Evaluation of thrombospondin�gene polymorphisms in corneal allograft rejection in Asian Indian patients

Purpose: To evaluate the frequency and the association of Thrombospondin 1 (THBS1) gene single nucleotide polymorphisms (SNPs) in Asian Indian patients with optical full thickness corneal grafting surgery. Methods: Prospective case朿ontrol analysis of optical penetrating keratoplasty patients with and without immune rejection and controls for genotyping of 3 THBS1 gene SNPs (rs1478604 A>G; rs2228261 C>T; rs2228262 A>G) by Amplification Refractory Mutation System-Polymerase Chain Reaction (ARMS PCR). Results: Among 58 patients [45 with immune allograft rejection (DNA isolation was possible in 38 samples) and 13 without immune corneal allograft rejection] and 65 controls, allele frequencies observed for rs1478604 (A>G) are A: 69.7% and 72.6%, G: 30.2% and 27.3%; for rs2228261 (C>T) are T: 70.2% and 62.3%, C: 29.7% and 37.6%; and for rs2228262 (A>G) A: 97.4% and 98.4%; G 2.5% and 1.5% respectively. Genotype frequencies were rs1478604 (A>G) AA: 57.8% and 59.3%, AG 23.6% and 26.5%; GG 18.4% and 14%; for rs2228261 (C>T) TT: 40.5% and 33.8%, TC: 59% and 56.9%, CC: 0% and 9.2%; for rs2228262 (A>G) AA: 94.8% and 96.8%, AG: 5.1% and 3.1% in rejection and controls respectively. The allele and genotype frequency for the 3 described THSB1 SNPs did not show any difference between the corneal graft immune rejection patients and controls. Conclusion: Asian Indian population evaluated for THBS1 gene SNPs by ARMS PCR genotyping in Asian Indian population did not show any genetic association to immune rejection occurrence in our study.

Mitochondrial DNA content of peripheral blood mononuclear cells in ART untreated & stavudine/zidovudine treated HIV-1-infected patients

Background & objectives: Nucleoside reverse transcriptase inhibitors (NRTIs) are known to cause mitochondrial toxicity. This study was done to estimate mitochondrial DNA (mtDNA) content of peripheral blood mononuclear cells (PBMCs) among human immunodeficiency virus (HIV) infected, NRTI treated and antiretroviral therapy (ART)-naïve patients and evaluate the utility of mtDNA content as a biomarker of mitochondrial toxicity. Methods: mtDNA content in PBMCs of 57 HIV-infected ART untreated and 30 ART treated with stavudine (d4T) or zidovudine (AZT) containing regimen were compared against 24 low-risk healthy controls (LoRHC). Results: There was a significant (P=0.01) reduction in mtDNA content among HIV-infected (104; 80-135) compared to LoRHC (127; 110-167), and it was the same in both the treated (104.8; 88-130) and untreated patients (104.7; 78-142). mtDNA significantly (P=0.014) declined in ART treated patients symptomatic for toxicity (97; 74-111) than the asymptomatic patients (128; 103- 153). Interpretation & conclusions: mtDNA depletion in PBMCs was evident among HIV-infected individuals on ART. Moreover, as mtDNA content was reduced among the patients symptomatic for toxicity than the asymptomatic in both the HIV-infected groups, the current study supports mtDNA content of PBMCs to serve as a biomarker of mitochondrial dysfunction induced by NRTI and HIV. Longitudinal studies with a large sample need to be done to confirm these findings.

Comparative Study Of Pain Relief Between Using Traction And Pillow Support In Proximal Femur Fractures

INTRODUCTION: There is always confusion regarding preoperative stabilization modality for proximal femur fractures to achieve pain relief. Objectives: The aim of this study was to study the effects of skeletal traction and pillow support on pain in patients with proximal femur fractures. Patients and Methods: A total of 50 patients contributed in this randomized clinical trial. Patients were randomly allotted into two equal groups: the skeletal traction and pillow support groups. The severity of pain was recorded at admission and 30 minutes, 24, 48 hours after traction/pillow support application using Visual Analogue Scale (VAS). Results: The severity of pain was significantly decreased in skeletal traction group and pillow support group only at the end of the first day after traction application. The number of pain complaints which needs analgesics was the same between the two groups. Conclusions: even though skeletal traction and pillow support had no effect on analgesic consumption, both have significantly decreased the pain at the end of the first day. The application of skeletal traction and pillow support both are recommended in patients with proximal femur fractures.

A rapid and low-cost microscopic observation drug susceptibility assay for detecting TB and MDR-TB among individuals infected by HIV in South India.

Background: The converging epidemics of HIV and tuberculosis (TB) pose one of the greatest public health challenges of our time. Rapid diagnosis of TB is essential in view of its infectious nature, high burden of cases, and emergence of drug resistance. Objective: The purpose of this present study was to evaluate the feasibility of implementing the microscopic observation drug susceptibility (MODS) assay, a novel assay for the diagnosis of TB and multi-drug-resistant tuberculosis (MDR-TB) directly from sputum specimens, in the Indian setting. Materials and Methods: This study involved a cross-sectional, blinded assessment of the MODS assay on 1036 suspected cases of pulmonary TB in HIV-positive and HIV-negative patients against the radiometric method, BD-BACTEC TB 460 system. Results: Overall, the sensitivity, specificity, positive predictive value, and negative predictive value of the MODS assay in detecting MTB among TB suspected patients were 89.1%, 99.1%, 94.2%, 95.8%, respectively. In addition, in the diagnosis of drug-resistant TB, the MODS assay was 84.2% sensitive for those specimens reporting MDR, 87% sensitivity for those specimens reporting INH mono-resistance, and 100% sensitive for specimens reporting RIF mono-resistance. The median time to detection of TB in the MODS assay versus BACTEC was 9 versus 21 days (P < 0.001). Conclusion: Costing 5 to 10 times lesser than the automated culture methods, the MODS assay has the potential clinical utility as a simple and rapid method. It could be effectively used as an alternative method for diagnosing TB and detection of MDR-TB in a timely and affordable way in resource-limited settings.

Evaluation of two indigenous rapid and two ELISA assays for the diagnosis of HIV infection India.

Purpose: Human immunodeficiency virus (HIV) diagnostic tests are being used extensively in India. However, the evaluation data on these assays are very limited. The present study evaluates indigenous HIV test kits manufactured in India. Materials and Methods: A total of 200 characterised specimens were assayed with Comb AIDS - RS Advantage HIV 1+2 Immunodot Test, Enzaids HIV 1+2 ELISA test, Enzaids Duet HIV Antigen+antibody ELISA test and Signal HIV Flow Through HIV 1+2 test kits. Performance characteristics of these assays were calculated. Results: Sensitivity, specificity, positive predictive value, negative predictive value and efficiency of all the assays were 100% except for Signal HIV Flow Through HIV 1+2 test kit. The specificity, positive predictive value and efficiency of the Signal HIV Flow Through HIV 1+2 test kit were 98.9%, 98.9% and 99.4%, respectively. The Enzaids Duet HIV kit was found to be extremely sensitive in detecting p24 Ag with the sensitivity of 1.5 pg/mL. Conclusions: To conclude, selection of better diagnostic assay is very much important to resolve discrepancies in HIV diagnosis. All these assays under evaluation in this report have got excellent performance characteristics and much suitable to use in serial testing algorithms in use for resources limited settings.

Establishment of T-lymphocyte subset reference intervals in a healthy adult population in Chennai, India.

Estimation of CD4+ T-lymphocytes continues to be an important aspect for monitoring HIV disease progression and response to antiretroviral therapy. Most of the diagnostic laboratories often rely on western text books for CD4+ T-lymphocyte reference values, which could, often be unreliable for usage in local settings. Therefore, we attempted to establish the reference values for T-lymphocyte subsets among healthy adults in a cross-sectional study carried out at the YRG Centre for AIDS Research and Education (YRG CARE) in Chennai, south India, in 213 (84 female and 129 male) healthy, HIV-1/2 seronegative adults as volunteers. Whole blood specimens were processed for CD4+, CD8+ T-lymphocyte estimation and haematological parameters. The established range of CD4+ T-lymphocyte counts for men and women were 383-1347 cells/microl (mean 865 and median 845 cells/microl) and 448-1593 cells/microl (mean 1021 and median 954 cells/microl), respectively. Women had significantly higher absolute CD4+ Tlymphocyte counts (P<0.001) and CD4+:CD8+ T-lymphocyte ratio as compared to men. The established normal range of CD4+ T-lymphocyte % was 21-59 (mean 40.2 and median 40.1). The influence of age was not observed in any of the parameters except CD4+/CD8+ T-lymphocyte ratio with the >45 yr age group. Further studies with greater sample size may be required to define the staging of HIV disease in relation to the normal CD4 T-lymphocyte count in the general population.

HIV-1 western blot assay: What determines an indeterminate status?

BACKGROUND: The Western blot assay is the gold standard for the detection of antibodies to human immunodeficiency virus type 1 (HIV-1). However, indeterminate Western blot reactivity to HIV-1 proteins may occur in individuals, who may not be infected with HIV. AIM: This retrospective study was aimed to determine the diagnostic value of the interpretation criteria in relation to commercial kits for HIV-1 diagnosis. METHODS AND MATERIALS: A total of 556 serum/plasma specimens collected from high-risk population attending our HIV clinic from 2000-2004 were tested by three different western blot kits: NEW LAV BLOT I (n=244), HIV BLOT 2.2; (n=112), Genetic Systems HIV-1 (n=237). And the results of western blot strips were analyzed using the various interpretation criteria: WHO/NACO, CDC/ ASTPHLD, ARC, FDA, CRSS and JHU. Some specimens were run on more than one kit. RT-PCR assay was performed on 5 specimens, which were indeterminate with LAV BLOT I. RESULTS: The discrepancy in LAV BLOT I positive results were between 157(64)-176(72), and indeterminate results were between 44(18) to 63(25). No such variations were observed in genetic systems. There are some HIV negative (by PCR) specimens were indeterminate in LAV BLOT I revealing the kit more sensitive and less effective for diagnostic purpose. CONCLUSION: The genetic systems kit is superior to other kits we analyzed and its results are concordant with HIV-1 PCR results. To report, the choice of western blot commercial kit is paramount important than the use of particular interpretation criteria for the diagnosis of HIV-1.

Serodiagnosis of syphilis in a community: An evaluatory study.

PURPOSE: To analyse the prevalence of syphilis in the apparently healthy population and to provide data for implementation of the joint STD/HIV control programme, a population based study was undertaken by using 'probability proportional to size' cluster survey method in three randomly chosen districts of Tamil Nadu, India namely Dindigul, Ramnad and Tanjore. METHODS: Blood samples were collected from adults (n=1873) aged 15-45 years, from the selected households enrolled in this study. The sera were tested parallelly by rapid plasma reagin (RPR) and Treponema pallidum haemagglutination (TPHA) tests. Reactive samples by RPR and/or TPHA were later analysed by fluorescent treponemal antibody absorption (FTA-ABS) test. RESULTS: The prevalence of syphilis in the community of Tamil Nadu as per RPR positivity was 2.7% (50/1873) as against 0.7% by TPHA (13/1873). FTA-ABS positivity was observed in only 12 out of 48 (25%) RPR/TPHA reactive samples tested. By taking the positivity by two of the three tests, the community prevalence of acute ongoing syphilis in Tamil Nadu was determined as 1.1% (20/1873). CONCLUSIONS: The results confirmed that no single serological test for syphilis can act as the marker of ongoing acute infection in an apparently healthy population. The study suggests that for specific diagnosis of ongoing syphilis, the FTA-ABS test may be performed along with RPR and TPHA.

Asymptomatic malarial parasitaemia in Tamil Nadu.

AIM: The aim of the study was to determine the community prevalence of asymptomatic malarial parasitaemia in the state of Tamil Nadu. METHODS: Free medical camps were organised in three randomly selected districts of Tamil Nadu, namely Dindigul, Ramnad and Thanjavur districts in November, 1997. Proportionate to population size cluster survey method was followed to collect peripheral blood smear by finger prick from 30 clusters in each district. Fifteen households were randomly selected from each district with the target age group of 15-45 years. Peripheral blood smears were stained by Leishman's stain and the slides were examined end to end by two independent experts to diagnose malarial parasites. RESULTS: The male:female ratio of the population studied was 1:1.6. Asymptomatic malaria was identified in 17 out of 569 individuals screened with a positive rate of 2.9% (CI 1.5-4.3). Out of the 17 malarial positive peripheral smears 15 were P. vivax and only two were P. falciparum with the predominance of gametocyte stage. CONCLUSION: This study reaffirms the prevalence of asymptomatic malaria in Tamil Nadu especially with P. vivax.

Identification of Pneumocystis carinii in induced sputum of AIDS patients in Chennai (Madras).

Induced sputum samples were collected from 32 AIDS patients with respiratory ailments. Pneumcystis carinii was demonstrated in 9 out of 32 AIDS cases by Indirect Immunofluorescence technique (HF). Four cases were positive by all the three techniques namely Giemsa staining, Toluidine blue staining and IIF, three were positive by both toluidine blue and IIF, and two were positive only by IIF. Among other microbial pathogens, acid fast bacilli was demonstrated in all the P carinii positive cases and Candida albicans in 53% AIDS cases from the induced sputum sample.