Assessment of genetic divergence of mango genotypes using multivariate techniques

Aim: The study was carried out to assess the overall genetic variability of 60 mango (Mangifera indica) genotypes for important fruit quality and yield characteristics to select better parents for mango breeding programme. Methodology: A total of 17 variables were scored and subjected to multivariate analysis. ANOVA, descriptive statistics, cluster analysis and principal component analysis were used to investigate variability among the genotypes for the selected traits. Results: Significant phenotypic variability was observed for studied traits. Cluster analysis classified all genotypes into two major groups according to shared similarity. Principal component analysis revealed that traits like fruit weight, fruit diameter, pulp content and pulp: stone ratio contributed positively, while fruit shape index, TSS:acid ratio, stone and peel traits contributed negatively for a large proportion of the observed variability. Interpretation: Pusa Arunima, Pusa Shresth, Pusa Lalima, Mallika, Ramkela, Amrapali, Extreema, Neelum, Gulab Jamun, S.B. Alibagh, Tommy Atkins, Primor-de-Amoreira genotypes were found unique for fruit and yield attributing traits, thus making them potential donor parent for fruit weight, fruit color, fruit diameter, fruit shape, pulp and pulp: stone traits in mango hybridization programme.

A Study of Prevalence of Aerobic Bacteria and Fungi in Sputum Specimens of Patients with Post Tubercular Bronchiectasis

Background: Patients with Post TB bronchiectasis colonize many aerobic bacteria and fungi and lead to an increase in exacerbations and decrease in quality of life. Objective: To study the prevalence of aerobic bacteria and fungi in sputum specimens of patients with post tubercular bronchiectasis and to find out the local antibiotic sensitivity and resistance patterns. Materials and Methods: This prospective observational study was carried out over a period of one year comprising of 50 patients with post Tubercular Bronchiectasis who were above 18 yrs, completed ATT smear negative and HRCT showing bronchiectactic changes. Identification of organisms from sputum samples were done by classical aerobic microbial staining and culture methods. Descriptive and inferential statistical analysis was carried out in this study. Results: This study comprises of the 50 patients with a larger group of individuals under the age group of 51-60 (30%), along with bronchial colonization of aerobic bacteria being 78% and growth of fungi being 2%.Among the 39 patients,16 patients (32%) had grown Pseudomonas aeruginosa, 12 patients(24%) had grown Klebsiella pneumoniae, 8 patients(16%) had grown Streptococcus species and rest 3 patients(6%) had grown staphylococcus species. The Antibiotic resistance noted highest being Amikacin (56.4%), piperacillin-tazobactam showing the least (2.6%) and highest sensitivity with imipenem(100%). Conclusion: This study signifies that prevalence of aerobic bacteria, especially Pseudomonas aeruginosa and Klebsiella pneumonia was common in patients with post tubercular bronchiectasis. The Antibiotic resistance noted highest being Amikacin (56.4%) and sensitivity with imipenem(100%). In patients colonizing Pseudomonas aeruginosa, there was significant decline in lung function.

Poland's syndrome with unusual hand and chest anomalies: A rare case report.

Poland’s syndrome is a rare congenital anomaly consisting of a unilateral absence of the pectoralis major, ipsilateral muscle, hand anomaly and occasionally associated other malformations of the chest wall and breast. Many structural and functional abnormalities have been described in association with this syndrome. We report an incidentally diagnosed case in a 27-year-old male patient who presented to us with symbrachydactyly. In addition to this, anterior depression of 2nd, 3rd and 4th ribs and bifid (forked) 5th rib was present on radiological investigations. The body of sternum was short and deformed on the right side with absence of xiphoid process. All middle phalanges were absent on right hand. It is a rare variant of Poland’s syndrome.

Simultaneous Gastric and Ileal Trichobezoars Causing Small Bowel Obstruction: A Case report.

Bezoars are conglomerates of food or fiber in the alimentary tract of humans and certain animals, mainly ruminants. A trichobezoar represents a mass of accumulated hair. Trichobezoars may present as an isolated gastric mass, as an extension into the small intestine, or as an independent fragmented mass in the small intestine. The presence of discrete coexisting gastric and ileal trichobezoars has been reported only rarely in the literature. This is a case report of a 10-year-old girl presenting with small-bowel obstruction secondary to synchronous trichobezoars in the stomach and ileum. The case highlights the role of imaging and importance of complete evaluation of the gastrointestinal tract at the time of surgical evacuation.

Diagnosis of tuberculosis in an era of HIV pandemic: A review of current status and future prospects.

HIV and tuberculosis co-infection interact in fundamentally important ways. This interaction is evident patho-physiologically, clinically and epidemiologically. There are several differences between HIV-infected and HIV-uninfected patients with tuberculosis (TB) that have practical diagnostic implications. TB is more likely to be disseminated in nature and more difficult to diagnose by conventional diagnostic procedures as immunosuppression progresses. As TB rates continue to increase in HIV-endemic regions, improved diagnostic techniques merit consideration as TB-control strategies. There is a need to develop more user friendly techniques, which can be adapted for use in the high-burden and low-income countries. This review focuses on the diagnostic challenges in HIV-TB co-infection with an update on the current techniques and future prospects in an era of HIV pandemic.

Factors Influencing Blood Donations and the Rational Use of Blood.

A multicentric quasi-experimental study was conducted in Delhi, from March 2007 to September 2007, on i) the factors which stimulate the donors to donate blood, ii) major barriers and myths associated with blood donation and iii) clinicians perception of the rational use of blood. The study design included a face-to-face survey, with a pre-tested questionnaire paper in two leading blood banks of Delhi and by relevant interviewers from the community and medical fraternity. The sample size was 240-blood donors from two different blood banks and the control group included 100 potential donors from community and 50 clinicians from various hospitals. The data generated was analyzed using excel sheet and Epi-Info software. The study revealed the factors which influence the blood donation included replacement credit and family/peer pressure. Regarding myths and barriers, among potential donors, about a quarter of them felt that it is time consuming, and 20% felt it could lead to sexual impairment or is not rewarding. A total of 10% were not aware about the blood donation while 15% said that donation time was inconvenient. Of the 50 clinicians, a quarter of them were not aware of the rational use of blood.

Characterization of RPO B gene for detection of rifampicin drug resistance by SSCP and sequence analysis.

Purpose: Because of the emergence of multidrug-resistant tuberculosis in recent times, the rapid detection of resistance to the first-line anti-tuberculosis drug rifampicin was felt worldwide. Accordingly, this study was conducted to evaluate the diagnostic potential of polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) for checking its utility as a rapid screening test for determination of rifampicin drug resistance. Materials and Methods: A total of 34 isolates of Mycobacterium tuberculosis ( M. tuberculosis ) (22 rifampicin resistant, 11 rifampicin sensitive and one control H37Rv) strains were analysed by PCR-SSCP and DNA sequencing within the 157-bp region of the rpo B gene (Ala 500 -Val 550 ). Results: Rifampicin resistance was detected successfully by PCR-SSCP in 20/22(90.90%) of rifampicin-resistant strains showing a total of nine different mutations in seven codon positions: codon 513 (CAA→CCA), 516 (GAC→GTC), 507 (GGC→GAC), 526 (CAC→GAC, TAC), 531 (TCG→TTG, TGG), 522 (TCG→TGG) and 533 (GTG→CCG). Two rifampicin-resistant strains showed an identical PCR-SSCP pattern with the wild type H37Rv; 77.27% rifampicin-resistant strains showed a single point mutation and 9.09% had no mutation. Three rifampicin-resistant strains showed characteristic double mutations at codon positions 526 and 531. Sensitivity and specificity were calculated as 90.90% and 100%. Conclusions: Rifampicin-resistant genotypes were mainly found in codon positions 516, 526 and 531. PCR-SSCP seems to be an efficacious method of predicting rifampicin resistance and substantially reduces the time required for susceptibility testing from 4 to 6 weeks to a few weeks.

Congenital CMV infection; diagnosis in symptomatic infants.

Background: Samples from babies exhibiting clinical symptoms suggestive of congenital infection are referred regularly to NICD, New Delhi,, from Government Hospitals located in Delhi and a home for abandoned children (Palna), for the diagnosis of etiological agents like toxoplasma, rubella, CMV and herpes. Blood samples of mothers of most of the affected babies are also received. Objective: Evaluation of rapid and accurate technique for the diagnosis of congenital CMV infection. Materials and Methods: One hundred and twenty five blood samples suggestive of symptomatic congenital CMV infection were selected from samples received at NICD during the period June 2005-March 2007. A request to collect and send the urine samples of the selected babies was sent to the respective hospitals. Serum samples of the babies were tested for CMV-IgM antibodies using µ-capture ELISA. Mothers' serum samples were subjected to CMV-IgM and IgG class antibodies assay by commercial ELISA kits. DNA isolation and amplification was performed in urine samples and some of the serum samples using a commercial PCR kit for detection of HCMV. Blood and urine samples from 20 normal babies were included in the study. Results: Twenty Seven serum samples (21.6%) of infants, of the 125 tested, were positive for CMV-IgM antibodies. Twenty five samples (20%) showed amplification of CMV -DNA. All 25 samples positive for PCR were positive for CMV IgM antibodies. Sera of 73 mothers, out of 75 tested (97.3%), were positive for CMV IgG antibodies. However, none of them was positive for CMV IgM antibodies. Mothers of all 27 positive babies were positive for CMV-IgG antibodies. Serum and urine samples from 20 normal babies were negative for ELISA and PCR. Conclusion: µ-capture ELISA technique was found to be more sensitive than PCR (92.6%) for detection of congenital CMV infection. ELISA is also rapid, less cumbersome and cost effective for diagnosis of CMV infection.

Chikungunya fever: a re-emerging viral infection.

Chikungunya (CHIK) fever is a re-emerging viral disease characterized by abrupt onset of fever with severe arthralgia followed by constitutional symptoms and rash lasting for 1-7 days. The disease is almost self-limiting and rarely fatal. Chikungunya virus (CHIKV) is a RNA virus belonging to family Togaviridae, genus Alphavirus. Molecular characterization has demonstrated two distinct lineages of strains which cause epidemics in Africa and Asia. These geographical genotypes exhibit differences in the transmission cycles. In contrast to Africa where sylvatic cycle is maintained between monkeys and wild mosquitoes, in Asia the cycle continues between humans and the Aedes aegypti mosquito. CHIKV is known to cause epidemics after a period of quiescence. The first recorded epidemic occurred in Tanzania in 1952-1953. In Asia, CHIK activity was documented since its isolation in Bangkok, Thailand in 1958. Virus transmission continued till 1964. After hiatus, the virus activity re-appeared in the mid-1970s and declined by 1976. In India, well-documented outbreaks occurred in 1963 and 1964 in Kolkata and southern India, respectively. Thereafter, a small outbreak of CHIK was reported from Sholapur district, Maharashtra in 1973. CHIKV emerged in the islands of South West Indian Ocean viz. French island of La Reunion, Mayotee, Mauritius and Seychelles which are reporting the outbreak since February, 2005. After quiescence of about three decades, CHIKV re-emerged in India in the states of Andhra Pradesh, Karnataka, Maharashtra, Madhya Pradesh and Tamil Nadu since December, 2005. Cases have also been reported from Rajasthan, Gujarat and Kerala. The outbreak is still continuing. National Institute of Communicable Diseases has conducted epidemiological, entomological and laboratory investigations for confirmation of the outbreak. These have been discussed in detail along with the major challenges that the country faced during the current outbreak.

Small intestinal bacterial overgrowth in North Indian patients with celiac disease.

BACKGROUND: Occasionally celiac patients continue to experience gastro-intestinal symptoms even with a gluten free diet. In these cases, small intestinal bacterial overgrowth may be one of the causes of the lack of response. Therefore, this prospective study was planned to determine the prevalence of small intestinal bacterial overgrowth in celiac patients. MATERIALS AND METHODS: We studied 87 confirmed cases of celiac disease from North India and 87 age and sex matched controls. Celiac disease was confirmed by positive IgA antitissue transglutaminase on ELISA. 80 g glucose hydrogen breath test (non-invasive test) was performed to establish small intestinal bacterial overgrowth. Rise of more than 10 ppm in hydrogen concentration over baseline value within two hours was considered suggestive of small intestinal bacterial overgrowth. RESULTS: Out of 87 patients with celiac disease, 49 were male and 38 were female.The mean (+/-SD) age for male patients was 26.3 +/- 16.3 years (range 14-59 years) and for female patients was 28.4 +/- 15.6 years (range 16-58 years). Amongst the controls, 52 (59.8%) were male and 35 (40.2%) were female. The mean (+/- SD) age for male controls was 27.6 +/- 14.5 years (range 15-57 years) and for female controls was 29.3 +/- 16.5 years (range 18-59 years). Hydrogen breath test was suggestive of bacterial overgrowth in 18 of the 87 (20.7%) celiac disease patients but not in any of the apparently healthy controls. CONCLUSION: This study indicates that a large number of celiac patients from North India suffer from bacterial overgrowth which can be accordingly treated with antibiotics.

Development and evaluation of an in vitro isolation of street rabies virus in mouse neuroblastoma cells as compared to conventional tests used for diagnosis of rabies.

In vitro isolation of rabies virus using mouse neuroblastoma cells (MNA) was evaluated. The sensitivity and reliability of in vitro procedure was performed in comparison with mouse inoculation test (MIT), the in vivo method of virus isolation, direct fluorescent antibody test (FAT) and Sellers staining. Of the 33 animal brain samples tested, 24 (72.72%) were positive by MIT. Sensitivity of Sellers stain, FAT and rapid tissue culture infection test (RTCIT) was found to be 54.16, 100 and 91.6% respectively. Concordance of Sellers stain, FAT, RTCIT with MIT was found to be 66.6, 100 and 93.93% respectively. Two samples which were positive by FAT and MIT showed gross contamination in cell lines, which is one of the drawbacks of RTCIT. However, rabies virus could be isolated in MNA cells from two of the eight human cerebrospinal fluid (CSF) samples from clinico-epidemiologically suspected cases of rabies. Both MIT and FAT showed negative results in the two CSF samples. RTCIT appears to be a fast and reliable alternative to MIT and holds promise in antemortem diagnosis of rabies, which is otherwise, a challenging task for a reference laboratory.

Diagnostic potential of IS6110, 38kDa, 65kDa and 85B sequence-based polymerase chain reaction in the diagnosis of Mycobacterium tuberculosis in clinical samples.

PURPOSE: The correlation between the presence of specific gene sequence of M. tuberculosis and specific diagnosis of clinical tuberculosis is not known. This study compared the results of polymerase chain reaction (PCR) amplification of M. tuberculosis specific DNA sequences (IS6110, 65kDa, 38kDa and mRNA coding for 85B protein) from different clinical samples of pulmonary and extrapulmonary tuberculosis. METHODS: One hundred and seventy-two clinical samples from suspected tuberculosis patients were tested for smear examination, culture (LJ and rapid BACTEC 460 TB system) and PCR. PCR was performed with specific primers for the targets: IS6110, 65 kDa, 38 kDa and 85 B. RESULTS: Each PCR test was found to have a much higher positivity than conventional test and BACTEC culture (P < 0.05). Smear positive samples (56) and the samples (36) showing positive results by conventional methods (smear and LJ medium culture) and BACTEC were found to be positive by all PCR protocols. No significant difference was found between the four PCR protocols (P> 0.05). The primer specific for amplifying the 123bp IS6110 fragment gave the highest positivity (83%), followed by 65kDa, 38kDa and 85B RT-PCR in descending order. CONCLUSIONS: These data suggest that the presence of IS6110 correlates more closely with the diagnosis of clinical tuberculosis than that of 65kDa, 38kDa and 85B proteins.

An outbreak of dengue fever in Veerannapet village, Cherial Mandal, of Warangal district, Andhra Pradesh.

An epidemiological and entomological investigation was carried out in Veerannapet village, Cherial Mandal of Warangal district, Andhra Pradesh. The study showed that all age groups and both the sexes were affected with the disease. Fever and bodyache was the first presenting feature, which was self-limiting and lasted for 4-5 days. Of the 19 Serum Samples tested, 17 showed high titre to Dengue antigen with 10 showing diagnostic titre. Five samples were positive for IgM antibodies to dengue virus. Larval surveys indicates high Breteau index (30.40%), House index (23.20%) and Container index (9.17%). The clinico-epidemiological and entomological investigation indicates present episode of fever outbreak is due to dengue fever. Strengthening and intensification of surveillance along with educating the community is recommended for prevention of outbreak.

Pediatric HIV infection.

OBJECTIVE: The present cross sectional study was undertaken to study clinical profile of HIV infection in children in Northern India. METHODS: 64 children from newborn to eighteen years, presenting for confirmation of diagnosis of HIV infection or monitoring of CD4-CD8 counts in confirmed cases, were evaluated. Children were categorized as per CDC classification of Pediatric HIV. The diagnosis was confirmed by serological tests or PCR assay. CD4-CD8 counts were done by FACS Count. RESULTS: Majority of the children were between 18 months to 5 years. Adolescents comprised 24% of the case. 51.5% children were infected through the mode of mother to child transmission. 39% of the case was transfusion-mediated. Unsafe medical injections probably contributed to 6.2% and heterosexual promiscuity led to 3.1% cases. Clubbing, not described in Indian studies so far, was seen in 9.3% cases. CONCLUSIONS: HIV infection is a chronic childhood disease extending into adolescence, and contaminated blood and unsafe medical injections are still important routes of HIV transmission in India.

Comparison of various microbiological tests including polymerase chain reaction for the diagnosis of osteoarticular tuberculosis.

PURPOSE: To evaluate the utility of the polymerase chain reaction (PCR) test for diagnosing osteoarticular tuberculosis (TB). METHODS: Clinical samples (synovial tissue and synovial fluid) obtained from 23 cases of suspected osteoarticular tuberculosis were subjected to Ziehl Neelsen (ZN) smear examination, radiometric BACTEC culture and PCR test for tuberculosis by amplifying 65 kDa antigen coding region of Mycobacterium tuberculosis (M.tb) genome. RESULTS: PCR test was found to be much sensitive than the ZN smear examination and BACTEC culture (p<0.05) in the diagnosis of osteoarticular TB. In synovial fluid samples, PCR was positive in 73.9%, ZN smear examination in 17.39% and BACTEC culture in 39.13% of cases. The positivities were relatively lower with synovial tissue samples, the corresponding figures being 60.8, 8.6 and 26.08% respectively. Moreover, on combining the results of synovial fluid and tissues, the corresponding figures further increased to 78.2, 21.7 and 43.3% respectively. Further, sensitivity and specificity for PCR employing BACTEC culture as the "gold standard" was 100% respectively. Using BACTEC culture, the earliest positivity was seen in three days using synovial tissue specimen and 13 days with synovial fluid, the average detection times being 23.2 days and 32.6 days respectively. On the other hand, PCR test gave a positive result within 24 hours. CONCLUSIONS: PCR test was shown to be much more sensitive than ZN smear examination and BACTEC culture test for diagnosing osteoarticular tuberculosis.

Entomological indices of Aedes aegypti at some international airports and seaports of southern India--a report.

Entomological surveys were undertaken at some of the international airports/seaports (Bangalore, Calicut, Chennai, Cochin, Thiruvanathapuram and Vishakapatnam) to find out the breeding prevalence of dengue vector mosquito in diverse breeding containers from 1998 to 2004. Three vector indices (House index, Container index and Breateu index) were used to assess the breeding potential at each airport/seaport. International Health Regulations urged national governments to keep all the international airports/seaports and peripheral areas up to 400 meters free from Aedes aegypti mosquitoes. However, surveys revealed high vector indices at all the airports and seaports. Health authorities of airports/seaports need to take cognizance of these facts and develop action plan for appropriate control measures with emphasis on vector surveillance.