Perinatal mental health in India: protocol for a validation and cohort study.

BACKGROUND: Common mental disorders (CMD) are among the largest contributors to global maternal morbidity and mortality. Although research on perinatal mental health in India has grown in recent years, important evidence gaps remain, especially regarding CMD. Our study aims to improve understanding of CMD among perinatal and non-perinatal women of reproductive age across two settings in India: Bangalore (Karnataka) and Tanda (Himachal Pradesh). METHODS: The study is embedded within the Maternal and Perinatal Health Research Collaboration India (MaatHRI). This mixed-methods observational study comprises three consecutive phases: (i) focus group discussions and individual interviews to explore women's knowledge and seek feedback on CMD screening tools; (ii) validation of CMD screening tools; and (iii) prospective cohort study to identify CMD incidence, prevalence and risk factors among perinatal and non-perinatal women. Results of the three phases will be analyzed using inductive thematic analysis, psychometric analysis and multivariable regression analysis, respectively. CONCLUSION: Improving understanding, detection and management of CMD among women is key to improving women's health and promoting gender equality. This study will provide evidence of CMD screening tools for perinatal and non-perinatal women in two diverse Indian settings, produce data on CMD prevalence, incidence and risk factors and enhance understanding of the specific contribution of the perinatal state to CMD.

Production and characterization of monoclonal antibodies to Newcastle Disease Virus.

Newcastle Disease (ND) is one of the major causes of economic loss in the poultry industry. Newcastle Disease Virus (NDV) is a single-stranded, negative-sense enveloped RNA virus (Fam. Paramyxoviridae; Order Mononegavirales). In the present study three monoclonal antibodies (MAbs) were produced by polyethyleneglycol (PEG)-mediated fusion of lymphocytes sensitized to NDV Bareilly strain and myeloma cells. NDV possesses ability to agglutinate erythrocytes of avian species. All the three MAbs designated as 2H7, 3E9 and 3G6 caused hemagglutination inhibition of NDV by specifically binding to NDV. The reactivity for all the 3 MAbs on indirect ELISA was found to be significantly higher than the antibody and antigen controls. On flowcytometry of HeLa cells infected with NDV using the MAbs as primary antibodies, there was a significant difference in the percentage of cells showing positive fluorescence compared to the mock control. One of the MAbs (3E9) was found to react with hemagglutinin-neuraminidase (HN) protein on western blot.

Characterization and evaluation of apoptotic potential of double gene construct pVIVO.VP3.NS1.

Viral gene oncotherapy, targeted killing of cancer cells by viral genes, is an emerging non-infectious therapeutic cancer treatment modality. Chemo and radiotherapy in cancer treatment is limited due to their genotoxic side effects on healthy cells and need of functional p53, which is mutated in most of the cancers. VP3 (apoptin) of chicken infectious anaemia (CIA) and NS1 (Non structural protein 1) of Canine Parvovirus-2 (CPV-2) have been proven to have oncolytic potential in our laboratory. To evaluate oncolytic potential of VP3 and NS1 together these genes needed to be cloned in a bicistronic vector. In this study, both these genes were cloned and characterized for expression of their gene products and its apoptotic potential. The expression of VP3 and NS1 was studied by confocal microscopy and flowcytometry. Expression of VP3 and NS1 in pVIVO.VP3.NS1 transfected HeLa cells in comparison to mock transfected cells indicated that the double gene construct expresses both the products. This was further confirmed by flowcytometry where there was increase in cells expressing VP3 and NS1 in pVIVO.VP3.NS1 transfected group in comparison with the mock control group. The apoptotic inducing potential of this characterized pVIVO.VP3.NS1 was evaluated in human cervical cancer cell line (HeLa) by DNA fragmentation assay, TUNEL assay and Hoechst staning. This double construct was observed to induce apoptosis in HeLa cells.

Development of dog mammary tumor xenograft in immunosuppressed Swiss albino mice.

Development and study of dog mammary tumour xenograft in immunosuppressed Swiss Albino Mice adds a new dimension in cancer research as dog tumors have many similarities with human tumors regarding progression, histopathology, molecular mechanism, immune response and therapy. Failure of the immune system to recognize and eliminate cancer cells leads to cancer progression and the fight between immune cells and cancer cells has a great role in understanding the mechanism of cancer progression and elimination. Rejection and acceptance of tumour xenograft depends on efficiency of CD4+, CD8+ and NK cell populations. In the present investigation, dog mammary tumor xenograft in cyclosporine-A and gamma-irradiated, immunosuppressed Swiss Albino mice was developed and the immune cell status of graft accepted and rejected mice was assessed. It was observed that all the major immune cells (CD4+, CD8+ and NK cells) play an equal role in tumour rejection.

Classical swine fever in the pygmy hog.

The pygmy hog is a rare, small and highly endangered mammal belonging to the Suidae family, and it is presently found only in the Assam state of India. While investigating the cause of death of pygmy hogs housed at a conservation centre for captive breeding and research at Basistha, Assam, it was confirmed that they were susceptible to and died as a result of contracting classical swine fever (CSF), caused by CSF virus (CSFV), which is a highly infectious endemic disease of domestic pigs in India. The post-mortem findings and serum CSFV-specific antibody titres, along with the isolation of CSFV from two pygmy hogs, and further confirmation by CSFV genomic E2 and 5' untranslated region (UTR) gene amplification in PCR (polymerase chain reaction), clearly established the cause of death of the pygmy hogs. Further, on phylogenetic analysis, the pygmy hog CSFV 5' UTR sequences were grouped in the genotype 1.1 cluster of Indian CSFVs, and hence the strains causing infection were closely related to CSFV isolates circulating in domestic pigs. Therefore, the occurrence of CSF in this endangered species may pose a potent threat to their existence unless properly controlled, and thus it needs urgent attention. To the authors' knowledge this is the first report on CSF in pygmy hogs.

In vitro expression studies of non structural 1 protein of Canine Parvo virus 2 by polyclonal antiserum raised against CPV2-NS1 protein expressed in Escherichia coli as an antigen.

The canine Parvovirus 2, non-structural 1 (NS1) is a novel candidate tumor suppressor gene. To confirm the expression of the NS1 in HeLa cells after transfection there was a need to raise antiserum against CPV2- NS1. Therefore, this study was carried out to express and purify the recombinant NS1 (rNS1), and characterize the polyclonal serum. CPV2-NS1, complete coding sequence (CDS) was amplified, cloned in pET32a+ and expressed in BL21 (DE3) (pLysS). SDS-PAGE analysis revealed that the expression of the recombinant protein was maximum when induced with 1.5 mM IPTG. The 6 x His tagged fusion protein was purified on Ni-NTA resin under denaturing conditions and confirmed by western blot using CPV2 specific antiserum. The rabbits were immunized with the purified rNS1 to raise anti-NS1 polyclonal antiserum. The polyclonal serum was tested for specificity and used for confirming the expression of NS1 in HeLa transfected with pcDNA.cpv2.ns1 by indirect fluorescent antibody test (IFAT), flow cytometry and western blot. The polyclonal antiserum against NS1 could be very useful to establish functional in vitro assays to explore role of NS1 in cancer therapeutics.

Development and in vitro characterization of a bivalent DNA containing HN and F genes of velogenic Newcastle disease virus.

Newcastle disease (ND) is highly contagious, economically important viral disease affecting most of avian species worldwide. Newcastle disease virus (NDV) has single stranded negative sense RNA genome which encodes for six structural and two non-structural proteins. Envelope glycoproteins i.e. hemagglutinin-neuraminidase (HN) and the fusion (F), elicit protective immune response. In this study, HN and F genes of velogenic (virulent) strain were amplified and cloned at multiple cloning sites A and B, respectively into pIRES bicistronic vector for use as bivalent DNA vaccine against ND. The recombinant plasmid was characterized for its orientation by restriction enzyme digestion and PCR. Expression of HN and F genes was assessed in transfected Vero cells at RNA level using RT-PCR in total RNA as well as protein level using IFAT, IPT and western blot using NDV specific antiserum. All these experiments confirmed that HN and F genes cloned in recombinant pIRES.nd.hn.f are functionally active. The recombinant construct is being evaluated as DNA vaccine against ND.

Rathke's cleft cyst: clinicopathological and MRI findings in 22 patients.

AIM: To evaluate clinical findings and magnetic resonance imaging (MRI) characteristics of Rathke's cleft cyst (RCC) in 22 patients. MATERIALS AND METHODS: Twenty-two patients were imaged using non-enhanced MRI and 17 underwent an additional contrast-enhanced MRI examination. Fifteen patients received an additional non-enhanced computed tomography (CT) examination, and amongst these, two underwent contrast-enhanced CT. Two radiologists read the images retrospectively. The imaging data were studied with regards to location, size, margin, signal intensity, enhancement characteristics, haemorrhage, and presence of calcifications. Clinical data, such as presenting signs and symptoms, physical findings, and medical histories, were collected. Histopathological studies were performed and analysed by two pathologists. RESULTS: Nine lesions were located in the intrasellar region, 12 in both the intra- and suprasellar regions and one in the suprasellar region. The maximum diameter of the RCCs varied from 0.7 to 4 cm, with an average size of 1.7+/-0.7 cm. MRI features of RCC were divided into three groups based on T1-weighted imaging (T1WI): hypo- (n=6), iso- (n=9), and hyperintensity group (n=7). Patients in the latter two groups were statistically younger than that in the former group. The lesion size in the iso- and hyperintensity groups was significantly less than that in the hypointensity group (F=6.421, p=0.007). Only two cases showed enhancement after contrast injection in the cohort. One lesion with haemorrhage was found as were two cases with intracystic nodules. CONCLUSION: Although MRI features of RCCs are variable, RCCs should be suspected when the following conditions occur: lesions located in the intrasellar region or involving both intra and suprasellar regions, less than 1.5 cm in diameter, iso- or hyperdense on T1WI and no signal enhancement after contrast injection. In addition, the first case of a RCC with a markedly enhanced intracystic nodule is reported.

5'-UTR-based phylogenetic analysis of Classical swine fever virus isolates from India.

Classical swine fever (CSF) caused by Classical swine fever virus (CSFV) is a globally significant disease of pigs. Genetic typing of CSFV isolates can help in understanding the epidemiology of disease and trace down the source of outbreak. 5'-UTR sequence analysis and subsequent genetic classification of nine CSFV field isolates from India indicated that 3 isolates belonged to genotype 2.1 and were closely related to European CSFV strains. The remaining 6 isolates belonged to genotype 1 that contained old and new strains. However, the genotype 2.1 group consisted of recent field isolates only. The study showed circulation of both genotypes 1 and 2.1 in north-eastern part of India.

Association of Childhood Attention Deficit Hyperactivity Disorder Symptoms with Academic and Psychopathological Outcomes in Indian College Students: a Retrospective Survey.

OBJECTIVE: To survey the prevalence of retrospectively recalled clinically significant symptoms of attention deficit hyperactivity disorder (ADHD) in childhood and determine the association of ADHD symptoms in childhood with current academic achievement and psychopathological outcomes among college students in the state of Kerala, India. METHODS: A self-administered questionnaire was distributed to 5784 students from 58 colleges selected by cluster random sampling. The Barkley Adult ADHD Rating Scale-IV was used for recollection of childhood ADHD symptoms; a total score of ≥60 (indicating the 99 percentile) was taken as the cut-off for clinically significant ADHD symptoms in childhood. The Alcohol, Smoking and Substance Involvement Screening Test was used to assess lifetime use of alcohol and tobacco. The Kessler Psychological Distress Scale was used to assess non-specific psychological distress. Lifetime suicidality and exposure to sexual abuse were assessed by asking relevant questions. Students who recalled having clinically significant ADHD symptoms in childhood were compared with those who did not. RESULTS: Of 5784 students, 639 (11.5%) did not complete the questionnaire. Of the remaining 5145 students, 1750 (34.8%) were men and 3395 (65.2%) were women, with a mean age of 19.4 years. 143 (2.8%) students reported clinically significant ADHD symptoms in childhood. Childhood ADHD symptoms were significantly more common in men and in those living in urban areas. In the bivariate analysis, those with clinically significant ADHD symptoms in childhood had significantly higher odds of poorer academic performance, alcohol use, tobacco use, psychological distress, suicidal thoughts, suicidal attempts, and contact and non-contact sexual abuse, after adjusting for sex and residence. CONCLUSIONS: Clinical evaluation and appropriate management may be warranted for adults who retrospectively recall clinically significant ADHD symptoms in childhood.

Sequence analysis of VP7 gene of Indian bluetongue virus serotype-23 shows its close phylogenetic relationship to Australian and Chinese serotypes.

Bluetongue, an arthropod borne viral disease of wild and domestic ruminants, causes heavy economic losses throughout the world. In the present study, full-length VP7 gene of Indian bluetongue virus (BTV) serotype 23 was sequenced and compared with prototype strains of BTV reported from different countries. Nucleotide sequence analysis of VP7 gene revealed Indian BTV serotype 23 to have 1154 nucleotides with the deletion of two nucleotides at 3' non-coding region and a unique amino acid change 211S-N. The Indian virus also demonstrated a maximum similarity of 94.2% with Australian serotype 1 and a minimum similarity of 67.4% with Australian serotype 15. However, at deduced amino acid level, it had maximum similarity of 99.7% and a minimum of 82.5% with Chinese serotypes 1, 2 and 4 and Australian serotype 15, respectively. Deduced amino acid sequence analysis of putative receptor binding domain (121-249) revealed all the nine hydrophilic domains to be conserved across the serotypes. Functional motifs present in VP7 protein were also conserved in almost all the BTV serotypes including Indian serotype 23. Phylogenetic analysis based on VP7 gene sequence revealed Indian BTV serotype 23 segregating into a monophyletic group along with Australian serotype 1 and Chinese serotypes 1, 2 and 4, indicating its close evolutionary relationship with these Australian and Chinese serotypes.

Phylogenetic analysis of glycoprotein B gene sequences of bovine herpesvirus 1 isolates from India reveals the predominance of subtype 1.1.

AIM: This study was conducted for the isolation and molecular characterization of bovine herpesvirus 1 (BoHV-1) isolated from the nasal and vaginal swabs collected from naturally infected cattle showing clinical symptoms of the respiratory disease. MATERIALS AND METHODS: Isolation of BoHV-1 virus performed on clinical samples collected from 65 cattle from five states of India. The BoHV-1 isolates were further confirmed by polymerase chain reaction (PCR) using primers specific for glycoprotein B (gB) genomic region. PCR amplification was performed using previously published gB gene-specific primer pairs. gB PCR amplicons obtained from all isolates were sequenced, and phylogenetic analysis was performed using software. RESULTS: A total of 12 samples were found positive in cell culture isolation. 11 isolates showed the visible cytopathic effect on Madin-Darby bovine kidney after 72 h. Partial sequence analysis of gB gene of all isolates revealed 99.0-100% homology between them. All isolates showed 99.2-99.8% homology with Cooper stain. CONCLUSION: BoHV-1.1 is the predominant circulating subtype of BoHV in India, and all isolates have homology with Cooper stain.

Pressure-jump studies of the folding/unfolding of trp repressor.

The dimeric protein, trp apo-repressor of Escherichia coli has been subjected to high hydrostatic pressure under a variety of conditions, and the effects have been monitored by fluorescence spectroscopic and infra-red absorption techniques. Under conditions of micromolar protein concentration and low, non-denaturing concentrations of guanidinium hydrochloride (GuHCl), tryptophan and 8-anilino-1-naphthalene sulfonate (ANS) fluorescence detected high pressure profiles demonstrate that pressures below 3 kbar result in dissociation of the dimer to a monomeric species that presents no hydrophobic binding sites for ANS. The FTIR-detected high pressure profile obtained under significantly different solution conditions (30 mM trp repressor in absence of denaturant) exhibits a much smaller pressure dependence than the fluorescence detected profiles. The pressure-denatured form obtained under the FTIR conditions retains about 50 % alpha-helical structure. From this we conclude that the secondary structure present in the high pressure state achieved under the conditions of the fluorescence experiments is at least as disrupted as that achieved under FTIR conditions. Fluorescence-detected pressure-jump relaxation studies in the presence of non-denaturing concentrations of GuHCl reveal a positive activation volume for the association/folding reaction and a negative activation volume for dissociation/unfolding reaction, implicating dehydration as the rate-limiting step for association/folding and hydration as the rate-limiting step for unfolding. The GuHCl concentration dependence of the kinetic parameters place the transition state at least half-way along the reaction coordinate between the unfolded and folded states. The temperature dependence of the pressure-jump fluorescence-detected dissociation/unfolding reaction in the presence of non-denaturing GuHCl suggests that the curvature in the temperature dependence of the stability arises from non-Arrhenius behavior of the folding rate constant, consistent with a large decrease in heat capacity upon formation of the transition state from the unfolded state. The decrease in the equilibrium volume change for folding with increasing temperature (due to differences in thermal expansivity of the folded and unfolded states) arises from a decrease in the absolute value for the activation volume for unfolding, thus indicating that the thermal expansivity of the transition state is similar to that of the unfolded state.

Review of eye plaque dosimetry based on AAPM Task Group 43 recommendations. American Association of Physicists in Medicine.

PURPOSE: AAPM Task Group 43 recently revised the dosimetry recommendations for 125I seeds. We reviewed these guidelines and studied the effects of the recommendations on the prescription absorbed dose for patients who have undergone eye plaque therapy in our clinic. METHODS AND MATERIALS: 95 consecutive patients were chosen for this study. Absorbed doses at various points of clinical interest were computed based on conventional dose calculation algorithm (3, 4, 7) and TG-43 recommendations. For three representative plaques chosen, the seeds are approximated by isotropic point and line sources, respectively, and absorbed doses were calculated at all points on the central axis of the plaque. RESULTS: For apical heights shorter than 5 mm, treatment plans using model 6711 seeds delivered 10-13% lower absorbed doses than that calculated previously. For lesions with apical heights 5 mm or larger, the absorbed dose was 6-12% lower than prescribed. Calculations for model 6702 seeds indicated that TG-43 recommendations would produce 0-6% lower absorbed doses. Point doses calculated along the central axis of the plaque and isodose distributions at various levels showed that point source approximation of the seeds was clinically acceptable. CONCLUSIONS: TG-43 recommendations, if implemented, would result in lower absorbed doses unless the dose prescription is modified. The clinicians need to be aware of the dosimetric implications of these recommendations. The seeds may be approximated by isotropic point sources.

Killing of Histoplasma capsulatum by macrophage colony stimulating factor-treated human monocyte-derived macrophages: role for reactive oxygen intermediates.

The interaction of human macrophages with the yeast-form of Histoplasma capsulatum was studied. The use of culture and a short-term assay period instead of microscopy gave direct evidence of the fungicidal activity of human macrophages. The present study reports the novel finding of fungicidal activity of macrophages derived from monocytes in the presence of macrophage colony-stimulating-factor (MCSF). The induction of fungicidal activity by this cytokine was dose dependent. MCSF at 10,000 U/ml was optimal with 73(SD3)% killing. Inhibition of macrophage killing by superoxide dismutase (SOD), but not catalase (CAT) or N-monomethyl-L-arginine (NMMA), established the role of the superoxide anion in the killing mechanism. The fungistatic activity of MCSF-derived human macrophages in a 24-h assay was also dose dependent and was not inhibited by SOD, CAT or NMMA. MCSF at 10,000 U/ml produced optimal macrophage fungistatic activity, 34.6(SD4)%.

Differential detection of Newcastle disease virus strains by degenerate primers based RT-PCR.

Degenerate primers based RT-PCR (previously described by [Avian Dis 26 (1997) 837]) has been used for the detection and differentiation of Newcastle disease (ND) viruses. Two sets of primers (A+B and A+C), with common forward primer and distinct reverse degenerate primers, designed from fusion protein gene encoding for cleavage site, could differentiate virulent and avirulent Newcastle disease viruses (NDV). Both sets of primers amplified "F" gene sequence of virulent (velogenic and mesogenic) viruses, whereas in avirulent strains, amplification was only with primer set A+C. Total 10 NDV isolates and two clinical samples including both known and unknown pathotypes, were checked. Based on amplification results 5 viruses were found to be virulent type and 6 as avirulent with one of the two clinical samples, earlier positive by RT-PCR using non-degenerate "F" gene specific primers was found negative in this study. The technique has been found to be a simple and quick for the detection and differentiation of virulent and avirulent NDV, which is important for control of the disease in the events of the outbreaks.

Extraction and separation studies of tellurium(IV) with tris-(2-ethyl hexyl) phosphate.

A method is proposed for the extraction of microgram levels of tellurium(IV) from halide media with tris-(2-ethyl hexyl) phosphate dissolved in toluene as extractant. The optimum conditions have been evaluated from a critical study of acid concentration, extractant concentration, period of equilibration and effect of solvent. Tellurium ion from the organic phase is stripped with water and determined spectrophotometrically with stannous chloride. The method affords binary separation of tellurium from copper, bismuth, gold and selenium and is applicable to the analysis of alloy samples and synthetic mixtures. The method is fast, accurate and precise.

Field evaluation of formalin inactivated Kyasanur forest disease virus tissue culture vaccine in three districts of Karnataka state.

A formalin inactivated Kyasanur forest disease (KFD) virus tissue culture vaccine produced by the health department of the State Government of Karnataka at Shimoga was administered in Shimoga, Uttar Kannada and Chikmangalur districts during 1990-92 KFD epidemic seasons. The selection of places for vaccination was based on the prevalence of KFD activity in previous years; villages adjacent to KFD affected areas and the villages from which mortality in monkeys was reported. A total of 284 villages was covered under vaccination; 26850 individuals received one dose whereas, 61302 received two doses of vaccine. No untoward reaction was observed in any of the vaccinees. In the 72 KFD affected villages there were 14 patients among 9072 and 10 among 21083 vaccinees receiving one and two doses respectively, whereas 325 patients were reported among 37373 unvaccinated individuals during the same period. In 1990-91 the number of males patients was more than females whereas, in 1991-92 the ratio was reserved. On analysis indicated that the vaccine has a highly significant protective effect.

Indoor radon exposure and active and passive smoking in relation to the occurrence of lung cancer.

Exposure to indoor radon and radon daughters is currently attracting great interest as a possible cause of lung cancer. This concern is supported by several studies, most of them relatively small in numbers or weak in the assessment of exposure. This study encompasses 177 persons with lung cancer and 677 noncancer referents, all deceased and with 30 years or more of residency in the same house in an area with radon-leaking alum shale deposits in the central part of southern Sweden. Exposure categories based on building material, type of house, and ground conditions were created, but measurements of the indoor radon daughter concentration were also made for 142 cases and 264 referents. Active and passive smoking was ascertained through questionnaires sent to the next-of-kin. Overall, the lung cancer risk was approximately twofold with regard to the categories of assumed radon daughter exposure for the rural sector of the population but not for the same categories of the urban sector, possibly because of less precise exposure assessment and influence from other factors. Occasional and passive smokers, as well as passive smokers alone, had a particularly increased risk of lung cancer in association with the increased exposure categories.

The changing pattern of surgical pathology of the thyroid gland in Zambia.

Review of reports of thyroid tissue sent for histopathology to the Department of Pathology at the University Teaching Hospital, Lusaka (UTH) by one of the five general surgical units at the institution during the period January 1981-December 1990 shows a 20pc decrease in the incidence of colloid goitre and doubling of the incidence of the adenoma and carcinoma when compared to the study done at the Central Hospital, Kitwe in the late sixties. In our study for the general pathology of the thyroid gland female to male ratio is 7.4:1; 1.25:1 when only thyroid cancers are considered. The incidence of papillary carcinoma is substantially lower than in the west. Recently, there has been an increased incidence of thyroid abscesses associated with HIV infection. Knowledge of the local pattern of surgical pathology of an organ is important for planning the operative management more effectively but in many developing countries, such as ours, this information is not readily available. Goitre is common in Zambia and to our knowledge to date there is only one comprehensive report done during the period January 1966, to March 1971. This deals with the Northern and Western regions of the country. This study was undertaken to look into the surgical pathology of the thyroid gland seen at the University Teaching Hospital, Lusaka, which is in the central part of the country.