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Biomedical subjects

E Herbert

Publications and source records attributed to E Herbert.

At least 19 recordsLinked to original sources
PubMed1996

Vav: function and regulation in hematopoietic cell signaling.

Vav, a 95 kDa proto-oncogene product expressed specifically in hematopoietic cells, was originally isolated as a transforming human oncogene. Vav contains an array of functional domains that are involved in interactions with other proteins and, possibly, with lipids. These include, among others, a putative guanine nucleotide exchange domain, a cysteine-rich region similar to the phorbol ester/diacylglycerol-binding domain of protein kinase C, a pleckstrin-homology domain, and Src-homology 2 and 3 (SH2 and SH3, respectively) domains. The presence of these domains, the transforming activity of the vav oncogene, and the rapid increase in tyrosine phosphorylation of Vav induced by triggering of diverse receptors indicate that it plays an important role in hematopoietic cell signaling pathways. Such a role is supported by recent studies using "knockout" mice and transiently transfected T cells, in which Vav deletion or overexpression, respectively, had marked effects on lymphocyte development or activation. The presence of a putative guanine nucleotide exchange domain, the prototype of which is found in the dbl oncogene product, implies that Vav functions as a guanine nucleotide exchange factor (GEF) for one (or more) members of the Ras-like family of small GTP-binding proteins. In support of such a role, Vav preparations were found in some (but not other) studies to mediate in vitro-specific GEF activity for Ras. Additional studies are required to identify the physiological regulators and targets of Vav, and its exact role in hematopoietic cell development and signaling.

Amino Acid Sequence↗
PubMed1994

Hematologic correlates of spontaneous echo contrast in patients with atrial fibrillation and implications for thromboembolic risk.

Spontaneous echo contrast has been observed in conditions of low blood flow velocity, such as rheumatic mitral stenosis and atrial fibrillation (AF). The phenomenon has been attributed to increased echogenicity due to aggregation of blood cells at low shear rates. The aim of this study was to determine whether abnormalities of blood composition also might contribute to spontaneous echo contrast formation by promoting cellular aggregation. Transesophageal echocardiography was performed in 185 patients with AF (31 with valvular and 154 with nonvalvular AF). The left atrium was examined for thrombus and spontaneous echo contrast, which was graded from 0 (nil) to 4+ (severe) by 2 independent observers. Forty milliliters of venous blood was obtained from each patient for hematologic analysis. Spontaneous echo contrast was observed in 46% of patients (74% with valvular and 41% with nonvalvular AF). In linear regression analysis, positive correlations were found between grade of spontaneous echo contrast and erythrocyte sedimentation rate (p < 0.001), low-shear blood viscosity (p < 0.001) and anticardiolipin antibody (p = 0.02) in the total study population, and in patients with nonvalvular AF. Spontaneous echo contrast correlated with mitral valve area (p < 0.01) and gradient (p = 0.03), but not with hematologic parameters in patients with valvular AF. Left atrial thrombus was present in 6 patients, all of whom had spontaneous echo contrast. Age (< 0.01), spontaneous echo contrast (p = 0.03) and the fibrinogen concentration (p = 0.03) correlated with previous embolic events.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

Tyrosine phosphorylation and activation of Vav GTP/GDP exchange activity in antigen receptor-triggered B cells.

Ag receptor triggering in B cells stimulates the activity of receptor-associated tyrosine protein kinases (TPK), leading to tyrosine phosphorylation of several cellular substrates, one of which is the Vav proto-oncogene product. We have recently determined that Vav is a TPK-regulated guanine nucleotide exchange factor for Ras in T cells. Here, we show that B cell extracts or Vav immunoprecipitates contain a Ras GDP/GTP exchange activity that is stimulated upon surface Ig (slg) triggering. The receptor-mediated stimulation of Vav exchange activity was blocked by the TPK antagonist, herbimycin A. Furthermore, immunodepletion of Vav from the B cell extracts removed approximately 80% of the Ras GDP/GTP exchange activity. These findings indicate, first, that B cell-derived Vav possesses GDP/GTP exchange activity for Ras; second, that the exchange activity of Vav is accelerated by a slg-triggered, herbimycin A-sensitive TPK and, third, that Vav accounts for most of the receptor-stimulated Ras GDP/GTP exchange activity. Thus, Vav may serve as a critical component in slg-mediated signal transduction pathways by coupling receptor-associated TPK to the activation of Ras proteins.

Animals↗
PubMed1994

Construction and characterization of lck- and fyn-specific tRNA: ribozyme chimeras.

Two src-family protein tyrosine kinases (PTKs), p56lck, and p59fyn, are thought to play an important role in the antigen-specific T cell receptor (TCR)/CD3-initiated signaling pathway, but their relative contribution to these events is not clearly defined. Here, we have explored the potential of catalytic RNA molecules, or ribozymes, as tools for selectively inhibiting expression of the corresponding target genes in T cells. Several lck- or fyn-specific hammerhead ribozymes were synthesized, cloned into a bacterial transcription vector, and found to display specific catalytic activity in vitro. In order to achieve stable high-level ribozyme expression in intact cells, selected ribozymes were subsequently cloned into a retroviral vector (DC-T5T) immediately downstream of a tRNA(met) transcription unit. Upon retroviral transduction of a human leukemic T cell line (Jurkat), two out of four chimeric tRNA:ribozymes, fyn-1 and lck-1, were stably expressed at levels of approximately 10,000 or approximately 25,000 copies/cell, respectively. Ribozyme expression was associated with a reduction of up to 80% (lck) or 61% (fyn) in endogenous target mRNA by comparison to the corresponding transcript levels in control clones transfected with vector alone. By contrast, expression of the corresponding target proteins was not reduced, suggesting a post-transcriptional compensatory mechanism that increases translation or stability of the p56lck and/or p59fyn proteins.

Base Sequence↗
PubMed1993

Effect of T-cell receptor antagonism on interaction between T cells and antigen-presenting cells and on T-cell signaling events.

T-cell receptor (TCR) antagonism induced by complexes of antigen analogue with major histocompatibility complex (MHC) molecules results in efficient inhibition of antigen-dependent T-cell responses. We have investigated some of the possible mechanisms by which TCR antagonists bound to the MHC molecules of antigen-presenting cells (APCs) can inhibit T-cell activation. Using a nonstimulatory analogue of the antigenic peptide influenza hemagglutinin-(307-319), we showed that MHC/antagonist complexes completely inhibit very early intracellular events of antigen-dependent T-cell activation, such as inositol phosphate turnover and Ca2+ influx. In a parallel series of experiments, the effect of TCR antagonist peptide on membrane-related activation events was also investigated. It was found that MHC/antagonist complexes on the surface of APCs did not induce stable conjugates with T cells and, most interestingly, did not inhibit antigen-induced conjugate formation. Thus, our data suggest that antagonistic peptides do not interfere with the cellular events that are required for stable T-cell/APC conjugate formation but do inhibit early biochemical events required for T-cell proliferation. The data are discussed with respect to the role of surface receptor clustering in TCR antagonism.

Animals↗
PubMed1990

Declining serum cholesterol levels prior to diagnosis of colon cancer. A time-trend, case-control study.

We report the results of a 10-year, time-trend, case-control study in which serum cholesterol level was determined at several points in time preceding the diagnosis of colon cancer in a population of individuals who sought general checkups at an ambulatory care screening facility. Each of the 69 patients with colon cancer (32 men and 37 women) was matched with a control patient who was randomly selected. At the time of diagnosis, the patients with colon cancer had significantly lower serum cholesterol values than control patients (5.56 +/- 0.31 mmol/L [SEM] vs 6.47 +/- 0.34 mmol/L). This difference did not vary with sex or Dukes' stage of the cancer. The percent of matched pairs in which the cancer patient had a lower serum cholesterol level increased from 42% at 10 years prior to cancer diagnosis to 77% at diagnosis. The ratio of serum cholesterol at each period to the level at time of diagnosis demonstrated an average decline of 13% during the 10 years prior to diagnosis for case patients vs an average rise of 2% in the same period for control patients. We conclude that individuals in whom colorectal cancer develops share the same level of serum cholesterol as the general population initially, but during the 10 years preceding the cancer demonstrate a decline in serum cholesterol level that is opposite to the rising level seen with age in the general population.

Aged↗
PubMed1988

Comparison of a carboxypeptidase E-like enzyme in human, bovine, mouse, Xenopus, shark and Aplysia neural tissue.

Several diverse species contain an enzyme with many properties in common with those of bovine carboxypeptidase E (CPE), a neuropeptide processing carboxypeptidase B-like enzyme. This enzyme has been designated EC 3.4.17.10, and is also known as enkephalin convertase and carboxypeptidase H. All tissues that are known to contain bioactive peptides also contain CPE-like enzymatic activity. In Xenopus laevis, enzyme activity is highest in the brain and pituitary, lower in the skin, and undetectable in liver and gut. In Aplysia californica, enzyme activity is highest in the atrial gland, but is also present in moderate amounts in the various neural tissue. CPE extracted from human, bovine, mouse, Xenopus, shark, and Aplysia neural tissue is substantially purified using substrate affinity chromatography and concanavalin A sepharose columns. The partially purified enzyme from all species examined possess very similar enzymatic properties. These properties include a pH optimum of 5.6, a stimulation by cobalt chloride, and an inhibition by chelating agents (1,10-phenanthroline). Arginine-derived active site-directed inhibitors show similar inhibition constants (Ki's) towards enzyme from the various species, whereas lysine-derived inhibitors are substantially less potent towards the Aplysia carboxypeptidase than towards enzyme isolated from the other species. The similar properties of the carboxypeptidase isolated from the various species suggests that a CPE-like is involved in the biosynthesis of many peptide neurotransmitters and hormones in a wide range of organisms.

Animals↗
PubMed1988

Lithium increases dynorphin A(1-8) and prodynorphin mRNA levels in the basal ganglia of rats.

The aim of this study was to understand the possible influence of the antimanic drug, lithium, and the neuroleptic, haloperidol, alone or in combination, on the regulation of dynorphin biosynthesis in the striatum. The study was done using male Fisher-344 rats subjected to a regimen of subchronic administration of lithium chloride (4 mEq/kg/day for 1,2,4 or 6 days, i.p.) or a regimen of chronic oral administration of a diet containing lithium carbonate (1.5 g/kg of the diet). Subchronic administration of lithium increased striatal dynorphin A(1-8)-like immunoreactivity (DN-LI) in a time-related fashion. Immunocytochemistry revealed an increase in DN-LI in fibers and cells clustered in 'patches' throughout striatum. The increase in DN-LI was reversible on cessation of lithium administration. Concurrent administration of lithium and an opiate antagonist, naltrexone, or a dopamine receptor antagonist, haloperidol, did not influence the changes induced by lithium. Chronic oral administration of lithium for 21 days led to an increase in DN-LI in the striatum. Co-administration of haloperidol with the 21 day regimen of lithium administration failed to affect the increase in DN-LI. The prodynorphin mRNA abundance in the striatum was quantitated by a molecular hybridization procedure using a prodynorphin 32P-cRNA probe generated from the Riboprobe system. Evidence from the Northern blot analysis reveals that lithium increases the prodynorphin mRNA abundance in the striatum. These results indicate that lithium affects the dynamics of prodynorphin biosynthesis in the striatum, presumably increasing transcription and/or translational processes.

Animals↗
PubMed1988

Cell-type specific processing of neuroendocrine precursor proteins using vaccinia recombinants.

The cell-type specific processing of human pro-enkephalin was determined using a vaccinia recombinant (VV:hPE). The results show that all cell types infected with VV:hPE efficiently synthesize pro-enkephalin following cleavage of the signal peptide after Ala24. In addition, pro-enkephalin is shown to undergo N-linked glycosylation as well as other post-translational modifications. However, only one cell line. AtT-20, was able to efficiently cleave pro-enkephalin to smaller peptides including Met-enkephalin. Some results have been previously reported (Science 232, 1641-1643).

Amino Acid Sequence↗
PubMed1988

Regulation of hypothalamic magnocellular neuropeptides and their mRNAs in the Brattleboro rat: coordinate responses to further osmotic challenge.

A paradigm was developed for the chronic osmotic stimulation of homozygous diabetes insipidus rats of the Brattleboro strain, a strain that fails to synthesize vasopressin. This study examines the adaptation of 2 sets of coexisting peptide hormone magnocellular neurons in the hypothalamoneurohypophyseal system (HNS) of Long Evans (LE), Brattleboro heterozygote (HZ), and Brattleboro homozygote (DI) rats: (1) the arginine8-vasopressin (AVP)/dynorphin (DYN) neurons, and (2) the oxytocin (OT)/cholecystokinin (CCK8) neurons of the paraventricular and supraoptic nuclei, which project to the posterior pituitary. The regimen of chronic intermittent salt-loading (CISL) involved the replacement of 2% saline for normal drinking water for 18 hr/d. This protocol effectively increased plasma levels of AVP and OT in LE and HZ rats, oxytocin in DI rats, and maintained the posterior pituitary in a state depleted of AVP, OT, CCK, and peptides derived from pro-dynorphin: DYN A 1-17, DYN A 1-8, and DYN B 1-13. The ratio of pituitary DYN A 1-17 to DYN A 1-8 content in DI rats or in LE, HZ, and DI rats following 6 d of CISL suggests a preferential release of DYN A 1-17 during periods of chronic secretory activity. In response to chronic secretory activity, mRNAs for AVP, OT, DYN, and CCK increased 1.5-2-fold in all 3 AVP rat strains, with mRNAs for coexisting peptide hormones displaying parallel increases. Mutant AVP mRNA in the DI rat was expressed at very low levels and DYN mRNA in very high levels, with each of these mRNAs continuing to be regulated by CISL in a normal manner. These results suggest a regulatory relationship between AVP and OT neurons, in which vasopressin neurons are feedback-regulated by AVP, most likely via plasma osmolarity, and that oxytocin neurons are modulated by peptides derived from pro-dynorphin.

Animals↗
PubMed1987

Patient response to sigmoidoscopy. A randomized, controlled trial of rigid and flexible sigmoidoscopy.

Sigmoidoscopy could aid in the control of large bowel cancer by early detection of the 55% of colorectal cancers that develop in the rectosigmoid and by the identification and eradication of significant rectosigmoid adenomas. Rigid sigmoidoscopy has had poor patient acceptance and therefore has not been successful. The present study is a prospective randomized trial to evaluate patient response to flexible as compared with rigid sigmoidoscopy. Patients reported significantly less discomfort (10.1% versus 29.7%), anxiety (9.8% versus 27.6%) and embarrassment (5.2% versus 12.8%) during flexible as compared with rigid sigmoidoscopy. Flexible sigmoidoscopy appears to have better patient acceptance than rigid sigmoidoscopy (P less than 0.01). This could enhance its value as a cancer-control instrument. This article addresses the feasibility of sigmoidoscopy. Its validity also needs to be addressed within the framework of a long-term trial, evaluating mortality for rectosigmoid cancer.

Clinical Trials as Topic↗
PubMed1987

Two mammalian genes transcribed from opposite strands of the same DNA locus.

This report describes the characterization of a genomic locus in the rat that encodes overlapping genes occupying both strands of the same piece of DNA. One gene (strand) encodes gonadotropin-releasing hormone (GnRH). A second gene, SH, is transcribed from the other DNA strand to produce RNA of undefined function. The RNAs transcribed from each DNA strand are spliced and polyadenylated, and share significant exon domains. GnRH is expressed in the central nervous system while SH transcripts are present in the heart. Thus, the genome of a mammalian organism encodes two distinct genes by using both strands of the same DNA.

Animals↗
PubMed1987

A solution hybridization assay for the quantitation of prodynorphin mRNA.

We have developed a RNA-RNA solution hybridization assay to quantitate the mRNA coding for prodynorphin precursor. This assay is extremely sensitive and highly specific. Using this assay we have measured the prodynorphin mRNA in various brain regions and reproductive tissues of rat. When we compared the distribution of prodynorphin mRNA with the dynorphin related peptides in these tissues, we found a general parallelism and a few noteworthy exceptions.

Animals↗
PubMed1987

The rat corticotropin-releasing hormone gene.

In this paper we have described the isolation and characterization of the rat corticotropin releasing hormone gene. Nucleotide sequence comparisons with the human CRH gene have demonstrated several interesting regions of homology and suggest that the gene was highly conserved through evolution. Additionally we have demonstrated the tissue-specific expression of the rat CRH gene. The regional distribution of expression parallels previously documented immunocytochemical demonstrations and supports the hypothesis that CRH peptides have multiple roles in different tissues. In the peripheral tissues that express CRH mRNA it will be very interesting to document the specific cell type of synthesis by using combined immunocytochemical and in situ histochemical techniques. Additionally we have described initial studies using gene transfer techniques to examine the cAMP responsiveness of the rat CRH gene. We are presently constructing other fusion genes (CRHCAT plasmids) in order to more carefully localize the DNA sequence in the rat CRH gene which mediates this effect, and compare it to the previously reported cAMP-responsive "consensus sequence." Similarly, we also plan to utilize the CRHCAT constructs to examine regulation of the rat CRH gene by glucocorticoids and several other hormone-mediated regulatory pathways. Through these CAT fusion studies we hope to gain a better understanding of the role of certain conserved sequences in the 5' flanking DNA for transcriptional control of the rat (and human) CRH genes.

Animals↗