Umbilical cord blood (UCB) has been shown to be a suitable source of haematopoietic stem cells (HSCs) for haematopoietic reconstitution. An increase in the number of UCB transplants indicates an expansion of utility in a broad spectrum of disease conditions. Along with the advantages, UCB also has limitations, and hence several investigators are working to further optimize UCB for this use. Beyond haematopoietic transplantation, additional potential applications of UCB include immunotherapy, tissue engineering and regenerative medicine. UCB banking has improved with time largely due to involvement of professional organizations and their published standards. However, accreditation of these organizations remains voluntary, and in India three of ten banks are public with the remaining being private. Only one public and one private bank are American Association of Blood Banks (AABB) accredited in India. Government agencies need to provide regulatory and safety oversight, which is lacking in serveral countries. Public policy regarding UCB is in its infancy throughout most of the world. Ethical issues, including access to UCB banking and use as therapy for diseases other than haematological and metabolic disorders are in the early phase of trials and remain speculative.

Background & objectives : Diagnosis for Mycoplasma pneumoniae usually relies on serological tests. PCR technology has some advantages but also limitations. The optimal selection for these tests still needs discussion. This paper reviews the overall diagnostic accuracy of PCR versus serological assays for diagnosis of M. pneumoniae infections and to identify factors associated with heterogeneity of results. Methods : MEDLINE and Embase databases were searched. Articles meeting the selection criteria were retrieved for data collection and analysis. Studies were assessed for methodological quality using QUADAS. Hierarchial summary receiver operating characteristic (HSROC) model was used to estimate summary ROC curve. Results : Initial meta-analysis showed a summary estimate of sensitivity (SEN) 0.62 (95% CI, 0.45-0.76), and specificity (SPE) 0.96 (95% CI, 0.93-0.98). Subgroup analyses were performed to identify factors associated with heterogeneity. For different gene targets, reference standards, subjects (children or adults) and different PCR types, these aspects can generate results of heterogeneity. The 16s rDNA target and adult subjects and real-time PCR may have better test results for PCR. Interpretation & conclusions : Commercial PCR tests generated consistent results with high specificity but a lower and more variable sensitivity. The findings suggest commercial PCR tests having superiorities in diagnosing M. pneumoniae infections but still cannot replace serology. PCR plus serology could be good screening tests for reliable and accurate diagnosis of M. pneumoniae.

Antibiotic resistance, a global concern, is particularly pressing in developing nations, including India, where the burden of infectious disease is high and healthcare spending is low. The Global Antibiotic Resistance Partnership (GARP) was established to develop actionable policy recommendations specifically relevant to low- and middle-income countries where suboptimal access to antibiotics - not a major concern in high-income countries - is possibly as severe a problem as is the spread of resistant organisms. This report summarizes the situation as it is known regarding antibiotic use and growing resistance in India and recommends short and long term actions. Recommendations aim at (i) reducing the need for antibiotics; (ii) lowering resistance-enhancing drug pressure through improved antibiotic targeting, and (iii) eliminating antibiotic use for growth promotion in agriculture. The highest priority needs to be given to (i) national surveillance of antibiotic resistance and antibiotic use - better information to underpin decisions on standard treatment guidelines, education and other actions, as well as to monitor changes over time; (ii) increasing the use of diagnostic tests, which necessitates behavioural changes and improvements in microbiology laboratory capacity; (iii) setting up and/or strengthening infection control committees in hospitals; and (iv) restricting the use of antibiotics for non-therapeutic uses in agriculture. These interventions should help to reduce the spread of antibiotic resistance, improve public health directly, benefit the populace and reduce pressure on the healthcare system. Finally, increasing the types and coverage of childhood vaccines offered by the government would reduce the disease burden enormously and spare antibiotics.

Background & objectives : Multiple drug resistance in epilepsy is a common problem and one third of epilepsy patients remain non responsive to antiepileptic drug (AED) therapy. In this study we aimed to investigate the relationship between the genetic polymorphism of cytochrome P450 genes, namely CYP2C9 and CYP2C19 with multiple drug resistance in epilepsy patients. Methods : A total of 402 patients with epilepsy were enrolled in this study; 128 were drug resistant and 274 were drug responsive. The peripheral blood samples of the patients with epilepsy were collected. Drug compliance was confirmed in 20 per cent patient population using HPLC. Genotyping of CYP2C9 (*2 and *3), and CYP2C19 (*2 and*3) was carried out by PCR-RFLP. Results : The genotype frequencies of CYP2C9 430 C>T (*2 variant) and CYP2C9 1075 A>C (*3 variant) did not differ significantly in drug resistant versus responsive patients. After combining CYP2C9 *2 and CYP2C9 *3, the frequency of CYP2C9*1/*3 was significantly lower in drug resistant as compared to drug responsive epilepsy patients (P=0.03, OR=0.53, 95%CI=0.30-0.95). Similarly, combined frequency of all the slow and poor metabolizer variants (2C9 *1/*2, *1/*3 and *2/*3) was also lower as compared to drug resistant group (P=0.03, OR=0.60, 95% CI 0.38-0.96). There was no significant differences in genotypic or allelic distribution of CYP2C190*2 while CYP2C19*3 was monomorphic in northern Indian population. Interpretation & conclusions : Our results demonstrated significant involvement of CYP2C9 genetic variants in the modulation of epilepsy pharmacotherapy confirming the important role of CYP2C9 mutants preventing epilepsy patients from developing drug resistance.

Background & objectives : MMR vaccine in a two dose schedule has successfully eliminated measles, mumps and rubella from many developed countries. In India, it is not a part of national immunization programme but is included in the State immunization programme of Delhi as a single dose between 15-18 months. This prospective study was carried out to assess the extent of seroprotection against these three diseases in immunized children and to study the immune response to a second dose of MMR. Methods : Consecutive children aged 4-6 yr, attending the immunization clinic of a tertiary care hospital in Delhi for routine DT vaccination, were enrolled. Second dose of MMR was given and pre- and post-vaccination antibody levels were compared. Results : The pre-vaccination percentage seropositivity observed in the 103 children recruited, was 20.4 per cent for measles, 87.4 per cent for mumps and 75.7 per cent for rubella. Amongst the 84 children who were followed up after the second dose, the percentage seroprotection for measles rose from 21.4 (18/84) to 72.6 per cent (61/84) and 100 per cent became seroprotected to mumps and rubella. Interpretation & conclusions: The percentage of children protected against measles was found to be alarmingly low which needs to be investigated. Though the observed protection against mumps and rubella was adequate, its durability was not known. The need for re-appraisal of the current MMR immunization policy is called for by carrying out longitudinal studies on a larger sample.

Background & objectives : Peak bone mass, a major determinant of osteoporosis is influenced by genetic, nutritional, lifestyle and hormonal factors. This study was designed to evaluate the impact of sports training on dietary intake and bone mineral and metabolic parameters in young healthy Indian females. Methods: Healthy female college going students (N=186, sportswomen, 90; controls 96) in the age group of 18-21 yr, residing in New Delhi (India) were evaluated for anthropometry, biochemistry (serum total and ionic calcium, phosphorus, total alkaline phosphatase, 25-hydroxyvitamin D & parathyroid hormone), diet, physical activity and lifestyle. Bone mineral density (BMD) at hip, forearm and lumbar spine were studied using central DXA. Results: Sports related physical activity (3 vs. 0 h/day, P<0.001) and direct sunlight exposure (120 vs. 30 min/day, P<0.001) were significantly higher in sportswomen than in controls with sedentary lifestyle. Significantly higher intake of all macronutrients (energy, protein, carbohydrates and fat) and dietary calcium was noted in the diets of sportswomen. Mean serum 25(OH)D levels were significantly higher (53.0±18.9 vs. 12.9±7.7 nmol/l; P<0.001) while PTH (35.3±17.6 vs. 51.7±44.9 pg/ml; P<0.001) and ALP levels (194.0±51.0 vs. 222.1±51.4 IU/l; P<0.001) were significantly lower in sportswomen when compared to controls. No significant difference was found in ionized calcium and inorganic phosphorus in the two groups. Significantly higher (P<0.001) total BMD and BMD at all sites except femur neck were found in sportswomen than controls (P<0.001). Interpretation & conclusions: Physical activity, optimal nutrition and adequate sun exposure are vital for attaining peak bone mass.

Background & objectives : Hyperprolactinaemia affects testicular functions by influencing hypothalamo-pituitary-testicular (HPT) axis at various levels. Available literature on the level of defect, time course of improvement of gonadal functions and its relation with decline in prolactin levels is scanty. We carried out this study to evaluate the HPT axis in patients with macroprolactinomas, before and six months after cabergoline therapy. Methods: Fifteen men with macroprolactinomas underwent gonadotropin and testosterone response to their respective stimuli before and after six months of cabergoline therapy. Results: Serum prolactin levels decreased after six months of therapy. Pretreatment, mean lutenizing and follicle stimulating hormones (LH and FSH) levels were 2.0 ± 0.4 and 1.4 ± 0.2 IU/l, respectively. However, LH and FSH responses to GnRH were preserved in majority of the patients and LH peaked to 12.1 ± 2.3 IU/l (P<0.01), while FSH to 2.9 ± 0.4 IU/l suggesting the influence of hyperprolactinaemia at the level of hypothalamus with preserved gonadotrope reserve. After cabergoline therapy, there was an increase in basal as well as stimulated LH and FSH levels, though these were not statistically significant when compared to respective pretherapy levels. Basal testosterone (T) levels significantly improved after therapy, but peak T response to hCG was similar at both pre- and post treatment. A significant correlation was observed between peak LH and peak T at baseline (r=0.53, P<0.01) and it further strengthened after therapy (r=0.70, P<0.01). After cabergoline therapy, there was significant improvement in seminal volume, sperm count and motility and sperm count correlated with peak FSH response (r=0.53, P<0.05). Interpretation & conclusions: Hyperprolactinaemia affects testicular functions probably by influencing at the level of hypothalamus resulting in subnormal basal secretion of gonadotropins required for optimal testicular functions.

Background & objectives : WNIN/GR-Ob is a mutant obese rat strain with impaired glucose tolerance (IGT) developed at the National Institute of Nutrition (NIN), Hyderabad, India, from the existing 80 year old Wistar rat (WNIN) stock colony. The data presented here pertain to its obese nature along with IGT trait as evidenced by physical, physiological and biochemical parameters. The study also explains its existence, in three phenotypes: homozygous lean (+/+), heterozygous carrier (+/-) and homozygous obese (-/-). Methods: Thirty animals (15 males and 15 females) from each phenotype (+/+, +/-, -/-) and 24 lean and obese (6 males and 6 females) rats were taken for growth and food intake studies respectively. Twelve adult rats from each phenotype were taken for body composition measurement by total body electrical conductivity (TOBEC); 12 rats of both genders from each phenotype at different ages were taken for clinical chemistry parameters. Physiological indices of insulin resistance were calculated according to the homeostasis model assessment for insulin resistance (HOMA-IR) and also by studying U ^[14] C 2-deoxy glucose uptake (2DG). Results: WNINGR-Ob mutants had high growth, hyperphagia, polydipsia, polyurea, glycosuria, and significantly lower lean body mass, higher fat mass as compared with carrier and lean rats. These mutants, at 50 days of age displayed abnormal response to glucose load (IGT), hyperinsulinaemia, hypertriglyceridaemia, hypercholesterolaemia and hyperleptinaemia. Basal and insulin-stimulated glucose uptakes by diaphragm were significantly decreased in obese rats as compared with lean rats. Interpretation & conclusions : Obese rats of the designated WNIN/GR-Ob strain showed obesity with IGT, as adjudged by physical, physiological and biochemical indices. These indices varied among the three phenotypes, being lowest in lean, highest in obese and intermediate in carrier phenotypes thereby suggesting that obesity is inherited as autosomal incomplete dominant trait in this strain. This mutant obese rat model is easy to propagate, and can easily be transformed to frank diabetes model by dietary manipulation and thus can be used for screening anti-diabetic drugs.

Background & objectives : WNIN/Ob (obese and euglycaemic) and WNIN/GR-Ob (obesity with impaired glucose tolerance), were isolated and established from Wistar rat stock (WNIN). Both strains showed physical, physiological and biochemical indices related to obesity. We present here haematology, histology and pathophysiological changes between the phenotypes of these strains, lean (+/+), carrier (+/-) and obese (-/-). Methods: A total of 72 animals of equal gender consisting of three phenotypes were used for the study. Haematology, organ weights were measured and histopathology of the tissues studied using standard procedures. In 12 lean and obese rats (equal gender) of WNIN/GR-Ob group morphometry of pancreatic islets was done immunohistochemically (IHC). Results: Obese rats of both the strains showed normal haematology (except low platelet count), but exhibited changes in the organ weights and in histopathology in organs like liver, kidney, brain and testis/ovary. Hyperplasia of adipocytes was seen in obese rats as compared to lean and carrier. IHC of obese rat pancreas showed that both islet density and volume were significantly (P<0.05) increased compared to lean littermates. Interpretation & conclusions : The histological and pathophysiological changes seen in these mutants were in tune with obese phenotype exhibited by these animals.

Background & objectives : The research on the association of metabolic syndrome (MS) and substance abuse is scanty. The present research aimed to study the prevalence and correlates of MS among the inpatients at a Drug De-addiction Centre in north India. Methods: Consecutive male subjects (N=110) admitted to a drug de-addiction centre during July to December 2009 with a primary diagnosis of alcohol or opioid dependence were evaluated for the presence of MS as per the International Diabetes Federation (IDF) criteria. Results: The prevalence of MS was 24.6 and 29.3 per cent in alcohol and opioid dependent groups, respectively. MS showed a significant association with the age and body mass index (BMI) in the opioid dependent group. Co-morbid tobacco use was not associated with MS in either group. Interpretation & conclusions : The prevalence of MS in our sample of alcohol and opioid dependent male inpatients was greater than the prevalence of MS in general population, however it was comparable to that reported in physical and other psychiatric disorder populations. Even though the absence of any comparative study limits the generalizability of our findings, results indicate towards a need for screening of the patients with substance dependence especially for those aged above 30 years and/or having a high BMI for MS.

Background & objectives : Three countries, Bangladesh, India and Nepal, set out to eliminate kala-azar by 2015. This study was aimed to document the knowledge and practices in kala-azar case management of public and private health providers in these three countries. Methods: A health care provider survey was conducted in 2007 at 4 study sites, viz., Muzaffarpur and Vaishali districts in India, Mahottari district in Nepal, and Rajshahi district in Bangladesh. Interviews were conducted with formal and informal health care providers at their home or practice. Results: About half of the providers in India and Nepal knew the rapid diagnostic test rK39 recommended by the elimination initiative, but this was not in Bangladesh. Knowledge of the recommended first-line drug, miltefosine, was good in India and Nepal but less so in Bangladesh. Interpretation & conclusions : Innovative tools for VL care have not yet been fully taken up by private for profit care providers in the three countries that launched a VL elimination initiative. The elimination initiative needs to address these gaps in private providers' knowledge, given their substantial share in the care of VL patients.

Background & objectives : The unique immunological functions of γδ T lymphocytes to contribute immunity against Mycobacterium tuberculosis0 attracted interest of researchers. However, little is known about the specificity of γδ Τ cell in tuberculosis patients and the lack of exact tuberculosis antigen recognized by γδ T cells limited its application. The analysis of complementary determinant region (CDR)3 sequence characteristic in γδ T cells of tuberculosis patients would contribute to understand the distribution specificity of γδ T cell. In present study, we investigated the diversity of the γ9/δ2 T cell immunorepertoire and analysed the specificity of the expressed CDR3 in pulmonary tuberculosis patients. Methods: The total RNA in peripheral blood mononuclear cell of 50 pulmonary tuberculosis patients and 10 healthy controls was extracted. The polymerase chain reaction was used to specifically amplify the CDR3 region of γ9 and δ2 chain. The PCR products were ligated into the pGEM-T easy vector. The plasmid DNA was sequenced using the ABI3700 and the T7 primer. Results: Our findings showed that predominant CDR3 sequence of δ2 chain in pulmonary tuberculosis patients was CACDTLVSTDKLIFGKG. The sequence specifically exists in almost all pulmonary tuberculosis patients. The conserved hydrophobic acid residue in 97 positions is present in the γδ T cell reactive to M. tuberculosis. The length of δ2 CDR3 in pulmonary tuberculosis patients has no relation with the disease progress. Interpretation & conclusions: Our results suggest that γδ T cells appear to use CDR3 sequence to recognise M. tuberculosis antigen. γδ T cells reactive to M. tuberculosis were diverse and polyclonal.

Background & objectives : Emergence and spread of drug resistant Mycobacterium tuberculosis is a serious threat to tuberculosis (TB) control programme. Therefore, the objective of this study was to genotype drug-resistant M. tuberculosis strains isolated from patients in Sichuan, China, using Mycobacterial Interspersed Repetitive Units (MIRU) for epidemiological analysis. Methods: Drug-resistance testing of M. tuberculosis isolates from pulmonary TB patients was confirmed by proportion method. Twelve MIRU loci were analyzed on 80 drug-resistant and 9 susceptible isolates by polymerase chain reaction and agarose gel electrophoresis. Hunter-Gaston discriminatory index (HGI) values were determined for each 12 MIRU loci for the evaluation of their discrimination power. Results: Among 12 MIRU loci examined, polymorphic bands could be generated on 11 loci. Sixty five isolates had distinct MIRU patterns, while other 24 belonged to 8 clusters and resistant to at least one anti-TB drug tested. The association between the MIRU patterns and the mutation patterns of drug-resistance relevant target genes was not significant among the drug-resistant isolates. Interpretation & conclusions : The results showed that with a satisfactory discrimination power exhibited, the 12 loci based MIRU typing could be a valuable tool for epidemiological studies in M. tuberculosis isolates from Sichuan.

Background & objectives: The mature fruits of Solanum nigrum contains steroidal glycosides. These are often used as vegetable and there are evidences on tribal use of these fruits as an oral contraceptive. The present study was carried out to evaluate the estrogenic potential of S. nigrum fruits by in vitro and in vivo assays. Methods: Defatted methanol extract of dried S. nigrum fruits was column fractionated and the glycoside positive fractions pooled. Definite concentrations of the fraction were used for in vitro and in vivo assays. The effect on cell viability was analyzed in MCF-7 cell lines by MTT assay followed by in vitro evaluation of estrogenicity by hydroxy apatite (HAP) binding assay. The results were further evaluated in vivo by performing uterotrophic assay in ovariectomized mouse models. Results: At low concentration (40 μg/ml), SNGF induced a dose-dependent increase in MCF-7 cell proliferation, while higher extract concentrations (80-320 μg/ml) caused progressive cell growth inhibition. The competitive binding assay using ^[3] H-E ₂ suggests that this effect is mediated by estrogen receptor. Mouse uterotrophic assay revealed a classical uterotrophic response in ovariectomized mice in response to S. nigrum glycoside fraction (SNGF). SNGF at a dose of 100 mg/kg of body wt induced the maximum height of luminal epithelial cells which indicated an increase of 30.8 per cent over control (P<0.01) with a correlated increase in uterine wet wt (150% increase over control). Higher doses (250 and 500 mg/kg body wt) of SNGF did not induce any uterotrophic effect. Interpretation & conclusions : Our preliminary data demonstrate the hormone like activity of Solanum glycosides both in vitro and in vivo in mouse, which needs to be further explored to evaluate the possible mechanism and clinical implications.

Background & objectives : Mosquito control is facing a threat due to the emergence of resistance to synthetic insecticides. Insecticides of plant origin may serve as suitable alternative biocontrol techniques in the future. The purpose of the present study was to assess the ethyl acetate, acetone and methanol extracts of Andrographis paniculata, Eclipta prostrata and Tagetes erecta leaves tested for oviposition-deterrent, ovicidal and repellent activities against malaria vector, Anopheles subpictus Grassi (Diptera: Culicidae). Methods: The dried leaves of the three plants were powdered mechanically and extracted with ethyl acetate, acetone and methanol. One gram of crude extract was first dissolved in 100 ml of acetone (stock solution). From the stock solution, test solution concentrations of 31.21- 499.42 mg/l for oviposition- deterrence assay and repellency and 15.60 - 998.85 mg/l were used in ovicidal assay. The percentage oviposition- deterrence, hatching rate of eggs and protection time were calculated. One-way analysis of variance was used for the multiple concentration tests and for per cent mortality to determine significant treatment differences. Results: The percentage of effective oviposition repellency was highest at 499.42 mg/l and the lowest at 31.21 mg/l in ethyl acetate, acetone and methanol extracts of A. paniculata, E. prostrata and T. erecta. The oviposition activity index (OAI) value of ethyl acetate, acetone and methanol extracts of A. paniculata, E. prostrata and T. erecta at 499.42 mg/l were -0.91, -0.93, -0.84, -0.84, -0.87, -0.82, -0.87, -0.89 and -0.87, respectively. Mortality (no egg hatchability) was 100 per cent with ethyl acetate and methanol extracts of A. paniculata, E. prostrata and T. erecta at 998.85 mg/l. The maximum adult repellent activity was observed at 499.42 mg/l in ethyl acetate extracts of A. paniculata, E. prostrata and methanol extracts of T. erecta, and the mean complete protection time ranged from 120 to 150 min with the different extracts tested. Interpretation & conclusions : The acetone extract of A. paniculata, methanol extract of E. prostrata and T. erecta showed good oviposition-deterrent, ovicidal and repellent activities respectively. These results suggest that the leaf extracts of A. paniculata, E. prostrata and T. erecta may have the potential to be used as an ideal eco-friendly approach for the control of the An. subpictus.

Background & objectives: Majoon Suranjan (MS) is a polyherbal formulation used in Unani system of medicine for the treatment of rheumatoid arthritis (RA). The present study evaluates the antiarthritic efficacy of this formulation in three different experimental models. Methods: The anti-inflammatory activity of MS (in doses of 450, 900 and 1800 mg/kg body wt) was evaluated using the turpentine oil induced paw oedema model and the antiarthritic efficacy was evaluated using the formaldehyde and complete Freund's adjuvant (CFA) induced arthritis models. Aspirin (100 mg/kg body wt) was used as the standard drug in all the models. In order to assess the safety of the test drug, oral acute and 28 day toxicity studies were also carried out. Results: MS produced a dose dependent protective effect in all the experimental models. Its antiarthritic efficacy was comparable to aspirin in formaldehyde induced arthritis and was superior to aspirin in turpentine oil induced paw oedema and CFA induced arthritis. MS also inhibited the delayed increase in joint diameter as seen in control and aspirin treated animals in CFA induced arthritis. Oral LD ₅₀ of MS was found to be >5000 mg/kg in rats. Chronic administration did not produce any significant physiological changes in the tested animals. Interpretation & conclusions : Results of the present study suggest that the antiarthritic activity of MS was due to the interplay between its anti-inflammatory and disease modifying activities, thus supporting its use in traditional medicine for the treatment of RA.

Background & objectives: During an outbreak of influenza like illness throat swab culture and virus isolation are done to identify causal virus, and it may cause a significant delay in diagnosis and isolation. There is a need to determine a sensitive laboratory parameter which could play a major role in identifying H1N1 infection among patients presenting with influenza like symptoms. We undertook this study to determine a laboratory parameter to identify swine influenza infection amoung patients presenting with influenza like symptoms while awaiting throat swab culture and virus isolation reports. Methods: This retrospective study was conducted on patients presenting to A.P TB and Chest Hospital, Hyderabad, with swine influenza symptoms during September-October 2009. They were divided into swine flu positive (SW+) and swine flu negative (SW-) groups, based on their throat swab culture reports and laboratory data. Neutrophils to lymphocyte (N/L) ratio was calculated for every patient and also mean N/L ratio for two groups. Mean WBC count was also noted for both groups. Results: There were 55 patients in each group. The WBC count in SW+ group varied from 3000 to 10600 cells/mm ³ (mean 5714), while in SW- group it varied from 4500 to 14300 cells/mm ³ (10252). The N/L ratio was less than two for 92.7 per cent of patients in SW+ group and for 3.63 per cent patients in SW- group. N/L ratio < 2 as an indicator for swine flu had a sensitivity of 92.7 per cent and specificity of 96.36 per cent. It has a positive predictive value of 96.22 per cent and a negative predictive value of 92.8 per cent. Interpretation & conclusions : N/L < 2 along with a decrease in WBC count can be used as a screening tool in patients presenting with influenza like symptoms, while awaiting throat swab culture reports for confirmation.