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Biomedical subjects

M Varma

Publications and source records attributed to M Varma.

At least 55 records · Page 3Linked to original sources
PubMed1995

Enhanced specificity of truncated transmembrane protein for serologic confirmation of human T-cell lymphotropic virus type 1 (HTLV-1) and HTLV-2 infections by western blot (immunoblot) assay containing recombinant envelope glycoproteins.

Immunoassays based on the highly immunogenic transmembrane protein of human T-cell lymphotropic virus type 1 (HTLV-1) (protein 21c) are capable of detecting antibodies in all individuals infected with HTLV-1 and HTLV-2. However, because of antigenic mimicry with other cellular and viral proteins, such assays also have a large proportion of false-positive reactions. We have recently identified an immunodominant epitope, designated GD21-I located within amino acids 361 to 404 of the transmembrane protein, that appears to eliminate such false positivity. This recombinant GD21-I protein was used in conjunction with additional recombinant HTLV type-specific proteins and a whole virus lysate to develop a modified Western blot (immunoblot) assay (HTLV WB 2.4). The sensitivity and specificity of this assay were evaluated with 352 specimens whose infection status was determined by PCR assay for the presence or absence of HTLV-1/2 proviral sequences. All HTLV-1-positive (n = 102) and HTLV-2-positive (n = 107) specimens reacted with GD21-1 in the HTLV WB 2.4 assay, yielding a test sensitivity of 100%. Furthermore, all specimens derived from individuals infected with different viral subtypes of HTLV-1 (Cosmopolitan, Japanese, and Melanesian) and HTLV-2 (IIa0, a3, a4, IIb1, b4, and b5) reacted with GD21-I in the HTLV WB 2.4 assay. More importantly, HTLV WB 2.4 analysis of 81 PCR-negative specimens, all of which reacted to recombinant protein 21e in the presence or absence of p24 and p19 reactivity in the standard WB assay, showed that only two specimens retained reactivity to GD21-I, yielding an improved test specificity for the transmembrane protein of 97.5%. None of 41 specimens with gag reactivity only or 21 HTLV-negative specimens demonstrated reactivity to GD21-I. In an analysis of additional specimens (n = 169) from different geographic areas for which PCR results were not available, a substantial increase in the specificity of GD21-I detection was demonstrated, with no effect on the sensitivity of GD21-I detection among specimens from seropositive donors. Thus, the highly sensitive, GD21-I-based HTLV WB 2.4 assay eliminates the majority of false-positive transmembrane results, thereby increasing the specificity for serologic confirmation of HTLV-1 and HTLV-2 infections.

Amino Acid Sequence↗
PubMed1993

Evaluation of plasma insulin-like growth factor-binding protein-3 as a potential predictor of preeclampsia.

OBJECTIVE: Elevated growth factor activity in women with preeclampsia is caused by an acid- and heat-labile protein complex with a native molecular mass of 150,000. The major plasma insulin-like growth factor-binding protein-3 has identical molecular characteristics. We hypothesized that increased endothelial cell release or decreased proteolytic degradation of insulin-like growth factor-binding protein-3 in preeclampsia would result in higher circulating levels of insulin-like growth factors and hence in increased mitogenic activity. STUDY DESIGN: Plasma insulin-like growth factor-binding protein-3 concentrations were determined by iodine-125 insulin-like growth factor-II radioligand blots of nonreducing gels in 16 normal and 16 preeclamptic patients collected from a prospective, case-control study. Enzyme-linked immunosorbent assays also were performed with a specific anti-insulin-like growth factor-binding protein-3 antiserum. RESULTS: Insulin-like growth factor-binding protein-3 concentrations were not different between women with normal versus preeclamptic pregnancies (p = 0.23). Enzyme-linked immunosorbent assays confirmed these results in a subset of patients. CONCLUSIONS: The findings indicate that insulin-like growth factor-binding protein-3 concentrations do not account for the elevated mitogenic activity observed in plasma from women with preeclampsia and are not a useful screening test for preeclampsia risk.

Adult↗
PubMed1993

In vitro BrdUrd incorporation of colorectal tumour tissue.

This study describes a novel in vitro method for the incorporation of the thymidine analogue, bromodeoxyuridine (BrdUrd), in fresh colorectal tumour tissue. Disaggregation by pronase, collagenase and DNAse resulted in high cell yields of viable single cell suspensions, representative of the original tumour, which could be infiltrated with BrdUrd. A modified ELISA identified optimal incubation times and BrdUrd concentrations. This technique has been used in preliminary studies to investigate two important areas intrinsic in the analysis of BrdUrd colorectal cell proliferation data: 1) to determine the effects of the individual constituents of the cell culture media, in particular glutamine, on BrdUrd incorporation in suspensions of colorectal cells and 2) to examine the denaturation step. This method will have wide applicability in investigations of cell proliferation status in both normal and diseased tissue.

Bromodeoxyuridine↗
PubMed1993

Platelet serotonin kinetics in acute myocardial infarction before and after thrombolysis.

Platelet count, platelet serotonin uptake and platelet serotonin content were analysed in 21 subjects consisting of 11 patients of evolving acute myocardial infarction (AMI) and 10 matched healthy controls before and after administration of streptokinase. Platelet counts were significantly reduced in AMI with a subsequent rise following thrombolysis. Platelet 5-HT uptake was also significantly increased in AMI and following thrombolysis, it showed a trend towards normalization. Platelet 5-HT content was significantly increased in AMI with no further significant change following thrombolysis. The results suggest that platelet activation as revealed by reduced platelet count and altered platelet serotonin kinetics, occurs in AMI and this activation is inhibited following thrombolysis. Further, it is also apparent that it is not the reperfusion but the thrombolytic agent that is responsible for inhibition of platelet activation.

Blood Platelets↗
PubMed1993

Eales' disease with progressive spastic paraparesis.

Eales disease is a disease of unknown aetiology characterised by ophthalmic changes and often neurological involvement as in multiple sclerosis. We are reporting a case of Eales' disease in a 27 yr old male with chronic progressive non compressive motor myelopathy.

Adult↗
PubMed1992

Nuclear type II sites and malignant cell proliferation: inhibition by 2,6-bis-benzylidenecyclohexanones.

Methyl-p-hydroxyphenyllactate (MeHPLA) is a bioflavonoid and/or tyrosine metabolite which may regulate cellular growth and proliferation through interactions with nuclear type II sites. Our current studies suggest that type II sites may function as MeHPLA receptors which are localized on the nuclear matrix, and occupancy of this binding site by MeHPLA directly correlates with the inhibition of normal and malignant cell proliferation. This ligand is inactivated by MeHPLA esterase in mammary tumors, resulting in a deficiency in MeHPLA, high quantities of unoccupied type II sites, and uncontrolled cellular proliferation. For these reasons we synthesized 2,6-bis((3,4-dihydroxyphenyl)methylene)-cyclohexanone (BDHPC) and 2,6-bis((3-methoxy-4-hydroxyphenyl)-methylene)cyclohexanone (BMHPC) for assessment as nuclear type II site and cell growth antagonists. These two esterase stable cyclohexanone derivatives, which bind to nuclear type II sites with high affinity (Kd 1-7 nM), mimic MeHPLA as cell growth-regulating agents. Dose-dependent occupancy of type II sites in MCF-7 human cells by BDHPC and BMHPC directly correlated with the inhibition of cell proliferation, and administration of BDHPC by silastic implant inhibited mouse mammary tumor growth in vivo. These findings demonstrate that esterase-stable type II antagonists such as BDHPC and BMHPC inhibit mammary cancer cell proliferation in vitro and in vivo and support earlier studies demonstrating that MeHPLA and functionally related compounds may regulate malignant cell proliferation at the level of this binding site.

Animals↗
PubMed1992

Apoptosis and DNA fragmentation precede TNF-induced cytolysis in U937 cells.

The hypothesis that activation of apoptosis and DNA fragmentation is involved in TNF-mediated cytolysis of U937 tumor cells was investigated. Morphological, biochemical, and kinetic criteria established that TNF activates apoptosis as opposed to necrosis. Within 2-3 h of exposure to TNF, U937 underwent the morphological alterations characteristic of apoptosis. This was accompanied by cleavage of DNA into multiples of nucleosome size fragments. Both of these events occurred 1-2 h prior to cell death as defined by trypan blue exclusion or 51Cr release. DNA fragmentation was not a non-specific result of cell death since U937 cells lysed under hypotonic conditions did not release DNA fragments. The percentage of cells undergoing apoptosis depended on the concentration of TNF and was augmented by the addition of cycloheximide. A TNF-resistant variant derived from U937 did not undergo apoptosis in response to TNF, even in the presence of cycloheximide. Furthermore, TNF could still activate NFkB in this variant, suggesting that this pathway is not involved in TNF-mediated cytotoxicity. Two agents known to inhibit TNF-mediated cytotoxicity, ZnSO4 and 3-aminobenzamide, were shown to inhibit TNF-induced apoptosis. Taken altogether, these data support the hypothesis that activation of apoptosis is at least one essential step in the TNF lytic pathway in the U937 model system.

Benzamides↗
PubMed1992

Platelet aggregation in migraine.

This study was conducted on forty male patients aged 17-35 years, suffering from migraine. Platelet aggregation was evaluated in them twice, once in the aura and headache phase of migraine and the other during the period in which patient was asymptomatic. We observed significant increase in platelet aggregation during the aura and headache phase of the migrainous attack in comparison to the headache-free interval. Platelet hyperaggregability may be an important factor in the pathogenesis of migraine.

Adult↗
PubMed1991

Selective effects of preeclamptic sera on human endothelial cell procoagulant protein expression.

Current concepts of preeclampsia suggest that dysfunction of maternal vascular endothelium in vivo is a central pathogenetic feature of this syndrome. This hypothesis is suggested by the activation of the coagulation cascade associated with preeclampsia and evidence for a role of endothelium in maintaining thromboresistance. Previous in vitro studies with monolayers of human umbilical vein endothelial cells demonstrated direct cytotoxic effects of sera from preeclamptic parturients. In the current studies, we have examined the in vitro expression of three procoagulant protein activities regulated by endothelial cells: cellular fibronectin, an important mediator of platelet aggregation known to be elevated in preeclamptic women in vivo; tissue factor, the most potent endogenous procoagulant activity; and von Willebrand factor, a major component of coagulation factor VIII. Monolayer cultures of human umbilical vein endothelial cells were incubated with pregnancy sera for 24 hours before these proteins and activities were quantified. Exposure of identical endothelial cell cultures to predelivery preeclamptic sera caused significantly greater release of cellular fibronectin than postdelivery preeclamptic or predelivery or postdelivery normal pregnancy sera (p less than 0.05). However, neither tissue factor activity nor von Willebrand factor expression appeared to be increased preferentially by preeclamptic sera. The data indicate that sera from women with preeclampsia induce a selective, but not a generalized, activation of endothelial cell procoagulant protein production.

Adult↗
PubMed1991

Platelet norepinephrine efflux in thrombotic stroke.

Twenty CT proven patients of thrombotic stroke who were non-diabetic, non-hypertensive and non-hyperlipidemic were evaluated in acute phase (within 7 days of onset of neurological deficit) for platelet count and spontaneous platelet norepinephrine (NE) efflux, measured as the fraction (of initial total content) of 3H-NE released from platelets in 30 and 60 min. NE efflux was significantly greater (P less than 0.001) in stroke patients (34.09 +/- 4.92% at 30 min and 50.45 +/- 7.1% at 60 min) as compared to controls (23.27 +/- 4.16% at 30 min and 39.49 +/- 3.27% at 60 min). This excessive efflux may result in increased concentrations of NE in local plasma, which by causing enhanced platelet aggregation and damage to vascular endothelium and vasospasm, may contribute towards the development and progression of cerebral thrombosis.

Blood Platelets↗
PubMed1990

Methyl p-hydroxyphenyllactate and nuclear type II binding sites in malignant cells: metabolic fate and mammary tumor growth.

Previous studies in our laboratory demonstrated that methyl p-hydroxyphenyllactate (MeHPLA) is an important cell growth-regulating agent which binds to nuclear type II binding sites in normal and malignant cells. Furthermore, this compound is deficient in a variety of rat and mouse mammary tumors and human breast cancer preparations, and this deficiency correlates with the loss of regulatory control. The present studies were performed to examine the metabolic fate of [3H]MeHPLA in mouse mammary tumors. Stable analogs of this compound such as 4,4'-dihydroxy benzylidene acetophenone were also assessed for nuclear type II site binding affinity and their ability to inhibit mammary cancer cell growth and proliferation in vitro and in vivo. The results demonstrate that mouse mammary tumors contain esterase activity which hydrolyzes MeHPLA to p-hydroxyphenyllactic acid, and this was the only major metabolite detected in these tumor preparations in vitro or in vivo. 4,4'-Dihydroxy benzylidene acetophenone, an esterase-stable MeHPLA analog, was found to bind with high affinity to nuclear type II sites but not the estrogen receptor, was capable of occupying type II sites in cultured MCF-7 cells, and inhibited the proliferation of these cells in concentrations which directly correlated with type II binding site occupancy. Similarly, 4,4'-dihydroxy benzylidene acetophenone administration by silastic implant or injection resulted in a dose-dependent inhibition of the growth of transplantable mammary tumors in mice, suggesting that this stable analog mimicks MeHPLA as a cell growth-regulating agent. Taken together, these results suggest esterase hydrolysis of MeHPLA in mammary tumors may result in a deficiency in this compound which correlates with a loss of regulatory control.

Animals↗
PubMed1990

Women with preeclampsia have higher plasma endothelin levels than women with normal pregnancies.

Endothelin, a newly discovered endothelium-derived peptide, has potent vasoactive properties in vivo and in vitro. The actions of endothelin in clinical conditions of hypertension have not yet been defined. This study examined the possible role of endothelin in the vasospasm and hypertension associated with a well-defined syndrome of gestational hypertension, preeclampsia. Our results indicate that the concentration of immunoreactive endothelin is elevated significantly in plasma obtained from women with preeclampsia and rapidly returns to a normal pregnancy value within 48 hours of delivery, as predicted by the prompt clinical resolution of this disorder. The findings suggest that endothelin may contribute to the vasospasm associated with this syndrome and lend further support to the involvement of endothelial cells in the pathophysiology of preeclampsia.

Adult↗
PubMed1989

Use of azygos vein as interposition graft for surgical unifocalization of pulmonary blood supply.

Hemodynamic repair in patients with pulmonary atresia, ventricular septal defect, hypoplastic pulmonary arteries with arborization defects, and major aortopulmonary collaterals necessitates prior unifocalization of pulmonary artery blood supply. When direct anastomosis between these collateral vessels and the central pulmonary arteries is not technically feasible, an interposition graft is required. Dacron, polytetrafluoroethylene, and pericardium have been used for this purpose. We describe our experience with the use of azygos vein as an interposition graft in 3 patients who underwent unifocalization of the pulmonary blood supply to the right lung. Our experience suggests that the azygos vein is a useful graft for use in this specific form of unifocalization procedure.

Aorta, Thoracic↗