Divergent strains of human immunodeficiency virus type 1 circulating in India, subtyped by heteroduplex mobility assay.

Genomic variations in HIV-1 represent a major problem in understanding disease progression, studying drug resistance and developing effective vaccines. Heteroduplex Mobility Assay (HMA) was used for analyzing HIV-1 subtypes resulting from genetic similarity or divergence of C2 -V3 -V5 region of envelope gene between HIV-1 strains obtained from clinical samples in a tertiary care center at Pune. DNA from the PBMCs of infected individuals was amplified by nested PCR. Heteroduplexes were then formed by denaturing DNA from the unknowns with DNA from the reference strains. The results were analyzed by polyacrylamide gel electrophoresis. Out of 177 samples analyzed, 170 were of subtype C (96%). Four samples were found to be of subtype B (2.2%); in three samples, no definitive assignment of subtype was possible by HMA and these perhaps could be circulating recombinant forms (CRFs) of HIV-1. These findings may have significant implications toward development of a candidate vaccine for India.

Early detection of HIV-1 in infants by PCR.

Infants of HIV-infected mothers are at great risk of becoming infected with HIV during childbirth. Many infants acquire HIV during labor and delivery. Others can acquire HIV through the mixing of fetal and maternal blood as the placenta separates. The duration of membrane rupture, acute chorioamnionitis and invasive delivery techniques that increase the baby's contact with the mother's blood have been associated with higher risks of MTCT during labor and delivery. HIV is present in breast milk and risk of its transmission during breastfeeding depends on several factors, including: infant age, pattern of breastfeeding, breastfeeding duration, breast health, maternal viral load and maternal immune status. Infants born to HIV infected mothers carry their mother's antibodies in their blood into the second year of life, even if the infants themselves are not infected. For this reason, standard HIV antibody tests cannot reliably confirm HIV infection in infants until after the maternal antibodies have disappeared. Tests that can diagnose pediatric HIV infection accurately during the first year of life include HIV-PCR, CD4/CD8 counts, P24 antigen tests, and viral cultures. PCR offers an effective tool to reliably diagnose HIV in a pediatric age group. Nineteen infants born by normal delivery to HIV-1 seropositive mothers were studied by PCR for the HIV1 env gene. Thirteen babies (68.5%) were negative whereas 6 babies were found to be positive (31.5%). Although PCR is one of the most useful tests for this clinical situation, it is not definitive. Therefore, PCR should be interpreted with caution and repeated at regular defined intervals, preferably lasting until the HIV antibody status of the infant is resolved.

Toxoplasmic encephalitis in acquired immunodeficiency syndrome.

OBJECTIVES: To study the clinical course and outcome of toxoplasmic encephalitis (TE) in patients with acquired immunodeficiency syndrome (AIDS). METHODS: Patients infected with human immunodeficiency virus (HIV) and neurological abnormality compatible with diagnosis of TE were enrolled in the study. These patients were treated with combination of trimethoprim/sulfamethoxazole and pyrimethamine. Response to therapy was assessed by clinical examination and repeat CT/MRI scan done after three weeks of starting treatment. Those showing response were put on prophylactic therapy. RESULTS: A total of 451 patients of HIV infections were admitted to this centre during the study period, of these 11 patients were diagnosed to have TE. The common presenting symptoms were fever (80%), seizures (45%), headache (45%) and altered sensorium (25%). Focal neurological deficit was present in 80% of cases. Nine cases had ring-enhancing lesions on CT scan while in the remaining two patient's ring lesions were seen on MRI. These were either multiple (55%) or solitary (45%). Antitoxoplasma antibody was detected in 10 patients. It was absent in one patient. Ten patients had clinical and radiological improvement with trimethoprim/sulfamethoxazole and pyrimethamine within 10 +/- 3 days of starting therapy. One patient died within 10 days of starting therapy. CONCLUSION: Toxoplasmosis is a common opportunistic infection of the central nervous system in patients with AIDS. Majority of patients with cerebral toxoplasmosis present with focal neurological abnormality in presence of characteristic neuroradiological abnormality and positive antitoxoplasma antibody titer. Response to empirical therapy helps to confirm the diagnosis, lifelong prophylaxis there after prevents relapse of potentially fatal and easily treatable condition.

A probable explanation for mild extra-articular manifestations in Indian patients of rheumatoid arthritis: a preliminary study.

One hundred four Rheumatoid factor (RF) positive Rheumatoid Arthritis (RA) patients fulfilling ARA criteria were screened for extraarticular manifestations. Rheumatoid nodules were present in 2.8% cases. Other extraarticular manifestations such as pulmonary, cardiac, occular, renal or GI involvement were absent in these cases. However, circulating immune complexes (CICs) were highly significantly raised in all the RA patients (P < 0.001). From amongst these cases a limited number (8 cases) have been subjected to qualitative and quantitative analysis of CICs to look for whether there could be any relationship between these and mild extraarticular manifestations that were being noticed in our groups of patients. Finding showed IgG-IgG CICs in five, IgG-IgM in two and IgG-IgA in one case. Quantitative analysis revealed mean IgG 4.97 +/- 1.7 IU/ml, IgM 14.58 +/- 5.53 IU/ml and IgA 5.08 +/- 1.53 IU/ml on LD Solugen plates. Serum concentration of C3 was not reduced (94.1 +/- 8.9 mg/dl). Low IgM contents of CICs and no reduction in complement level is the likely explanation for less severe inflammatory manifestations seen in our study. The conclusion and findings have been discussed in the light of observation reported by the Western workers.

HLA class-I and class-II antigen association in rheumatoid arthritis at Varanasi, India.

Clinical presentation of rheumatoid arthritis (RA) and its severity differs in different races. Genetic factors play a significant role in its predeliction. The present study was undertaken to find out association of HLA class I and class II antigens with rheumatoid arthritis prevalent in Asian Indians residing at Varanasi. Ninety rheumatoid arthritis patients strictly fulfilling American Rheumatism Association criteria were screened for prevalent HLA class I and class II antigen by Terasaki Microlympho-cytotoxicity test. Results were compared with 100 healthy controls and 35 Seronegative Spondyloarthritides cases (SSA). Rheumatoid arthritis patients showed increased frequency of HLA-A2 and B40 antigens compared to healthy controls (p < .001). SSA patients showed significantly increased Phenotype frequency (PF) of HLA-B27 (p < .0001) and B40 (p < .001). Significant detection of HLA-A2 exclusively in RA patients suggests a more positive association of A2 in rheumatoid arthritis at Varanasi. HLA-B40 could not be attributed absolute significance of association with SSA or RA as it showed increased frequency in both diseases.

Evaluation of cellular immunity in diabetic uraemics by cutaneous response to recall antigens and 2:4 dinitrochlorobenzene and T-cell rosette formation.

Twenty diabetic uraemics and twenty two healthy matched controls comprised the material for this study. Cell mediated immunity was assessed by estimation of T-cell rosette percentage and cutaneous response to recall antigens--purified protein derivative, candida antigen and 2:4 dinitrochlorobenzene. Results analysis revealed depressed cell mediated immunity in diabetic uraemics in the form of impaired cutaneous response to recall antigens and reduction in T cell rosette percentage, in comparison to controls.

Effect of levamisole as an immunomodulating agent in trophoblastic lesions.

Both cell mediated immunity and humoral immunity was assessed in 16 patients of hydatidiform mole and 6 patients of choriocarcinoma. Fifty percent patients of choriocarcinoma and 11 patients of vesicular mole were given levamisole (LVM) trial and were followed for 2 months. Absolute lymphocyte count (ALC) was significantly increased in vesicular mole after levamisole treatment but in choriocarcinoma no effect was obtained. Marked improvement of T cell rosette count was also seen in LVM treated patient of both vesicular mole (p < 0.001) and choriocarcinoma. Cutaneous DTH response to 2:4 DNCB in vesicular mole was also increased after LVM. Before treatment only 31.25% patients had strong cutaneous response but after treatment 53.35% patients had strong response, while cases of choriocarcinoma were unaffected. LVM also raised all the serum immunoglobulins (IgG, IgM, IgA) in both vesicular mole and choriocarcinoma. Hence, levamisole therapy was found to have a beneficial effect on both cellular and humoral immunity in the lesions of trophoblasts.

Tropical myositis. A clinical immunological and histopathological study.

Thirty cases of tropical myositis, (22 suppurative, 8 non-suppurative) aged 11 to 65 years were seen in a period of one year. There were 22 males and 8 females. There was a total of 78 muscular lesions in 22 suppurative cases and 19 muscular lesions in 8 non-suppurative cases. The most common presentation was localised myalgia (100%), fever (96.7%) generalized myalgia (56.7%), arthralgia (40%), pain in abdomen (33.3%) and breathlessness (30%). Extramuscular complications were present in 50% cases. Twenty four muscle biopsies were taken. Sixteen showed changes of suppurative myositis i.e. non-specific acute inflammatory reaction, muscle necrosis with myocytolysis, vacuolation of cytoplasm and loss of striations. Cell mediated immunity was found to be suppressed in patients of non-suppurative myositis in comparison with the suppurative group. IgG, IgA and IgM were significantly raised in patients in comparison to controls (p less than 0.05). The intact humoral immunity indicates good response to acute phase reaction and increased levels of IgG, IgA and IgM (specially IgG) can be taken as good prognostic parameter.

Pre-cancerous lesions of stomach.

Total 39 cases of carcinoma stomach were noticed out of 142 malignant tumours of GIT (27.46 percent). Histologically maximum cases were of diffuse type (56.41 percent) followed by intestinal type (35.89 percent) and indolent mucoid carcinoma (7.69 percent) of the stomach. The surrounding epithelium showed lot of changes in the intestinal type of carcinoma stomach. About 78.57 percent showed intestinal metaplasia, 14.28 percent of these cases showed chronic gastric ulcer and severe dysplasia (carcinoma in situ) and another 14.28 percent revealed villous adenoma with carcinoma in situ. In contrast to this, in diffuse variety, only 13.63 percent cases revealed intestinal metaplasia, 27.27 percent showed basal cell hyperplasia, stratification of the epithelium of crypts and diffuse infiltration of mucosa by malignant cells and 4.54 percent showed atrophic gastritis also. In mucoid carcinoma all cases had basal cell hyperplasia and stratification of crypts. Hence these conditions should be taken as premalignant lesions of stomach and should be cured in proper time.