Subclinical circulation of Chicken infectious anaemia virus- A Sero-molecular study.

Chicken infectious anaemia-an important immunosuppressive viral disease of chicken-gained much attention in the recent past. Based on huge mortality and production loss observed in the fast-growing poultry sector, the present study aimed to find out the current status of the chicken infectious anaemia virus (CIAV), among chicken flocks in the Punjab state of India by sero-molecular study. The sera from the blood samples were tested for anti-CIAV antibodies by indirect ELISA and also compared with haematological parameters. DNA from sero-positive samples underwent PCR amplification, sequencing and phylogenetic analysis of the most conserved genomic region (VP3 gene) to detect viraemia in asymptomatic birds. The serological study using indirect ELISA showed a high sero-positivity of 77.27% in chicken flocks. Additionally, the present study also revealed the high molecular evidence (72.54%) of CIAV in apparently healthy birds. Genetic analysis showed that all CIAVs have conserved VP3 genes without any nucleotide substitutions, indicating presence of CIAV and its subclinical circulation among apparently healthy flocks. The wide distribution of CIAV among birds may be the reason for huge mortality and production loss. Further, it is suggested that studies be conducted to find out the co-involvement of CIAV with other immunosuppressive microbial agents and the immunosuppressive effect of CIAV in apparently healthy birds. Also, its role in vaccine failure and outbreaks of various other avian diseases needs to be explored.

Applicability of new sustainable and efficient green metal-based nanoparticles for removal of Cr(VI): Adsorption anti-microbial, and DFT studies.

Artemisia absinthium leaves were utilized as a reducing agent for green synthesis of Zinc oxide nanoparticles (particle size 17 nm). Synthesized green-ZnO (g-ZnO) were characterized by SEM/EDX, FTIR, XRD, UV, and BET analyses and then further used as an adsorbent to remove Cr(VI) ions from simulated wastewater. Optimal pH, temperature and adsorbent dosage were determined through batch mode studies. High removal efficiency and adsorption capacity were observed at pH 4, 0.25 g L-1 dosage, and 25 mg L-1 concentration of Cr(VI). Experimental data were modelled with different adsorption kinetics (Elovich model, PFO, PSO, IDP model) and isotherms (Langmuir, Freundlich, and Temkin), and it was found the adsorption process was well fitted to Langmuir with an R2 value greater than>0.99. Computational calculation showed that the g-ZnO nanoparticles became ∼14 times more dynamic with delocalized surface states making them a relevant platform to adsorb Cr with greater work function compatibility supporting the experimental findings. The Qmax adsorption capacity of g-ZnO was 315.46 mg g-1 from Langmuir calculations. Thermodynamic calculations reveal that the Cr (VI) adsorption process was spontaneous and endothermic, with a positive ΔS value representing the disorder at the solid-solution interface during the adsorption. In addition, the present study has demonstrated that these g-ZnO nanoparticles show strong antibacterial activities against P. aeruginosa (MTCC 1688) and E. coli (MTCC 1687). Also, the novel g-ZnO adsorbent capacity to remove Cr(VI) from simulated water revealed that it could be reused at least six times with higher removal rates during regeneration experiments. The results obtained from adsorption and antimicrobial activities suggest that g-ZnO nanoparticles could be used effectively in real-time wastewater and agricultural safety applications.

Mechanistic and recent updates in nano-bioremediation for developing green technology to alleviate agricultural contaminants.

The rise in environmental pollutant levels in recent years is mostly attributable to anthropogenic activities such as industrial, agricultural and other activities. Additionally, these activities may produce excessive levels of dangerous toxicants such as heavy metals, organic pollutants including pesticide and herbicide chemicals, and sewage discharges from residential and commercial sources. With a focus on environmentally friendly, sustainable technology, new technologies such as combined process of nanotechnology and bioremediation are urgently needed to accelerate the cost-effective remediation process to alleviate toxic contaminants than the conventional remediation methods. Numerous studies have shown that nanoparticles possess special qualities including improved catalysis and adsorption as well as increased reactivity. Currently, microorganisms and their extracts are being used as promising, environmentally friendly catalysts for engineered nanomaterial. In the long term, this combination of both technologies called nano-bioremediation may significantly alter the field of environmental remediation since it is more intelligent, safe, environmentally friendly, economical and green. This review provides an overview of soil and water remediation techniques as well as the use of nano-bioremediation, which is made from various living organisms. Additionally, current developments related to the mechanism, model and kinetic studies for remediation of agricultural contaminants have been discussed.

Prevalence and molecular characterization of canine parvovirus.

BACKGROUND AND AIM: Canine parvovirus (CPV) belonging to family Parvoviridae causes hemorrhagic gastroenteritis in dogs and heavy mortality in young dogs. The virus has three structural (VP1, VP2 and VP3) and two non-structural proteins (NS1 and NS2), VP2 being highly immunogenic. This study aims to study molecular epidemiology of CPV by sequence analysis of VP2 gene to determine the prevailing antigenic type(s) in the northern regions of India. MATERIALS AND METHODS: A total of 118 rectal swabs collected from dogs exhibiting clinical signs of CPV infection were processed for the isolation of DNA and subjected to polymerase chain reaction (PCR) and nested PCR (NPCR). A total of 13 NPCR products selected randomly were subjected to sequence analysis of VP2 gene. RESULTS: The percent positivity of CPV was found 28% and 70% by PCR and NPCR, respectively. Dogs with vaccination history against CPV too were found positive with a percent positivity of 24.10%. Gene sequencing and phylogenetic analysis of VP2 gene from these isolates revealed that most samples formed a clade with CPV-2a isolates. CONCLUSION: Sequence analysis and phylogenetic analysis of VP2 gene in the studied regions of northern India revealed that CPV-2a was the most prevalent antigenic type.

Detection and molecular characterization of chicken infectious anaemia virus in young chicks in Punjab region of north-western India.

Chicken infectious anaemia (CIA) is an important viral disease of chicken causing significant immunosuppression and severe anaemia worldwide. Occurrence of severe disease and mortality is noticed in young chicks (2-3 weeks). Vertical mode of transmission increases chance of infection and persistence of virus among the infected flocks. The current study was conducted in Punjab state for confirmation and genetic characterization of CAV among chicken flocks of various poultry farms. DNA was extracted from the tissue samples and subjected to polymerase chain reaction (PCR) of VP1 gene and whole genome. PCR products were further sequenced for confirmation of chicken infectious anaemia virus (CIAV) genome in the clinical samples. PCR amplification of DNA from the tissue samples yielded expected product size of 1350 bases of VP1 gene and 2.3 kb of whole genome. Out of 16 commercial poultry farms, 11 were confirmed with presence of CIAV, and out of 65 birds, 39 were found positive (60%) for CIAV genes. Among the various organs, the presence of viral gene was detected at highest level in thymus when compared with other organs. It is concluded that chicken infectious anaemia virus detected from Punjab state is closely related to other Indian isolates and neighbouring countries which necessitates need of more intensive studies with a greater number of samples for implementing effective control measures.

Corticosteroid-associated lupus pancreatitis: a case series and systematic review of the literature.

BACKGROUND: Acute pancreatitis is an uncommon complication that occurs in 0.85% to 4% of patients with systemic lupus erythematosus (SLE). In some patients, it occurs within days to weeks of starting medium-to-high dose corticosteroids. The authors have used the term 'corticosteroid-associated lupus pancreatitis' for these patients, and they report a case series and perform a systematic review of previously published reports. METHODS: For the purpose of this study, corticosteroid-associated lupus pancreatitis was defined as occurrence of acute pancreatitis in patients with SLE (fulfilling the 1997 ACR), within 3 weeks of starting therapy with medium-to-high dose corticosteroids - either newly initiated or escalated from a lower dose. All patients with SLE admitted in the last 2.5 years in a North Indian university hospital were reviewed, and those with pancreatitis who fulfilled the above criteria were included in the case series. For the systematic review, a PUBMED search using the keywords 'lupus' and 'pancreatitis' was performed, and reports in English were reviewed for an association with corticosteroids. RESULTS: Among 420 admissions of SLE patients, six patients (1.4%) fulfilled criteria for corticosteroid-associated lupus pancreatitis. All were female, with mean age and disease duration of 19.7 ± 3.3 and 3.8 ± 2.5 years respectively. All had active disease and developed acute pancreatitis within 48-72 hours of newly initiating medium-to-high dose corticosteroids (in three patients) or escalating them to medium-high dose (in three patients). After the development of pancreatitis, corticosteroids were continued in all except one patient. In addition, two patients received pulse methylprednisolone, two received pulse cyclophosphamide and one was started on azathioprine. Three patients died during hospitalization, all with severe pancreatitis. On systematic review, among 451 cases of lupus pancreatitis reported, 23 (5%) fulfilled criteria for 'corticosteroid-associated lupus pancreatitis'. A majority of them had pancreatitis within 3 days of starting treatment with medium-to-high dose corticosteroids. The mortality in these patients was 37.5%. CONCLUSION: In a small but substantial proportion of patients with lupus who develop pancreatitis, it occurs within days to weeks of starting medium-to-high dose corticosteroids. Many of these patients continue to receive corticosteroids, and some receive more aggressive immunosuppression. However, they have significant mortality, and further studies are required to identify appropriate treatment in this subgroup of patients.

Cumulative Risk of Metabolic Syndrome Correlated with the Coexistence of (-1306C/T) and Altered Circulating MMP2 level.

BACKGROUND: Interindividual genetic variations and environmental factors both play pivotal roles in the pathogenesis of metabolic syndrome (MetS). The rationale of this study conducted was to analyze the association of Matrix Metalloproteinase (MMP) gene variants, MMP-1 (-1607 1G/2G) and MMP-2 (-1306 C/T) with susceptibility to MetS and its effect on serum MMP level. METHODS: Study involved 370 subjects with 1:1 distribution of cases and controls. Patients were recruited according to modified NCEP-ATP III criteria for MetS. Clinical, biochemical analysis, PCR-RFLP and ELISA methods were employed for genotyping and estimation of serum MMP level. RESULTS: Significantly (p<0.001) higher Serum MMP-2 (39.13±19.96 ng/ml) was detected in cases as compared to controls. The MMP-2 (-1306 C/T) was significantly associated with the risk of MetS. The variant genotype TT was significantly associated with increased risk of MetS. (p=0.032; OR=2.31; 95%CI=1.07-4.97). No significant association of MMP-1(-1607 1G/2G) was found with risk of MetS. CONCLUSION: Our study concluded that presence of MMP-2 (-1306 C/T) might be associated the risk of MetS. Serum MMP2 level was significantly higher in patients and correlated with clinical parameters of MetS. Clinical implication of the work may help to identify the individuals with high risk of MetS and further complications.

Phylogenetic analysis of VP2 gene of canine parvovirus and comparison with Indian and world isolates.

Canine parvovirus (CPV) causes hemorrhagic enteritis, especially in young dogs, leading to high morbidity and mortality. It has four main antigenic types CPV-2, CPV-2a, CPV-2b and CPV-2c. Virus protein 2 (VP2) is the main capsid protein and mutations affecting VP2 gene are responsible for the evolution of various antigenic types of CPV. Full length VP2 gene from field isolates was amplified and cloned for sequence analysis. The sequences were submitted to the GenBank and were assigned Acc. Nos., viz. KP406928.1 for P12, KP406927.1 for P15, KP406930.1 for P32, KP406926.1 for Megavac-6 and KP406929.1 for NobivacDHPPi. Phylogenetic analysis indicated that the samples were forming a separate clad with vaccine strains. When the samples were compared with the world and Indian isolates, it was observed that samples formed a separate node indicating regional genetic variation in CPV.

Multiplex real-time PCR for identification of canine parvovirus antigenic types.

Canine parvovirus (CPV) is an important disease causing gastroenteritis and/or haemorrhagic gastroenteritis in dogs. There are four antigenic types of CPV reported worldwide viz. CPV 2, CPV 2a, CPV 2b and CPV 2c. The diagnosis of CPV with the identification of the antigen type responsible remains problematic. In the present study, identification as well as antigenic typing of CPV was done using a de novo multiplex real time PCR to combat the problem of antigenic type identification. From the study it could be concluded that the here developed multiplex real time PCR assay could be used for rapid detection of CPV as well as typing of its three antigenic types.

Isolation of Canine parvovirus with a view to identify the prevalent serotype on the basis of partial sequence analysis.

AIM: The aim of this study was to isolate Canine parvovirus (CPV) from suspected dogs on madin darby canine kidney (MDCK) cell line and its confirmation by polymerase chain reaction (PCR) and nested PCR (NPCR). Further, VP2 gene of the CPV isolates was amplified and sequenced to determine prevailing antigenic type. MATERIALS AND METHODS: A total of 60 rectal swabs were collected from dogs showing signs of gastroenteritis, processed and subjected to isolation in MDCK cell line. The samples showing cytopathic effects (CPE) were confirmed by PCR and NPCR. These samples were subjected to PCR for amplification of VP2 gene of CPV, sequenced and analyzed to study the prevailing antigenic types of CPV. RESULTS: Out of the 60 samples subjected to isolation in MDCK cell line five samples showed CPE in the form of rounding of cells, clumping of cells and finally detachment of the cells. When these samples and the two commercially available vaccines were subjected to PCR for amplification of VP2 gene, a 1710 bp product was amplified. The sequence analysis revealed that the vaccines belonged to the CPV-2 type and the samples were of CPV-2b type. CONCLUSION: It can be concluded from the present study that out of a total of 60 samples 5 samples exhibited CPE as observed in MDCK cell line. Sequence analysis of the VP2 gene among the samples and vaccine strains revealed that samples belonged to CPV-2b type and vaccines belonging to CPV-2.

Correlation of serum phosphate with carotid intimal-medial thickness in chronic kidney disease patients.

While increased serum phosphate concentration is a significant risk factor for vascular calcification, it is unclear whether serum phosphate is also a risk factor for increased arterial wall thickness in chronic kidney disease (CKD) patients. Using B-mode ultrasonography, we examined carotid intimal-medial thickness (CIMT) of CKD patients and analyzed risk factors for increased IMT with regard to the effect of serum phosphate. One hundred patients were enrolled (73 patients without diabetes, 27 patients with diabetes; 57 men, 43 women; age, 46.2 ± 15.3 years). CIMT of patients with diabetes was significantly greater than that of patients without diabetes (0.78 ± 0.250 versus 0.66 ± 0.178 mm; P < 0.0001). For the group of all patients, CIMT correlated strongly and significantly with serum phosphate (r = 0.911; P < 0.001). In multiple regression analysis serum phosphate level (ß = 0.356; <0.0001) was found to be a significant independent risk factor for increased CIMT, in addition to other independent risk factors, including advanced age, higher systolic blood pressure, urinary albumin and the presence of diabetes (R2 = 0.956; P < 0.00001). In conclusion, high serum phosphate level is a significant and independent factor associated with advanced arteriosclerosis in CKD patients with and without diabetes in addition to advanced age.

Antigenic typing of canine parvovirus using differential PCR.

Canine parvovirus (CPV) is an enteric pathogen causing hemorrhagic enteritis in pups of 3-6 months of age and is mainly transmitted via feco-oral route. In the present study, a total of 85 animals rectal swabs suspected of CPV were tested using a PCR, nested PCR and a newly designed differential PCR. Using PCR 7 (8.23 %) animals were positive whereas 39 (45.88 %) were positive by using nested PCR and 40 (47.05 %) were positive for either one or more than one antigenic types of CPV using differential PCR. Using differential PCR it was found that CPV-2a and CPV-2b were the most prevailing antigenic types. Also it was found that dogs that were vaccinated too yielded positive CPV indicating a possible presence of additional CPV antigenic types. Thus, the primers used in differential PCR can be used in a single PCR reaction to detect various antigenic types of CPV.

Non-specific sex-differential effect of DTP vaccination may partially explain the excess girl child mortality in Ballabgarh, India.

OBJECTIVE: To test the hypothesis that a gender differential exists in the effect on child mortality of BCG, DTP, measles vaccine as administered under programme conditions in Ballabgarh HDSS area. METHODS: All live births in 28 villages of Ballabgarh block in North India from 2006 to 2011 were followed until 31 December 2011 or 36 months of age whichever was earlier. The period of analysis was divided into four time periods based on eligibility for vaccines under the national immunisation schedule (BCG for tuberculosis, primary and booster doses of diphtheria-tetanus-pertussis and measles). Cox proportional hazards regression was used to assess the association between sex and risk of mortality by vaccination status using age as the timescale in survival analysis and adjusting for wealth index, access to health care, the presence of a health facility in the village, parental education, type of family, birth order of the child and year of birth. RESULTS: 702 deaths (332 boys and 370 girls) occurred among 12,142 children in the cohort in the 3 years of follow-up giving a cumulative mortality rate of 57.5 per 1000 live births with 35% excess girl child mortality. Age at vaccination for the four vaccines did not differ by sex. There was significant excess mortality among girls after immunisation with DTP, for both primary (HR 1.65; 95% CI:1.17-2.32) and DTPb (2.21; 1.24-3.93) vaccinations. No significant excess morality among girls was noted after exposure to BCG 1.06 (0.67-1.67) or measles 1.34 (0.85-2.12) vaccine. CONCLUSION: This study supports the contention that DTP vaccination is partially responsible for higher mortality among girls in this study population.

Boning up on glypicans--opportunities for new insights into bone biology.

Bone formation is remarkable for the convergence in the activity of four major signalling pathways, the bone morphogenetic protein (BMP), fibroblast growth factor (FGF), hedgehog (HH) and wingless-integrated (WNT) pathways. These pathways cooperate in morphogenetic, proliferative and differentiative processes that underpin the development, growth and repair of skeletal structures. They are regulated by pathway-specific modulators and by another class of molecules, the glypicans. Glypicans are proteoglycans located on the cell surface, where they act as coreceptors to promote or inhibit signalling by ligands of the BMP, FGF, HH and WNT pathways, through protein-protein and protein-carbohydrate interactions. In this review, we discuss glypican structure, expression and function in the context of bone development and growth, with emphasis on the long bone growth plate where five of the six glypicans are expressed in overlapping patterns in the chondrogenic zone. Analyses of gene knockout models and the human conditions of Simpson-Golabi-Behmel syndrome and omodysplasia, which arise from mutations in glypican 3 (GPC3) and GPC6, respectively, highlight both subtle and striking effects of glypicans on bone growth. We draw attention to challenges and areas of opportunity, where the actions of glypicans on BMP, FGF, HH and WNT signalling might be profitably studied to help illuminate the complex interplay of signalling that drives bone growth.

Toxicopathological studies on the effects of aflatoxin B(1), ochratoxin A and their interaction in New Zealand White rabbits.

New Zealand White rabbits, divided into 4 groups were fed with feed containing aflatoxin B(1) (AFB(1)) @ 0.5 ppm (group I), ochratoxin A (OA) @ 1 ppm (group II), AFB(1) and OA @ 0.5 ppm and 1 ppm, respectively (group III) and standard feed (group IV) for 60 days. Mortality and decrease in body weight were highest in the interaction group. Superoxide dismutase, catalase and malondialdehyde levels were increased in all the toxin fed groups with maximum elevation in group III. Significant decrease in the antibody titre to sRBC was observed in groups I and II. Significant reduction of HI and CMI responses was observed in group III. Grossly, liver was the most affected organ in AFB(1) treated animals. Microscopically, vacuolar degeneration of hepatocytes, bile duct epithelial hyperplasia and hypertrophy and peribiliary fibrosis were consistently observed. Ultrastructurally, hepatocytes revealed varying degrees of degeneration, swollen endoplasmic reticulum and pleomorphism of mitochondria. OA produced significant nephrotoxicity with the pale, soft and enlarged kidneys showing discoloured foci over the surface. Microscopically, kidneys revealed degeneration of the proximal convoluted tubules and the testes were atrophic. Ultrastructurally, disruption of the mitochondrial membrane and swelling of the endoplasmic reticulum were observed. In group III, gross and histopathological changes were observed both in liver and kidneys and were of greater severity as compared to those of groups I and II. Ultrastructurally, hepatocytes revealed nuclear distortion, marginated heterochromatin, chromatolysis, electron opaque mitochondria with vacuolations and disarray of cristae and loss of cytoplasmic organelles. The results suggested an additive interaction of AFB(1) and OA in rabbits.

Fetomaternal outcome in pregnancy with severe thrombocytopenia.

OBJECTIVES: STo study the feto-maternal outcome in pregnancy with severe thrombocytopenia. MATERIALS AND METHODS: It was an observational study involving 1150 pregnant women with term gestation in labour, who were screened for thrombocytopenia. Ninety-four subjects (8.17%) were found to have thrombocytopenia i.e. platelet count < 1,50,/000/mm3, out of which 47 subjects (group A) had platelet count of less than 50.000/mm3 Simultaneously, 47 term pregnant women (group B) having a normal platelet count i.e. > 1.5 lac/mm3 formed the control group. All the subjects were followed during labour and postpartum period for any feto-maternal outcome. RESULTS: Significant history of bleeding tendencies like prolonged bleeding from wound site, easy bruisability and menorrhagia (p = 0.023) was evident in the study population. Abruptio placentae and early onset pregnancy induced hypertension (PIH) in previous gestations was more commonly found in the study population. Anemia and PIH were significantly more in group A. Incision site oozing during cesarean section was significantly more in group A. Moderate thrombocytopenia was more in neonates of study group (p = 0.014), but without any bleeding complications in neonates. CONCLUSIONS: Careful surveillance is required in these high risk patients for earlier detection and treatment of complications so as to decrease the fetomaternal morbidities.

Studies on apoptotic changes in combined toxicity of citrinin and endosulfan in pregnant wistar rats and their fetuses.

BACKGROUND: Citrinin (mycotoxin) and endosulfan (pesticide) both environmental contaminants easily enter the food chain and are caoomon causes of various toxicities. MATERIALS AND METHODS: In the present investigation, citrinin (CIT) (10 mg/kg feed) and endosulfan (1 mg/kg body weight) were administered orally alone and in combination to pregnant Wistar rats from gestational day 6 to 20 to study their effect to cause apoptosis in the pregnant Wistar rats and their fetuses. Apoptosis was assessed in dams by agarose gel electrophoresis, flow cytometry and electron microscopy, while in the fetuses it was assessed by flow cytometry only. RESULT: Citrinin and endosulfan in the combination group caused apoptosis in an additive manner as there was increased number of apoptotic cells as compared to the individual toxin and control groups. The fetuses also showed increased number of apoptotic cells in the combination groups, which also indicated that both the toxins crossed the placental barrier. CONCLUSION: So it was concluded that apoptosis played a significant role in the pathogenesis of endosulfan and citrinin toxicity.

Herbal medicines used in the treatment of diabetes mellitus in Arunachal Himalaya, northeast, India.

ETHNOPHARMACOLOGICAL RELEVANCE: Medicinal plants have played an important role in treating and preventing a variety of diseases throughout the world. Khampti tribal people living in the far-flung Lohit district of the Eastern Arunachal Himalaya, India still depend on medicinal plants and most of them have a general knowledge of medicinal plants which are used for treating a variety of ailments. This survey was undertaken in Lohit district in order to inventory the medicinal plants used in folk medicine to treat diabetes mellitus. MATERIALS AND METHODS: Field investigations were conducted in seventeen remote villages of Lohit district starting from April 2002 to May 2004 through interviews among 251 key informants who were selected randomly during our household survey. To elucidate community domains and determine differences in indigenous traditional knowledge of medicinal plants with anti-diabetic efficacy, we repeated our field survey starting from April 2008 to May 2010 with one hundred traditional healers locally called as "Chau ya" in Khampti of Lohit district. "Chau ya" traditional healers who know and use medicinal plants for treating diabetes mellitus were interviewed using a semi-structured questionnaire. RESULTS: This study reports an ethnobotanical survey of medicinal plants in Lohit district of Arunachal Pradesh reputed for the treatment of diabetes mellitus. Forty-six plant species were identified in the study area to treat diabetes mellitus by the Khamptis "Chau ya" traditional healers. Comparative published literature survey analysis of this study with other ethnobotanical surveys of plants used traditionally in treating diabetes mellitus suggests that eleven plant species make claims of new reports on antidiabetic efficacy. These plant species are Begonia roxburghii, Calamus tenuis, Callicarpa arborea, Cuscuta reflexa, Dillenia indica, Diplazium esculentum, Lectuca gracilis, Millingtonia hortensis, Oxalis griffithii, Saccharum spontaneum, and Solanum viarum. Some of the plants reported in this study have an antidiabetic effect on rodent models but none have sufficient clinical evidence of effectiveness. CONCLUSIONS: The wide variety of medicinal plants that are used to treat diabetes mellitus in this area supports the importance of plants in the primary healthcare system of the rural people of Lohit district of Arunachal Pradesh. The finding of new plant uses in the current study reveals the importance of the documentation of such ethnobotanical knowledge.

Determinants of neonatal mortality in rural Haryana: a retrospective population based study.

OBJECTIVE: To identify the determinants of neonatal mortality. STUDY DESIGN: Nested case-control study. STUDY SETTING: 28 villages under the intensive field practice area of Comprehensive Rural Health Services Project, Ballabgarh, Haryana serving a population of 87,016, as on 31st December 2009. The study period was from 2005 to 2009. METHODS: The data were obtained from Health Management Information System and analyzed using multivariate logistic regression analysis. A hierarchical approach was used to analyze the factors associated with neonatal deaths, using community level factors, socio-economic status and biological determinants. The population attributable fractions were estimated for significant variables. RESULTS: The total live births during the study period were 10392 and neonatal deaths were 248. The infant and neonatal mortality rates during the study period were 45.6 and 23.8 per 1000 live births, respectively. Socio-economic determinants (Low educational status of parents [OR 2.1, 95% CI; 1.4, 3.3]; fathers occupation [OR 1.8, 95% CI; 1.0, 3.0]; Rajput caste [OR 2.0, 95% CI; 1.2, 3.4] appeared to explain a major fraction (45.7%) of neonatal deaths. Community level factors (villages with no health facility [OR 1.5, 95% CI; 1.0, 2.1]; villages with population >6000 [OR 1.7, 95% CI; 1.2, 2.5]) were associated with 27.3% of all neonatal deaths. Proximate determinants (early childbearing age of mother (<20 years) [OR 2.0, 95% CI; 1.2, 3.2]) were least important. All the three level of variables seemed to act independently with little mediation among them. CONCLUSION: Neonatal mortality is affected by socioeconomic, community level and proximate biological determinants.

Potential allergens of green gram (Vigna radiata L. Millsp) identified as members of cupin superfamily and seed albumin.

BACKGROUND: No systematic study on allergenicity of green gram seed proteins have been performed so far, although incidences of IgE-mediated reaction to green gram seedlings have been reported. OBJECTIVE: We sought to investigate the allergenic potential of green gram, followed by identification and characterization of its relevant allergens using proteomic approaches. METHODS: BALB/c mice were sensitized intraperitoneally with green gram proteins, and levels of specific Igs, Th2 cytokines, histamine, anaphylactic symptoms and histopathological responses were studied. Twelve naso-bronchial allergic patients with a history of sensitization to green gram were selected on the basis of positive skin prick test and elevated specific IgE levels. Green gram allergens were identified and characterized by their ability to endure pepsin, by IgE immunoblot of two-dimensional (2D) gels in combination with mass spectrometry and by bioinformatics approaches. RESULTS: Increased specific IgE, IgG1, Th2 cytokine and histamine levels, high anaphylactic scores and histological changes in lungs and spleen of green gram crude protein extract-treated mice are indicative of its sensitization ability. Four proteins (molecular weights: 52, 50, 30 and 18 kDa) showed pepsin resistance and IgE-binding capability with sensitized human and mice sera. The four proteins tentatively named as Vig r2 (52 kDa, pI 5.7), Vig r3 (50 kDa, pI 5.8), Vig r4 (30 kDa, pI 6.6) and Vig r5 (18 kDa, pI 5.5) showed significant sequence similarity with known allergens of soybean, lentil, pea, lupin, etc. Mass spectrometric analysis identified Vig r2 as 8S globulin ß-isoform precursor, Vig r3 as 8S globulin α-isoform precursor and Vig r4 as seed albumin. CONCLUSION AND CLINICAL RELEVANCE: Green gram seeds contain at least four clinically relevant allergenic proteins, namely Vig r2, Vig r3, Vig r4 and Vig r5 that were capable of inducing strong IgE-mediated reactions. One of the most important steps towards diagnostic and therapeutic approaches to deal effectively with food allergy is continued identification of newer food allergens and their characterization. The significance of this study can be enormous as the data generated may work as basic biology data in developing a green gram species modified genetically that may have reduced allergenicity.