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N Ramachandran

Publications and source records attributed to N Ramachandran.

At least 19 recordsLinked to original sources
PubMed2006

Current status of transrectal ultrasound-guided prostate biopsy in the diagnosis of prostate cancer.

In contemporary practice, most prostate cancers are either invisible on ultrasound or indistinguishable from concurrent benign prostatic hyperplasia. Diagnosis therefore rests on prostate biopsy. Biopsies are not simply directed at ultrasonically visible lesions, as these would miss many cancers; rather the whole gland is sampled. The sampling itself is systematic, using patterns based on prostate zonal anatomy and the geographical distribution and frequency of cancer. This review explains the evolution of the prostate biopsy technique, from the classical sextant biopsy method to the more recent extended biopsy protocols (8, 10, 12, >12 and saturation biopsy protocols). Extended protocols are increasingly being used to improve diagnostic accuracy, especially in those patients who require repeat biopsy. This trend has been facilitated by the ongoing improvement in safety and acceptability of the procedure, particularly with the use of antibiotic prophylaxis and local anaesthesia. The technical details of these extended protocols are discussed, as are the current data regarding procedure-related morbidity and how this may be minimized.

Age Factors↗
PubMed2004

Mortality patterns in dairy animals under organized herd management conditions at Karnal, India.

Mortality patterns of two Zebu cattle breeds, Sahiwal and Tharparkar, and two crossbred strains, Karan Swiss and Karan Fries, maintained at the National Dairy Research Institute, Karnal were studied. Nine-year (1989--90 to 1997--98) data on mortality were analysed for year, season, age and cause effects on mortality rate. The overall mortality was 14.17%. The mortality in Sahiwal, Tharparkar, Karan Swiss and Karan Fries averaged 14.35%, 7.21%, 17.12% and 13.46%, respectively. The breed mortality rate did not vary significantly between years, seasons, age categories and causes of disease. However, the trends indicated appreciable difference in mortality rates. The mortality was highest in the year 1994--95 (19.53%) and lowest in 1991--92 (8.56%). There was very little variation in seasonal mortality rate and mortality rate averaged 4.53%, 4.81% and 4.84% in hot-dry (March-June), hot-humid (July-October) and cold (November-February) seasons, respectively. The mortality up to 2 months of age accounted for a major share (50-60% or higher) in different breed groups. Digestive problems followed by respiratory disorders together accounted for 70-80% of total deaths.

Age Distribution↗
PubMed2001

Fluorophore-labeled S-nitrosothiols.

A series of fluorophore-labeled S-nitrosothiols were synthesized, and their fluorescence enhancements upon removal of the nitroso (NO) group were evaluated either by transnitrosation or by photolysis. It was shown that, with a suitable alkyl linker, the fluorescence intensity of dansyl-labeled S-nitrosothiols could be enhanced up to 30-fold. The observed fluorescence enhancement was attributed to the intramolecular energy transfer from fluorophore to the SNO moiety. Ab initio density functional theory (DFT) calculations indicated that the "overlap" between the SNO moiety and the dansyl ring is favored because of their stabilizing interaction, which was in turn affected by both the length of the alkyl linker and the rigidity of the sulfonamide unit. In addition, one of the dansyl-labeled S-nitrosothiols was used to explore the kinetics of S-nitrosothiol/thiol transnitrosation and was evaluated as a fluorescence probe of S-nitrosothiol-bound NO transfer in human umbilical vein endothelial cells.

Cell Membrane↗
PubMed2001

Mechanism of transfer of NO from extracellular S-nitrosothiols into the cytosol by cell-surface protein disulfide isomerase.

N-dansylhomocysteine (DnsHCys) is quenched on S-nitrosation. The product of this reaction, N-dansyl-S-nitrosohomocysteine, is a sensitive, direct fluorogenic substrate for the denitrosation activity of protein disulfide isomerase (PDI) with an apparent K(M) of 2 microM. S-nitroso-BSA (BSA-NO) competitively inhibited this reaction with an apparent K(I) of 1 microM. The oxidized form of DnsHCys, N,N-didansylhomocystine, rapidly accumulated in cells and was reduced to DnsHCys. The fluorescence of DnsHCys-preloaded human umbilical endothelial cells and hamster lung fibroblasts were monitored as a function of extracellular BSA-NO concentration via dynamic fluorescence microscopy. The observed quenching of the DnsHCys fluorescence was an indirect measure of cell surface PDI (csPDI) catalyzed denitrosation of extracellular S-nitrosothiols as decrease or increase in the csPDI levels in HT1080 fibrosarcoma cells correlated with the rate of quenching and the PDI inhibitors, 5,5'-dithio-bis-3-nitrobenzoate and 4-(N-(S-glutathionylacetyl) amino)phenylarsenoxide inhibited quenching. The apparent K(M) values for denitrosation of BSA-NO by csPDI ranged from 12 microM to 30 microM. Depletion of membrane N(2)O(3) with the lipophylic antioxidant, vitamin E, inhibited csPDI-mediated quenching rates of DnsHCys fluorescence by approximately 70%. The K(M) for BSA-NO increased by approximately 3-fold and V(max) decreased by approximately 4-fold. These findings suggest that csPDI catalyzed NO released from extracellular S-nitrosothiols accumulates in the membrane where it reacts with O2 to produce N(2)O(3). Intracellular thiols may then be nitrosated by N2O3 at the membrane-cytosol interface.

Animals↗
PubMed1999

N-dansyl-S-nitrosohomocysteine a fluorescent probe for intracellular thiols and S-nitrosothiols.

The fluorescence emission spectrum of N-dansyl-S-nitrosohomocysteine was enhanced approximately 8-fold upon removal of the NO group either by photolysis or by transnitrosation with free thiols like glutathione. The fluorescence enhancement was reversible in that it could be quenched in the presence of excess S-nitrosoglutathione. Attempts were then made to utilize N-dansyl-S-nitrosohomocysteine as an intracellular probe of thiols/S-nitrosothiols. Fluorescence microscopy of fibroblasts in culture indicated that intracellular N-dansyl-S-nitrosohomocysteine levels reached a maximum within 5 min. N-Dansyl-S-nitrosohomocysteine fluorescence was directly proportional to intracellular GSH levels, directly determined with HPLC. N-Dansyl-S-nitrosohomocysteine preloaded cells were also sensitive to S-nitrosoglutathione uptake as the intracellular fluorescence decreased as a function of time upon exposure to extracellular S-nitrosoglutathione.

Animals↗
PubMed1998

A distinct subgroup of small DRG cells express GDNF receptor components and GDNF is protective for these neurons after nerve injury.

Several lines of evidence suggest that neurotrophin administration may be of some therapeutic benefit in the treatment of peripheral neuropathy. However, a third of sensory neurons do not express receptors for the neurotrophins. These neurons are of small diameter and can be identified by the binding of the lectin IB4 and the expression of the enzyme thiamine monophosphatase (TMP). Here we show that these neurons express the receptor components for glial-derived neurotrophic factor (GDNF) signaling (RET, GFRalpha-1, and GFRalpha-2). In lumbar dorsal root ganglia, virtually all IB4-labeled cells express RET mRNA, and the majority of these cells (79%) also express GFRalpha-1, GFRalpha-2, or GFRalpha-1 plus GFRalpha-2. GDNF, but not nerve growth factor (NGF), can prevent several axotomy-induced changes in these neurons, including the downregulation of IB4 binding, TMP activity, and somatostatin expression. GDNF also prevents the slowing of conduction velocity that normally occurs after axotomy in a population of small diameter DRG cells and the A-fiber sprouting into lamina II of the dorsal horn. GDNF therefore may be useful in the treatment of peripheral neuropathies and may protect peripheral neurons that are refractory to neurotrophin treatment.

Age Factors↗
PubMed1993

Expression of the mature and the pro-form of human sterol carrier protein 2 in Escherichia coli alters bacterial lipids.

Sterol carrier protein 2 (SCP2) is a protein that is believed to be involved in the intracellular transport of cholesterol and phospholipids. Expression in mammalian COS cells of a cDNA encoding SCP2 revealed that the mature protein is synthesized as a pro-form containing a 20-residue amino-terminal leader sequence. The function of this presequence is currently not known, and pro-SCP2 is generally not detected in tissues. In order to obtain large quantities of pro-SCP2 as well as the mature form of human SCP2, Escherichia coli expression plasmids were constructed. Both proteins were produced in high yield (10-30% of the total cell protein) and were found in the supernatant fraction after cell lysis. Recombinant human SCP2 and pro-SCP2 were purified to homogeneity by acid precipitation followed by ion-exchange chromatography. Both recombinant human SCP2 and pro-SCP2 had sterol exchange activity similar to that seen with SCP2 purified from rat liver. In addition, the lipid content of SCP2- and pro-SCP2-producing E. coli was analyzed. Acidic lipids were significantly increased in the transfected cells. Specifically, fatty acids were increased 2-3-fold, phosphatidylglycerol was increased 2-fold, and lipid A was increased 3-4-fold, while neutral lipids were decreased 2-3-fold as compared to control cells. This alteration of the lipid composition of E. coli expressing SCP2 or pro-SCP2 is consistent with the proposed role for SCP2 in intracellular lipid movement.

Amino Acid Sequence↗
PubMed1993

Antiplatelet efficacy and specificity of DMP728, a novel platelet GPIIb/IIIa receptor antagonist.

The present study was undertaken to define the platelet GPIIb/IIIa affinity and specificity of DMP728, the cyclic [(D-2-aminobutyrate-N-methyl-L-arginyl-glycyl-L-aspartyl)-3-aminomethyl- benzoic acid] methane sulfonate. DMP728 demonstrated similar potency (IC50 = 0.046 +/- 0.002 microM) in inhibiting human platelet aggregation induced by various agonists or combination of agonists as assessed either by light transmittance aggregometry or impedance techniques. Similarly, DMP728 inhibited (IC50 = 2.3 +/- 0.8 nM) with equipotency in inhibiting 125I-fibrinogen binding to human gel-purified platelets regardless of the agonist used. In purified human GPIIb/IIIa ELISA, DMP728 demonstrated a competitive high affinity binding (Ki = 0.4 nM). Additionally, a high binding affinity (Kd = 0.1 nM) of 3H-DMP728 was demonstrated in human platelets. Furthermore, a platelet deaggregatory efficacy was shown. DMP728 demonstrated a high degree of specificity for platelet GPIIb/IIIa (alpha 2/beta 3) as compared to other integrins on endothelial cells (vitronectin receptors), platelets GPIb/1X, alpha v/beta 3, and other integrins on leukocytes or nonintegrin-related systems. In conclusion, DMP728 is a novel antiplatelet agent with high affinity and specificity for platelet GPIIb/IIIa.

Animals↗
PubMed1987

Penicillin allergy.

Explore the source record for details and available documents.

Drug Hypersensitivity↗
PubMed1981

Re-evaluation of molecular weight of pig heart NAD-specific isocitrate dehydrogenase.

NAD-specific pig heart isocitrate dehydrogenase was earlier reported, on the basis of gel filtration experiments, to have a molecular weight of approximately 340,000. In the present study, the enzyme is shown by equilibrium ultracentrifugation to have a weight average molecular weight of approximately 224,000 which can be attributed to a rapidly associating-dissociating protein system. The results of light-scattering measurements are consistent with the lower value of molecular weight. The enzyme exhibits an average frictional ratio, f/f0, of 1.39 as determined from ultracentrifuge experiments, and this deviation from typical proteins may account for the previous high molecular weight estimates. An average Stokes radius of 6.0 nm was calculated from the present gel filtration experiments. By use of this value and a sedimentation coefficient of 9.1 S, an average molecular weight of 245,000 has been calculated. Previous studies (Ramachandran, N., and Colman, R. F. (1980) J. Biol. Chem. 255, 8859-8864) have indicated that the enzyme is composed of three different subunits, present in the ratio 2:1:1, each of which has a molecular weight of about 40,000. These results, together with the present observations, lead to the conclusion that, under stabilizing conditions in solution, the NAD-dependent isocitrate dehydrogenase predominantly exhibits a minimum, molecular weight of 160,000 but behaves as a mixture of oligomeric species with an average Mr of about 224,000.

Animals↗
PubMed1980

Chemical characterization of distinct subunits of pig heart DPN-specific isocitrate dehydrogenase.

Pig heart DPN-dependent isocitrate dehydrogenase is heterogeneous on isoelectric focusing in 6 M urea. Under these conditions, three types of subunits (termed alpha, beta, and gamma), which have isoelectric points of about 5.7, 6.6, and 7.2, respectively, can be separated. On the basis of densitometric scans of analytical isoelectric focused gels stained with Coomassie blue, it is estiated that the subunits are present in the whole enzyme in the approximate ratio of 2 alpha:1 beta: 1 gamma. The three isolated subunits have distinct amino acid compositions and the amino acid composition of the total enzyme, when expressed as residues per average polypeptide chain of 40,000 daltons, is consistent with contributions of alpha, beta, and gamma subunits in the ratio of 2:1:1. Each isolated subunit yields a readly distinguishable tryptic peptide map which is much simpler than that of the total enzyme, and is consistent with the number of peptides expected from the lysyl plus arginyl residues for the subunit. The alpha and beta chains both have alanine as the NH2-terminal amino acid, whereas phenylalanine is the NH2-terminal residue of the gamma subunit. Since the alpha subunit exhibits a molecular weight of 39,000 and the beta and gamma subunits have indistinguishable molecular weights of 41,000, the two-band pattern observed on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate is understandable. These results suggest that a complete DPN-specific isocitrate dehydrogenase would have a minimum molecular weight of 160,000.

Amino Acids↗
PubMed1978

Evidence for the presence of two nonidentical subunits in NAD-dependent isocitrate dehydrogenase of pig heart.

The NAD-dependent isocitrate dehydrogenase [threo-D(S)-isocitrate:NAD(+) oxidoreductase (decarboxylating); EC 1.1.1.41] from pig heart is a multisubunit enzyme with a molecular weight of approximately 340,000. Electrophoresis of the enzyme in 10% polyacrylamide gels containing sodium dodecyl sulfate reveals two discrete bands with molecular weights of 41,000 and 39,000. The two bands exhibit approximately equal intensity when stained with Coomassie Blue, Amido Black, and Bromophenol Blue, suggesting that these polypeptide chains are present in equimolar quantities in the native enzyme. The same two-band pattern is observed when the sulfhydryl groups of the enzyme are blocked by alkylation with iodoacetate prior to electrophoresis, indicating that sulfhydryl oxidation is not responsible for the observed heterogeneity. Each of the subunits appears as a single band when eluted from the gel and again subjected to electrophoresis under the same conditions. Isocitrate dehydrogenase contains a total of 41 lysine and arginine residues per average subunit of 40,000 daltons. The observation of approximately 80 peptides upon paper chromatography and high voltage electrophoresis of tryptic digests of the enzyme is consistent with the existence of two distinct polypeptide chains. Dansylation yields two NH(2)-terminal amino acid derivatives: dansyl-phenylalanine and dansyl-alanine. It is concluded that the NAD-specific isocitrate dehydrogenase is composed of equal numbers of two nonidentical subunits.

Amino Acid Sequence↗
PubMed1977

Evidence for a critical glutamyl and an aspartyl residue in the function of pig heart diphosphopyridine nucleotide dependent isocitrate dehydrogenase.

The pH dependence of the maximum velocity of the reaction catalyzed by diphosphopyridine nucleotide (DPN) dependent isocitrate dehydrogenase indicates the requirement for the basic form of an ionizable group in the enzyme-substrate complex with a pK of 6.6. This pK is unaltered from 10 to 33 degrees C, suggesting the ionization of a carboxyl rather than an imidazolium ion. The enzyme is inactivated upon incubation with 1-cyclohexyl-3-(2-morpholinoethyl)carbodiimide in the presence of glycinamide or glycine ethyl ester. This inactivation is dependent on pH and the rate constant (k) increases as the pH is decreased in the range 7.3 to 6.25. A plot of 1/(H+) vs. 1/k suggests that the enzyme is inactivated as a result of the modification of a single ionizable group in this pH range. The coenzyme DPN and substrate alpha-ketoglutarate do not affect the rate of inactivation. In contrast, manganous ion (2 mM) and isocitrate (60 mM) produce a sevenfold decrease in the rate constant. The allosteric activator ADP (1 mM) does not itself influence the rate of inactivation; however, it reduces the concentration of Mn2+ (1 mM) and isocitrate (20 mM) required to produce the same decrease in the inactivation constant. These observations imply that the modification occurs at the substrate-binding site. Experiments employing [1-14C]glycine ethyl ester show a net incorporation of 2 mol of glycine ethyl ester per subunit (40 000), concomitant with the complete inactivation of the enzyme. The radioactive modified enzyme, after removal of excess reagent by dialysis, was exhaustively digested with proteolytic enzymes. High voltage electrophoretic analyses of the hydrolysate at pH 6.4 and 3.5 yield two major radioactive spots with approximately equal intensity, which correspond to gamma-glutamylglycine and beta-aspartylglycine, the ultimate products of reaction with glutamic and aspartic acids, respectively. Modification in the presence of manganous ion and isocitrate results in significant reduction in the incorporation of radioactivity into the two dipeptides. These results suggest that carbodiimide attacks one glutamyl and one aspartyl residue per subunit of the enzyme and that the integrity of these residues is crucial for the enzymatic activity.

Animals↗