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Biomedical subjects

U Kumar

Publications and source records attributed to U Kumar.

At least 91 records · Page 5Linked to original sources
PubMed1994

Mitochondria from Alzheimer's fibroblasts show decreased uptake of calcium and increased sensitivity to free radicals.

Many of the biological abnormalities seen in samples of the brain and other tissues from patients with Alzheimer's disease, are consistent with an excessive action of oxygenic free radicals. Mitochondria from fibroblasts grown in tissue culture from skin samples taken at autopsy from patients with histologically confirmed Alzheimer's disease, take up significantly less calcium than do fibroblast mitochondria from age matched non-demented controls. Following exposure to oxygenic free radicals generated by adding ferrous ions to the incubation mixture, calcium uptake is increased to a greater extent in Alzheimer's mitochondria than in controls. At the doses used, pretreatment with the lazaroid antioxidant drug U-74500A, or with the iron chelator deferoxamine, prevents the free radical induced increase in calcium uptake by control mitochondria, but provides only partial protection for Alzheimer's mitochondria. These observations suggest that Alzheimer's fibroblast mitochondria have impaired calcium transport processes and show increased sensitivity to oxygenic free radicals.

Adult↗
PubMed1994

Serum HCV RNA levels in chronic HCV hepatitis measured by quantitative PCR assay; correlation with serum AST.

A quantitative PCR assay is described for serum HCV RNA which is based on a recombinant competitive titrating RNA template. This template is derived from the 5' non-coding region of the genome and generates a shorter PCR product (93 bp) than that from the serum-derived wild type genomes (279) bp from the same sets of primers. By using this assay we have found serum HCV RNA concentrations ranging between 3.3 x 10(2) and 6.6 x 10(8) viral genomes/ml of serum in 15 samples from 10 different patients, 9 cases of chronic hepatitis and one case of fulminant hepatitis. Comparison of viraemias with serum transaminase levels has shown a good correlation for individual patients between these two parameters during chronic HCV hepatitis.

Adult↗
PubMed1994

Metalloporphyrin speciation by liquid chromatography and inductively coupled plasma-mass spectrometry.

Inductively coupled plasma-mass spectrometry is investigated for the detection of metalloporphyrins (cobalt protoporphyrin, hemin, and zinc protoporphyrin) separated by liquid chromatography. A Hypersil SAS C1 column with mobile phase containing 68% methanol at a pH of 4.5 is used. The detection limits obtained are in the nanogram range for cobalt and zinc protoporphyrins, whereas for hemin, detection limits are in the microgram range. The accuracy of the method is evaluated by determining zinc protoporphyrin from the whole blood of a lead-poisoned patient.

Chromatography, High Pressure Liquid↗
PubMed1994

The acute neurotoxic effect of beta-amyloid on mature cultures of rat hippocampal neurons is attenuated by the anti-oxidant U-78517F.

The neuropathological characteristics of Alzheimer's disease include the selective loss of neurons and the development of senile plaques and neurofibrillary tangles. These plaques and tangles are invariably associated with deposits of an insoluble protein, beta-amyloid, and the hypothesis that the beta-amyloid is responsible for the neuronal loss in Alzheimer's disease has received considerable support. The neurotoxic action of beta-amyloid has been demonstrated in primary cultures of cortical and hippocampal neurons and in PC-12 cells. The present study reports that the neurotoxicity of beta-amyloid is reduced by the antioxidant drug U-78517F. On the basis of this observation, we suggest that the neurotoxic effect of beta-amyloid is mediated by oxygen free radicals. The clinical use of antioxidant interventions may reduce neurodegeneration and the progression of the symptoms in patients with Alzheimer's disease.

Amyloid beta-Peptides↗
PubMed1993

Sequence variation in the large envelope glycoprotein (E2/NS1) of hepatitis C virus during chronic infection.

Sequence variation in the putative large surface glycoprotein (E2/NS1) of hepatitis C virus (HCV) was analyzed over 18 months in 4 patients with chronic relapsing non-A, non-B liver disease, of whom 2 were treated with lymphoblastoid interferon-alpha. Sequence analysis showed marked heterogeneity between isolates from different patients at the hypervariable region mapping to the amino-terminus of E2/NS1 (amino acid position, 386-411) and the extended sequence analyzed again confirmed the existence of two types, HCV1 and HCV2. Sequential nucleotide analysis of the hypervariable region over 1 year for each patient showed 0-9 amino acid changes, more common in the untreated than in the interferon-alpha-treated patients (3 and 9 vs. 1 and 0). Peaks of raised serum transaminases in individual patients showed no consistent association with mutations within this region. However, results do not exclude the possibility that viral persistence may be due to the emergence of "escape mutants" in the hypervariable region.

Adult↗
PubMed1993

Pendred's syndrome.

Explore the source record for details and available documents.

Audiometry, Evoked Response↗
PubMed1992

Quantitative dot-blot assay for proteins using enhanced chemiluminescence.

A sensitive non-radioactive method for detection of specific proteins on Western blots is commercially available. The protein is immobilized on nitrocellulose membrane and immunolabelled with HRP-conjugated secondary antibody. HRP catalyzes the oxidation of luminol, a cyclic diacylhydrazide, resulting in the emission of light which is recorded on film. Using dot blot, we have shown that the signal generated by this system is proportional to the amount of protein loaded onto the membrane. Standard curves were linear (r2 greater than 0.95) over a 10-50-fold range. Linearity was also achieved with tissue extracts probed for a specific antigen. The sensitivity of the method is such that less than 10 fmol protein can be measured. The sensitivity and range are comparable to a previously reported dot-blotting assay based on the use of 125I-protein A, but the method does not require the handling of radioactive compounds. This method was used to estimate the level of chromogranin A in a mixture of proteins extracted from human brain.

Adrenal Glands↗
PubMed1992

Serological response and detection of viraemia in acute hepatitis C virus infection.

The serological response during acute hepatitis C virus (HCV) infection was examined by enzyme-linked immunosorbent assay (ELISA) in sequential serum samples from 13 haemophiliacs following their first exposure to factor VIII concentrates contaminated with HCV. The commercially available C100-3 peptide and a new 22 kDa recombinant protein (p22) encoded by the nucleocapsid region of the viral genome were used for antibody detection, whilst a nested polymerase chain reaction (PCR) method was used for the detection of viraemia. In addition, eight sporadic cases of acute HCV infection were studied. The results in haemophiliacs demonstrated that seroconversion to the C100-3 antigen occurred in only one-third of the patients within 12 weeks of disease onset, but all of the patients had a diagnostic serological response to p22 during this phase of the disease. The new test was positive in all the sporadic cases at a time when the commercially available test was negative. Although PCR offers a sensitive method for the detection of recent HCV infection, the complex methodology makes it unsuitable for diagnostic laboratories. The new ELISA test with p22 may therefore have a useful diagnostic role in acute disease.

Adult↗
PubMed1992

Cloning and sequencing of the structural region and expression of putative core gene of hepatitis C virus from a British case of chronic sporadic hepatitis.

We report the cloning and sequencing of the putative structural region of the hepatitis C virus (HCV) genome (2229 nucleotides) from an isolate derived from a British case of chronic sporadic non-A, non-B hepatitis. The overall sequence shows a higher similarity with one type of HCV, HCV1 (92%), than with HCV2 (80%), is very highly conserved at the 5' end (99%) preceding the long open reading frame, is well conserved also in the putative core region (90 to 97%), but shows marked variation in the putative envelope region, particularly in the envelope 2/non-structural 1 region (70%). The putative core gene was cloned in pJ3 omega under the early simian virus 40 promoter and expressed in human hepatoma cells. A predominantly cytoplasmic 22K polypeptide was expressed which was antigenically reactive with serum from chronically infected HCV patients.

Amino Acid Sequence↗
PubMed1992

Hepatic changes during short-term haemodialysis.

Twenty five patients with end stage renal disease were studied to find out the biochemical and hepatic morphological changes during short term haemodialysis. Asymptomatic hepatomegaly was seen in 32% of cases, transaminase elevation in 28% and positive Australia antigen in 12% of cases. Histopathological changes were observed in all the 12 patients in whom liver biopsy was done. Ten patients (82%) exhibited multiple abnormalities. The commonest findings were fatty change (8 patients); hepatic congestion, focal necrosis, Kupffer cell hyperplasia (6 patients each); and portal triaditis (5 patients).

Adult↗
PubMed1990

Development of cholinergic markers in the neuroblastoma, N1E-115.

The neuroblastoma, N1E-115, was grown for 9 days after subcultivation. The development of acetylcholinestrase (AChE), choline acetyltransferase (CAT), QNB-binding, choline uptake, and acetylcholine release was measured on days 1, 3, 6, and 9. In parallel experiments the irreversible AChE inhibitor soman was added to neuroblastoma on day 1 and the development of the above parameters except for release, was followed. AChE activity in the normal cells was found to develop also after confluency, whereas CAT activity and QNB-binding followed the development of most of the cellular proteins, i.e., ceased to develop at confluency. Both choline uptake and acetylcholine release appeared independent of cellular development. In the soman-treated cells the development of AChE was inhibited for up to 6 days and thereafter developed with the same rate as in the normal cells. CAT and QNB-binding developed as in the normal cells, but at a significantly reduced level. The development of choline uptake was not significantly different in soman-treated and normal neuroblastoma. It is concluded that the development of the cholinergic marker QNB-binding is intimately associated with that of the "presynaptic" marker CAT, whereas the development of AChE seems to be unrelated to these cholinergic parameters. The choline transport and the acetylcholine release seem to be equally well expressed on all the days studied.

Acetylcholine↗
PubMed1990

Changes in striatal cholinergic, gabaergic, dopaminergic and serotoninergic biochemical markers after kainic acid-induced thalamic lesions in the rat.

Kinetic parameters of 3H-choline, 3H-GABA and 3H-dopamine (DA) uptakes in striatal homogenates containing nerve endings were determined 2 to 3 weeks after kainic acid injection into the ipsilateral "centre médian"-parafascicular complex area of the thalamus in the rat. Results showed a marked decrease in 3H-choline uptake concomitant with a selective decrease in Vmax. Data also showed a large decrease in 3H-GABA uptake resulting from a decreased affinity of uptake sites for their substrate. These data were associated with the previously described decrease in choline acetyltransferase and increase in glutamic acid decarboxylase apparent activity, respectively. An apparent marked increase in 3H-DA uptake was likewise measured, mainly related to an increase in Vmax. Determination of serotonin (5HT) and 5-hydroxyindole acetic acid (5HIAA) endogenous contents showed in the deafferented striatum a decrease in 5HT concentrations associated with an increase in 5HIAA levels. Taken together, all these changes in neurotransmitter markers suggest that, directly through the thalamostriatal pathway or indirectly, the thalamus can exert a complex influence on striatal cholinergic and GABAergic neuronal functions as well as on the activity of dopaminergic and serotoninergic striatal afferent fibers.

Animals↗
PubMed1988

gamma-Aminobutyric acid function in the rat striatum is under the double influence of nigrostriatal dopaminergic and thalamostriatal inputs: two modes of regulation?

Glutamic acid decarboxylase (GAD), gamma-[3H]-aminobutyric acid [( 3H]GABA) high-affinity uptake into synaptosomes, and endogenous GABA content were measured in the rat striatum 2-3 weeks following 6-hydroxydopamine injection in the ipsilateral substantia nigra to destroy the nigrostriatal dopaminergic pathway and after kainic acid injection into the centromedial-parafascicular complex of the ipsilateral thalamus to lesion the thalamostriatal input. Both lesions resulted in apparent GAD increase concomitant with a decreased [3H]GABA uptake into striatal synaptosomes. GABA content was increased selectively following the dopaminergic lesion. Kinetic analysis of the uptake process for [3H]GABA showed selectively a decreased Vmax following the dopaminergic lesion; in animals with thalamic lesion, however, the change only concerned the Km, which showed a decreased affinity of the transport sites for [3H]GABA. Determination of Km and Vmax for GAD action on its substrate glutamic acid showed an increased affinity of GAD for glutamic acid in the case of the dopaminergic lesion without any change in Vmax, whereas the thalamic lesion resulted in GAD increase concomitant with a selective increase in Vmax. These data suggest that striatal GABA neurons are under the influence of nigrostriatal dopaminergic neurons which may reduce the GABA turnover, whereas the exact nature of the powerful control also revealed on these neurons following thalamic lesion remains to be determined. Both lesions induced adaptive neurochemical responses of striatal GABA neurons, possibly reflecting in the case of the dopaminergic deprivation an increased GABA turnover.

Animals↗