Search PubMed⌕ Search

Biomedical subjects

J Burton

Publications and source records attributed to J Burton.

At least 55 records · Page 3Linked to original sources
PubMed1994

Validation of a simple technique for the detection of abnormal mucosal cell replication in humans.

Abnormal intestinal crypt cell proliferation is considered to be an important early risk marker for colorectal cancer but measurement of the rate and spatial distribution of cell division by histochemical localization of DNA synthesis is labour-intensive and expensive. We developed and evaluated a simpler technique for measurement of these parameters using direct visual analysis of mitotic figures in microdissected crypts. The direct crypt analysis technique was applied to colorectal biopsies from patients with ulcerative colitis or no mucosal abnormality. A characteristic shift of cell division toward the intestinal lumen was detected in patients with ulcerative colitis. The direct method was validated using rats fed diets containing cellulose, or guar gum to stimulate mucosal cell proliferation. The crypt cell proliferation rate (CCPR) was measured by the metaphase-arrest technique and the results were compared with direct crypt analysis. There was a fivefold range of CCPR values at three sampling sites across the dietary groups. An excellent linear correlation between the results by the two techniques was obtained (r = 0.98; P < 0.001). In a second experiment the spatial distribution of dividing cells between five zones in colonic crypts, determined by the new method or by staining with BrdU, was compared. Good agreement was again achieved. Visual analysis of intact crypts is a valid technique for the measurement of crypt cell cytokinetics and it is particularly suited for use in a clinical environment.

Animals↗
PubMed1994

Pharmacological characterization of rabbit corpus cavernosum relaxation mediated by the tissue kallikrein-kinin system.

1. The roles of the tissue kallikrein-kinin system and nitric oxide (NO) release in Phoneutria nigriventer venom-induced relaxations of rabbit corpus cavernosum (RbCC) smooth muscle have been investigated by use of a bioassay cascade. 2. Phoneutria nigriventer venom (10-30 micrograms), porcine pancreatic kallikrein (100 mu), rabbit urinary kallikrein (10 mu), bradykinin (BK, 0.3-3 nmol), acetylcholine (ACh, 0.3-30 nmol) and glyceryl trinitrate (GTN, 0.5-10 nmol) caused relaxations of the RbCC strips. Captopril (1 microM) substantially potentiated Phoneutria nigriventer venom- and BK-induced RbCC relaxations without affecting those elicited by GTN. 3. The bradykinin B2 receptor antagonist, Hoe 140 (D-Arg-[Hyp3,Thi5,D- Tic7,Oic8]-BK, 50 nM), aprotinin (10 micrograms ml-1) and the tissue kallikrein inhibitor, Pro-Phe-Aph-Ser-Val- Gln-NH2 (KIZD-06, 1.3 microM) significantly inhibited Phoneutria nigriventer venom-induced RbCC relaxations, without affecting those provoked by GTN and ACh. The B1 receptor antagonist, [Leu9]des Arg10BK (0.5 microM) and soybean trypsin inhibitor (SBTI, 10 micrograms ml-1) had no effect on Phoneutria nigriventer venom-induced RbCC relaxations. 4. The relaxations induced by Phoneutria nigriventer venom, porcine pancreas kallikrein, BK and ACh were significantly inhibited by N omega-nitro-L-arginine methyl ester (L-NAME, 10 microM) but not by D-NAME (10 microM). L-NAME did not affect GTN-induced relaxations. L-Arginine (300 microM), but not D-arginine (300 microM), significantly reversed the inhibitory effect of L-NAME. 5. Our results indicate that Phoneutria nigriventer venom activates the tissue kallikrein-kininogen-kinin system in RbCC strips leading to NO release and suggest a functional role for this system in penile erection.

Adrenergic beta-Antagonists↗
PubMed1994

Prevention and regression of coronary atherosclerosis. Is it safe and efficacious therapy?

OBJECTIVE: Atherosclerotic coronary heart disease (CHD) continues to be the dominant disease in Western society. A large body of evidence directly linking serum cholesterol levels and CHD risk has stimulated population treatment strategies designed to reduce cholesterol and CHD risk. Data indicating a relation between low cholesterol and non-CHD risk have, however, suggested that cholesterol reduction may not always be desirable. The primary goal of this evaluative review of the available evidence was to answer the following question: Is prevention/regression therapy for CHD safe and effective? DATA SOURCES: Three lines of evidence were reviewed: epidemiologic studies; primary and secondary prevention trials with clinical end points; and secondary prevention trials with quantitative coronary angiography as a surrogate end point for clinical CHD. STUDY SELECTION: Original studies and meta-analyses were reviewed. The principal selection criteria for the epidemiologic studies were large size and prolonged follow-up; for the trials, randomization and viable clinical (CHD events, CHD mortality, total mortality) or angiographic end points. DATA EXTRACTION: The data were initially extracted by a single reviewer using common qualitative guidelines. The data were then evaluated by all authors acting as a data interpretation team. DATA SYNTHESIS: Overall, the epidemiologic data revealed excess risk of fatal and nonfatal CHD events was directly related to total cholesterol and low-density lipoprotein (LDL) cholesterol levels, for both men and women and for both younger (< 65 years) and older (> or = 65 years) patients, over a wide range of serum cholesterol levels. The predictive value was higher in younger men than older men and women, although part of this quantitative interaction may be due to fewer studies, with fewer end points, in the older and female populations. The CHD events and CHD mortality, but not total mortality, were consistently reduced in trials of cholesterol-lowering therapy. The regression trials, predominantly in CHD patients with high cholesterol values (mean 7.1 mmol/L), demonstrated improvement in angiographic atherosclerosis in every study. The evidence for elevated risk of non-CHD death at very low levels of cholesterol is uncertain and controversial. The most likely possibilities for this apparent relationship are unknown confounding variables and the play of chance. CONCLUSIONS: Serum cholesterol levels are directly associated with CHD risk, and there is no threshold level below which there is no risk. Reduction of high serum cholesterol levels reduces CHD risk. Whether lipid-lowering and adjunctive antiatherosclerotic therapies are effective and safe in the majority of CHD patients who have desirable or borderline cholesterol levels remains undetermined.

Aged↗
PubMed1994

Does IL-2 receptor expression and secretion in chronic B-cell leukemia have a role in down-regulation of the immune system?

Recent evidence of cell membrane expression of interleukin-2 receptors (IL-2R) by malignant B cells in hairy cell leukemia (HCL) and B-chronic lymphocytic leukemia (B-CLL) has lead to speculation that growth factors, such as IL-2, may play a role in the pathophysiology of these diseases. However, to date, it is not clear that IL-2 is a consistent growth factor in vitro or in vivo for malignant B cells. What then is the potential significance of membrane IL-2R on the malignant B-cell membrane? Laboratory analysis indicates that the malignant cells are the source of elevated serum levels of soluble Tac protein (sIL-2r alpha) in both diseases. Indeed, these cells spontaneously secrete sIL-2R alpha into culture medium. We speculate that the presence of an expanding mass of malignant B cells possessing high and low affinity membrane IL-2R may contribute significantly to the associated immunodeficiency seen in B-CLL. In particular, it is the cell associated high affinity IL-2R that have the greatest potential for reducing the levels of free IL-2 available to normal immune cells.

B-Lymphocytes↗
PubMed1993

A mammalian guanine-nucleotide-releasing protein enhances function of yeast secretory protein Sec4.

Small GTP-binding proteins of the ras superfamily are important for exocytosis from eukaryotic cells. GTP-binding proteins can exist in two different conformations depending on whether they are bound to GDP or GTP, and are thought to function as molecular switches that regulate a variety of cellular processes. The GTP-GDP cycle is controlled by accessory proteins that promote the exchange of bound GDP or the hydrolysis of GTP. The protein Sec4, a member of the Sec4/Ypt1/Rab branch of the Ras superfamily, is involved in a late stage of the secretory pathway in yeast. Here we report the isolation of a mammalian complementary DNA, mss4, encoding a GDP-releasing protein that enhances Sec4 function. The Mss4 protein also stimulates GDP release from Ypt1 and from the mammalian protein Rab3a, but not from Ras2. Mss4 shows sequence similarity to Dss4, a yeast protein with similar biochemical properties.

ATP-Binding Cassette Transporters↗
PubMed1993

SPK1 is an essential S-phase-specific gene of Saccharomyces cerevisiae that encodes a nuclear serine/threonine/tyrosine kinase.

SPK1 was originally discovered in an immunoscreen for tyrosine-protein kinases in Saccharomyces cerevisiae. We have used biochemical and genetic techniques to investigate the function of this gene and its encoded protein. Hybridization of an SPK1 probe to an ordered genomic library showed that SPK1 is adjacent to PEP4 (chromosome XVI L). Sporulation of spk1/+ heterozygotes gave rise to spk1 spores that grew into microcolonies but could not be further propagated. These colonies were greatly enriched for budded cells, especially those with large buds. Similarly, eviction of CEN plasmids bearing SPK1 from cells with a chromosomal SPK1 disruption yielded viable cells with only low frequency. Spk1 protein was identified by immunoprecipitation and immunoblotting. It was associated with protein-Ser, Thr, and Tyr kinase activity in immune complex kinase assays. Spk1 was localized to the nucleus by immunofluorescence. The nucleotide sequence of the SPK1 5' noncoding region revealed that SPK1 contains two MluI cell cycle box elements. These elements confer S-phase-specific transcription to many genes involved in DNA synthesis. Northern (RNA) blotting of synchronized cells verified that the SPK1 transcript is coregulated with other MluI box-regulated genes. The SPK1 upstream region also includes a domain highly homologous to sequences involved in induction of RAD2 and other excision repair genes by agents that induce DNA damage. spk1 strains were hypersensitive to UV irradiation. Taken together, these findings indicate that SPK1 is a dual-specificity (Ser/Thr and Tyr) protein kinase that is essential for viability. The cell cycle-dependent transcription, presence of DNA damage-related sequences, requirement for UV resistance, and nuclear localization of Spk1 all link this gene to a crucial S-phase-specific role, probably as a positive regulator of DNA synthesis.

Base Sequence↗
PubMed1992

Mapping the binding site of tissue kallikrein: preparation and testing of all possible substrate analog inhibitors homologous with the sequence of kininogen between Ser386 and Gln392.

Programs aimed at converting peptide inhibitors of proteolytic enzymes into more traditional drug structures require an understanding of the role played by the individual amino acid residues in the inhibitor. To this end, all possible substrate analogues occurring within the sequence Ser386-Pro-Phe-Arg-Ser-Val-Gln392 from bovine kininogen were synthesized and tested as inhibitors of tissue kallikrein (EC 3.4.21.35, beta-PPK). Of the 21 sequences which can be formed from the heptapeptide, 11 have inhibitory constants which could be measured in the chromogenic assay employed in these studies. No dipeptide and only one tripeptide, Ac-Phe-Arg-Ser-NH2 (Ki = 718 microM), measurably inhibits the enzyme. All longer peptides inhibit beta-PPK. The heptapeptide Ac-Ser-Pro-Phe-Arg-Ser-Val-Gln-NH2 is the most effective inhibitor in this series (Ki = 101 microM). Each amino acid residue in the sequence appears to alter binding in a relatively independent manner. The N-terminal seryl residue (P4) and the prolyl residue (P3) slightly improve the Ki of the various inhibitors. The phenylalanyl residue at P2 appears to have a more pronounced effect on Ki. The arginyl residue at P1 and the seryl residue at P1' appear to be the most important residues in the inhibitory sequence. They contribute approximately one-third and one-fourth of the binding energy to the interaction between the substrate analogues and beta-PPK, respectively. The valyl residue at P2', and the C-terminal glutaminyl residue improve Ki of each of the peptides tested. Almost 80% of the binding energy of the substrate analogue inhibitors comes from the core sequence Phe-Arg-Ser which occurs between P2 and P1'. Molecular models developed from the Chen-Bode coordinates of the aprotinin-beta-PPK complex have been used to interpret the results of these studies.

Amino Acid Sequence↗
PubMed1992

Studies on the design of orally active peptide analogues.

Converting peptide inhibitors of tissue kallikrein into drugs requires, among other things, that the compounds be made orally active. Measurement of the rate at which peptides diffuse across model epithelial membranes that mimic the human gut has allowed a quantitative relationship relating lipophilicity and % absorption to be developed. The absorption of simple peptides from the human gut may be predicted with this equation.

Administration, Oral↗
PubMed1992

A systematic approach for determining minimum inhibitory sequence and contribution of individual residues in binding of kininogen fragments to tissue kallikrein.

A systematic approach to evaluate the contribution of individual residues occurring within the sequence Ser386-Pro-Phe-Arg-Ser-Val-Gln392 from bovine kininogen towards binding to tissue kallikrein is developed. Of the 21 sequences which can be formed, no dipeptide and only one tripeptide measurably inhibits the enzyme. Almost 80% of the binding energy of the substrate analogue inhibitors comes from the core sequence Phe-Arg-Ser which occurs between P2 and P1'. Molecular models developed from the Chen-Bode coordinates of the aprotinin--beta-PPK complex have been used to interpret the results of these studies.

Amino Acid Sequence↗
PubMed1992

Activity distribution of seven digestive enzymes along small intestine in calves during development and weaning.

Ten groups of calves were used to study the changes in activity levels and distribution of seven hydrolases in the intestinal mucosa during development and weaning. The calves in the first group were sacrificed at birth while those in the remaining nine groups were either milk-fed until slaughter on days 2, 7, 28, 56, 70, and 119; or weaned between days 28 and 56 and then slaughtered on days 56, 70, and 119, respectively. The small intestine was immediately cut off and divided into five segments, ie, duodenum, proximal jejunum, median jejunum, distal jejunum, and ileum. In the milk-fed animals, the activity levels of aminopeptidases A and N, alkaline phosphatase, lactase, and isomaltase were maximum at 2 days of age, and then declined sharply between days 2 and 7 but did not change significantly thereafter. By contrast, the maltase activity increased between days 7 and 119, while no sucrase activity was detected. Weaning resulted in a decrease in the activity of lactase and an increase in that of aminopeptidase N, maltase, and isomaltase. The distribution of all these enzymes along the small intestine was slightly influenced by age but not at all by weaning.

Animals↗
PubMed1992

Plasma and tissue levels of digestive regulatory peptides during postnatal development and weaning in the calf.

Changes in the concentrations of cholecystokinin, gastric inhibitory peptide, gastrin, motilin, pancreatic polypeptide, secretin, somatostatin, and vasoactive intestinal peptide in calf plasma and antral, duodenal and/or pancreatic tissues were assessed by radioimmunoassay during postnatal development and after weaning in 50 male Holstein-Friesian calves (randomly distributed into 10 groups of 5 animals each). The calves in the first group were killed at birth while those in 6 other groups were colostrum-fed for 2 days and then milk-fed until 7, 28, 56, 70 or 119 days of age. Those in the remaining 3 groups were given the same diets until day 28, were then weaned between day 29-56, and slaughtered on days 56, 70 or 119. In milk-fed animals, changes in plasma and tissue concentrations of almost all digestive regulatory peptides were observed during the 1st month of postnatal life, especially at day 2. Weaning was accompanied by variations in the plasma concentrations of somatostatin, secretin, gastrin, pancreatic polypeptide and gastric inhibitory peptide but not by any apparent change in peptide tissue concentrations (except VIP in the duodenum). Thus, the variations in tissue concentrations are primarily age-related, while plasma concentrations were modified by age and weaning.

Aging↗
PubMed1992

Gastric and pancreatic enzyme activities and their relationship with some gut regulatory peptides during postnatal development and weaning in calves.

Changes in the activities of three gastric and nine pancreatic enzymes plus colipase were determined during postnatal development and weaning in calves. In calves exclusively milk-fed for 2, 7, 28, 56, 70 and 119 d, the enzyme activities per kilogram of empty live weight increased with age for chymotrypsin, elastase, carboxypeptidases A and B, ribonuclease and alpha-amylase, decreased for chymosin, lysozyme and colipase but showed no change in the case of pepsin, trypsin, lipase and phospholipase A2 compared with animals at birth. The greatest increase was that in alpha-amylase activity (about 50-fold between d 2 and 119). In calves weaned between d 28 and 56, all the activities were higher than in milk-fed animals, except that of chymosin (which was slightly lower) and that of colipase (which did not change). At 119 d of age, chymotrypsin, carboxypeptidase A, alpha-amylase and lipase were 1.6- to fourfold higher in ruminants than in preruminants. Thus, most enzyme activities were modified first by colostrum and milk intake, and again upon weaning by development of the forestomachs and ingestion of solid food. These ontogenic patterns might be under the control of many gut regulatory peptides, the plasma concentrations of which changed simultaneously. Some gastric and pancreatic enzymes were correlated to plasma concentrations of these gut regulatory peptides.

Abomasum↗
PubMed1992

Comparison of plasma prekallikrein levels in human skin blister and chamber fluids from volunteer subjects and psoriatic patients.

Suction blisters were successfully raised on lesional, perilesional and non-lesional skin of psoriatic patients. Similar blisters were created on the skin of volunteer subjects. A specific method has been established for the measurement of plasma prekallikrein, and validated for blister fluid, skin chamber fluid, plasma and synovial fluid. Combined levels of plasma prekallikrein and its active form were compared in blister fluids obtained from psoriatic patients and the volunteers. Significantly higher values for plasma prekallikrein were found in the non-lesional blister fluids of psoriatic patients when compared to the volunteer subjects.

Blister↗