviruses and support the need for the surveillance of RVAs in animals and humans with a "one health" approach. IMPORTANCE Rotaviruses are globally prevalent diarrheal pathogens in young animals including foals, piglets, calves, goats, sheep, cats, and dogs along with humans. The genome of rotaviruses consists of 11 segments, which enables them to undergo reshuffling by reassortment of segments from multiple species during mixed infections. In this study, the prevalence of equine rotaviruses was 32.11% in organized equine farms of North India. The complete genome analysis of two ERVA isolates revealed an unusual genomic constellation, which was previously reported only in a bat RVA strain. A segment-wise phylogenetic analysis revealed that most segments of both isolates were highly similar either to bat or to bat-like human rotaviruses. The occurrence of unusual bat-like rotaviruses in equines emphasizes the need of extensive surveillance of complete genomes of both animal and human rotaviruses with a "one health" approach.

Sorption behavior studies of Cs and its migration in soil samples around Visakhapatnam, India.

Meeting the requirement of high specific activity of radioisotopes and carrying out comprehensive research and development activities in the nuclear field, different nuclear facilities, including their waste disposal facilities, are going to be operational at Visakhapatnam, India. Due to environmental processes, the engineered disposal modules may lose their structural integrity and may release some radioactivity to the geo-environment. The subsequent migration of radionuclides reaching the geological environment will be governed by the distribution coefficient (Kd). Cs was chosen for the sorption study in two soil samples (soil-29 and 31) and to estimate the Kd in all the 40 soil samples through the laboratory batch method at the new campus of DAE, Visakhapatnam, India. Different soil chemical parameters like pH, organic matter, CaCO3, and cation exchange capacity were determined in 40 soil samples and their effect on Cs sorption was investigated. The effect of solution pH and initial concentration of Cs on sorption was also studied. The results show that the sorption of Cs increases with increasing pH. The Cs sorption was well explained by Freundlich and Dubinin-Radushkevich (D-R) isotherm models. Site-specific distribution coefficients (Kd) were also estimated and the values were found to vary from 75 ± 1 to 540 ± 12 L kg-1. The observed wide variation in Kd could be due to large variations in the physico-chemical properties of collected soil. The competitive ions effect study suggests that K+ has higher interference for Cs+ sorption as compared to Na+. The present study results will help assess the environmental impact due to Cs release in any unforeseen scenario and in planning effective remediation strategies.

Diagnosis of osteoarticular tuberculosis by immuno-PCR assay based on mycobacterial antigen 85 complex detection.

Diagnosis of osteoarticular tuberculosis (OATB) exhibits serious challenges owing to paucibacillary nature of specimens and localization of disease at sites that are difficult to access. We recently developed indirect immuno-PCR (I-PCR) and real-time I-PCR (RT-I-PCR) assays for the detection of mycobacterial antigen 85 complex (Ag85) in OATB patients. Detection limits for the purified Ag85 protein were found to be 1 and 41 fg ml-1 by I-PCR and RT-I-PCR, respectively, which were at least 105 -fold lower than respective ELISA. While spiking synovial fluids of non-TB control subjects with the purified Ag85 protein, LODs of 100 and 120 fg ml-1 were obtained by I-PCR and RT-I-PCR, respectively, thus demonstrating the sample matrix effect. Sensitivities of 87·5 and 70·5% were observed in bodily fluids of confirmed (n = 8) and clinically suspected (n = 51) OATB cases, respectively, by I-PCR, with a specificity of 93·9% (n = 33). Markedly, the sensitivities obtained by I-PCR/RT-I-PCR were significantly higher (P < 0·05-0·01) than ELISA and GeneXpert assay (n = 30). However, no substantial difference in sensitivity was observed between the I-PCR and RT-I-PCR assays. After further improving the accuracy of I-PCR, this test may lead to development of an attractive diagnostic kit.

Influence of food availability on mate-guarding behaviour of ladybirds.

A recent study on ladybird, Menochilus sexmaculatus (Fabricius) demonstrates that males perform post-copulatory mate guarding in the form of prolonged mating durations. We investigated whether food resource fluctuation affects pre- and post-copulatory behaviour of M. sexmaculatus. It has not been studied before in ladybirds. For this, adults were subjected to prey resource fluctuations sequentially at three levels: post-emergence (Poe; 10 days), pre-mating (Prm; 24 h) and post-mating (Pom; 5 days; only female). The food resource conditions at each level could be any one of scarce, optimal or abundant. Pre-copulatory and post-copulatory behaviour, and reproductive output were assessed. Post-emergence and pre-mating nutrient conditions significantly influenced the pre-copulatory behaviour. Males reared on scarce post-emergence conditions were found to require significantly higher number of mating attempts to establish mating unlike males in the other two food conditions. Under scarce post-emergence and pre-mating conditions, time to commencement of mating and latent period were high but opposite result was obtained for mate-guarding duration. Fecundity and per cent egg viability were more influenced by post-mating conditions, with scarce conditions stopping oviposition regardless of pre-mating and post-emergence conditions. Present results indicate that pre- and post-copulatory behaviour of ladybird is plastic in nature in response to food resource fluctuations.

Resource partitioning in a ladybird, Menochilus sexmaculatus: function of body size and prey density.

In the present study, resource partitioning by natural conspecific size variants (small and large) of ladybird, Menochilus sexmaculatus (Fabricius) females, in response to varying prey densities was assessed using functional and numerical responses as measures of prey density. The prey provided was small (second) and large (fourth) instars of Aphis craccivora Koch. Results revealed that under choice condition, small and large females of M. sexmaculatus consumed higher number of small and large instars, respectively. Small females exhibited a modified Type II functional response on small aphid instars and a Type II functional response on fourth aphid instars. Large females exhibited a Type II functional response when provided either second or fourth aphid instars. Numerical response in terms of numbers of eggs laid by both the females increased with increase in the density of either of the aphid instars. However, in small females, oviposition had a positive correlation with the numbers of small and large aphid instars consumed; being strong for the small aphid instars. While in large females, oviposition was positively correlated with the numbers of large aphid instars consumed and not small aphid instars. It therefore seems that intraspecific resource partitioning in M. sexmaculatus occurs prominently in large females than the small females.

Subperiosteal inferior maxillectomy in mucormycosis patients: case series.

OBJECTIVE: Mucormycosis is a rapidly progressive and fulminant fungal infection mainly affecting the nose and paranasal sinuses and often requiring aggressive surgical debridement, which commonly includes inferior maxillectomy. Conventional inferior maxillectomy involves removal of the bony hard palate and its mucoperiosteum. This can lead to formation of an oroantral fistula and thereby increase the morbidity in these patients leading to prolonged rehabilitation. Subperiosteal inferior maxillectomy involves sparing of the uninvolved mucoperiosteum of the hard palate. This flap is used for closure of the oroantral fistula, which preserves the functional capabilities of the patient, such as speech, mastication and deglutination. METHOD: This case series describes the experience of using the technique of mucosa-preserving subperiosteal inferior maxillectomy in five patients with mucormycosis. RESULTS: With the technique used in this study, complete oronasal separation was achieved in all six patients. The overall surgery time was also decreased when compared with free tissue transfer. Patients also did not have to bear the weight of prosthesis. CONCLUSION: Mucoperiosteal palatal flap-preserving subperiosteal inferior maxillectomy is an excellent approach for all patients with mucormycosis and healthy palatal mucosa.

Characterization of phi112, a Molecular Marker Tightly Linked to the o2 Gene of Maize, and Its Utilization in Multiplex PCR for Differentiating Normal Maize from QPM.

Quality Protein Maize (QPM) contains higher amounts of essential amino acids lysine and tryptophan. The QPM phenotype is based on regulating zein protein synthesis by opaque2 transcription factor. Many gene modifiers act to optimize the amino acid content and agronomic performance. An SSR marker, phi112, is present upstream of the opaque2 DNA gene. Its analysis has shown the presence of transcription factor activity. The functional associations of opaque2 have been determined. The putative transcription factor binding at phi112 marked DNA was identified through computational analysis. The present study is a step towards understanding the intricate network of molecular interactions that fine-tune the QPM genotype to influence maize protein quality. In addition, a multiplex PCR assay for differentiation of QPM from normal maize is shown, which can be used for Quality Control at various stages of the QPM value chain.

Study of neurocognitive endophenotypes in drug-naïve obsessive-compulsive disorder patients, their first-degree relatives and healthy controls.

OBJECTIVE: Obsessive-compulsive disorder (OCD) is a debilitating heritable neuropsychiatric condition. Attempts to delineate genetic contributions in OCD have met with limited success, with an ongoing search for neurocognitive endophenotypes. In this study, an attempt has been made to study and compare the neurocognitive functioning of patients with OCD, their first-degree relatives (FDRs) and healthy controls. METHOD: A cross-sectional design study was carried out with thirty dyads of patients with OCD, their FDRs and thirty matched healthy controls and screened using Mini International Neuropsychiatric Interview, Verbal Adult Intelligence Scale, Yale Brown Obsessive-Compulsive Scale, Montgomery Åsberg Depression Rating Scale, International Personality Disorder Examination (Anankastic personality scale).Tests of National Institute of Mental Health and Neurosciences Neuropsychological Battery were used to assess domains of attention, verbal memory, visual memory, set-shifting, response inhibition, planning and visuoconstructive abilities. spss version 14.0 was used for descriptive and analytical data analysis. RESULTS: There were no statistically significant differences between patients with OCD and their FDRs on neurocognitive domains of delayed verbal recall, set-shifting, response inhibition and visuoconstructive abilities (P > 0.05) which were impaired compared with healthy controls. Significant differences (P < 0.05) on domains of attention, planning time and visual memory were noted between FDRs and patients. CONCLUSION: The present study supports set-shifting and inhibitory control and proposes visuoconstructive abilities and delayed verbal recall as potential endophenotypes for OCD.

Outcomes of Mechanical Thrombectomy in the Early (<6-hour) and Extended (≥6-hour) Time Window Based Solely on Noncontrast CT and CT Angiography: A Propensity Score-Matched Cohort Study.

BACKGROUND AND PURPOSE: Current stroke care recommendations for patient selection for mechanical thrombectomy in the extended time window demand advanced imaging to determine the stroke core volume and hypoperfusion mismatch, which may not be available at every center. We aimed to determine outcomes in patients selected for mechanical thrombectomy solely on the basis of noncontrast CT and CTA in the early (<6-hour) and extended (≥6-hour) time windows. MATERIALS AND METHODS: Consecutive mechanical thrombectomies performed for acute large-vessel occlusion ischemic (ICA, M1, M2) stroke between February 2016 and August 2020 were retrospectively reviewed. Eligibility was based solely on demographics and noncontrast CT (ASPECTS) and CTA, due to the limited availability of perfusion imaging during the study period. Propensity score matching was performed to compare outcomes between time windows. RESULTS: Of 417 mechanical thrombectomies performed, 337 met the inclusion criteria, resulting in 205 (60.8%) and 132 (39.2%) patients in the 0- to 6- and 6- to 24-hour time windows, respectively. The ASPECTS was higher in the early time window (9; interquartile range = 8-10) than the extended time window (9; interquartile range = 7-10; P = .005). Propensity score matching yielded 112 well-matched pairs. Equal rates of TICI 2b/3 revascularization and symptomatic intracranial hemorrhage were observed. A favorable functional outcome (mRS 0-2) at 90 days was numerically more frequent in the early window (45.5% versus 33.9%, P = .091). Mortality was numerically more frequent in the early window (25.9% versus 17.0%, P = .096). CONCLUSIONS: Patients selected for mechanical thrombectomy in the extended time window solely on the basis of noncontrast CT and CTA still achieved decent rates of favorable 90-day functional outcomes, not statistically different from patients in the early time window.

COPD exacerbation rates are higher in non-smoker patients in India.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is common among non-smokers exposed to solid fuel combustion at home. Different clinical characteristics in these patients may have significant therapeutic and prognostic implications.METHODS: We used medical record review and a questionnaire among COPD patients at 15 centres across India to capture data on demographic details, different types of exposures and clinical characteristics. Chest radiography and pulmonary function testing were performed in all 1984 cases; C-reactive protein and exhaled breath nitric oxide were measured wherever available.RESULTS: There were 1388 current or ex-smokers and 596 (30.0%) non-smokers who included 259 (43.5%) male and 337 (56.5%) female patients. Sputum production was significantly more common in smokers with COPD (P < 0.05). The frequency of acute symptomatic worsening, emergency visits and hospitalisation were significantly higher (P < 0.05) in non-smokers with COPD; however, intensive care unit admissions were similar in the two groups. There was no significant difference with respect to the use of bronchodilators, inhalational steroids or home nebulisation among smoker and non-smoker patients. The mean predicted forced expiratory volume in 1 sec in smokers (43.1%) was significantly lower than in non-smokers (46.5%).CONCLUSION: Non-smoker COPD, more commonly observed in women exposed to biomass fuels, was characterised by higher rate of exacerbations and higher healthcare resource utilisation.

Concurrent infection of Bluetongue and Peste-des-petits-ruminants virus in small ruminants in Haryana State of India.

Bluetongue (BT) and peste-des-petits-ruminants (PPR) are major transboundary diseases of small ruminant, which are endemic in India. Testing of bluetongue virus (BTV) and peste-des-petits-ruminants virus (PPRV) from recent outbreaks (2015-2016) in different regions of Haryana State of India revealed that 27.5% of the samples showed the presence of dual infection of BTV and PPRV. Analysis of Seg-2 of BTV (the serotype-determining protein) showed the presence of BTV-12w in several isolates. However, analysis of N gene fragment amplicons showed that viruses belong to lineage IV were most closely related to a pathogenic strain of PPRV from Delhi. This is the first report of co-circulation of PPRV lineage IV and bluetongue virus serotype 12 in the state.

Hepatic calcification following dengue virus-induced fulminant hepatic failure.

Hepatic calcification can be seen with various infectious and neoplastic conditions. We report a 32-year- old man who developed massive calcification in the right lobe of liver following recovery from dengue virus-associated fulminant liver failure.

Prevalence of tobacco use among school going youth in North Indian States.

BACKGROUND: Tobacco use, which is the cause of several respiratory diseases, generally starts in the teens. Global Youth Tobacco Survey (GYTS) is an international initiative to investigate the tobacco use in school going youth of 13 to 15 years of age. This report describes the GYTS findings in the North Indian region. METHODS: A two-stage cluster sample was used with selection of schools on probability proportional to enrollment size followed by systematic equal probability sampling with a random start of classes from each school. The 85-item questionnaire included 'core GYTS' and other additional questions. Data analysis was performed using EpiInfo 3.2 software and the results accounted for the complex sampling design and weighting factors in the data set. RESULTS: The sample consisted of 9319 students out of the total eligible population of 30488 from 100 schools. The prevalence of ever-use of tobacco varied between 2.9 to 8.5% in boys and 1.5 to 9.8% in girls. The prevalence was highest in Chandigarh and lowest in Punjab. Between 16 to 46% of students were exposed to the habit of tobacco among parents or friends. There were 10 to 34% students who were passively exposed to environmental tobacco smoke. CONCLUSION: Tobacco use is present in up to 10 percent of school going youth in the region. A majority of them had desired to quit. Tobacco control and cessation programmes therefore, remain important health issues.

Arthroscopic reconstruction of the anterior cruciate ligament using bone-patellar tendon-bone autograft: experience of the first 100 cases.

PURPOSE: To review the results of patients who underwent arthroscopic reconstruction of the anterior cruciate ligament (ACL) via a single incision technique using a bone-patellar tendon-bone autograft. METHODS: Patients with ACL-deficient knees who were symptomatic and wanted to maintain an active lifestyle or continue sporting activities were included. ACL reconstruction using the bone-patellar tendon-bone graft was performed on 100 patients. One-year follow-up was completed in 78 patients who were then reviewed. The mean age of patients reviewed was 26.8 years (range, 21-39 years), of whom 35 (44.9%) were aged between 26 and 30 years. There were 73 men and 5 women (ratio, 14.6:1). Injuries on the right side outnumbered those on the left (44 versus 34). Sports injuries accounted for 66.7% (n = 52) of patients, motor vehicle accidents and household injuries accounted for 30.8% (n = 24) and 2.6% (n = 2), respectively. RESULTS: Excellent and good-to-excellent results were achieved in 7 (9%) and 61 (78.2%) of patients. Residual anterior knee pain (n = 18) was the most common complication, followed by difficulty in regaining full range of motion (n = 10) and divergence of femoral screw (n = 9). CONCLUSION: This procedure provides consistent and reproducible results in carefully selected patients and allows them an early return to sporting activities with minimal residual morbidity.

Characterization of cytopathogenicity of classical swine fever virus isolate induced by Newcastle disease virus.

Classical swine fever virus (CSFV), the causative agent of classical swine fever, belongs to the family Flaviviridae and genus Pestivirus. Some pestiviruses exhibit cytopathic effect in cell culture but exact phenomenon is unknown. Over expression of NS2-3 gene, presence of defective interfering particle and exaltation of Newcastle disease virus (END) phenomenon could be the reasons of cytopathogenicity. In the present study, a CSFV isolate exhibiting cytopathic effect (CPE) in Madin-Darby Canine Kidney (MDCK) cell line was characterized. To characterize cytopathogenicity of such isolate, END test was carried out. Interference of Newcastle disease virus (NDV) in MDCK adapted CSFV was confirmed by RT-PCR and virus neutralization test. Absence of CPE and NDV specific nucleic acid after neutralization confirmed the induction of CPE by NDV. Further, identity of the CSFV isolate in MDCK cell line by immunoperoxidase test, immunoblotting and RT-PCR post NDV neutralization established the virus replication without CPE (non-cytopathic isolate). Findings suggest that, there could be a chance of mixed infection of both CSFV and NDV in the piglet from which the sample was collected for virus isolation.

Establishment and characterization of a continuous cell line from thymus of striped snakehead, Channa striatus (Bloch 1793).

The establishment and characterization of a continuous cell line from the thymus of air-breathing fish Channa striatus are described. The cell line, designated C. striatus thymus (CST), has been subcultured over 71 times and shows optimal growth at 28°C in Leibovitz's-15 (L-15) medium supplemented with 20% fetal bovine serum. The CST cells exhibited low plating efficiency which improved with increase in seeding density. The karyotype analysis revealed that CST cells have a normal diploid karyotype with 2n = 40. Partial amplification and sequencing of two mitochondrial genes, viz. 16S ribosomal RNA (rRNA) and cytochrome oxidase I, confirmed that the cell line originated from C. striatus. CST cells were successfully transfected indicating their potential application for expression of recombinant proteins. In immunocytochemical staining, CST cells showed characteristics of epithelial cells. These cells were sensitive to extracellular products of Vibrio cholerae MTCC 3904 as well as to heavy metal mercuric chloride. The CST cell line would be a useful tool in functional genomic studies such as RNA interference and gene knockout as well as for cytotoxicity studies.

Plasma plasminogen activator inhibitor-1 activity in normoglycemic hypertriglyceridemic north Asian Indian subjects: a preliminary case-control study.

BACKGROUND: Recent evidence suggests that increased activity of plasma plasminogen activator inhibitor-1, an important component of the insulin resistance syndrome, plays a crucial role in the pathogenesis of atherosclerosis. METHODS AND RESULTS: In this case-control study, relationships between plasma plasminogen activator inhibitor-1 activity, serum triglyceride levels and hyperinsulinemia were explored in 40 non-diabetic patients with primary hypertriglyceridemia (Group 1) and 40 non-diabetic normotriglyceridemic controls (Group 2) matched for potential confounders like smoking and physical activity. Mean values of fasting serum insulin levels were increased in Group 1 (p>0.05). Hyperinsulinemia was observed in 14 (17.5%) individuals in Group 1 and 11 (13.8%) individuals in Group 2. Mean plasma plasminogen activator inhibitor-I activity in Group 1 (9.8+/-8.4 IU) was higher than in Group 2 (7.0+/-7.7 IU), though the difference was not significant (p>0.05). However, when only subjects with elevated levels of plasma plasminogen activator inhibitor-1 activity were taken into account, mean values were significantly higher in Group 1 (p<0.05). The plasma plasminogen activator inhibitor-1 activity was higher in subjects with body mass index >25 in both the groups, significantly so in males (p=0.05). Hyperinsulinemic subjects with a body mass index >25 and raised serum triglyceride levels had higher mean values of plasma plasminogen activator inhibitor-1 activity (18.42+/-11.15 IU) than subjects with similar characteristics and normal triglyceride levels (14.22+/-8.20 IU, p<0.05). CONCLUSIONS: Though in the current study a trend for hyperinsulinemia and high plasma plasminogen activator inhibitor-1 activity was observed in hypertriglyceridemic subjects, a larger study is needed to achieve significant differences and correlations. Obese male subjects, irrespective of their lipid profile, are at risk for thrombotic events in view of their significantly higher plasma plasminogen activator inhibitor-1 values. Procoagulant tendency is further enhanced if hypertriglyceridemia and hyperinsulinemia are added on to obesity.

Pneumocystis carinii pneumonia in a patient with active untreated systemic lupus erythematosus.

Pneumonias due to Pneumocystis carinii (PCP) commonly occur in immunocompromised hosts. Although a treatable infection, it is associated with high mortality. A case of PCP presenting in an untreated case of systemic lupus erythematosus is reported, in view of the rarity of this association.

Studies on the development of an insulin resistant rat model by chronic feeding of low magnesium high sucrose diet.

Magnesium deficiency and excess sucrose in the diet have been shown to play an important role in the development of insulin resistance. In the present study we have looked at the combined effect of a low magnesium high sucrose diet on basal glucose and insulin levels, erythrocyte insulin receptors and lipid profile in rats. For this purpose rats were divided into four groups and fed control, low magnesium, high sucrose and low magnesium high sucrose diets respectively for three months. The biochemical analysis showed a significant increase in blood glucose and triglyceride levels after one, two and three months of feeding in both the high sucrose and the low magnesium high sucrose groups, while rats fed a low magnesium diet showed a significant increase in blood glucose and triglyceride levels only after the second month. Insulin levels increased significantly in low magnesium, high sucrose and low magnesium high sucrose groups by the end of the study period. Compared to control rats, the binding of insulin to the erythrocyte insulin receptors was reduced significantly in the high sucrose and the low magnesium high sucrose groups. Cholesterol levels were found to increase significantly in the high sucrose group at the end of one month and three months of feeding. HDL-cholesterol decreased significantly in the low magnesium high sucrose group by the end of the study. Serum and RBC magnesium levels demonstrated a significant decrease in the low magnesium and the low magnesium high sucrose groups. The post heparin plasma lipoprotein lipase activity was decreased significantly in low magnesium, high sucrose and low magnesium high sucrose groups compared to control rats. These findings suggest that feeding a diet low in magnesium and high in sucrose causes insulin resistance in rats.