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Biomedical subjects

R Sharma

Publications and source records attributed to R Sharma.

At least 37 records · Page 2Linked to original sources

Evidence of immunosuppression by bovine respiratory syncytial virus.

Respiratory syncytial virus (RSV) is a major respiratory pathogen in human infants and calves. Calves and lambs infected with bovine RSV show mild clinical signs but they are more susceptible to secondary infection with Pasteurella haemolytica. Lambs infected with P. haemolytica 6 days after experimental infection with bovine RSV had significantly greater magnitudes of fever, higher disease and lesion scores and higher mortality rates than those infected with P. haemolytica or bovine RSV alone (P less than 0.05). Experimental infection with bovine RSV is characterized by alterations in lymphocyte subpopulations and down-regulation of some of their functions. For example, the number of T helper cells is significantly reduced during the first week of infection and peripheral blood mononuclear cells obtained from bovine RSV-infected lambs were less responsive to the mitogen phytohaemagglutinin but more susceptible to P. haemolytica cytotoxin than those obtained from control lambs. Infection with bovine RSV does not significantly affect the humoral immune responses of lambs against P. haemolytica cytotoxin. Bovine RSV does not appear to affect the capacity of alveolar macrophages to present antigens in vitro.

Animals

Vasoactive substances induce cytoskeletal changes in cultured rat glomerular epithelial cells.

Angiotensin II (ANG II), atrial natriuretic peptide III (ANP), and sodium nitroprusside (SNP) alter capillary hydraulic conductivity in isolated glomeruli. These agents also affect cyclic nucleotide levels of glomerular epithelial cells (GEC). ANG II increases cAMP, whereas ANP and SNP increase cGMP. The effects of these vasoactive substances on GEC cytoskeleton were tested by incubating cells from primary cultures or an established cell line with each agent. Changes in the cytoskeleton were assessed by staining for F-actin with Bodipy phallacidin and for tubulin with NBD-colcemid. Control cells exhibited short bundles of F-actin or stress fibers near the base of the cells. These were frequently arranged in parallel and occasionally appeared to radiate from the center of the cell. Microtubules were arranged in a fine network throughout the cell with increased density adjacent to the nucleus and within the nucleolus. Incubation of GEC with 10(-7) M ANG II, cholera toxin, or 8Br-cAMP for 2 at 37 degrees C resulted in rearrangement of F-actin into distinct stellate patterns with a decrease in the relative intensity of the peripheral staining, all concurrent with a fivefold increase in intracellular cAMP. The incubation of GEC with 10(-6) M ANP or 10(-7) M SNP for 2 h at 37 degrees C resulted in apparent disassembly of stress fibers, sparse and more diffuse fluorescence, and some increase of fluorescence at the periphery of the cells, all concurrent with a 10-fold increase in intracellular cGMP. Cytochalasin D incubation led to complete disassembly of actin filaments.(ABSTRACT TRUNCATED AT 250 WORDS)

Actin Cytoskeleton

Measurement of albumin reflection coefficient with isolated rat glomeruli.

Macromolecular permeability of the glomerular capillary has been inferred from the clearance of endogenous protein or infused macromolecules. Permeability is increased after treatment with polycations as well as after renal injury. It has previously been shown that the capillaries of glomeruli isolated from normal mammals expand or collapse in response to transcapillary albumin gradients and that the magnitude of the changes in capillary volume and in total glomerular volume are directly proportional to the applied oncotic gradients. In the experiments presented here, the volume responses of control glomeruli and of glomeruli treated with protamine (100 to 600 micrograms/mL for up to 60 min) were used to calculate the albumin reflection coefficient, sigma albumin, and the convectional permeability, P convectional albumin = (1 - sigma albumin), of the capillary wall. Sigma albumin for normal glomeruli was about 1 (P convectional albumin = 0); sigma albumin fell to a minimum of 0.2 +/- 0.1 (P convectional albumin = 0.8 +/- 0.1) after incubation with protamine sulfate (600 micrograms/mL) for 30 min. Retraction and fusion of podocyte foot processes and denudation of the underlying matrix was seen on scanning electron micrographs of protamine-treated glomeruli. These results confirm that it is possible to study macromolecular permeability of the glomerular capillary in vitro and to calculate sigma albumin independent of hemodynamic and systemic humoral influences. This method will permit the assessment of the effects of individual mediators of glomerular injury and the study of glomeruli from kidneys affected by experimentally induced or naturally occurring renal diseases.

Albumins

Intradural spinal cavernomas.

Intradural cavernomas are rare vascular lesions of the spinal cord. Four cases of histologically verified cavernomas of the cord are reported, of which two were extramedullary and two were intramedullary in location. Progressive neurological deficit was the presenting feature in three cases while one patient had a rapid evolution of neurological deficits and was found at surgery to have had bled from the extramedullary lesion. All the patients were subjected to surgery and total excision of the cavernomas was carried out in each case. While two patients improved after surgery the other two remained static. The available literature on spinal cord cavernomas is reviewed.

Adult

Immunocytochemical and electrophysiological investigations of central neurones in the parasitic nematode Ascaris suum.

Here we report on preliminary investigations into the morphological, neurochemical and electrophysiological characteristics of central neurones in the parasitic nematode Ascaris suum. The neurones do not display all or none action potentials and apparently have low resting membrane potentials. Neurones have been shown to respond to acetylcholine. Immunoreactivity to a diverse range of neuropeptides has been demonstrated.

Acetylcholine

Low concentrations of neuroactive peptides modulate cholinergic transmission and cyclic AMP levels in Helix aspersa.

1. FMRFamide and the Catch relaxing peptide (CARP) at 0.01-0.1 nM modulate acetylcholine (ACh) induced currents of identified neurons of Helix aspersa, but have no direct effect on membrane current and conductivity. 2. Both FMRFamide and CARP noncompetitively inhibit ACh Cl- responses while having either no effect or increasing ACh Na+/K+ response. 3. The inhibitory effect of FMRFamide and CARP on the ACh Cl- response was eliminated following pretreatment with forskolin (20 microM), an activator of adenylate cyclase. 4. Ascaris peptide (ASC) and a synthetic CARP analogue NORL-CARP, in which the amino acid methionine is replaced by either leucine or norleucine, at 1-10 microM, showed no effect on responses to ACh. 5. FMRFamide and CARP, at 10 nM, increased cAMP levels to 240% and 148% respectively above resting basal cAMP levels, while ASC and NORL-CARP had no significant effective. 6. Our results suggest that FMRFamide and CARP, in low concentrations, modulate ACh responses of Helix neurons, possibly through changes in cAMP levels. They also indicate the importance of the presence of methionine in these neuroactive peptides.

Acetylcholine

Age-specific development of malate-aspartate shuttle in the liver and kidney of mice.

The activities of malate-aspartate shuttle enzymes were measured in the liver and kidney of 15-, 30-, and 60-day old mice. The results indicate that the activities (U/mg protein) of both isoenzymes (cytosolic and mitochondrial) of both malate dehydrogenase and aspartate aminotransferase are significantly higher in the liver of 15-day old mice than in the liver of 30- and 60-day old animals. However, the shuttle enzymes showed a peak value in the kidney of 30-day old mice. In vitro reconstitution of malate-aspartate shuttle showed a similar pattern of activity in the tissues studied. These findings suggest that the activity of malate-aspartate shuttle is expressed differentially in these tissues of mice at different postnatal ages.

Aging

Angiomyolipoma of kidney.

Angiomyolipoma is an uncommon benign tumour accounting for less than 1% of all surgically excised tumours of the kidney. It may present as an isolated lesion or in association with tuberous sclerosis. An impalpable tumour is often detected incidentally during investigation of a case for some urinary tract pathology. CT scan has proved to be a useful investigation for pre-operative diagnosis; it may however, be of no value if the fat content of the tumour is low. Here a case of angiomyolipoma of left kidney is reported where pre-operative diagnosis was suggested by CT scan and histopathology confirmed it.

Female

Effect of pregnancy associated hypertension on immunoglobulin levels in newborns.

In a prospective study of 124 neonates born to mothers with normal pregnancy and pregnancy associated hypertension (PAH), serum IgG, IgA and IgM were estimated by single radial immunodiffusion technique. Significantly low levels of IgG were found in mothers having PAH, as compared to normal pregnancy (p less than 0.001), whereas IgA and IgM showed no difference in the two groups. There was no statistical difference in maternal and cord blood IgG in either the control or study group. IgG was significantly higher (p less than 0.001) in cord blood of babies born by vaginal route as compared to forceps (via vaginal route) or cesarean section. IgA and IgM levels did not vary with mode of delivery.

Adult

Nucleophilic scission of thioester linkages and conformational changes in human plasma low density lipoproteins.

A comparison between conformations of native and methylamine modified human plasma low density lipoproteins (hydrated density 1.032-1.043 g/ml) has been presented. Near UV circular dichroism and difference absorption spectra of modified low density lipoprotein have suggested substantial differences in the local environments of several aromatic amino acid side chains. Relatively lower ellipticity at 222 nm of modified lipoprotein indicated alterations in the secondary structures of its protein moiety. Nucleophilic reaction of methylamine did not cause the peptide bond scission but it brought conformational transition such that some of the buried hydrophobic domains of the protein moiety got exposed to the aqueous environment.

Apolipoproteins B

Pregnancy in a rudimentary uterine horn.

A case of pregnancy reaching term in a rudimentary non-communicating horn of the uterus is presented. The pregnancy resulted in the delivery of a live child by cesarian section. The case is being reported for the rarity with which the foetus survives in this condition.

Adult

Purification and characterization of glutathione S-transferases from rat pancreas.

Glutathione S-transferases (GSTs) of rat pancreas have been characterized and their interrelationship with fatty acid ethyl ester synthase (FAEES) has been studied. Seven GST isozymes with pI values of 9.2, 8.15, 7.8, 7.0, 6.3, 5.9 and 5.4 have been isolated and designated as rat pancreas GST suffixed by their pI values. Structural, immunological and kinetic properties of these isozymes indicated that GST 9.2 belonged to the alpha class, GST 7.8, 7.0, 6.3 and 5.9 belonged to the mu class, whereas GST 8.15 and 5.4 belong to pi class. The N-terminal sequences and pI values of the mu class isozymes suggested that rat GST subunits 3, 4 and 6 may be expressed in pancreas. N-Terminal sequences of both the pi class isozymes, GST 8.15 and 5.4, were similar to that of GST-P, but there were significant differences in the substrate specificities of these two enzymes. Results of peptide finger print studies also indicated minor structural differences between these two isozymes. None of the GST isozymes of rat pancreas expressed FAEES activity. Rat pancreas had a significant amount of FAEES activity, but it segregated independently during the purification of GST indicating that these two activities are expressed by different proteins and are not related as suggested previously.

Acyltransferases

Independent segregation of glutathione S-transferase and fatty acid ethyl ester synthase from pancreas and other human tissues.

Glutathione S-transferase (GST) isoenzymes of human pancreas were purified, characterized and evaluated for their possible role in the metabolism of ethanol. Human pancreas has at least two GST isoenzymes belonging to the Alpha class (pI 8.8 and 8.1), one belonging to the Mu class (pI 6.4) and one belonging to the Pi class (pI 4.9). During the purification of GSTs from pancreas as well as from heart, liver, lung, brain and muscle, the fatty acid ethyl ester synthase (FAEES) activity was monitored in order to evaluate the role of GSTs in metabolism of ethanol, as suggested in earlier studies. Both t.l.c. and h.p.l.c. were used to identify ethyl oleate in reaction mixtures to monitor FAEES activity. During the purification of GSTs with the use of affinity chromatography on GSH linked to epoxy-activated Sepharose 6B, FAEES and GST activities from each of these tissues segregated independently. Purified GST isoenzymes from these tissues did not exhibit any FAEES activity. Antibodies raised against Pi-class GST, as expected, immunoprecipitated most of the GST activity of brain and heart without precipitating FAEES activity. These results suggest that human GST isoenzymes belonging to the Alpha, Mu and Pi classes do not express FAEES activity. The independent segregation of GST and FAEES activities was further demonstrated by monitoring GST activity during the purification of FAEES from pancreas. It was found that purified FAEES had no GST activity towards 1-chloro-2,4-dinitrobenzene and a number of other electrophilic substrates. Results of these studies demonstrate that FAEES and GSTs are distinct proteins.

Acyltransferases

The anionic conjugates of bilirubin and bile acids stimulate ATP hydrolysis by S-(dinitrophenyl)glutathione ATPase of human erythrocyte.

These studies demonstrate that bilirubin-ditaurate (an analog of bilirubin-diglucuronide), lithocholic acid 3-O-sulfate, and lithocholic acid 3-O-glucuronide, which are believed to be transported from liver into bile through an active transport process stimulate ATP hydrolysis by purified dinitrophenylglutathione ATPase of human erythrocytes. The Km and Vmax values of the enzyme for these substrates are similar to those for dinitrophenylglutathione indicating the transport mechanisms for bilirubin conjugates, and anionic bile acid-conjugates from hepatocytes to bile and transport of GSH-conjugates from erythrocytes may be mediated by similar mechanisms.

Adenosine Triphosphatases

Purification and characterization of an ATPase from human liver which catalyzes ATP hydrolysis in the presence of the conjugates of bilirubin bile acids and glutathione.

An ATPase has been purified from the membrane fraction of human liver which catalyzes ATP in the presence of bilirubin ditaurate, lithocholic acid 3-O-sulfate and lithocholic acid 3-O-glucuronide as well as dinitrophenylglutathione and other glutathione conjugates. Its subunit Mr value (38,000) and immunological properties are similar to dinitrophenylglutathione ATPase of human erythrocytes. Kinetic constants of the enzyme for the conjugates of glutathione, bile acids and bilirubin are comparable indicating that this ATPase may mediate active transport of all these anionic conjugates in liver.

Adenosine Triphosphatases