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Biomedical subjects

P Saxena

Publications and source records attributed to P Saxena.

At least 37 records · Page 2Linked to original sources
PubMed1999

Blood transfusion practices--a case study.

Blood transfusion practices of the treating doctors in a district hospital in Haryana were studied through retrospective study of blood bank records for the years 1992 to 1994 and interview of the clinicians of various specialities in the hospital. It was found that utilization of the whole blood was 90%, 89% and 81% respectively of the total blood units utilized during this period. Single unit transfusions out of the total transfusions done were 87% in 1992 and 1993 while these were 89.9% in 1994. Blood was often requested for volume replacement in acute haemorrhage. Only homologous blood transfusions were done at the hospital. This study has highlighted that there is scope for improvement of blood transfusion practices by strictly following the indications for use of blood, promoting the preparation and use of blood components, use of plasma expanders for acute blood loss, avoiding single unit transfusions and promoting the use of autologous blood during routine surgery.

Acute Disease↗
PubMed1999

5-HT receptors mediating external carotid vasoconstriction in vagosympathectomized dogs.

One specific example reflecting the complexity of cardiovascular responses induced by serotonin (5-hydroxytryptamine; 5-HT) and the progress achieved in the pharmacological characterization of the receptors involved can be illustrated by the effects of 5-HT on the canine external carotid artery bed. Within this framework, it has been shown that the external carotid vasoconstrictor response to 5-HT in the dog is mediated by '5-HT1-like' receptors, which being blocked by the 5-HT1B/1D receptor antagonist GR127935, resemble 5-HT1B/1D (previously called 5-HT1D beta/1D alpha) receptors. It was proposed that these receptors could belong to the 5-HT1B, rather than the 5-HT1D, subtype on the basis of their resistance to blockade by a high dose of ritanserin (a potential 5-HT1D receptor antagonist) and the presence of mRNA for 5-HT1B(5-HT1D beta) receptors, but not for 5-HT1D(5-HT1D alpha) receptors, in vascular smooth muscle. With the advent of subtype-selective antagonists it was subsequently shown that external carotid vasoconstriction to 5-HT and sumatriptan is dose-dependently antagonized by the selective 5-HT1B receptor antagonist SB224289 (2,3,6,7-tetrahydro-1'-methyl-5-[2'-methyl-4' (5-methyl-1,2,4-oxadiazol-3-yl) biphenyl-4-carbonyl] furo [2,3-f] indole-3-spiro-4'-piperidine hydrochloride), whereas the selective 5-HT1D receptor antagonist BRL15572 (1-(3-chlorophenyl)-4-[3,3-diphenyl (2-(S,R) hydroxypropanyl) piperazine] hydrochloride) was ineffective. These findings represent the first in vivo evidence showing that vascular constriction induced by 5-HT and sumatriptan is mediated primarily via 5-HT1B, but not 5-HT1D receptors. The pharmacological profile of these receptors could be similar (isolated human temporal artery and porcine carotid arteriovenous anastomoses) to other putative 5-HT1B receptors mediating vasoconstrictor responses. In view of the putative pathophysiologic role of external carotid (and extracerebral) vasodilation in migraine, the constriction of these blood vessels by sumatriptan via 5-HT1B receptors may be, at least partly, responsible for its therapeutic efficacy in migraine.

Animals↗
PubMed1998

Mutational analysis of domain II beta of bacteriophage Mu transposase: domains II alpha and II beta belong to different catalytic complementation groups.

This study examines the contribution of domain II beta of bacteriophage Mu transposase (A protein), a subdomain of the central catalytic domain II, to the transposition reaction. The properties of several point mutations implicate a role for this domain in facilitating metal-assisted assembly of the synaptic complex, as well as in intramolecular DNA strand transfer. Point mutations as well as deletions in domain II beta can be complemented by those in domain II alpha but not those in domain III alpha. Thus, residues within subdomains II alpha and II beta belong to different catalytic complementation groups.

Bacteriophage mu↗
PubMed1997

Probing Flp: a new approach to analyze the structure of a DNA recognizing protein by combining the genetic algorithm, mutagenesis and non-canonical DNA target sites.

A topological and functional overview of a DNA recognition protein with unknown structure can be achieved by combining three different, but complementary approaches: modeling by the genetic algorithm, functional analysis of mutated variants, and testing the target DNA using non-canonical oligonucleotides. As an example we choose the Flp protein, a site-specific recombinase from Saccharomyces cerevisiae. We derive the topological outline including the DNA binding cleft, examine DNA binding regions by deletional and mutational analysis, and analyze the DNA binding site using 7-deazaadenine, 7-deazaguanine, inosine and 4-O-methylthymine as probes. The combined data offer a comprehensive sketch of a plausible protein architecture for Flp. The structure is detailed enough to verify the prediction accuracy for different peptide regions from pre-existing data and by new experimental design.

Binding Sites↗
PubMed1997

Computer simulation of neuronal toxicity in the spinal cord.

The use of computers to model biological systems is a relatively new research tool. For example, it is possible to write mathematical systems to model neuronal activity involved in memory and learning and to model blood flow in any organ such as the brain. There is also an interest in designing computer-controlled machines to simulate human activities such as hand movements and vision. One of the most important uses of computer modeling is as a research tool to test hypotheses and aid in formulating new hypotheses. This enables the investigator to apply preliminary tests on several experimental strategies and select for animal experimentation the ones that are most likely to produce unambiguous and interpretable results. In the following article, we describe a computer model of neuron toxicity in the mammalian spinal cord.

Dynorphins↗
PubMed1996

Purification and characterization of a membrane-bound deglycating enzyme (1-deoxyfructosyl alkyl amino acid oxidase, EC 1.5.3) from a Pseudomonas sp. soil strain.

Searching for novel approaches for uncoupling glycation from hyperglycemia as a cause of diabetic complications, a Pseudomonas sp. soil strain containing a membrane-bound enzyme that deglycates amino acids under release of free fructosamine was isolated (Gerhardinger, C., Marion, S. M., Rovner, A., Glomb, M., and Monnier, V. M. (1995) J. Biol. Chem. 270, 218-224). This enzymatic activity was found to be very sensitive to inactivation by most detergents. From the plasma membrane ( approximately 3 mg/ml protein concentration), the enzyme could be solubilized in active form using 10 mM 3-[(3-chlolamidopropyl) dimethylammonio]-2-hydroxy-1-propanesulfonate aided by 2 M NaCl and 10% glycerol (27% optimal solubilization yield). The supernatant from a 55% saturation (NH4)2SO4 cut was fractionated onto a phenyl-Superose HR 5/5 column and enzymatic activity was eluted with a inverse gradient of (NH4)2SO4. Following removal of (NH4)2SO4 with PD-10 columns and fractionation with a Mono Q HR 5/5 column, a sharp peak of enzyme activity was eluted. Analysis on sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a major band at 106 kDa and, on isoelectrofocusing gel, a pI of 5.1. The activity was completely inhibited by CN- and N3-, suggestive of copper as a likely cofactor. Identification of the protein was confirmed by affinity labeling with 14CN- and isoelectrofocusing. The "amadoriase" activity was also inhibited by Hg2+, Ag2+, Cu2+, and Zn2+ and had Km and Vmax values of 0.14 mM and 0.48 unit/ml (16 units/mg of protein), respectively, for epsilon-(1-deoxyfructosyl) aminocaproate. Significant activity was noted toward many glycated amino acids (highest with epsilon-fructosyl lysine) but not with glycated proteins. The sequence of the first 16 NH2-terminal amino acids and a search in various data bases revealed that this amadoriase enzyme is a novel protein. Based on its properties, this deglycating enzyme, which degrades Amadori products oxidatively into free fructosamine, is classified as fructosyl aminocaproate:oxygen oxidoreductase (EC 1.5.3).

Amino Acid Oxidoreductases↗
PubMed1996

High galactose levels in vitro and in vivo impair ascorbate regeneration and increase ascorbate-mediated glycation in cultured rat lens.

In contrast to conventional view that glucose is the sole glycating agent, ascorbate has now emerged as a potential precursor of advanced glycation products in lenses during cataractogenesis, owing to the high concentration present in human lens. The effects of high hexose environment in vitro and in vivo on the disruption of redox equilibrium of ascorbate (ASA) to dehydroascorbate (DHA), which is required for ascorbate-mediated crystallin modification by the Maillard reaction during cataractogenesis were examined. Organ culture experiments were performed with rat lenses that were first exposed to high galactose levels in vitro and in vivo and then incubated with 1-14C-labeled ASA, DHA or DKG (2,3-diketogulonic acid). Formation of ASA degradation products as a function of time was assessed by radiometric TLC method. Upon incubation with ASA or DHA, an elevated level of the degradation product, DKG, was detected in lenses exposed to galactose in vivo and in vitro. ASA uptake was significantly enhanced in the galactosemic lenses as compared to controls (P = 0.01). Regeneration of ASA from DHA in both galactose treated and galactosemic lenses was impaired when compared to control lens which completely converted DHA from the medium into ASA. Surprisingly, the galactose exposed lenses showed enhanced permeability to DKG which was picked up readily from the medium in contrast to normal healthy lenses which remained impermeable to DKG. Galactose exposed lenses both in vitro and in vivo showed a 5-9-fold increase in crystallin bound Schiff base-linked radioactivity when incubated with 1-14C-labeled ASA or DHA. As a preamble to the question of whether lens pigmentation predisposes towards ascorbate oxidation, lens homogenate from normal young and old pigmented cataractous lenses were incubated with [1-14C]ASA. After 2 days, ASA levels were found to have decreased by 74% and DKG levels increased by 48% in brunescent lens as compared to the young lens. These data demonstrated that profound abnormalities in ASA metabolism exist in lenses exposed to a high sugar environment suggestive of a breakdown of the redox equilibrium of ASA to DHA and a loss of membrane permeability barrier for DKG. The latter would further contribute toward a ASA-catalysed Maillard reaction in the redox impaired lens.

2,3-Diketogulonic Acid↗
PubMed1994

Role of tyrosine phosphorylation-dephosphorylation in copy number control of the yeast plasmid 2 micron circle.

A key feature of the copy control in the 2 micron circle plasmid of Saccharomyces cerevisiae is its ability to amplify when the copy number drops below the steady state value. The Flp protein encoded by the plasmid is an essential component of the amplification mechanism. A central regulatory event in amplification involves the phosphorylation/dephosphorylation of Tyr-343 of Flp. Tyrosine phosphorylation is achieved by a transesterification mechanism involving a specific phosphodiester within the 2 micron circle. The dephosphorylation is also a transesterification reaction that uses a specific 5'-OH (generated during tyrosine phosphorylation) as the phosphoryl acceptor. A sum of four phosphorylation/dephosphorylation reactions, coordinated in sets of two, is thought to invert the relative directions of a pair of replication forks. This allows more than one copy of the plasmid to be made from a single replication initiation event. In this paper we discuss the structural features of the Flp active site that control and coordinate the transesterification reactions required for amplification.

Base Sequence↗
PubMed1993

Expression of the Escherichia coli dnaX gene.

The Escherichia coli dnaX gene encodes both the tau and gamma subunits of DNA polymerase III. This gene is located immediately downstream of the adenine salvage gene apt and upstream of orf12-recR, htpG, and adk. The last three are involved in recombination, heat shock, and nucleotide biosynthesis, respectively. apt, dnaX, and orf12-recR all have separate promoters, and the first two are expressed predominantly from those separate promoters. However, use of an RNase E temperature-sensitive mutant allowed the detection of lesser amounts of polycistronic messengers extending from both the apt and dnaX promoters through htpG. Interestingly, transcription of the weak dnaX promoter is stimulated 4- to 10-fold by a sequence contained entirely within the dnaX reading frame. This region has been localized; at least a portion of the sequence (and perhaps the entire sequence) is located within a 31-bp region downstream of the dnaX promoter.

Base Sequence↗
PubMed1992

Expression of argU, the Escherichia coli gene coding for a rare arginine tRNA.

The Escherichia coli argU gene encodes the rare arginine tRNA, tRNA(UCUArg), which decodes the similarly rare AGA codons. The argU promoter is, with two exceptions, a typical, strongly expressed stable RNA gene promoter which is stimulated by an upstream activator sequence. Unlike other tRNA operons, however, argU expression is severely inhibited by sequences downstream of the transcription start point. In vivo, nucleotides +2 to +45 inhibited expression by 25- to 100-fold when measured by fusion of argU promoter regions to the chloramphenicol acetyltransferase reporter gene or by quantitative primer extension analysis. In vitro, linearized argU promoter fragments on which the argU region ended at +1 supported 5- to 10-fold-more transcription than when the argU region ended at +45. This difference in degree of inhibition between in vivo and in vitro conditions suggests that several factors, some of which could be absent in vitro, might limit expression in vivo. Alternatively, one mechanism might limit expression both in vivo and in vitro but function more efficiently in vivo. A second difference from strongly expressed stable RNA promoters is the fact the argU gene is relatively insensitive to growth rate regulation, at least when assayed on a multicopy plasmid.

Base Sequence↗
PubMed1980

Weak estrogenic activity of lindane in rats.

Administration of lindane (20 mg/kg . d) for 30 d to ovariectomized rats caused no significant change in the weight of the uterus, cervix, or vagina. Histological changes in these organs were comparable to those in ovariectomized control rats. Treatment with estradiol dipropionate alone and in combination with lindane induced a significant increase in the weights of these organs. Microscopically, these organs appeared normal. Lindane induced a significant increase in the glycogen content of the uterus, cervix, and vagina of ovariectomized rats compared to ovariectomized control rats, whereas a two- to fourfold increase in glycogen was observed in ovariectomized rats treated with estradiol dipropionate or estradiol dipropionate plus lindane. There was a significant increase in total erythrocytes and hemoglobin in ovariectomized rats treated with lindane, estradiol dipropionate, or lindane plus estradiol dipropionate. An increase in neutrophils with a corresponding decrease in lymphocytes was also observed in rats treated with lindane or lindane plus estradiol dipropionate.

Animals↗
PubMed1976

Euryblepharon with associated ocular anomalies.

Two cases of euryblepharon with other associated ocular abnormalities have been described. Case one showed flattening of the bridge of the nose and an abnormally large distance between the two medial canthi. Case two showed congenital ptosis in one eye and concomitant alternating divergent squint.

Adolescent↗