Search PubMed⌕ Search

Biomedical subjects

M Srinivasan

Publications and source records attributed to M Srinivasan.

At least 127 records · Page 7Linked to original sources
PubMed1992

Expression and release of neuroregulators during development: monoamines and neuropeptides.

The turnover of catecholamines (CA) was determined in the adrenal medulla and brain of rat fetuses and pups. In general we found a considerable increase soon after birth. The expression of mRNA for CA-synthesizing enzymes was also considerably enhanced in the adrenals shortly after birth. Furthermore, we demonstrated increased expression of neuropeptides after birth, increased synthesis of mRNA encoding for neuropeptide Y in the adrenals 24 h after birth; and considerable activation of the substance P gene in a respiratory nuclei of rabbit pups which had been breathing for 2 h as compared with fetuses at term.

Adrenal Medulla↗
PubMed1992

Production and regulation of a thermostable protease by Pseudomonas sp. B45.

A Pseudomonas sp. produced an extracellular thermostable protease which required induction by peptone. Growth of the organism and the production of protease was optimum at 30 degrees C. The enzyme was subjected to catabolite repression by glucose. Both chloramphenicol and rifamycin completely abolished protease production indicating de novo synthesis of the enzyme. Leucine, lysine, histidine and glycine enhanced the protease production considerably and they were the most effective when added during the active period of production. Glucose repression could not be relieved by addition of leucine.

Amino Acids↗
PubMed1991

Isolation, characterization, and in vitro expression of a cDNA that encodes the kidney isoenzyme of the mitochondrial glutaminase.

A cDNA that encodes the kidney isoenzyme of the mitochondrial glutaminase (pGA) was generated by recombination of two cDNAs that were isolated from a random-primed rat brain lambda gt11 library. pGA encodes 674 amino acids which includes an N-terminal sequence of 16 residues that should form an amphipathic helix, typical of a mitochondrial targeting sequence. Residues 73-90 correspond to the N-terminal sequence of the more abundant 65-kDa glutaminase peptide. In vitro transcription and translation of pGA yields a 72-kDa peptide that is immunoprecipitated with glutaminase-specific antibodies. Incubation of the glutaminase precursor with isolated mitochondria yields the 68- and 65-kDa peptides that are characteristic of the mature glutaminase. Thus, the two mature glutaminase peptides are synthesized from a single precursor. The complete 3' nontranslated region of the GA mRNA was characterized by sequencing a GA cDNA (pGA12) that was isolated from an oligo(dT)-primed rat kidney lambda gt10 library. This segment contains numerous AU-rich regions, four potential stem-loop structures, and a 48 base pair repeat of CA dinucleotides. Such domains may contribute to the increased stability of the GA mRNA that occurs in response to metabolic acidosis.

Amino Acid Sequence↗
PubMed1991

Characterization of naturally processed antigen bound to major histocompatibility complex class II molecules.

Helper T lymphocytes recognize peptide fragments of antigen bound to major histocompatibility complex (MHC) class II molecules presented on the surface of antigen-presenting cells (APCs). Previous studies showed that the MHC class II, I-Ek molecules purified from APCs that had processed Drosophila melanogaster cytochrome c (DMc) contained functional, processed antigen-I-Ek complexes. This was demonstrated by the ability of purified I-Ek, incorporated into liposomes, to stimulate DMc-specific T cells in the absence of any additional antigen. Here we describe the isolation and characterization of the processed antigen bound to I-Ek. This was accomplished using DMc radiolabeled across its entire length by reductive methylation of its lysine residues, allowing an analysis of the totality of processed antigen bound to MHC class II molecules. After processing, only about 0.2% of the APC I-Ek molecules contained processed DMc (approximately 800 per cell), yet these were sufficient to stimulate specific T cells. The DMc peptides isolated from the I-Ek molecules showed only two predominant radioactive peaks as analyzed by reverse-phase chromatography. Less processed antigen was bound to purified I-Ak molecules, and these peptides were distinct from those bound to I-Ek. The association of processed DMc with the I-Ek and I-Ak molecules appears highly specific in that no radiolabeled peptides were isolated from purified MHC class I molecules, Kk and Dk, or from the B-cell differentiation antigen B220. The majority of processed antigen-I-Ek complexes migrated more slowly than the majority of the I-Ek protein as analyzed by SDS/PAGE under nonreducing conditions without heating of the sample. This form of I-Ek may be analogous to the earlier described "floppy" form of MHC class II molecules [Dormair, K., Rothenhausler, B. & McConnell, H. M. (1990) Cold Spring Harbor Symp. Quant. Biol. 54, 409-416]. Since newly processed antigen binds nearly exclusively to this slow-migrating form, it may be of functional significance.

Amino Acid Sequence↗
PubMed1991

Enhanced in vivo release of substance P in the nucleus tractus solitarii during hypoxia in the rabbit: role of peripheral input.

In the adult, pentobarbitone-anaesthetized rabbit, the in vivo release of substance P-like immunoreactivity was measured in the nucleus tractus solitarii using microdialysis and radioimmunoassay. Increased 160 +/- 16%) extracellular concentrations of substance P-like immunoreactivity were observed during hypoxic provocations of 9% O2 in N2 which also resulted in an increase in phrenic nerve activity. In bilateral carotid sinus nerve-denervated animals no enhanced release of substance P was seen in response to hypoxic challenges (105 +/- 6%) and the phrenic nerve activity was not significantly affected. Perfusion of the nucleus tractus solitarii region with the dopamine agonist, apomorphine (10(-5) M) resulted in a significant decrease in the extracellular level of substance P. These results provide further evidence that substance P is involved in the mediation of the hypoxic drive inputs from the peripheral chemoreceptors. The interactions of apomorphine with substance P release might also suggest a presynaptic modulation of substance Pergic neurons by dopamine in the nucleus tractus solitarii.

Animals↗
PubMed1991

Analysis of the retention signals of two resident luminal endoplasmic reticulum proteins by in vitro mutagenesis.

Protein disulfide isomerase (PDI, ERp59), ERp72, and ERp61 are luminal proteins of the endoplasmic reticulum (ER) that are characterized by the presence of sequences corresponding to the active site regions of PDI. Each one of these proteins possesses a different COOH-terminal tetrapeptide ER retention signal. In order to investigate what other tetrapeptide sequences could serve as retention signals and to determine to what extent the function of the retention signal is modulated by the protein carrying the signal, we have constructed a set of mutants of two of these resident ER proteins, PDI and ERp72. In each of these proteins, the wild type tetrapeptide sequences were replaced by each member of the set of the 12 possible combinations consisting of (K,R,Q)-(D,E)-(D,E)-L. Analysis of the efficiency of retention of the variant proteins when each was transiently expressed in COS cells showed that the retention efficiencies vary with both the COOH-terminal sequence and with the protein that carries this sequence.

Amino Acid Sequence↗
PubMed1991

Chronic treatment with SCH-23390, a selective dopamine D1 receptor blocker decreases preprotachykinin-A mRNA levels in nucleus tractus solitarii of the rabbit: role in respiratory control.

Acute intravenous administration of the selective D1 receptor blocker SCH-23390 resulted in an enhanced respiratory motor output as evidenced by the phrenic nerve activity, whereas local perfusion into the region of nucleus tractus solitarii had no effect. The increase in phrenic nerve activity was accompanied by a concomitant increase in the release of substance P in the region of nucleus tractus solitarii as measured by in vivo microdialysis technique. Chronic administration of SCH-23390 via subcutaneously implanted Alzet mini osmotic pumps, significantly decreased the level of preprotachykinin-A mRNA in the region of respiratory relay neurons in nucleus tractus solitarii but was without effect in the ventral medullary surface structure, wherein the central chemoreceptors are thought to be located. A smaller, but significant decrease was also seen in the striatum. The results suggest that chronic treatment with SCH-23390 leads to a disinhibition of an inhibitory dopaminergic input to the neurons in nucleus tractus solitarii from a suprapontine level, which may account for a subsequent inhibition of tachykinin-containing neurons in the nucleus tractus solitarii, the relay station for respiratory reflexes.

Animals↗
PubMed1991

Hypoxia-mediated in vivo release of dopamine in nucleus tractus solitarii of rabbits.

A wide variety of neuroactive substances have been suggested to be involved in the respiratory depression observed in response to severe hypoxia. By use of the technique of microdialysis, the release of dopamine (DA) was measured in the nucleus tractus solitarii during severe hypoxic provocations (6% O2 in N2) in the adult pentobarbital-anesthetized rabbit. DA release was analyzed by high-performance liquid chromatography with electrochemical detection. Such hypoxic provocations caused pronounced phase of depression in the phrenic nerve activity and enhanced release of DA. After bilateral carotid sinus nerve denervation, acute severe hypoxia did not give rise to enhanced release of DA or to phrenic nerve depression. Mild hypoxic (9% or 12% O2 in N2) or hypercapnic (6% CO2) stimuli resulted in an increased phrenic nerve activity without any concomitant changes in DA release. Decerebration at the midcollicular level in rabbits prevented an enhanced release of DA in the nucleus tractus solitarii during severe hypoxia. The results suggest that 1) DA is involved in the central ventilatory response to severe hypoxia, 2) not only the initial excitatory but also the second depressive phase in response to severe hypoxia is mediated partially by the peripheral chemoreceptors, and 3) the depressive phase is dependent on intact connections from suprapontine structures.

Animals↗
PubMed1991

Birth-related activation of preprotachykinin-A mRNA in the respiratory neural structures of the rabbit.

The perinatal ontogeny of preprotachykinin-A gene expression was assessed in central respiration-related structures. On the day of birth, there was an enhanced expression of preprotachykinin-A mRNA in the region of the nucleus tractus solitarii, the primary relay station for respiratory and cardiovascular reflexes. This increased expression was also seen in the pups delivered by cesarean section and allowed to breathe for a couple of hours as compared with their littermates, which were not allowed to breathe at all. On the basis of this finding, we suggest that the commencement of continuous breathing at birth, unlike the episodic breathing of fetal life, is associated with the enhanced expression of preprotachykinin-A mRNA in the nucleus tractus solitarii.

Animals↗
PubMed1990

ERp72, an abundant luminal endoplasmic reticulum protein, contains three copies of the active site sequences of protein disulfide isomerase.

We have cloned, sequenced, and expressed full length cDNA clones encoding two abundant, luminal endoplasmic reticulum proteins (ERp), ERp59/PDI and ERp72. ERp59/PDI has been identified as the microsomal enzyme protein disulfide isomerase (PDI). An analysis of the amino acid sequence of ERp72 showed that it shared sequence identity with ERp59/PDI at three discrete regions, having three copies of the sequences that are thought to be the CGHC-containing active sites of ERp59/PDI. Thus, ERp72 appears to be a newly described member of the family of CGHC-containing proteins. ERp59/PDI has the sequence KDEL at its COOH terminus while ERp72 has the related sequence KEEL. Removal of the KDEL of ERp59/PDI or the KEEL of ERp72 by in vitro mutagenesis techniques and subsequent analysis of the mutants in transient expression assays, showed that both sequences are endoplasmic reticulum retention signals for their respective proteins. The most dramatic difference in secretion between the wild type and the mutant forms of the protein was seen in the case of ERp72.

Amino Acid Sequence↗
PubMed1990

Acanthamoeba keratitis in non-contact lens wearers.

Nine cases of Acanthamoeba keratitis not associated with contact lens wear were diagnosed between July 1987 and August 1989. Patients were treated with topical neomycin-polymyxin B-bacitracin (Neosporin) drops alone or in combination with either miconazole nitrate or ketoconazole drops. At the time of data collection four patients were available for follow-up for an average of 4 months; however, four patients were unavailable for follow-up and one is still undergoing treatment. In four patients corneal infiltrates cleared completely with topical medication (Neosporin, two patients; Neosporin plus miconazole, two patients). Simple laboratory methods were found to be adequate for the diagnosis of Acanthamoeba keratitis. Therapy with Neosporin drops can result in resolution of corneal infiltrates due to Acanthamoeba species.

Acanthamoeba↗
PubMed1990

Macular dystrophy of the cornea. A systemic disorder of keratan sulfate metabolism.

The serum of most patients with type 1 macular corneal dystrophy (MCD), the most prevalent subtype, lacks detectable antigenic keratan sulfate (KS), and it has been postulated that such individuals may lack antigenic KS in their cartilage as well. To test this hypothesis, we studied the cornea, serum, and nasal cartilage from an MCD patient using light and electron microscopy, immunohistochemistry, and a quantitative enzyme-linked immunosorbent assay (ELISA) which uses a monoclonal antibody against a sulfated epitope on the KS chain to measure KS content. Histologically, corneal deposits seen were characteristic of MCD. No abnormal deposits were noted in the cartilage. The lack of immunoreactivity in corneal sections with antibodies against sulfated epitope on KS and the absence of this epitope in serum showed that the patient had type 1 MCD. The cartilage specimen showed no immunoreactivity in the chondrocytes or extracellular matrix. Quantitative analysis by ELISA demonstrated that the antigenic KS content of the cornea and cartilage was at least 800 times lower than that in normal controls. This provided direct evidence that the abnormality in the sulfation of keratan in type 1 MCD involves the cornea and cartilage.

Aged↗
PubMed1990

Isolation of a functional antigen-Ia complex.

The helper T-cell recognition of globular protein antigens requires that the antigen be processed and presented by an I-region associated (Ia)-expressing antigen-presenting cell (APC). Processing involves the uptake of antigen into an intracellular, proteolytic, acidic compartment; release of peptide fragments containing the T-cell antigenic determinant; association of these peptides with Ia; and presentation of these complexes on the cell surface for recognition by the specific T cells. The molecular mechanisms by which processed antigenic peptides associate with Ia within the APC are poorly understood. To date, functional antigen-Ia complexes have not been isolated from cells that have processed native antigens, although the resolution of the structure of a major histocompatibility complex (MHC) class I protein indicates that peptide is bound in a groove between two alpha-helical regions of the molecule and synthetic peptides have been demonstrated to bind purified MHC both in detergent solution and incorporated into planar membranes, where the MHC-peptide complexes function to activate specific T cells. Here we demonstrate that Ia purified from APCs that have processed the native globular protein antigen cytochrome c, when incorporated into lipid membranes, stimulates cytochrome c-specific T cells in the absence of exogenous antigenic peptide. The T-cell response to Ia purified from cytochrome c-pulsed APCs shows the same MHC restriction and antigen fine specificity as the response to antigen-pulsed APCs. Indeed, T-cell recognition of pigeon cytochrome c (Pc) shows a well documented high-affinity heteroclitic cross-reaction to insect cytochromes c-namely, those of Drosophila melanogaster (DMc) and tobacco hornworm moth (THMc). The same heteroclitic response is observed when purified Ia from unpulsed cells, incorporated into lipid membranes, is used to present antigenic peptides of Pc and of THMc. Significantly, Ia purified from APCs that have processed DMc is approximately 50-fold more active in stimulating specific T cells compared to Ia purified from APCs that have processed Pc. The peptide-Ia complex isolated here may provide the necessary material for analysis of the physiochemical properties of the processed form of the antigen that is produced by the APC and associates with Ia.

Animals↗
PubMed1990

Factors influencing extracellular protease synthesis in an Aspergillus flavus isolate.

Our studies on the control of extracellular protease synthesis in an Aspergillus flavus strain isolated by us indicate that in a defined medium a protein must be present for enzyme to be produced. Soya bean protein and cotton seed protein were efficient inducers. The ability to induce enzyme synthesis was a characteristic property of the individual protein. Enzyme activity was not derepressed in the absence of a protein by limitation of nutrients. Cycloheximide blocked enzyme synthesis. Low levels (1%) of various carbohydrates did not repress enzyme synthesis, whereas most carbohydrates at 3% levels repressed enzyme synthesis. Addition of glucose to cultures actively producing the enzyme blocked further synthesis of enzyme. Addition of glucose to cultures producing the enzyme in the absence of sodium nitrate resulted in a decrease of enzyme activity.

Aspergillus flavus↗
PubMed1989

Fluorescein-conjugated lectin visualization of fungi and acanthamoebae in infectious keratitis.

The authors investigated the efficacy of two fluorescein-conjugated lectins (FCLs), concanavalin A (F-ConA) and wheat germ agglutinin (F-WGA), to visualize microorganisms from clinical specimens with documented mycotic and acanthamoebic keratitis. Corneal scrapings from 18 patients with culture-proven keratomycosis and deparaffinized histopathologic specimens from five culture-proven cases of acanthamoebic keratitis were evaluated. The F-ConA provided consistently bright staining of the mycotic structures in each of the corneal scrapings. Both F-ConA and F-WGA stained acanthamoebic trophozoites and cysts in the histopathologic specimens. In both the corneal scrapings and the histopathologic specimens, the microorganisms were easily differentiated from background corneal cells and tissue. These staining patterns correlate well with the results from experimental studies, and with the known cell wall carbohydrate compositions for fungi and acanthamoebae. This study provides further evidence that FCLs (particularly F-ConA) may eventually become effective first-line stains for the visualization of microorganisms in specimens from ocular infections.

Acanthamoeba↗
PubMed1989

Stomal varices: percutaneous transhepatic embolization.

Peristomal varices occasionally form in patients with chronic liver disease who have surgically created intestinal anastomoses and stomas. Hemorrhage from these varices carries an estimated mortality of 3%-4% per episode, as opposed to the 30%-40% mortality associated with gastroesophageal variceal bleeding. The cases of four patients who underwent transhepatic mesenteric vein catheterization with embolization of stomal varices for recurrent, intractable bleeding are presented. In three of the four patients there was no additional hemorrhage for at least 5 months. One patient was lost to follow-up. There were no complications. Since stomal variceal hemorrhage has a low mortality, transhepatic embolization is presented as a means of hemorrhage control when sclerotherapy fails and when shunt surgery presents an unacceptably high rate of morbidity and mortality relative to the underlying disease.

Aged↗
PubMed1989

A possible dopaminergic pathway mediating hypoxic depression in neonatal rabbits.

Previous studies show a differential effect of dopamine and its agonist apomorphine on ventilation, being depressant on adults and stimulatory on fetal respiratory breathing movements. In view of this we studied the effect of apomorphine, specific antagonists of dopamine receptors SCH 23390 (D1), sulpiride and domperidone (D2), and cis(z)-flupentixol (D1 + D2) on the typical ventilatory response to hypoxia in neonatal rabbit pups. This ventilatory response to hypoxia is characterized by an initial stimulatory (phase 1) followed by a declining (phase 2) pattern. Cis(z)-flupentixol markedly increased phase 1 and abolished phase 2. A similar effect was seen with SCH 23390. Administration of sulpiride or domperidone increased phase 1 but did not affect phase 2, thereby not altering the biphasic pattern of ventilation to hypoxia. The abolition of the biphasic response in neonatal pups on decerebration was also found to be restored by topical application of apomorphine in the fourth ventricle. Our results suggest that dopamine is acting centrally in mediating the declining phase of the biphasic response to hypoxia in rabbit pups possibly via the D1-receptors.

Animals↗