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Biomedical subjects

E Herbert

Publications and source records attributed to E Herbert.

At least 19 recordsLinked to original sources

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

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

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

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

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

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

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

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

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

Immunohistochemical demonstration of proGnRH and GnRH in the preoptic-basal hypothalamus of the primate.

An antiserum (ARK-1) specific to the gonadotropin-releasing hormone precursor (proGnRH) was produced by immunizing with a synthetic peptide (proGnRH 6-16; Gly-Leu-Arg-Pro-Gly-Gly-Lys-Arg-Asp-Ala-Glu) which bridges the proteolytic cleavage site of proGnRH. When used in the radioimmunoassay, ARK-1 bound 25% of the iodinated 5-16 fragment at a 1:30,000 dilution with a sensitivity of 1 pg/tube. Using immunohistochemical techniques, we observed that in serial and the same sections through the preoptic-basal hypothalamus (POA-BH), the precursor molecule was primarily present in the cell soma, whereas GnRH was found in the cell soma, nerve fibers, and terminals of the same neurons. These data indicate that the processing of proGnRH to biologically active peptides (e.g., GnRH) in the rhesus macaque and the baboon POA-BH primarily occurs in the cell soma.

Animals

Expression of the prodynorphin gene in male and female mammalian reproductive tissues.

Recent studies suggest that opioid peptides may be involved in modulating the hypothalamus-pituitary-gonadal axis at a variety of levels in both males and females. We report here the presence of mRNA coding for the opioid peptide precursor prodynorphin in rat ovary, uterus, and testis. Expression of this opioid peptide precursor gene is compared to expression of two other opioid peptide precursor genes, proenkephalin and proopiomelanocortin, in mammalian reproductive tissues. Immunohistochemical analysis reveals that in the rat testis, prodynorphin-derived peptides are present in Leydig cells. The distribution of dynorphin immunoreactivity in various reproductive tissues was determined. Male reproductive tissues of the rat, rabbit, and guinea pig as well as rat ovary and uterus all contain detectable levels of dynorphin immunoreactivity. These observations suggest that prodynorphin-derived peptides may exert paracrine and/or autocrine effects in mammalian reproductive tissues.

Animals

Rat corticotropin-releasing hormone gene: sequence and tissue-specific expression.

The rat corticotropin releasing hormone (CRH) gene has been isolated and characterized by DNA sequence analysis. The gene exhibits a structural organization similar to that of the human CRH gene. The nucleotide sequence encoding the entire rat CRH precursor is located on the second exon, while exon I encodes the 5'-untranslated region of the mRNA. Analysis of the nucleotide sequence homology between the human and rat CRH genes reveals several highly conserved regions including the CRH peptide-encoding sequence and the 5'-flanking sequence. RNA blot analysis demonstrates that CRH mRNA can be observed in numerous regions of the rat brain as well as the spinal cord, adrenal gland, pituitary, and testis.

Amino Acid Sequence

Expression and cell type--specific processing of human preproenkephalin with a vaccinia recombinant.

The posttranslational maturation of a complex precursor polyprotein, human proenkephalin, was assessed by infection of a wide spectrum of cell types with a recombinant vaccinia virus that expressed human proenkephalin. The infected cells rapidly produced both cellular and secreted Met-enkephalin immunoreactivity. Although each cell line could secrete intact proenkephalin, only a mouse pituitary line was capable of processing proenkephalin to mature enkephalin peptides. The quantity of intact proenkephalin secreted from BSC-40 cells (derived from African Green monkey kidney) was sufficient to establish the value of vaccinia virus as a mammalian cell expression vector.

Animals