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Biomedical subjects

R Abraham

Publications and source records attributed to R Abraham.

At least 73 records · Page 4Linked to original sources

Synthesis and secretion of lipoproteins by primary cultures of rat hepatocytes.

Synthesis and secretion of VLDL and LDL by primary cultures of rat hepatocytes maintained in serum free medium have been studied. A time-dependent increase found in the [3H]leucine labelled lipoproteins which floated at a density of 1.006 g/ml indicate the secretion of VLDL into the medium. That the hepatocytes also secrete. LDL is shown by floatation of [3H]leucine labelled lipoproteins by sequential centrifugation at a density range of 1.006-1.06 g/ml. Electrophoretic and immunoprecipitation analysis show that about 60% and 65% respectively of 3H-radioactivity is associated with apoB in the two fraction of lipoproteins. At about 12hr 70-75% lipoproteins in the culture medium is in the VLDL density range and 25-30% is in the LDL density range. Conversion of secreted VLDL to LDL has also been shown by incubating hepatocytes with pre-labelled lipoproteins when there is a decrease in the fraction of VLDL range with a corresponding increase in the fraction of the LDL density range. Addition of glycosaminoglycans such as hyaluronic acid, chondroitin sulphate, and heparin into the medium cause significant increase in the synthesis and secretion of [3H]apoB into the medium indicating a possible secretory control of apoB by local reuptake.

Animals↗

Mucosal microenvironment and mucosal response.

Considerable investigative effort is currently being directed towards the use of oral immunization for the prevention of mucosal infections, including otitis media, in infancy and childhood. The development of immune response to mucosally introduced vaccines or environmental antigens is significantly influenced by the mucosal microflora, enzymatic activity, factors influencing epithelial permeability and the nature of vaccine antigens administered. Studies carried out during the past several years have suggested that antigen uptake, antigen processing, and immune response to environmental antigens in the respiratory and intestinal mucosa are greatly altered by coexisting mucosal infections. High levels of ovalbumin or ragweed antigens were often observed in the serum associated with increased IgE-specific antibody responses following concurrent infection with respiratory syncytial virus, or rotavirus, respectively. The influence of the mucosal enzymatic environment has been recently evaluated after oral immunization with replicating poliovaccine or parenteral immunization with inactivated poliovaccines. High levels of neutralizing and VP3-specific antibody response and antibody activity against antigenic determinants generated in the intestine were observed characteristically after oral immunization with replicating virus. Such responses were conspicuously absent after parenteral immunization. These observations suggest that diverse elements of the mucosal microenvironment play an important role in the outcome of infections and development of immune responses at mucosal surfaces.

Animals↗

Synthesis and secretion of apo B containing lipoproteins by primary cultures of hepatocytes isolated from rats fed atherogenic diet.

The effect of experimentally induced atherosclerosis on the synthesis and secretion of lipoproteins in the density range of very low density lipoproteins (VLDL) and low density lipoproteins (LDL) have been studied using primary cultures of rat hepatocytes. Rats fed atherogenic diet showed higher levels of lipids associated with serum VLDL and LDL fraction, aorta and liver when compared with animals fed normal diet. Incorporation of [3H]leucine into apo B associated with the cell layer and secreted by hepatocytes from rats fed atherogenic diet was significantly more when compared with normal hepatocytes. [14C]Acetate incorporation studies showed that the synthesis of cholesterol was lower in hepatocytes from atherogenic diet fed rats, but more of the newly synthesised cholesterol was found in the secreted VLDL; secretion of lipids, particularly triglycerides, unesterified cholesterol and cholesterol in the lipoproteins in the density range of VLDL and LDL was significantly more in these hepatocytes. The relative distribution of [3H]-radioactivity in the LDL density range was 57% in hepatocytes from atherogenic diet fed animals as compared with 28% in controls, suggesting a relatively higher production of lipoproteins in the LDL density range than VLDL by these cells. These results indicate that the hypercholesterolemia in atherogenic diet fed animals may among other factors be caused by increased synthesis of apo B by liver cells and resultant increase in the secretion of apo B containing lipoproteins.

Acetates↗

Shedding of virulent poliovirus revertants during immunization with oral poliovirus vaccine after prior immunization with inactivated polio vaccine.

Fecal shedding of virulent revertant polioviruses was examined in isolates from infants previously immunized with > or = 1 dose of orally administered live attenuated polio vaccine (OPV) alone, enhanced-potency inactivated polio vaccine (EIPV) alone, or a combination of both. After administration of OPV alone, vaccine poliovirus serotypes were recovered in feces within 1 week and for as long as 31-60 days in 30%-80% of subjects after 1 or 2 doses and in 30%-50% after immunization with > or = 3 doses. No revertant poliovirus shedding was observed after OPV challenge in subjects immunized previously with > or = 3 doses of OPV. However, fecal shedding of revertant poliovirus after OPV challenge was observed in 50%-100% of subjects previously immunized with > or = 3 doses of the EIPV. These findings suggest that prior immunization with EIPV does not prevent fecal shedding of revertant polioviruses after subsequent reexposure to OPV.

Feces↗

Rapid detection of poliovirus by reverse transcription and polymerase chain amplification: application for differentiation between poliovirus and nonpoliovirus enteroviruses.

This report describes a rapid method of detection of poliovirus from viral isolates of clinical specimens using a single set of primers selected from the conserved 5' noncoding region of the poliovirus genome. Of the 144 clinical viral isolates examined, 81 were positive for polioviruses and 50 were positive for nonpoliovirus enteroviruses by tissue culture neutralization and infectivity. All 81 (100%) of the viral isolates identified as poliovirus by tissue culture infectivity were also positive by polymerase chain reaction. Of 50 nonpoliovirus enterovirus isolates found to be negative for poliovirus by tissue culture neutralization and infectivity, 48 were also negative by polymerase chain reaction. The high sensitivity (100%) and specificity (96%) of the primer set indicate that this assay has potential clinical applicability in the diagnosis of nonpoliovirus enterovirus infection.

Base Sequence↗

Delay in the diagnosis of rupture of the uterus due to epidural anesthesia in labor.

A case report of uterine rupture in labor with epidural anesthesia is presented. The woman had good analgesia on the left side, but complained of severe labor pais on her right side. Uterine rupture occurred which was manifested by sudden vaginal bleeding, fainting, low blood pressure and fetal distress. She did not feel any pains typical of uterine rupture. Rupture of the left uterine wall, with a large hematoma in the left parametrium was seen at surgery. It seems the unilateral anesthesia of the left side concealed the early signs of rupture.

Adult↗

Synthesis and secretion of VLDL by rat hepatocytes--modulation by cholesterol and phospholipids.

The synthesis and secretion of apoB, the major protein component of very low density lipoprotein (VLDL) and low density lipoprotein (LDL), were studied using rat hepatocytes maintained in primary culture. Supplementation of hepatocytes with rat serum VLDL and LDL increased the production of apoB while delipidated lipoproteins had no significant effect, suggesting a role for lipids in the production of apoB. Addition of cholesterol to the culture medium also increased the production of apoB in a concentration-dependent manner. Pulse labelling followed by chase in presence of cholesterol indicated enhancement in apoB secretion. Mevinolin which inhibits cholesterol synthesis significantly reduced the secretion of apoB. The presence of phosphatidylcholine and phosphatidylethanolamine in the culture medium also increased the secretion of apoB into the medium. These data suggest that availability of lipids, particularly cholesterol, is an important determinant of apoB synthesis and secretion as VLDL.

Animals↗

Neuropathy in chronic obstructive pulmonary disease: a multicentre electrophysiological and clinical study.

The incidence and type of neuropathy in patients with chronic obstructive pulmonary disease (COPD) were assessed. In a selected group of 89 patients, abnormal nerve conduction studies were found in 44%. Electrophysiological signs of a generalized peripheral neuropathy were found in 5-18%, depending on diagnostic criteria. Lesions which were thought to be due to compression or other forms of trauma were present in a further 24%. In the patients with peripheral neuropathy, the changes were distally predominant, affected mainly sensory fibres, and were consistent with an axonal type of neuropathy. There was a significant correlation between age and the incidence of peripheral neuropathy. Electrophysiological evidence of neuropathy was three times as common as clinical evidence. Much of the variation in the reported incidence of neuropathy in COPD is probably due to imprecise diagnostic criteria.

Adult↗

Metabolism of very low density lipoproteins--effect of sardine oil.

The effect of feeding fish oil on the metabolism of lipoproteins was studied in rats. Rats were fed diet containing 10% sardine or groundnut oil for 6 weeks. There was a significant decrease in the total cholesterol, phospholipids and triglycerides as well as the amount of the lipids associated with VLDL and LDL in serum in fish oil-fed rats. The synthesis and secretion of lipoproteins particularly apoB containing lipoproteins by primary cultures of hepatocytes from these rats were studied by 14(C)-acetate or 3(H)-leucine labelling. Primary cultures of hepatocytes derived from sardine oil-fed rats showed reduced incorporation of 3(H)-leucine into apoB containing lipoproteins secreted into the medium when compared to those fed groundnut oil, indicating a decreased synthesis and secretion of apoB. This was further confirmed by significantly lower incorporation of 14(C)-radioactivity into total and individual lipids of VLDL secreted into the medium, as well as that associated with different lipids in cell layer. The activity of lipoprotein lipase in adipose tissue and aorta was significantly higher in rats fed sardine oil which may cause an increased clearance of triglyceride-rich lipoproteins from circulation. These results indicate that the fish oil exerts hypolipidemic effect particularly by decreasing the synthesis and secretion of VLDL by liver and possibly by an increased clearance of triglyceride-rich lipoproteins from circulation.

Administration, Oral↗

The influence of periodate oxidation on monoclonal antibody avidity and immunoreactivity.

Eight monoclonal antibodies of different classes and isotypes and rabbit IgG were oxidized under a variety of conditions with 5-50 mM periodate. The number of aldehyde groups generated per immunoglobulin were measured by reduction with tritiated sodium borohydride or coupling of fluoresceinthiosemicarbazide. There were up to 25.5 aldehyde groups detected on the periodate-oxidized antibody 96.5, measured by borohydride reduction whereas the same conditions led to only 9.6 aldehydes per IgG on the antibody L6 of the same IgG2A isotype. Fluoresceinthiosemicarbazide bound to oxidized antibodies but not to the same extent as tritium. On mildly oxidized IgMs it was possible to generate more than 200 aldehyde groups per antibody molecule. Depending on the conditions and the antibody used periodate oxidation could lead to antibody crosslinking. The avidities of the modified antibodies were determined by Scatchard analyses and inhibition assays. A new mathematical method to evaluate the immunoreactivities of modified antibodies relative to the unlabeled native antibody from inhibition binding data was established. Periodate concentrations higher than 50 mM decreased the avidities and immunoreactivities of all IgGs tested. This effect is more pronounced if the oxidation is performed at pH 5.6 and 25 degrees C instead of pH 4.6 and 0 degree C. The BR96 antibody is inactivated even under mild oxidation conditions.

Animals↗

Effect of prior immunity on the shedding of virulent revertant virus in feces after oral immunization with live attenuated poliovirus vaccines.

Groups of infants were immunized with one or two doses of orally inoculated live attenuated Sabin poliovirus vaccine (OPV group) or with one or two doses of enhanced-potency inactivated poliovirus vaccine (EIPV) administered parenterally followed by one or two doses of OPV (EIPV-OPV group). The fecal specimens from both groups were tested for poliovirus shedding 1-2 months after OPV. The virus isolates were examined for nucleic acid sequences in the 5' noncoding regions (bases 480, 481, and 472 for serotypes 1, 2, and 3, respectively) to determine whether the viruses shed represented nonattenuated revertants, attenuated parent vaccine strains (nonrevertants), or both. In the OPV group, 4 of the 6 virus isolates recovered 30-60 days after the first immunization dose and 1 of the 3 isolates obtained after the second dose were found to be nonrevertants (parent vaccine strain). In contrast, 11 of the 12 isolates in the EIPV-OPV group were of the nonvaccine revertant virus types. The frequency for reversion appeared to differ for different poliovirus serotypes. However, all revertant type 3 isolates were recovered from subjects previously immunized with EIPV.

Antibodies, Viral↗

Desensitization of muscarinic M1 receptors of murine neuroblastoma cells (clone N1E-115) without receptor down-regulation and protein kinase C activity.

Acute desensitization of M1 muscarinic receptor-mediated responses (cyclic GMP formation and inositol phosphate release) was studied in murine neuroblastoma cells (N1E-115 clone). After a 45-min incubation at 37 degrees of N1E-115 cells either in monolayer or in suspension, with the muscarinic agonist carbachol (1 mM), the receptor-mediated cyclic GMP response to carbachol was nearly completely lost. This loss was associated with greater than 80% loss of carbachol-mediated inositol phosphate release. The protein kinase C activator phorbol 12-myristate 13-acetate (PMA) inhibited both responses with similar potencies. Carbachol or PMA reduced by 30-40% the number of muscarinic receptor sites for antagonist and agonist on intact cells (determined in binding assays using [3H]N-methylscopolamine) only for cells in monolayer and not for those in suspension. PMA but not carbachol pretreatment of cells in monolayer or in suspension caused a translocation of [3H]phorbol 12,13-dibutyrate binding and protein kinase C activity. In addition, desensitization to carbachol occurred in cells largely depleted of protein kinase C by chronic exposure to PMA. Thus, agonist-mediated down-regulation is not needed for muscarinic M1 receptor desensitization, which may be a result of the activation of a receptor-activated kinase different from protein kinase C.

Animals↗

Immunoreactivity of boronated antibodies.

Boronated antibodies have already been evaluated as agents in neutron capture therapy. Because the boronation procedure may alter the properties of the antibody it is important to study the immunoreactivity of the conjugated antibody before in vivo use. In our studies of two dextran-boronated monoclonal antibodies, anti-glial fibrillary acidic protein antibody, and anti-hyaluronectin antibody, we have used ELISA and immunohistological methods to determine antibody activity and specificity. A ten-fold decrease in activity was observed for both antibodies in ELISA, and non-specific interactions were seen in both immunohistological and ELISA procedures. The boron compound used was shown to be at least partly responsible for these non-specific interactions.

Animals↗

Adipose fatty acid composition and the risk of serious ventricular arrhythmias in acute myocardial infarction.

The relation between subcutaneous adipose tissue fatty acid composition and serious ventricular arrhythmias during acute myocardial infarction was studied in 2 groups of patients. In group 1 (n = 42), studied retrospectively, patients with ventricular fibrillation or tachycardia had a higher concentration of long-chain saturated fatty acids than those without (32.5 +/- 0.8% vs 29.7 +/- 0.4% [mean +/- standard error of the mean], p less than 0.01). In a prospective study, patients with arrhythmias (n = 106) had higher levels of long-chain saturated fatty acids (32.1 +/- 0.5% vs 30.7 +/- 0.4%, p less than 0.05) and of stearic acid (4.9 +/- 0.2% vs 4.4 +/- 0.1%, p less than 0.02) and a lower concentration of palmitoleic acid (7.3 +/- 0.3% vs 8.1 +/- 0.2%, p less than 0.005). When peak plasma creatine kinase concentrations were included with the individual fatty acid levels in a multiple logistic regression, only creatine kinase correlated significantly with ventricular arrhythmias (p less than 0.01). Thus, saturated fatty acids in cardiac membranes may lead to greater vulnerability to ventricular arrhythmias, although infarct size is the only statistically significant predictor after multiple regression analysis.

Adipose Tissue↗

Gastroplasty for morbid obesity: technique, complications and results in 60 cases.

A series of 60 gastroplasty operations based on Mason's vertical banded procedure are described. Details of technique, modifications, complications and results are discussed. Operative treatment results in a reduction to within 40 per cent of ideal weight in 80 per cent of patients between 12 and 18 months after surgery. The constricting band should not be greater than 5 cm; if it is most procedures will fail after 18 months. Pouch size should be measured and made no greater than 20 ml. The importance of nutritional counselling is emphasized. The surgical treatment should form part of a multidisciplinary approach with careful selection and long-term follow-up.

Adolescent↗