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R Verma

Publications and source records attributed to R Verma.

At least 55 records · Page 3Linked to original sources
PubMed1999

A quantitative analysis of G-actin binding proteins and the G-actin pool in developing chick brain.

The large G-actin pool in individual actively motile cells has been shown to be maintained primarily by the actin sequestering protein thymosin beta four (Tbeta4). It is not clear whether Tbeta4 or an isoform also plays a primary role in neural tissue containing highly motile axonal growth cones. To address this question we have made a definitive analysis of the relative contributions of all the known G-actin sequestering proteins: Tbeta4, Tbeta10, profilin, and phosphorylated (inactive) and unphosphorylated (potentially active) forms of both ADF and cofilin, in relation to the G-actin pool in developing chick brain at embryonic days 13 and 17. From our measurements we estimate the intracellular concentration of G-actin as 30-37 microM and of Tbeta4 as 50-60 microM in an 'average' brain cell in embryonic chick brain. No other beta thymosin isoforms were detected in these brain extracts. The ratio of soluble, unphosphorylated ADF to Tbeta4 is only 1:7 at 13 embryonic days, but increases to 1:4 at 17 days. Profilin and cofilin concentrations are an order of magnitude lower than Tbeta4. Combining the contributions of Tbeta4, unphosphorylated ADF and unphosphorylated cofilin, we estimate a mean G-actin critical concentration of approximately 0.45 microM and approximately 0.2 microM, respectively, in day 13 and day 17 embryonic brain extracts, suggesting a significant developmental decrease. We conclude that (a) Tbeta4 is the major actin sequestering protein in embryonic chick brain and the only beta thymosin isoform present; (b) ADF may play a significant developmental role, as its concentration changes significantly with age; (c) the known G-actin binding proteins can adequately account for the G-actin pool in embryonic chick brain.

Actin Depolymerizing Factors↗
PubMed1999

Rapid detection of Mycobacterium bovis on its lipid profile by thin layer chromatography.

Sixteen Mycobacterium bovis (M. bovis) strains isolated from bovine tissues and one standard reference strain of M. bovis AN5 alongwith other species of mycobacteria for comparison were investigated for the presence of phenolic glycolipid (PGL) and phthiocerol dimycocerosate (PDIM) for rapid identification of M. bovis by thin-layer chromatography (TLC). The study indicated presence of PGL with an Rf value of 0.75 in chloroform-methanol solvent in all 17 M. bovis strains. The dimycocerostate A corresponding to spot A was the major constituent among all the three spots in M. bovis strains. TLC appeared to be a promising alternative to conventional biochemical methods for identification of M. bovis taking into consideration both PGL and PDIM lipids.

Animals↗
PubMed1999

Towards effective obstetric anaesthetic audit in the UK.

Obstetric audit is multidisciplinary, but maternal mortality data represent the only national obstetric anaesthetic audit currently available in the UK. Maternity and neonatal audit is progressing towards the collection of both numerator and denominator data in order to compare local, regional and national figures. Obstetric anaesthetists as a professional group play a significant role in maternity care and have in the past developed a minimum data set. Such a set now requires revision of items, agreement on definitions and integration with national projects. Since local and regional obstetric anaesthesia data collection systems are available, albeit in various manual or computerized forms, this is an achievable target. A standard maternity and neonatal data set which incorporates obstetric anaesthetic clinical items could offer a qualitative comparison of process variables and outcome, but should be under professional anaesthetic control. In addition, the process may enable professional standards to be defined and tested so that high quality obstetric anaesthetic care can be maintained.

Journal Article↗
PubMed1999

Nephrin localizes to the slit pore of the glomerular epithelial cell.

BACKGROUND: Recognition that mutation of the protein nephrin, encoded by the NPHS1 gene, singly results in the cellular alterations that result in foot process effacement, and nephrotic range proteinuria emphasizes the pivotal role that this protein plays in regulating glomerular filter integrity. This article reports the development of reagents necessary to study the biology of nephrin in mouse, and describes the initial characterization of the nephrin protein. METHODS: A cDNA including the full-length mouse nephrin open reading frame was cloned and sequenced. Immuno-affinity purified polyclonal antiserum directed against the cytoplasmic domain of mouse nephrin was developed. RESULTS: Nephrin identified in mouse glomerular extract was found to be a glycoprotein with an apparent molecular mass of 185 kDa. As detected by indirect immunofluorescence microscopy and immunogold electron microscopy, nephrin was located only in visceral glomerular epithelial cells, where it was targeted to intercellular junctions of mature podocyte foot processes. In developing glomeruli of newborn mouse, antinephrin immunolocalized to the earliest slit pore regions between differentiating podocytes, sites where slit diaphragms first become visible. CONCLUSION: As a putative cell adhesion molecule of the immunoglobulin superfamily, nephrin likely participates in cell-cell interactions between podocyte foot processes and may represent a component of the slit diaphragm.

Age Factors↗
PubMed1998

Antibody engineering: comparison of bacterial, yeast, insect and mammalian expression systems.

Engineered antibody molecules, and their fragments, are being increasingly exploited as scientific and clinical tools. However, one factor that can limit the applicability of this technology is the ability to express large amounts of active protein. In this review we describe the relative advantages and disadvantages of bacterial, yeast, insect and mammalian expression systems, and discuss some of the problems that can be encountered when using them. There is no 'universal' expression system, that can guarantee high yields of recombinant product, as every antibody-based molecule will pose its own problems in terms of expression. As a result the choice of system will depend on many factors, including the molecular species being expressed, the precise sequence of the individual antibody and the preferences of the individual investigator. However, there are general rules with regards to the design of expression vectors and systems which will help the investigator to make informed choices as to which strategy might be appropriate for their application.

Animals↗
PubMed1998

The intermittent ductus revisited: echocardiographic evidence and successful coil occlusion: a case report and review of literature.

Intermittent occurrence of a large ductal shunt by physical examination and Doppler echocardiography is reported. Cineangiography confirmed a tubular ductus arteriosus with an angulated, narrow, pulmonary end. Presumably this angulation intermittently caused functional closure of the ductus. Trans-arterial delivery of coils resulted in complete occlusion.

Blood Flow Velocity↗
PubMed1998

Haemorrhagic septicaemia vaccines.

Haemorrhagic septicaemia (HS), an economically important disease of cattle and buffaloes, is caused by Pasteurella multocida (6:B). Vaccination against this disease is widely practised. Plain broth bacterins, or alum precipitated and aluminium hydroxide gel vaccines are administered twice a year since these vaccines offer an immunity of 4-6 months. Many countries use oil adjuvant vaccine (OAV), which gives both a higher degree and a longer duration of immunity up to 1 year. A double emulsion and multiple emulsion vaccine consisting of a thin viscosity have also been experimentally developed that gave an immunity parallel to OAV. Recently, a live vaccine developed from a fallow deer strain (B:3,4) has been used in Myanmar that offers an immunity for more than a year but is not free from constraints. The present review provides information on HS vaccines developed from time to time using whole bacteria or their components. The kinetics and isotype of antibody and cell-mediated immune responses have also been poorly understood so far, and hence information on their role in protection against HS is reviewed.

Animals↗
PubMed1997

Phosphorylation of Sic1p by G1 Cdk required for its degradation and entry into S phase.

G1 cyclin-dependent kinase (Cdk)-triggered degradation of the S-phase Cdk inhibitor Sic1p has been implicated in the transition from G1 to S phase in the cell cycle of budding yeast. A multidimensional electrospray mass spectrometry technique was used to map G1 Cdk phosphorylation sites in Sic1p both in vitro and in vivo. A Sic1p mutant lacking three Cdk phosphorylation sites did not serve as a substrate for Cdc34p-dependent ubiquitination in vitro, was stable in vivo, and blocked DNA replication. Moreover, purified phosphoSic1p was ubiquitinated in cyclin-depleted G1 extract, indicating that a primary function of G1 cyclins is to tag Sic1p for destruction. These data suggest a molecular model of how phosphorylation and proteolysis cooperate to bring about the G1/S transition in budding yeast.

Amino Acid Sequence↗
PubMed1997

Protection, humoral and cell-mediated immune responses in calves immunized with multiple emulsion haemorrhagic septicaemia vaccine.

A multiple emulsion (ME), vaccine against Pasteurella multocida (P52) infection in cattle was prepared and the efficiency in terms of immunity to direct challenge, duration of this immunity for up to 1 year and the role of humoral and cell-mediated immune mechanisms were studied. ME vaccine was sterile, safe and was potent when tested in rabbits and calves. Nineteen calves were immunized with a single 4 ml dose of ME vaccine intramuscularly. Group of these calves were challenge infected with virulent P. multocida (P52) (10(-1) 18 h old broth culture) given by the subcutaneous route at 21 days, 3 months, 6 months, 9 months and 1 year. All the immunized calves withstood challenge infection and showed 100% protection. Humoral immune response was measured by indirect haemagglutination test (IHA) and enzyme-linked immunosorbant assay (ELISA). Statistically ELISA values were found to be superior to IHA values because of small coefficient of variance. A fall in mean antibody titres during 24 h, 48 h, post-challenge infection was recorded whereas a steady increase in the titre after 72 h up to 10 days was noticed. The prechallenge mean titre in animals correlated with survival of animals. Humoral antibodies were detected as early as 7 day post-immunization and persisted to 1 year after immunization. Leucocyte migration inhibition test showed > 20% migration inhibition during all pre- and post-challenge periods in animals suggesting an involvement of cell-mediated immune mechanism in protection. Our findings suggested that both humoral and cell-mediated immune responses contribute to protection in vaccinated animals. The results of these studies of ME vaccine showed that it can be successfully used for the effective control of haemorrhagic septicaemia.

Animals↗
PubMed1997

Renal osmotic stress-induced cotransporter: expression in the newborn, adult and post-ischemic rat kidney.

The renal osmotic stress-induced cotransporter (ROSIT), a new putative member of a family of organic solute transporters, is highly expressed in the kidney. Our in situ hybridization data now reveal that large amounts of ROSIT mRNA can be found in the S3 segment of the proximal tubule. In the developing kidney, ROSIT mRNA is expressed after the S-shaped body stage. Because the S3 segment is the major site of damage in the post-ischemic kidney, we evaluated alterations in ROSIT mRNA expression after ischemic acute tubular necrosis. Renal osmotic stress-induced cotransporter mRNA levels were already decreased eight hours post-ischemia. At seven days post-ischemia, ROSIT mRNA reappeared in a mosaic pattern in the regenerating S3 segment, being fully expressed three weeks after the insult except for focal areas. The exact localization of this putative osmolyte transporter in the kidney, together with that of other known osmolyte transporter will contribute to a better understanding of the mechanism of medullary osmolyte accumulation and its vectorial transport.

Age Factors↗
PubMed1997

SIC1 is ubiquitinated in vitro by a pathway that requires CDC4, CDC34, and cyclin/CDK activities.

Traversal from G1 to S-phase in cycling cells of budding yeast is dependent on the destruction of the S-phase cyclin/CDK inhibitor SIC1. Genetic data suggest that SIC1 proteolysis is mediated by the ubiquitin pathway and requires the action of CDC34, CDC4, CDC53, SKP1, and CLN/CDC28. As a first step in defining the functions of the corresponding gene products, we have reconstituted SIC1 multiubiquitination in DEAE-fractionated yeast extract. Multiubiquitination depends on cyclin/CDC28 protein kinase and the CDC34 ubiquitin-conjugating enzyme. Ubiquitin chain formation is abrogated in cdc4ts mutant extracts and assembly restored by the addition of exogenous CDC4, suggesting a direct role for this protein in SIC1 multiubiquitination. Deletion analysis of SIC1 indicates that the N-terminal 160 residues are both necessary and sufficient to serve as substrate for CDC34-dependent ubiquitination. The complementary C-terminal segment of SIC1 binds to the S-phase cyclin CLB5, indicating a modular structure for SIC1.

Anaphase-Promoting Complex-Cyclosome↗
PubMed1997

Transumbilical catheterization of cerebral arteries.

OBJECTIVE AND IMPORTANCE: For neonates requiring cerebral endovascular procedures, an alternative route of arterial access, the umbilical artery, is described. Transfemoral catheterization, with its attendant risks, can thus be avoided. CLINICAL PRESENTATION: Six neonates with severe cardiac failure secondary to aneurysmal malformations of the vein of Galen underwent transarterial embolization in an effort to reduce flow through the intracranial arteriovenous malformations and therefore improve control of the high output cardiac failure. TECHNIQUE: The transumbilical route was used in each case, with successful devascularization of some of the malformations in all patients. The femoral arteries were preserved for future staged embolizations. No complications related to the umbilical artery catheterization were encountered. CONCLUSION: Femoral artery cannulation in neonates is technically challenging and may result in stenosis or thrombosis of the vessel. The umbilical artery provides an alternative route of vascular access for cerebral artery catheterization and embolization in these patients, preserving the femoral arteries for future interventions.

Angiography, Digital Subtraction↗
PubMed1995

Intraaortic spring coil loops: early and late results.

OBJECTIVES: Our aim was to determine the late fate of intraaortic spring coil loops after embolization of aortopulmonary vessels. BACKGROUND: In some aortopulmonary collateral vessels and patent ductus arteriosi, the narrowest segment is close to the aorta; coils used to close such vessels will "straddle" the lesion, allowing one or more coil loops to protrude into the aortic lumen. The consequences of this procedure are unknown. METHODS: We reviewed the cineangiograms of all patients who had at least one aortopulmonary collateral vessel or patent ductus arteriosus closure between January 1, 1988 and August 31, 1993. From this group, 53 patients had multiple-plane angiographic evidence of intraaortic coil loops. All subsequent cineangiograms were reviewed to determine coil position or movement and evidence of recanalization or endothelial coverage of the coil loop. We also reviewed each hospital record or communicated directly with referring physicians to identify any subsequent complications such as emboli or endocarditis. RESULTS: Of the 53 patients with intraaortic coil loops, 49 patients had closure of one or more aortopulmonary collateral vessels (59 vessels), and 4 had closure of a patient ductus arteriosus (4 vessels). Patient follow-up ranged from 1 day to 66 months (median 20 months); follow-up was not available in 6 patients. Five of the 53 patients (9.3%; 95% confidence limits [CL] 3.1% to 20.7%) died at operation or of end-stage heart failure. Patients with late angiography had no residual flow in 31 of 35 aortopulmonary collateral vessels (88.6%; 95% CL 73.3% to 96.8%), and 0.5 mm separated the coil and aortic contrast column in all 12 coils with adequate angiography, suggesting endothelial coverage of the intraaortic coil loop. No episodes of stroke, embolic events, endocarditis or coil migration were reported. CONCLUSIONS: Although coil occlusion of aortopulmonary collateral vessels or patent ductus arteriosi may produce intraaortic coil loops, endothelialization appears routine. No late complications associated with intraaortic coil loops were observed.

Aorta, Thoracic↗