Correlation of partial env gene sequences with disease progression parameters in HIV-positive pregnant women from India.

Ever since the beginning of the epidemic of HIV, one of the poignant aspects of HIV infection is transmission of the virus from mother to child. It is not known whether pregnancy accelerates the progression of HIV infection from a clinically asymptomatic stage to a progressive clinical phase. Present study was carried out to understand disease progression in pregnant women from India. We studied co-receptor utilization (the major determinant of HIV disease progression), N-glycosylation sites, and sequence variability. Blood samples were collected from 25 HIV sero-positive patients, eleven from the antenatal risk group (experimental group), nine from heterosexual male, and five from heterosexual female risk group (control group). Partial env gene was amplified by PCR and sequenced. BLAST search and phylogenetic analysis were used to determine the subtype. The deduced amino acid sequence of the V3 region was used to predict co-receptor, determine sequence variability and N-glycosylation site. The experimental group comprising the antenatal risk group did not exhibit any difference in terms of co-receptor, N-glycosylation, and sequence variability when compared with the control, non-pregnant group. Pregnancy does not seem to accelerate the clinical course of HIV infection. The female body during the gestation phase possibly acquires certain strategies to impede or at least alleviate the disease progression during the crucial immune-compromised pregnancy phase, which would otherwise adversely affect the mother as well as the fetus during the infection.

The first dominant co-circulation of both dengue and chikungunya viruses during the post-monsoon period of 2010 in Delhi, India.

Independent outbreaks of dengue virus (DENV) infection and sporadic cases of chikungunya virus (CHIKV) have been recorded in the metropolitan city of Delhi on several occasions in the past. However, during a recent 2010 arboviral outbreak in Delhi many cases turned negative for DENV. This prompted us to use duplex reverse transcriptase-polymerase chain reaction (D-RT-PCR) to establish the aetiology of dengue/chikungunya through sequencing of CprM and E1 genes of dengue and chikungunya viruses. Interestingly, for the first time, both DENV and CHIKV co-circulated simultaneously and in equally dominant proportion during the post-monsoon period of 2010. DENV-1 genotype III and the East Central South African genotype of CHIKV were associated with post-monsoon spread of these viruses.

Genetic analysis of precore/core and partial pol genes in an unprecedented outbreak of fulminant hepatitis B in India.

We investigated an unprecedented outbreak of fulminant hepatitis B virus (HBV) that occurred in Modasa, Gujarat (India) in 2009. Genomic analysis of all fulminant hepatic failure cases confirmed exclusive predominance of subgenotype D1. A1762T, G1764A basal core promoter (BCP) mutations, insertion of isoleucine after nt 1843, stop codon mutation G1896A, G1862T transversion plus seven other mutations in the core gene caused inhibition of HBeAg expression implicating them as circulating precore/BCP mutant virus. Two rare mutations at amino acids 89 (Ile→Ala) and 119 (Leu→Ser) in addition to other mutations in the polymerase (pol) gene may have caused some alteration in either of four pol gene domains to affect encapsidation of pregenomic RNA to enhance pathogenicity. Sequence similarity among patients' sequences suggested an involvement of a single hepatitis B mutant strain/source to corroborate the finding of gross and continued usage of HBV mutant-contaminated syringes/needles by a physician which resulted in this unprecedented outbreak of fulminant hepatitis B. The fulminant exacerbation of the disease might be attributed to mutations in the BCP/precore/core and pol genes that may have occurred due to selection pressure during rapid spread/mutation of the virus.

Emergence of pandemic 2009 influenza A H1N1, India.

BACKGROUND & OBJECTIVES: Pandemic H1N1 caused deluge of cases from 74 countries and prompted World Health Organization to raise warning to phase 6. The present study was conducted on throat and nasal swab samples received and tested at National Centre for Disease Control, Delhi, India during 2009-2010 to collect epidemiological and clinical information on positive cases. METHODS: Throat and nasopharyngeal swabs from category C influenza A H1N1 patients during May 2009-September 2010 along with their clinico-epidemiological details were collected from identified hospitals from Delhi and other States. Samples were tested by Real time reverse transcriptase PCR using primers and probes developed at CDC, Atlanta for four influenza target genes. RESULTS: A total of 33,751 samples, both throat and nasal swab samples from each patient were tested for H1N1 influenza virus, of which, 7943 (23.5%) were positive for pandemic influenza A H1N1 and 3759 (11.1%) were positive for influenza A (seasonal flu). Maximum number of positive cases (N=2792, 35.1%) were from 20-39 yr age group, comprising 1790 (22.5%) males and 1182 (14.8%) females. Only 2620 (33%) positive cases were close contact of influenza A H1N1 positive patient. Majority cases presented (N=2792, 35.1%) with fever 7005 (88.1%), followed by 6133 cases (77.2%) exhibiting fever and cough, 377 (4.7%) complained of fever, cough, nasal catarrh and 362 (4.5%) cases had fever with shortness of breath. INTERPRETATION & CONCLUSIONS: The study showed a peak of cases of pandemic influenza A H1N1 in December 2009 and indicated predominance of H1N1 positive cases among 20-39 yr age group and among males compared to females.

Blood tests and histological correlates in children with eosinophilic oesophagitis.

AIM: To determine the relationship between blood tests and oesophageal histology in Eosinophilic oesophagitis (EoE). METHODS: All children diagnosed with EoE at one hospital from 2000 to 2009 were considered for inclusion in this study. Three blood test results were analysed, blood eosinophil count, serum total immunoglobulin E (IgE) and radioallergosorbent tests (RAST) to common food allergens. Oesophageal histology was prospectively re-reviewed, and mean eosinophil counts were enumerated. Blood test results were correlated with oesophageal eosinophil counts using Spearman's rank test. RESULTS: Forty children (70% boys) were included in this study, median age at diagnosis 6.5 years (range 0-15). At the time of diagnosis, 78% of children had a raised blood eosinophil count, 90% had a raised serum total IgE and 83% had one or more positive RAST tests. The mean oesophageal eosinophil count was significantly correlated with both blood eosinophil count (p=0.008) and serum total IgE level (p=0.008). CONCLUSION: This study shows that blood tests are often abnormal in children with EoE at the time of diagnosis. Our data demonstrate an association between histological abnormalities and blood test results in children with EoE.

Evaluation of nucleic acid probe for rapid identification of Mycobacterium tuberculosis complex in extra-pulmonary culture isolates.

Patients infected with Non-tuberculous mycobacteria (NTM) usually do not respond to conventional anti-tubercular treatment and are misdiagnosed as infection with multi-drug resistant strains of mycobacterium tuberculosis (M.tb) due to lack of correct species identification, particularly in the developing countries like India. One of the challenges faced by clinicians in the treatment of tuberculosis is the absence of an easy, reliable and rapid identification tool that can accurately differentiate disease caused by M.tb complex from NTM. Keeping this in consideration, the performance of species specific nucleic acid probe i.e. Accuprobe was assessed and compared with conventional niacin production, nitrate reductase assay techniques for identification of M.tb complex in 80 mycobacterial isolates obtained from different extra-pulmonary sites. Accuprobe identified 62 isolates (77.5%) as M. tuberculosis complex and remaining 18 isolates (22.5%) as NTM whereas 64 isolates (80%) were identified as M.tb and rest 16 (20%) were interpreted as NTM by conventional biochemical techniques. The overall agreement between both techniques was 96.9% The sensitivity, specificity, positive predictive value(PPV) and negative predictive value (NPV) shown by accuprobe were 96.9%, 100%, 96.9%, and 88.9% respectively. Thus, accuprobe has showed impressive sensitivity and specificity giving results in < 3 hrs from culture-positive isolates and have sure edge over conventional biochemical methods which are, nonetheless, labour intensive and cumbersome to perform thus delaying prompt mycobacterial identification.

Reduced quality of life in children with gastro-oesophageal reflux disease.

AIM: To assess self-reported Quality of life (QoL) in children with Gastro-oesophageal reflux disease (GORD) aged 5-18 and compare this with both disease and healthy control children in a prospective consecutive sample. METHODS: All children attending a tertiary paediatric gastroenterology clinic from February 2009 to May 2009 with GORD, chronic constipation and inflammatory bowel disease (IBD) were asked to complete the validated PedsQL generic QoL assessment (self-report) at their clinic appointment. The PedsQL considers physical, emotional, social and school domains and is scored from 0 to 100. Healthy children were also recruited from the same site. Groups were compared using the independent samples Student's t-test. RESULTS: A total of 184 children completed the assessment [103 (56%) male, mean age 10.7 years +/- 3.3] including 40 children with GORD, 44 with chronic constipation, 59 with IBD and 41 healthy children. QoL was significantly lower in the GORD group compared with both children with IBD (74 vs. 82) and healthy children (74 vs. 84), and was comparable to that of children with chronic constipation (74 vs. 74). CONCLUSIONS: Self-reported QoL in children with GORD attending a tertiary paediatric gastroenterology clinic is significantly reduced compared with both healthy children and children with IBD.

Concomitant TB and cryptococcosis in HIV-infected patients.

Four cases of concomitant tuberculosis and cryptococcosis infection in HIV-positive patients are described. As the HIV pandemic progresses and the proportion of patients with end-stage disease increases, a high suspicion of incidence and unusual forms of infections must always be kept in mind.

Histone as future drug target for malaria.

Malaria continues to be a major cause of mortality and morbidity in tropical countries and affecting around 100 countries of the world. As per WHO estimates, 300-500 million are being infected and 1-3 million deaths annually due to malaria. With the emerging knowledge about genome sequence of all the three counterparts involved in the disease of malaria, the parasite Plasmodium, vector Anopheles and host Homo sapien have helped the scientists to understand interactions between them. Simultaneous advancement in technology further improves the prospects to discover new targets for vaccines and drugs. Though the malaria vaccine is still far away in this situation there is need to develop a potent and affordable drug(s). Histones are the key protein of chromatin and play an important role in DNA packaging, replication and gene expression. They also show frequent post-translation modifications. The specific combinations of these posttranslational modifications are thought to alter chromatin structure by forming epigenetic bar codes that specify either transient or heritable patterns of genome function. Chromatin regulators and upstream pathways are therefore seen as promising targets for development of therapeutic drugs.

Effect of obesity and insulin resistance on diabetic control.

UNLABELLED: Diabetes Mellitus in obese and non-obese Indian individuals. AIMS: Effect of Obesity and insulin resistance on diabetic control. SETTINGS AND DESIGN: 50 each groups Diabetic individuals obese and non-obese. METHODS AND MATERIAL: On selected 50 each group diabetic patient and normal, following blood investigations has been performed-Plasma Glucose, Glycohemoglobin and Serum Insulin. STATISTICAL ANALYSIS USED: Individuals patient's results were analyzed and compared with the normal controls. RESULTS AND CONCLUSION: The changes in glycosylated haemoglobin are mainly proportional to the post lunch glucose level (r=0.773) (p<0.01) and not correlated to circulating insulin or the body mass index. However the levels were higher in obese diabetes (Type I and II both) than in non-obese. Mechanism of resistance in insulin receptor interactions due to obesity is well known. However, obesity does not seem to affect directly glycosylated haemoglobin. Under such circumstances, the reduction of weight for a diabetic person can improve sugar control by minimizing insulin resistance and thereby can improve glycosylated haemoglobin levels.

A Retrospective Analysis of Apheresis Donor Deferral and Adverse Reactions at a Tertiary Care Centre in India.

INTRODUCTION: With increasing demand of platelet component each day, blood bank plays a pivotal role in ensuring supply of safe blood as and when required. Plateletpheresis procedure is a relatively simple, safe and important adjunct to blood bank inventory. However, recruitment of healthy blood donors is a challenge that the health industry is facing today. AIM: To determine the reasons and rates of apheresis donor deferral along with investigation of adverse reactions encountered during the procedure. MATERIALS AND METHODS: Records of single donor apheresis were retrospectively analysed from 1st January 2010 to 31st December 2014. The study was carried out at Blood Bank, Safdarjung Hospital, New Delhi, India. The donor details that were studied included - age, sex, type of donation (voluntary/replacement/ repeat), reason for donor deferral and type of adverse reaction, if encountered during the procedure. RESULTS: Among the 478 donors screened for plateletpheresis procedure during a study period of 5 years, 134 (28.03%) were deferred. Temporary deferrals accounted for majority (93.28%) of the deferrals. Low platelet count (50.75%) was the main reason of donor deferral followed by low haemoglobin (20.89%). Amongst the 344 selected donors, 15 (4.36%) had some type of adverse reaction associated with the procedure. CONCLUSION: We suggest that the selection criteria for plateletpheresis donors should be revised to deal with shortage of apheresis donors. The criteria regarding minimum pre-procedure platelet count (above1.5 lac/µl) and haemoglobin (above 12.5 g/dl) need to be lowered so as to suit the Indian scenario. The lower adverse reaction rates, 14/344 (4.06%) associated with this procedure encourages safety of donors and is important in recruitment of new donors.

Medicinal plants of the family Caryophyllaceae: a review of ethno-medicinal uses and pharmacological properties.

Several species of the family Caryophyllaceae are widely used by many ethnic communities as traditional medicine throughout the world. The highest number of plants of the family are used in Chinese traditional medicine. The ethnopharmacologial studies of this family indicate that plants of the family possess anticancer, antibacterial, antifungal, antiviral, antioxidant, and anti-inflammatory properties. Other miscellaneous properties reported are ribosome inactivation properties, inhibition of prostatic enlargement in rats, and inhibition of intestinal enzyme carboxyelasterase in rats, cerebro-protective activity, and antiobesity in rats. Few reviews have been published yet, providing information regarding medicinal plants of the family and their biomedical properties. All published reviews have focused either on a particular taxa or a few species. The present review is focused on the traditional medicinal uses of the plants of the family Caryophyllaceae along with phytochemical and pharmacological studies of the family. A study of the literature revealed significant traditional medicinal importance of the family. Major chemical constituents of Caryophyllceae are saponins, Phytoecdysteroids, benzenoids, phenyl propanoids, and nitrogen containing compounds. The most important property of plants of the family is anticancer activity and is shown by the large number of plant species studied. This review of traditional medicinal and pharmacological uses of plants of the family, provide a ground for future research in the family.

Asymptomatic reproductive tract infections/sexually transmitted infections among HIV positive women.

This study aims to highlight the importance of screening all HIV positive women for various reproductive tract infections/sexually transmitted infections (RTIs/STIs) irrespective of symptoms and to determine its occurrence in asymptomatic HIV positive women. Relevant specimens were collected for diagnosis of various RTIs/STIs. STIs were diagnosed in nearly one-third of the HIV positive asymptomatic patients which is quite high. The national strategy for STIs/RTIs control misses out large number of asymptomatic RTIs/STIs in HIV positive women which is responsible for silently transmitting these infections in the community. So this strategy should be modified to include screening of all HIV positives women irrespective of symptoms of STIs/RTIs.

Rising trend of antimicrobial resistance among Neisseria gonorrhoeae isolates and the emergence of N. gonorrhoeae isolate with decreased susceptibility to ceftriaxone.

CONTEXT: Gonorrhoea is one of the most common sexually transmitted infections (STI) in developing countries and is a global health problem. AIMS: To analyze the trend of antimicrobial susceptibility of Neisseria gonorrhoeae isolates over the years, in a tertiary care hospital of North India. SETTINGS AND DESIGN: The study population comprised males with urethritis and females with endocervicitis attending STI clinic of our hospital. MATERIALS AND METHODS: In our STI laboratory, all gonococcal isolates are subjected to antimicrobial susceptibility testing by disc diffusion method as per CLSI guidelines. ß-Lactamase production is determined by chromogenic cephalosporin test. Minimum Inhibitory Concentration (MIC) for ceftriaxone is determined by E-test. STATISTICAL ANALYSIS USED: Data were expressed as percentages. The differences in percentages were tested for statistical significance by using χ2 test and P values were determined. RESULTS: The percentage of penicillinase producing N. gonorrhoeae (PPNG) increased from 8% in 1995-96 to 20% in 2004-05 and 88% in 2011-2013. Quinolone-resistant N. gonorrhoeae (QRNG) showed a significant increase from 12% in 1995-96 to 98.3% in 2004-05, while 84% isolates were found to be QRNG by 2011-2013. In January 2013 we detected our first gonococcal isolate with decreased susceptibility to third-generation cephalosporins; Ceftriaxone, Cefixime and Cefpodoxime (MIC for ceftriaxone = 0.19 µg/ml). CONCLUSIONS: The results of our study highlighted an alarming increase in the percentage of PPNG and QRNG strains over the years. Emergence of N. gonorrhoeae isolates with decreased susceptibility to third-generation cephalosporins is a cause of concern and thus emphasises the importance of antimicrobial susceptibility testing.

Complete nucleotide sequence of the 6 kb element and conserved cytochrome b gene sequences among Indian isolates of Plasmodium falciparum.

The malaria parasite contains a nuclear genome with 14 chromosomes and two extrachromosomal DNA molecules of 6 kb and 35 kb in size. The smallest genome, known as the 6 kb element or mitochondrial DNA, has been sequenced from several Plasmodium falciparum isolates because this is a potential drug target. Here we describe the complete nucleotide sequence of this element from an Indian isolate of P. falciparum. It is 5967 bp in size and shows 99.6% homology with the 6 kb element of other isolates. The element contains three open reading frames for mitochondrial proteins-cytochrome oxidase subunit I (CoI), subunit III (CoIII) and cytochrome b (Cyb) which were found to be expressed during blood stages of the parasite. We have also sequenced the entire cyb gene from several Indian isolates of P. falciparum. The rate of mutation in this gene was very low since 12 of 14 isolates showed the identical sequence. Only one isolate showed a maximum change in five amino acids whereas the other isolate showed only one amino acid change. However, none of the Indian isolates showed any change in those amino acids of cyb which are associated with resistance to various drugs as these drugs are not yet commonly used in India.

Whole blood samples as an alternative to serum for detection of immunity to measles virus by ELISA.

The aim of this study was to evaluate enzyme linked immunosorbent assay (ELISA) as a testing strategy for detection of antibodies against measles virus from microquantities of blood soaked onto filter paper. We studied 165 healthy children in the age group of 1 to 2 years, attending the outpatient department of pediatrics. Two sets of samples were collected from each child. One by venipuncture and the other on Whatman filter paper-3 discs of 20 mm size by finger or heel prick so that each strip is completely soaked with blood on both sides. These were tested for measles virus antibodies by ELISA using Melotest measles IgG commercial ELISA kit manufactured by Melotec S. A. (Barcelona, Spain). The resulting sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the filter paper (FP) ELISA compared to serum ELISA was 100, 90, 97.8, and 100%, respectively. The correlation coefficient r = 0.93% (p < 0.001) and the agreement between the two techniques was 98% as calculated by the Kappa statistical method. The present study has found filter paper testing by ELISA to be a promising qualitative technique for detection of immunity against measles.

Possible existence of a higher coverage quasi-one-dimensional phase of argon adsorbed on bundles of single-walled carbon nanotubes.

We present results of Ar adsorption isotherms at very low coverages in the first layer and, beyond monolayer completion, on bundles of close-ended single-walled carbon nanotubes. The low coverage results were used to determine the isosteric heat of adsorption and the binding energy of Ar in the groove sites in the first layer. The higher coverage results show evidence of the possible formation of a second-layer groove phase, beyond monolayer completion. Our results for higher coverages are compared with recent computer simulations for this system.

Comparison of two blood collection techniques for measles antibody detection in children.

BACKGROUND & OBJECTIVES: Though several seroprevalence and seroconversion studies have been conducted for measles using the filter paper haemagglutination inhibition (FPHAI), very few studies have compared the conventional serum HAI with the FPHAI. The present study was aimed at the evaluation of whole blood samples on filter paper as an alternative to serum specimens for detection of antibodies to measles virus. METHODS: Serum and whole blood samples soaked on filter paper were collected from 165 randomly selected healthy children in the age groups of 1-2 yr. HAI test was performed on both sets of samples and the results compared. RESULTS: Ninety samples that gave a titre of 8 or more by serum HAI also had titres of > or = 8 by FPHAI showing 100 per cent agreement between the two assays. Seventeen samples that had a serum HAI titre of 2 and 4 were missed by FPHAI due to the starting dilution of 8 of the latter. However, FPHAI gave no false positive results compared with serum HAI. INTERPRETATION & CONCLUSION: Collection of the FP samples by finger prick is more acceptable and requires less expertise than venepuncture. Thus, in spite of the small percentage of missed cases by the filter paper method, this technique of sampling was found to be a convenient and reliable alternative to venepuncture, for detection of measles virus antibody especially in large scale seroepidemiological studies.

Relationship of Xba1 and EcoR1 polymorphisms of apolipoprotein-B gene to dyslipidemia and obesity in Asian Indians in North India.

BACKGROUND: Genetic investigation of dyslipidemia and obesity prevalent in the Indian population form the basis of this study. METHODS AND RESULTS: The frequency of restriction fragment length polymorphisms (Xba1 and EcoR1) of the apolipoprotein-B gene was investigated in a case-control study of 30 hyperlipidemic and 40 normolipidemic subjects. By univariate analysis, old age, higher body mass index, waist-hip ratio and sum of four skinfolds were found to be significantly associated with hyperlipidemia. The frequencies of X- and E+ alleles of the apolipoprotein-B gene were significantly higher in North Indians in the state of New Delhi (0.83 and 0.91, respectively) as compared to the observations made in Caucasians in previous studies, but was similar to the frequency reported in Indians settled in Singapore and the UK. There were no significant differences in the allele or genotype frequencies of either Xba1 or EcoR1 polymorphisms between the hyperlipidemic and normolipidemic groups. On multiple logistic regression analysis considering body mass index, waist-hip ratio, percentage body fat and genotypes as independent variables, no association was observed between the apolipoprotein-B genotypes and serum lipid components. Further, there were no associations between apolipoprotein-B polymorphisms and generalized obesity (as assessed by body mass index, sum of four skinfolds, and percentage total body fat) and abdominal obesity (as measured by waist circumference and waist-hip ratio). CONCLUSIONS: We conclude that apolipoprotein-B (Xba1 and EcoR1) polymorphisms do not appear to influence serum lipid levels and parameters of generalized andregional obesity in the study sample.