Aruna Pawar

Venous thromboembolism in young patients from western India: a study.

The goal of this article is to study the association of known markers of thrombophilia with venous thrombosis in young patients (< 45 years) from the Western part of India. A prospective study of 432 patients (252 males and 180 females, age 1-45 years) was conducted between 1994 and 2000 (6 years). The diagnosis was confirmed in all the patients by ultrasound with Doppler or by a computed tomograph (CT) scan of the brain with or without contrast depending on the case. Detailed clinical examination, and family history was taken to establish recurrent thrombosis and familial occurrence of thrombosis. The markers studied were protein C, protein S, antithrombin (AT) III, factor V Leiden mutation, prothrombin gene G20210A polymorphism, and the thermolabile MTHFR variant C677T polymorphism, using appropriate techniques. Lupus inhibitor was tested in the first 72 patients using Dilute Russel Viper Venom Time (DRVVT) test, and anticardiolipin antibodies were tested by enzyme-linked immunosorbent assay. Protein C, protein S, and AT III deficiency was detected in 9.5%, 6.5%, and 2.6%, respectively, among the patients. Anticardiolipin antibody was present in 9.9% of the patients, whereas lupus anticoagulant was present in 8.3% of patients; factor V Leiden mutation was detected in 3% of patients; thermolabile variant of MTHFR C677T polymorphism was present in 14.9% of patients with 1.2% homozygotes. Prothrombin G20210A polymorphism was not detected in any sample in this population. One hundred and four patients of 432 (24.9%) had recurrent attacks of thrombosis without any proximate precipitating cause, whereas 7.5 % of the patients had another close member of the family with a history of deep venous thrombosis. Eighty-six members from 28 families (out of 32 families giving family history of thrombosis) were investigated and found to have protein C and protein S deficiency in seven each; factor V Leiden was present in 6, and MTHFR C677T polymorphism was present in 5 cases. Hence, 25 of 86 members (28%) from the family of patients with familial history deep venous thrombosis had positive markers for thrombophilia. Thus, we could show that in young patients presenting with thrombosis, at least 34% of them had a demonstrable cause for thrombophilia. Prothrombin gene polymorphism G20210A seems to be nonexistent in our population and AT III deficiency also appears to be low compared to other markers of thrombophilia. There is a high prevalence of variant MTHFR C677T in our series, but the incidence of MTHFR C677T in our general population is also high. Hence, the significance of this finding in our cases of deep venous thrombosis remains to be seen, but we did not see any homozygotes when we tested 70 randomly selected asymptomatic persons, whereas in the present series, 1.8% of the patients had homozygosity for the MTHFR C677T polymorphism.

Development of inhibitors in patients with haemophilia from India

Four hundred and seven patients (352 haemophilia A and 55 haemophilia B) were investigated for the presence of factor VIII and IX inhibitors. Twenty‐four out of 292 severe and two out of 36 moderate haemophilia A patients showed the presence of inhibitors. The mean age at development of inhibitors was 17.7 years (range 6–52 years). In 12 patients the inhibitors were detected due to suboptimal response to factor replacement therapy (symptomatic) and in the remaining 14 patients the inhibitors were detected during the routine screening of the patients’ samples for inhibitors. They had, however, responded well to the usual doses of factor concentrates and there was no suspicion in these patients that they had developed an inhibitor (asymptomatic). There were two families in which the inhibitors were detected in more than one family member. The level of inhibitors in symptomatic patients ranged from 2.2 Bethesda units (BU) mL–1 to 460.6 BU mL–1, and in asymptomatic patients it ranged from 0.8 BU mL–1 to 3.2 BU mL–1. The inhibitors persisted in all patients except one, who developed an inhibitor postoperatively for a brief period of 3 months. All these patients were followed up from first factor exposure and were tested for inhibitors at least twice a year. The mean number of exposure days before they developed inhibitors was 47.5 exposure days (range 17–98 exposure days). No inhibitors appeared after more than 100 exposure days in any of the patients. When 50 consecutive patients were investigated for intron 22 inversions of the factor VIII gene, 17 patients were found to be positive for inversions (10 proximal inversion; seven distal inversion) out of whom four patients developed inhibitors, three patients belonging to the same family. Out of 35 haemophilia B patients, only one patient developed an inhibitor. The overall prevalence of inhibitors was thus 8.2%, which is similar to the reports from western countries, prior to the introduction of highly purified factor concentrate therapy.

Potential of denaturing gradient gel electrophoresis for scanning of β‐thalassemia mutations in India

Over the last few years, substantial progress has been made in developing strategies for the detection and characterization of various mutations causing β‐thalassemia. The Indian population comprises of numerous endogamous caste groups and β‐thalassemia is seen in almost all of them. Knowledge of the spectrum of β‐thalassemia mutations in the population is a prerequisite for successful implementation of a prevention programme. Among the different approaches available today, Denaturing Gradient Gel Electrophoresis (DGGE) offers a valid technical approach which is applicable for screening of known mutants and polymorphisms as well as in locating regions of DNA bearing unknown mutations.

Prenatal Diagnosis of β-Thalassemia Among Indians using Denaturing Gradient Gel Electrophoresis

We have offered first trimester prenatal diagnosis to 55 couples at risk for β-thalassemia, originating from various parts of India, using polymerase chain reaction and denaturing gradient gel electrophoresis. Apart from the six common mutations, codon 30 (CAG→CAA), Cap site +1 (A→C), as well as three uncharacterized mutations were seen among the parents. in the majority of cases, the diagnosis was possible by scanning only one fragment (B) where most of the Indian mutations are situated. in 18 out of 55 cases, framework analysis could also have been used to offer prenatal diagnosis without characterizing the β-thalassemia mutations. in the two cases where the mutations were uncharacterized, prenatal diagnosis was done only on the basis of the anomalous denaturing gradient gel electrophoresis patterns seen in the parents and in previously affected children. This is the first attempt of prenatal analysis using denaturing gradient gel electrophoresis in the extremely diverse Indian population where the prof...

HLA involvement in nevirapine-induced dermatological reaction in antiretroviral-treated HIV-1 patients

114 Journal of Pharmacology and Pharmacotherapeutics | April-June 2011 | Vol 2 | Issue 2 asthmatics at Shiraz University of Medical Sciences. Iran: Ahvaz University of Medical sciences; School of Nursing and Midwifery; 1997. 6. Miranzadeh Omran H. Patient opinion about MDI use. Iran: Golestan University of Medical Sciences; 1997. 7. Farhadi K, Atai P. Difficulties with MDI use among asthmatics, bronchial asthma and influence of education. Iran: Kordestan University of Medical Sciences; 2001. 8. Scherer YK, Bruce S. Knowledge, attitudes and self-efficacy and compliance with medical regimen, number of emergency department visits, and hospitalization in adults with asthma. Heart Lung 2001;30:250-7. 9. Becker MH, Maiman LA. Sociobehavioral determinants of compliance with health and medical recommendations. Med Care 1975;13:10-24. 10. Scherer YK, Bruce S. Knowledge, attitudes and self-efficacy and compliance with medical regimen, number of emergency department visits, and hospitalization in adults with asthma. Heart Lung 2001;30:250-7.

Role of HLA class I (HLA-A, B) and HLA class II (HLA-DRB, DQB) in HIV-1 patients with and without pulmonary tuberculosis.

097. 642 | www.jaids.com q 2009 Lippincott Williams & Wilkins Letters to the Editor J Acquir Immune Defic Syndr Volume 51, Number 5, August 15, 2009

HLA-A and HLA-B alleles associated in psoriasis patients from Mumbai, Western India

Background: Psoriasis, a common autoimmune disorder characterized by T cell-mediated keratinocyte hyperproliferation, is known to be associated with the presence of certain specific Human Leukocyte Antigen (HLA) alleles. Aim: To evaluate distribution of HLA-A and HLA-B alleles and hence identify the susceptible allele of psoriasis from patients in Western India. Materials and Methods: The study design included 84 psoriasis patients and 291 normal individuals as controls from same geographical region. HLA-A and HLA-B typing was done using Serology typing. Standard statistical analysis was followed to identify the odds ratio (OR), allele frequencies, and significant P value using Graphpad software. Results: The study revealed significant increase in frequencies of HLA-A2 (OR-3.976, P<0.0001), B8 (OR-5.647, P<0.0001), B17 (OR-5.452, P<0.0001), and B44 (OR-50.460, P<0.0001), when compared with controls. Furthermore, the frequencies of HLA-A28 (OR-0.074, P=0.0024), B5 (OR-0.059, P<0.0001), B12 (OR-0.051, P=0.0002), and B15 (OR-0.237, P=0.0230) were significantly decreased in psoriasis patients. Conclusion: This study shows the strong association of HLA-A2, B8, and B17 antigens with psoriasis conferring susceptibility to psoriasis patients from Western India, while the antigens HLA-A28, B5, and B12 show strong negative association with the disease.

Human leucocyte antigen class II DRB1 and DQB1 associations in human immunodeficiency virus‐infected patients of Mumbai, India

The pathogenesis of human immunodeficiency virus (HIV) infection clearly involves immunoregulatory host factors and products of major histocompatibility complex class II genes, which present antigenic peptides to the T‐cell receptor on CD4+ cells, which in turn increase the production of specific antibodies and cytotoxic T lymphocytes. The main objective of this study was to determine the associations of human leucocyte antigen (HLA) DRB1 and DQB1 alleles and their haplotypes in 210 HIV‐1‐infected patients and compare them with 129 healthy normal individuals with same ethnic background. The HLA DRB1 and DQB1 alleles were genotyped using polymerase chain reaction product and sequence‐specific probes for reverse line hybridization, analysed with the Invitrogen Dynal PMP software. Our results revealed a highly significant increase of HLA DRB1*0902 [odds ratio (OR) = 17.12; P = 0.004], DQB1*030103 (OR = 53.53; P = 4.61E‐07) and DQB1*050201 (OR = 16.26; P = 0.0002) alleles while in contrast highly significant decrease in frequency of HLA DQB1*030101 (OR = 0.36; P = 0.0002), DQB1*050301 (OR = 0.22; P < 0.0001) and DQB1*060101 (OR = 0.43; P < 0.0001) among the HIV‐1‐infected patients when compared with the controls. The haplotype DRB1*0902‐DQB1*030103 (OR = 10.65; P = 0.06) was significantly increased in HIV1 patients, while haplotypes DRB1*150101–DQB1*060101 (OR = 0.386, P < 0.0001), DRB1*030101–DQB1*020101 (OR = 0.197, P = 0.004) and DRB1*070101–DQB1*0202 (OR = 0.167, P = 0.001) were significantly decreased. Our results indicate clearly that there are HLA class II alleles involved in the susceptibility to and protection from HIV‐1 infection in our study group and further they vary in different ethnic groups reported in literature.

Frequency distribution of antigens in the human platelet antigen-1 system in the western Indian population

BACKGROUND: Neonatal alloimmune thrombocytopenic purpura (NAITP) occurring because of fetomaternal incompatibility in the human platelet antigen‐1 (HPA‐1) system is increasingly being detected worldwide. Several studies have reported the frequency and distribution of HPA‐1 alleles in different countries and ethnic populations. A paucity of data regarding the frequency of the antigens in the HPA‐1 system in the Indian population prompted an undertaking of this study. The molecular method of genotyping the platelet antigens is preferred to serology. It will enable future prenatal diagnosis in mothers suspected to have NAITP so that they can be managed better.

Use of Immunofluorescence and Confocal Laser Scanning Microscopy in Identifying Rare Cases of Anti-Neutrophil Cytoplasmic Antibodies (ANCA) Showing Dual Specificities to Myeloperoxidase and Proteinase3

Anti-neutrophil cytoplasmic antibodies (ANCA) are the immunodiagnostic markers for idiopathic necrotizing crescentic glomerulonephritis affecting mainly medium to small sized blood vessels. The diagnosis of ANCA associated vasculitis (AAV) is mainly based on clinical and histopathological characteristics along with the serological evidence. Immunofluorescence microscopy (IIF) is considered as the “gold standard” for ANCA detection, and ANCA showing two major patterns ie, cytoplasmic (c-ANCA) and perinuclear (p-ANCA) react with different antigenic targets of neutrophils like Proteinase3 (PR3) and Myeloperoxidase (MPO). A third unusual and rare immunofluorescence pattern called as “X-ANCA” or atypical ANCA is also sometimes seen. The difficulty in identification of ANCA immunofluorescence patterns is mainly seen due to the rare dual patterns seen in the same sera and also the additional nuclear immunofluorescence seen due to presence of anti-nuclear antibodies. ANCA testing by immunofluorescence and Confocal Laser scanning microscopy, as well as by specific ELISAs for detection of anti-PR3 and anti-MPO antibodies have helped in improving the diagnosis. Patients having dual specificities to MPO and PR3 in a patient is a rare finding. Among 425 clinically and histopathologically proven cases of AAV, eight patients (1.9%) had dual specificities, of which five patients showed mixed immunofluorescence pattern and 3 patients showed X-ANCA pattern which was confirmed by both immunofluorescence and Confocal Laser scanning microscopy and the dual specificities to MPO and PR3 were detected by individual ELISAs.