Biomedical subjects
C E Day
Publications and source records attributed to C E Day.
Health of dairy herds converting to organic farming.
Explore the source record for details and available documents.
Frequent recombination in the human T-cell receptor beta gene complex.
Although individual TCRVBV gene segments exhibit limited polymorphism, human T-cell receptor beta (TCRB) haplotypes are characterized by multiple different combinations of allelic markers. This observation suggests that genetic recombination may have played a role in the generation of these haplotypes. Meiotic recombination in a region spanning approximately 250 kilobases (kb) at the 3' end of the TCRB gene complex was investigated by extended family studies and by analysis of single sperm. Segregation patterns of polymorphic TCRB markers in families allowed the assignment of TCRB alleles to parental haplotypes and detection of recombinants among the offspring. Among the 178 informative paternal meioses, four (approximately 2%) were recombinant, whereas no recombinants were found in the 199 maternal meioses. In addition, segregation of two allelic markers was examined in a total of 1101 individual sperm from two heterozygous donors to detect exchange events in this region. The results revealed a similar rate of recombination, approximately 1.3%, which, along with the family data, suggests that at, least in males, meiotic recombination in this 250 kb region may be six times higher than the "average" rate of 1% per 10(6) bases that has been estimated for the human genome.
Serum antioxidants as predictors of adult respiratory distress syndrome in patients with sepsis.
Adult respiratory distress syndrome (ARDS) can develop as a complication of various disorders, including sepsis, but it has not been possible to identify which of the patients at risk will develop this serious disorder. We have investigated the ability of six markers, measured sequentially in blood, to predict development of ARDS in 26 patients with sepsis. At the initial diagnosis of sepsis (6-24 h before the development of ARDS), serum manganese superoxide dismutase concentration and catalase activity were higher in the 6 patients who subsequently developed ARDS than in 20 patients who did not develop ARDS. These changes in antioxidant enzymes predicted the development of ARDS in septic patients with the same sensitivity, specificity, and efficiency as simultaneous assessments of serum lactate dehydrogenase activity and factor VIII concentration. By contrast, serum glutathione peroxidase activity and alpha 1Pi-elastase complex concentration did not differ at the initial diagnosis of sepsis between patients who did and did not subsequently develop ARDS, and were not as effective in predicting the development of ARDS. Measurement of manganese superoxide dismutase and catalase, in addition to the other markers, should facilitate identification of patients at highest risk of ARDS and allow prospective treatment.
Organization of human T-cell receptor beta-chain genes: clusters of V beta genes are present on chromosomes 7 and 9.
To ascertain the extent and organization of the germ-line human T-cell receptor (TCR) beta-chain gene repertoire, beta-chain variable region (V beta) genes were mapped by pulsed-field gel electrophoresis, cosmid cloning, and in situ hybridization. Probes derived from the 24 known V beta families were mapped to a total of six Sfi I fragments in DNA samples from multiple individuals representing all possible haplotypes of TCR V- and C (constant)-region insertion/deletion-related polymorphisms. Four of the Sfi I fragments were linked to one another to develop an extended map of the TCR beta-chain gene complex previously localized to chromosome 7q35. The remaining two Sfi I fragments, containing 6 V beta genes, could not be linked to the TCR beta-chain gene complex. Using human-hamster somatic cell hybrids and in situ hybridization, these orphon genes were localized to chromosome 9p. Nucleotide sequences of the orphon V beta genes, derived from cosmid clones, were 93-97% identical to V beta genes in the TCR beta-chain gene complex. Open reading frames in three of the orphon V beta genes were intact as were the recombination signal sequences. As expected, based on their orphon status, none of the V beta genes of chromosome 9 was detected in transcripts containing C beta. These results indicate that the functional germ-line V beta repertoire in humans is substantially (10%) smaller than previously estimated.
[Allelic polymorphism in the coding region of human T cell receptor V beta 12 gene].
Sequence substitutions in T cell antigen receptor (TCR) beta chains have been shown to have a profound impact upon the fine specificity of T lymphocytes. It is, therefore, of interest to characterize allelic variations in the TCR gene segments that assemble to encode TCR molecules. In the present report, the sensitive technique of single stranded conformational polymorphism (SSCP) was used to screen for allelic sequence variations in the coding region of TCR V beta 12 gene. TCR V beta 12 specific primers were used to PCR amplify and 32P label a 273bp segment of the variable (V) gene segment. Sequence differences between different allelic forms were detected by denaturation of the DNA and separation on a non-denaturing, high resolution acrylamide gel. Using this rapid approach, four alleles of the TCR V beta 12 gene were detected in a population of 90 ethnically diverse individuals. Correct Mendelian segregation of the four alleles was demonstrated in family studies (n = 23) and allele frequencies were 0.4722, 0.3166, 0.2056, and 0.0056 respectively. Sequence analysis revealed sequence differences of 1-3 nucleotides involving three separate codons. However, predicted amino acid sequences of all four alleles are identical. These data strongly suggest that the TCR V beta 12 protein sequence is highly conserved.
Erosion of veterinary influence.
Explore the source record for details and available documents.
Silent allelic variants of a T-cell receptor V beta 12 gene are present in diverse human populations.
Amino acid substitutions in variable regions of the T-cell receptor (TCR) can alter T-cell reactivity; however, relatively little is known about the extent of allelic variation in human TCR coding sequences. In the present studies, coding region variation in the human TCR V beta 12.2 gene was examined in detail. Virtually the entire V beta 12.2 coding region was screened for nucleotide substitutions by single-stranded conformational polymorphism analysis. Four alleles were identified in a sample population of 90 unrelated people from diverse genetic backgrounds. Three of the alleles were common, with estimated frequencies of 0.32, 0.47, and 0.20. Sequence analyses revealed that variation between the alleles was confined to three single-base differences in codons 24, 31, and 45; none of these changes altered the amino acid sequence. No evidence for other coding region differences in this gene were found. This analysis suggests that coding region variation in V beta 12.2 is limited, and amino acid sequence is highly conserved.
Increased serum catalase activity in septic patients with the adult respiratory distress syndrome.
Excessive hydrogen peroxide (H2O2) generation appears to contribute to the development of the adult respiratory distress syndrome (ARDS), but H2O2-combatting antioxidant defenses have not been evaluated. We found that serum from septic patients with ARDS scavenged more (p less than 0.05) H2O2 in vitro (82.7 +/- 3.8%) than did serum from septic patients without ARDS (56.9 +/- 3.1%) or control subjects (20.2 +/- 2.4%). Serum from septic patients with ARDS also had more (p less than 0.05) catalase activity (54.9 +/- 10.9 U/ml) than did serum from septic patients without ARDS (28.6 +/- 3.4 U/ml) or control subjects (7.3 +/- 0.8 U/ml). In contrast, serum from septic patients with or without ARDS and control subjects had the same glutathione peroxidase (GPX) activity. Serum H2O2 scavenging activity correlated with serum catalase (r = 0.77) but not GPX (r = 0.33) activity and was inhibitable (greater than 90%) by sodium azide, a catalase inhibitor. Increases in serum catalase activity did not appear to be derived from erythrocytes (RBC) because septic patients with or without ARDS and control subjects had similar RBC hemolysis in response to osmotic stress in vitro and serum haptoglobin concentrations. Serum from septic patients with ARDS also protected endothelial cells against H2O2-mediated damage (34.5 +/- 2.2% 51Cr release) better (p less than 0.05) than serum from septic patients without ARDS (47.3 +/- 7.4%) or control subjects (82.1 +/- 10.2%), but killing of bacteria by neutrophils in vitro was the same in serum from patients and control subjects. Our findings indicate that patients with sepsis and/or ARDS have increased serum catalase activity, which may alter H2O2-dependent processes.
Dimethyl sulfoxide decreases lung neutrophil sequestration and lung leak.
To investigate basic mechanisms of acute edematous lung injury (adult respiratory distress syndrome), the formylated tripeptide formyl-norleucyl-leucyl-phenylalanine (FNLP) was instilled intratracheally into hamsters. Intratracheal FNLP produced time-dependent and dose-dependent increases in neutrophils recoverable by lung lavage (neutrophil alveolitis) and leak of intravenously injected albumin into the extravascular lung space (lung leak). Treatment with dimethyl sulfoxide (DMSO) decreased (p less than 0.05) neutrophil alveolitis and lung leak in hamsters given FNLP intratracheally. The effect of DMSO on various neutrophil functions was also studied in vitro. Addition of DMSO at concentrations (about 0.20%) measured in plasma of hamsters given DMSO decreased (p less than 0.05) neutrophil chemotaxis but not neutrophil superoxide anion generation or adherence to cultured endothelial cell monolayers or nylon fiber in vitro. We conclude that intratracheal FNLP causes neutrophil alveolitis and lung leak and that DMSO treatment ameliorates these processes, possibly by inhibiting neutrophil chemotaxis.
Guar gum reduces serum cholesterol in sea quail fed a cholesterol diet.
Guar gum at 6% and 12% in the diet reduces total serum cholesterol by 31% and 51%, respectively, in male SEA quail fed a diet containing 0.5% cholesterol for a period of one week. Although the potency of hypocholesterolemic action of guar gum in cholesterol fed SEA quail is much less than that in cholesterol fed chickens, SEA quail are a reasonable alternative to chickens for evaluation of guar and similar products, based on other considerations such as smaller size and compound requirements and shorter duration of testing.
Activity of psyllium hydrophilic colloid for reducing serum cholesterol in sea quail fed a diet supplemented with cholesterol.
Groups of six SEA quail were fed a 0.5% cholesterol supplemented diet to which was added 3, 6, and 10% microcrystalline cellulose (controls) or the same concentrations of psyllium hydrophilic colloid for a period of 14 days. Total serum cholesterol was measured enzymically on each animal at the end of the treatment period. Cellulose control cholesterol values averaged approximately 1500 mg/dl overall. Psyllium hydrophilic colloid had no significant effect on total serum cholesterol at either 3% or 6% in the diet but did reduce cholesterol significantly by 52% at the 10% dietary level. These results demonstrate that SEA quail appear to be less sensitive to the hypocholesterolemic action of psyllium than are chickens.
Hypocholesterolemic activity of beta-sitosterol in cholesterol fed sea quail.
Male SEA quail were fed a 0.5% cholesterol supplemented diet, to which was added 0%, 1%, and 2% beta-sitosterol, for a period of seven days. Dietarily administered beta-sitosterol reduced total serum cholesterol levels by 62% and 72% at the 1% and 2% treatment doses, respectively. This hypocholesterolemic activity of sitosterol in cholesterol fed SEA quail was anticipated on the basis of the numerous earlier studies demonstrating similar activity in cholesterol fed chickens. Beta-sitosterol was tested in SEA quail to experimentally confirm its expected serum cholesterol lowering effects and to expand further the utility of the SEA quail model in cholesterol and atherosclerosis research.
Non-specific pruritus in dogs.
Explore the source record for details and available documents.