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Biomedical subjects

M A Thompson

Publications and source records attributed to M A Thompson.

At least 109 records · Page 6Linked to original sources

Characterization of variable-region genes and shared crossreactive idiotypes of antibodies specific for antigens of various influenza viruses.

Several syngeneic monoclonal anti-idiotypic antibodies were obtained against PY206, a monoclonal antibody specific for X-31 (H3N2) influenza virus hemagglutinin. This idiotype was found in the sera of BALB/c mice immunized with various influenza viruses. Adsorption experiments indicated that the PY206 Id was borne by antibodies specific for viral hemagglutinin (HA) and/or neuraminidase (NA). This idiotype was identified on other monoclonal antibodies specific for various influenza HAs (H3 and H1). Study of the variable-region (V) genes of these monoclonal antibodies showed that its expression is independent of variable kappa (VK)21 light-chains and that the heavy-chains of the strongly idiotype-positive hybridomas derive from either the variable heavy (VH) J558 or VH 7183 family. Finally, Western blot analysis demonstrated that PY206 idiotypic determinants are located exclusively on the heavy chain.

Animals↗

cDNA and deduced protein sequence for the beta B1-crystallin polypeptide of the chicken lens. Conservation of the PAPA sequence.

The nucleotide sequence of two cDNA clones corresponding to the beta B1-crystallin mRNA (formerly beta 35) of the chicken eye lens has been determined. The derived amino acid sequence of the chicken beta B1 polypeptide fits well with the two-domain, four "Greek Key" motif structure common to the beta gamma-crystallin superfamily of proteins. The calculated molecular weight of the encoded chicken beta B1 protein is 27,267. The beta B1 polypeptide has both an N- and C-terminal extension and is highly homologous to the mammalian beta B1-crystallin polypeptide. There is a 72% homology between the core regions of the chicken and bovine beta B1 polypeptides; by contrast, there is only 27% homology between the N-terminal extensions of these polypeptides. The N-terminal extension of chicken beta B1 contains a short alternating proline-alanine (PAPA) sequence, like that in the mammalian beta B1, and has some homology with the N-terminal region of histone H1.4, myosin light chain, prokaryotic outer membrane protein A, and adenovirus 24/28-kDa early protein. At the nucleic acid level, the chicken beta B1 crystallin gene has an atypical polyadenylation signal, AATTAAA. This appears to be associated with microheterogeneity of the polyadenylation site by comparison of two cDNA clones, suggesting an additional level at which diversity in crystallin gene expression may arise.

Amino Acid Sequence↗

Influence of ketanserin pretreatment on the haemodynamic responses to sternotomy.

The effect of intravenous ketanserin on the pressor response to sternotomy was studied in 36 patients undergoing coronary artery surgery. Two doses of the drug (10 mg and 20 mg) were compared with a placebo injection of saline. After induction of anaesthesia, haemodynamic variables were measured until the institution of cardiopulmonary bypass. Plasma and platelet 5-hydroxyindoles and 5-hydroxytryptamine were measured in a subset of 13 patients. Ketanserin induced a dose-dependent amelioration of the pressor response to sternotomy. Plasma 5-hydroxyindoles and platelet 5-hydroxytryptamine levels did not correlate with clinical response. The increased effectiveness of the higher dose of ketanserin may be due to an effect other than serotonin antagonism.

Drug Evaluation↗

Equol: a contributor to enigmatic immunoassay measurements of estrogen.

The efficacy of radioimmunoassay (RIA) for the measurement of estradiol-17 beta (E2) in murine plasma was investigated. When Sephadex LH-20 or celite column chromatography was used to separate E2 from estrone (E1) and other cross-reacting compounds, the results were erratic if small volumes of mouse plasma were resolved. Assay of a diethyl ether extract of plasma (500 microL) was the most practical method for estimating the concentration of estradiol-17 beta in mice. This method was used to determine the pattern of estrogen secretion during the estrous cycle, on the day of implantation and during pregnancy. No convincing change in estrogen secretion was observed in the diestrous/proestrous mouse. By comparison, estrogen levels were elevated during pregnancy. Taken together, these results implied that cross-reactive components in plasma masked low levels of endogenous estrogen. Further evaluation of mouse plasma and urine using a co-chromatography technique to examine estrogen elution from a reverse-phase HPLC system followed by GC/MS analysis indicated the presence of equol [7-hydroxy-3-(4-hydroxyphenyl)chroman], a phytoestrogen metabolite with a ring structure similar to estradiol-17 beta. Equol and possibly other cross-reactive components of plasma may account for the apparent lack of increased estrogen secretion during the mouse estrous cycle and on the day of implantation as determined by the radioimmunoassay of ether extracts of plasma.

Animals↗

HPLC separation of anti-estrogen and estrogen receptor binding components of mouse plasma.

Proestrous mouse plasma and urine were subjected to diethyl ether extraction, enzyme hydrolysis and HPLC separation of estrogen components. Radioimmunoassay of the treated proestrous samples with a broad spectrum anti-estrogen serum failed to detect estradiol-17 beta, estrone or estriol. HPLC chromatograms contained two peaks of immunoreactive and estrogen receptor binding material with polarities between those of estriol and estradiol-17 beta. Similar peaks were detected in HPLC chromatograms of urinary extracts from ovariectomized and ovariectomized-adrenalectomized mice. The least polar of the two peaks produced a mass spectrum identical to that of authentic equol [7-hydroxy-3-(4'-hydroxyphenyl)chroman], a phytoestrogen metabolite. The presence of significant quantities of circulating equol in all strains studied, combined with apparently low plasma levels of endogenous classical estrogens during proestrus, confound attempts to study estrogen secretion in the mouse.

Animals↗

Nucleotide sequence comparison of the Adh gene in three drosophilids.

The alcohol dehydrogenase (Adh) gene has been isolated from Drosophila simulans and D. mauritiana by screening lambda clone libraries of each with a previously cloned Adh gene from D. melanogaster. The isolated lambda clones were subcloned and partially sequenced to determine the relatedness of these species and to examine details of evolutionary change in the structure of the Adh gene. We report the sequence of the first 704 nucleotides of each gene as well as 127 bases in the 5' untranslated region. When these sequences are compared, D. melanogaster differs from D. simulans and D. mauritiana by 2.8% and 3.1%, respectively. D. simulans and D. mauritiana differ by only 1.8%, implying that they are more closely related to each other than either is to D. melanogaster. This is consistent with phylogenetic relationships established by a variety of genetic, biochemical, and morphological means and illustrates that DNA sequencing of a single gene may be used to assess the evolutionary relationships of species.

Alcohol Dehydrogenase↗

Identification of a non-steroidal estrogen, equol, in the urine of pregnant macaques: correlation with steroidal estrogen excretion.

Macaque urinary estrogens at late pregnancy were separated by high performance liquid chromatography and quantified, both with radioimmunoassay and an in vitro uterine estrogen receptor assay. Five estrogens were measured. Four were steroids: estriol, estrone, 17 beta-estradiol, and 16 alpha-hydroxyestrone. The fifth was a flavonoid, equol, a metabolite of plant isoflavonoids, formononetin and genistein. By mass, estrone and equol were the predominant urinary estrogens, with equol reaching levels of microgram/mg creatinine in three of 8 pregnancies studied. Both quality and quantity of urinary estrogen excretion in the rhesus (Macaca mulatta) was compared to those in 4 other species (Macaca fascicularis, Macaca nemestrina, Macaca radiata and Macaca silenus). All 5 estrogens present in the rhesus were also present in the other 4. Variability in mass of each estrogen excreted appeared no greater between species than within the rhesus. In a longitudinal study, urinary equol levels were most highly correlated with those of estrone, the predominant excretory steroid of macaque pregnancy. We conclude endogenous steroidal estrogen is related to production of equol in macaques, however, equol is not dependent on the feto-placental unit as low levels of equol were also present in male macaque urine.

Animals↗

Patterns in the distribution of REM sleep in normal human sleep.

Results are reported of four analyses of the distribution of REM sleep across nights of two subjects who slept for 50 consecutive nights on a regimented, but normal, sleep schedule. We found (a) a strong phase-setting effect of sleep onset on the distribution of REM sleep within nights, (b) no systematic change in the phase of the distribution of REM sleep across nights, and (c) a relationship between the nocturnal temperature minimum and the distribution of REM sleep within nights. Our results are consistent with the notion that the nightly distribution of REM sleep may be determined by an oscillatory process, the phase of which is reset at sleep onset, but which may be subject to other influences, such as the circadian temperature rhythm. These results are in general agreement with those found by investigators studying subjects on free-running or other abnormal sleep schedules.

Adolescent↗

Characterization of the estrogenic properties of a nonsteroidal estrogen, equol, extracted from urine of pregnant macaques.

Recent reports of substantial urinary levels of equol in pregnant macaques and humans pose a concern, because equol poisoning in the ovine is characterized by an often permanent failure of reproductive processes. Equol (Fig. 1), a metabolite of phytoestrogens, is thought to act through estrogen receptors. The present study made a direct comparison of the estrogenic activity of equol from macaque urine, (+/-) equol and 17 beta-estradiol (E2) in vitro and in vivo. Relative binding affinity of equol for rat uterine receptor was 1% that of E2, and the dissociation rate of equol from the receptor was very high. Consistent with equol's binding properties in vitro, it was ineffective in stimulating rat uterine weight gain and possessed limited ability to increase progesterone receptor. Uterine nuclear receptors after doses of equol sufficient to produce depletion and replenishment of cytosol estrogen receptor were not measurable by exchange assay. No antiestrogenic activity of equol could be demonstrated. Equol's weak potency and lack of antiestrogenic activity are difficult to reconcile with its ability to induce ovine infertility. We conclude species differences at some level other than classical estrogen receptor as defined in the rat model are responsible for variability in equol's impact.

Animals↗

Lorazepam in open-heart surgery--plasma concentrations before, during and after bypass following different dose regimens.

Thirty-six patients (29 males and 7 females) undergoing open-heart surgery received one of three different dose regimens of lorazepam. All received a weight-related oral dose (2 mg, 3 mg or 4 mg) pre-operatively for night sedation. Twenty-four patients had an additional weight-related dose (2 mg, 3 mg or 4 mg intravenously) either as part of the induction (12 patients) or just prior to connection of the heat-lung machine (12 patients). Plasma concentrations of lorazepam were measured 20 minutes after induction, immediately before bypass, 30 and 60 on bypass and 30 minutes after bypass. Only when additional intravenous lorazepam was given prior to connection to the heart-lung machine were plasma lorazepam concentrations obtained compatible with complete amnesia.

Adult↗

Testicular androgen and estrogen secretion and benign prostatic hyperplasia in the beagle.

The natural history of benign prostatic hyperplasia (BPH) in the dog is characterized by a slow progression through two phases. The early phase of the disease process is characterized by glandular hyperplasia and occurs as early as 2.5 yr of age. The late phase of the disease is characterized by cystic hyperplasia and occurs after 4 yr. The results of the present study are the first to compare Leydig cell structure and steroidogenic function in dogs with BPH with those in age-matched controls. It was discovered that glandular BPH can occur in young dogs (2.5 yr) in which Leydig cell mass and ultrastructure and maximally stimulated androgen secretion are indistinguishable from those in age-matched controls. These results support the concept that the early phase of BPH (glandular hyperplasia) is not related temporally to some defect in the Leydig cell. In contrast, the late phase of BPH (cystic hyperplasia) in beagles 6 yr of age is associated with diminished smooth endoplasmic reticulum in the Leydig cell and with diminished production of androgens by perfused testes in vitro. During the course of these studies, we discovered that the testes of young beagles with BPH, but not age-matched controls or old beagles with BPH, secrete an unidentified molecule (putative estrogen). This molecule was characterized partially in that it is extractable from testicular venous effluent with diethyl ether, elutes in a discrete fraction in several different high performance liquid chromatographic systems, reacts with an antibody that recognizes 17 beta-estradiol, estrone, and estriol, and competes with 17 beta-[3H]estradiol for the rat uterine estrogen receptor. Based on the elution volume from high performance liquid chromatography, the unknown molecule (putative estrogen) is not estriol, estradiol, or estrone.

Androgens↗

Complement activation during cardiopulmonary bypass: quantitative study of effects of methylprednisolone and pulsatile flow.

Forty four patients undergoing open heart surgery were divided into three groups. Group 1 (17 patients) underwent routine anaesthesia and surgery; group 2 (17 patients) received two doses of methylprednisolone (30 mg/kg), one during induction of anaesthesia and the other immediately before induction of cardiopulmonary bypass; and group 3 (10 patients) received pulsatile flow while undergoing pulsatile perfusion by the heart-lung machine. A modification of the previously described technique was used to detect and measure complement activation in plasma before and during the bypass period using crossed immunoelectrophoresis. About 45% of all patients showed measurable complement activation (greater than 4.5%) during cardiopulmonary bypass and the mean activation in this group was 6.4%. There was no significant difference between the three groups in complement activation. In group 2, however, women showed significantly more complement activation than men (p less than 0.05). It is suggested that neither corticosteroids nor pulsatile flow affect complement activation, but caution should be exercised in women receiving methylprednisolone.

Animals↗

Restricted adult clonal profiles induced by neonatal immunization. Influence of suppressor T cells.

The effects of neonatal antigen exposure on the adult B cell repertoire have been examined by characterizing the influenza hemagglutinin (HA)-specific response of adult BALB/c mice given antigen soon after birth. Ligand exposure during early life exerts a profound and lasting effect upon the B cell repertoire, characterized by the expansion and preservation of particular antigen-reactive clones and the apparent loss of others. The precise subset of clonotypes selectively preserved depends upon the age at which antigen is first encountered; and is predictable given a knowledge of the emerging primary pool's dynamics and composition. The preserved (secondary) B cells differ from their unprimed precursors with respect to (a) expression of the surface marker detected by the monoclonal antibody J11d, and (b) susceptibility to T cell-mediated suppression. These studies thus demonstrate a strong relationship between the heritable dynamics of the emerging primary B cell repertoire and the effect of ligand-driven events upon repertoire phenotype. In addition, they provide a mechanistic model for certain forms of antigen-induced oligoclonal dominance, especially the phenomenon of original antigenic sin.

Aging↗