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Biomedical subjects

J Pierce

Publications and source records attributed to J Pierce.

At least 37 records · Page 2Linked to original sources

Lower levels of cigarette consumption found in smoke-free workplaces in California.

OBJECTIVE: We examined the relationship between workplace smoking policies and smoking prevalence and cigarette consumption. METHODS: California residents were questioned by telephone with the 1990 California Tobacco Survey. All respondents (11,704) above age 18 years who were employed indoors were used. Respondents were asked about smoking status, workplace smoking policy, desire to quit, and smoking history. Logistic regression was used to determine the relationship of workplace smoking policy to smoking status, accounting for demographic variables. RESULTS: Prevalence of regular smokers was significantly lower in smoke-free workplaces than in those with no restrictions (13.7% vs 20.6%, P < .001). Continuing regular smokers in smoke-free workplaces smoked fewer cigarettes than those in workplaces with no restrictions (296 vs 341 packs per year, P < .001). More comprehensive smoking policies were associated with smokers more likely to contemplate quitting (P = .014). CONCLUSIONS: Employees in smoke-free workplaces have a lower smoking prevalence and, among continuing smokers, lower cigarette consumption than individuals working where smoking is permitted. We estimate cigarette consumption among employees indoors is 21% below that if there were no smoking restrictions in California workplaces. Furthermore, if all California workplaces were smoke-free, cigarette consumption among employees would be 41% below that if there were no workplace smoking restrictions, approximately a $406 million annual loss in sales to the tobacco industry. This study supports the hypothesis that smoke-free workplace policies are an effective public health measure for decreasing smoking prevalence and cigarette consumption among continuing smokers.

Adolescent

Genetic diversity from a limited repertoire of mutations on different common allelic backgrounds: alpha 1-antitrypsin deficiency variant Pduarte.

alpha 1-Antitrypsin (alpha 1AT) is one of the most polymorphic gene loci in the human genome. alpha 1AT variants are typically identified by their migration position in an isoelectric focusing gel at pH 4-5. Heterogeneity of the isoelectric point of alpha 1AT variants, hence variant migration, most often results from amino acid substitutions which alter the net charge of the molecule. We identified an individual heterozygous for an alpha 1AT variant migrating in the "P" variant region which differs from other known "P" variants. Using isoelectric focusing on an immobilized pH gradient at pH 4.50-4.85 the novel P allele, Pduarte, migrates between Pst. albans and Plowell. Densitometric analysis of normal "M" type alpha 1AT and the deficiency variant Plowell major bands separated by isoelectric focusing demonstrates that Pduarte contributes approximately 41% as much alpha 1AT to the total serum alpha 1AT concentration as the normal "M" alpha 1AT, similar to Plowell. Direct DNA sequencing of the proband's genomic DNA demonstrates that the Pduarte allele differs from the normal M1 (V213) allele by two amino acid substitutions, R101 (CGT)-->H(CAT) and D256 (GAT)-->V(GTT). Individually, these amino acid substitutions characterize the normal M4 allele (R101-->H) and the deficient Plowell allele (D256-->V). Thus the Pduarte allele differs from the Plowell allele only by the normal allelic background in which the V256 mutation occurs. Comparison of amino acid sequences among several alpha 1AT variants demonstrates that Pduarte is an example of a more general observation regarding diversity within the PI (protease inhibitor) system.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged

Stressors experienced by relatives of patients in an innovative rehabilitation program.

The social work component of this study was conducted for ethical reasons to monitor the impact on relatives of providing therapy to severely brain-injured patients on an innovative rehabilitation treatment program in an acute care hospital. The authors identify stressors experienced by the family caregivers. Findings are based on 450 interviews conducted by three research social workers during the 18-month study period with 78 family caregivers of the 37 patients. The results indicate that involvement in providing treatment was stressful, because of both the demands on the family and the unproven efficacy of the treatment. The authors recommend that a family's ability to provide treatment be clearly assessed before recruitment and that relatives' stress be monitored during their involvement. The findings emphasize the important role social workers can perform in identifying and monitoring such stress.

Brain Injuries

Restoration of spermatogenesis by follicle-stimulating hormone despite low intratesticular testosterone in photoinhibited hypogonadotropic Djungarian hamsters (Phodopus sungorus).

By exposing male Djungarian hamsters to short-day photoperiods, a hypogonadotropic condition resembling that of hypophysectomized animals can be achieved without resorting to surgery. By use of this animal model, differential effects of gonadotropins on spermatogenesis have been studied revealing that FSH selectively restimulates spermatogenesis up to the level of spermatids. In the present study, we investigated whether FSH (6 IU/day) also restores fertility in hamsters exposed to short-day photoperiods (8L:16D). Animals kept under long-day photoperiod (16L:8D) served as controls. After 8 wk under 8L:16D, gonads were involuted and the animals (n = 9-10 per group) were treated with FSH, or testosterone implants, or both for the following 33 days. An additional group received FSH for 33 days and, for the last 12 days only, FSH plus testosterone to induce copulatory and ejaculatory behavior. The implants caused no increase in intratesticular testosterone as compared to that in short-day controls. All males were paired with adult females for the last 5 days of treatment, and killed thereafter. Litter sizes were determined 22 days later. The examination of the testes by flow-cytometrical techniques revealed qualitatively normal spermatogenesis in animals receiving FSH, whereas testosterone caused an insignificant increase to a few round spermatids. In most FSH-treated animals, sperm were present in the epididymides. In hamsters receiving FSH along with testosterone, treatment resulted in restoration of full fertility as compared to long-day controls. Fertility was also restored in animals receiving testosterone only for the last 12 days of FSH treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

A mouse mammary tumor virus promoter element near the transcription initiation site.

Transcription from the promoter of mouse mammary tumor virus is subject to both positive and negative control by cellular factors, and proviral promoter elements that mediate a basal level of transcription must in some way respond to these cellular regulatory signals. Several such elements, including a TATA box, a region containing three octamer-related sequences, and a binding site for nuclear factor 1, have been previously defined. Additional promoter mutations have allowed a fourth basal promoter element to be identified near the transcription initiation site between +2 and +10. Sequence alterations within this element affect transcription both in vivo and in vitro. Gel electrophoresis mobility shift and DNase I footprinting assays define a nuclear protein, termed initiation site-binding protein, that specifically recognizes this region of the promoter. Optimal levels of transcription from the mouse mammary tumor virus promoter require initiation site-binding protein, as demonstrated by a correlation between protein affinity and transcriptional activity and by specific inhibition of transcription in vitro by an oligonucleotide capable of titrating the protein from transcriptionally active fractions.

Animals

First-year impact of the 1989 California cigarette tax increase on cigarette consumption.

We employed a time series design to evaluate the impact of the 1989 California cigarette tax increase on cigarette consumption in California. Adult per capita consumption data from 1980 to 1990 were analyzed for California and the United States. Trend data indicated a sharp drop in California cigarette consumption coincident with the tax increase. Time-series regression analyses support this observation, and suggest that a 5% to 7% decline in consumption is attributable to the tax increase.

California

The California Diabetes and Pregnancy Program: implementation of a multicenter experience with diabetic pregnancies.

The California Diabetes and Pregnancy Program (CDAPP) began in 1984 as a multicenter, collaborative project with support provided by the state Department of Health Services, Maternal and Child Health Branch. Between its inception and 1988, it expanded from three to eight perinatal regions, making the CDAPP model of care available to patients in 19 clinical affiliate sites. The care was provided by a multidisciplinary team composed of physicians, a diabetes educator, a registered dietician, a social worker, and appropriate consultants. The elements of this model of care included comprehensive patient education, active patient participation in care, maternal and perinatal medical assessment, and collection of standardized patient information adequate to allow a programmatic and medical evaluation of the CDAPP. Despite impressive growth of the program by December 1988, statewide implementation of CDAPP is incomplete.

Adolescent

Humoral and cell surface interactions during gamma-irradiation leukemogenesis in vitro.

Gamma-irradiation of plateau phase cultures of the clonal murine bone marrow stromal cell line D2XRII followed by cocultivation of a clonal interleukin 3 (IL-3) (granulocyte-macrophage colony-stimulating factor (GM-CSF)-dependent hematopoietic progenitor cell line FDC-P1JL26 results in a significant increase in "cobblestone islands" of attachment and emergence of subclonal factor-independent malignant sublines. Biochemical purification of conditioned medium from irradiated D2XRII cells yielded a 75,000-dalton glycoprotein termed leukemogenic stromal factor (LSF) that was neutralized by a polyclonal antiserum to murine macrophage colony-stimulating factor (M-CSF). A monoclonal antibody to the murine M-CSF receptor (c-fms) neutralized the biological activity of this molecule in a manner comparable to its effect on recombinant human or murine M-CSF. FDC-P1JL26 parent cells were positive for Ly5, MEL-14, mGR, VLA-4, PGP-1 (CD44), and Thy1.2. After culture in LSF, Thy1.2, MEL-14, and mGR became undetectable; however, significant cell surface MAC-1 antigen and c-fms (M-CSF receptor) were expressed. Neither line was positive for Ly6, Ly22, I-CAM-1, or B220 antigen. LSF-precultured FDC-P1JL26 cells transferred as single cells to microwell culture with 5000-cGy-irradiated D2XRII cells revealed a 60-fold increase in frequency of cobblestone island formation and evolution of factor-independent subclones compared to the parent line. Both parent and LSF-precultured cells became factor independent at a 100-fold lower frequency if kept in suspension in LSF in the absence of stromal cells. Antiserum to M-CSF or monoclonal antibody to the murine M-CSF receptor (c-fms) did not inhibit or displace cobblestone island formation by either clone of FDC-P1 on irradiated stromal cells indicating a mechanism of binding not involving the M-CSF receptor. However, anti-serum to the M-CSF receptor inhibited growth of one factor-independent subclone. In separate studies, a subclone of IL-3-dependent 32Dc13 cells, expressing the transfected murine c-fms protooncogene but not the parent 32Dc13 cell line or another subclone expressing the transfected gene for the human M-CSF receptor, showed adherence and became factor independent when cocultivated with irradiated D2XRII stromal cells. Thus, irradiated stromal cells bind M-CSF receptor-positive hematopoietic progenitor cells and induce c-fms-dependent factor-independent tumorigenic subclones. The cellular interactions in this model may be relevant to gamma-irradiation leukemogenesis in vivo.

Animals

Mouse platelet-derived growth factor receptor alpha gene is deleted in W19H and patch mutations on chromosome 5.

The mouse W19H mutation is an x-ray-induced deletion of more than 2 centimorgans on chromosome 5 encompassing the white spotting mutation W (encoded by the Kit protooncogene), patch (Ph), and recessive lethal (l) loci. The platelet-derived growth factor receptor alpha gene (PDGFRA) like Kit encodes a transmembrane receptor tyrosine kinase. By using mouse-Chinese hamster somatic cell hybrids and haplotype analysis in interspecific backcross mice, mouse Pdgfra was mapped to chromosome 5 in tight linkage with Kit. Hybridization of a PDGFRA probe to DNAs from W19H/ + heterozygous mice and patch heterozygous mice, and their wild-type littermates, demonstrated deletion of Pdgfra. Pulsed-field gel electrophoresis indicated that Kit and Pdgfra are linked on a 630-kilobase Mlu I DNA fragment. Thus the W19H deletion removes at least two receptor tyrosine kinases and the results suggest Pdgfra as a candidate for the Ph locus.

Animals

The kinase insert domain of colony stimulating factor-1 receptor is dispensable for CSF-1 induced phosphatidylcholine hydrolysis.

Mouse NIH 3T3 fibroblasts transfected with human colony stimulating factor-1 receptor produced diacylglycerol in response to CSF1 and this correlated with elevated phosphatidylcholine hydrolyzing activity measured in an in vitro assay. Treatment of cells with the isoflavone derivative genistein attenuated PC hydrolysis in vitro suggesting a role for CSF1R tyrosine kinase activity. A CSF1R mutant lacking 67 amino acids of the kinase insert domain, which may affect the association of receptor with certain substrates, stimulated PC hydrolysis in response to CSF1. Coupling to PC hydrolysis is likely a general property of CSF1R and the kinase insert domain is dispensable for this activity.

Animals

A mutational analysis of phosphatidylinositol-3-kinase activation by human colony-stimulating factor-1 receptor.

Colony-stimulating factor-1 (CSF1) is a cell lineage-specific hemopoietin required for the growth, differentiation, and survival of macrophages and their precursors. The human CSF1 receptor (CSF1R) is a 150-kDa transmembrane glycoprotein whose cytoplasmic tyrosine kinase domain is split by a kinase insert (KI) region of approximately 70 amino acids. We tested the ability of CSF1R KI domain deletion mutants to stimulate phosphatidylinositol-3-kinase (PtdIns-3-kinase), an enzyme whose activity is augmented by tyrosine kinase oncogenes and receptor tyrosine kinases, and to support mitogenesis in transfected cells. Receptor immunoprecipitates from CSF1-stimulated cells contained greater than 5-fold more PtdIns-3-kinase activity compared to nonstimulated cells. High performance liquid chromatography analysis of the PtdIns-3-kinase product scraped from thin layer chromatography plates indicated that PtdIns-3-P was produced. CSF1R KI domain deletion mutants retained tyrosine kinase activity in vitro. Receptor immunoprecipitates of two partially overlapping 28 and 30 amino acid KI deletion mutants of CSF1R retained some PtdIns-3-kinase activity, in contrast to immunoprecipitates of CSF1R lacking 67 amino acids of the KI domain. Each deletion mutant stimulated CSF1-dependent DNA synthesis in transfected cells at much reduced levels compared to wild-type receptor expressing cells. These data suggest a role for the CSF1R KI domain in PtdIns-3-kinase association and for CSF1-induced thymidine incorporation into DNA.

1-Phosphatidylinositol 4-Kinase

Quantification of aminofluorene adduct formation and repair in defined DNA sequences in mammalian cells using the UVRABC nuclease.

Using the UVRABC nuclease as a reagent coupled with DNA restriction and hybridization analysis we have developed a method to quantify N-acetoxy-2-acetylaminofluorene (NAAAF)-induced DNA damage in the coding and noncoding sequences of the dihydrofolate reductase (DHFR) gene in Chinese hamster ovary (CHO) cells. High performance liquid chromatography analysis shows that the only DNA adduct formed in NAAAF-treated CHO cells is N-(deoxyguanosine-C8-yl)-2-aminofluorene (dG-C8-AF). DNA sequencing analysis demonstrates that the UVRABC nuclease incises at all potential sites in which dG-C8-AF adduct may form in linear DNA fragments. We have found that the formation and removal of dG-C8-AF adducts in the coding and 3' downstream noncoding sequences of the DHFR domain are similar in cells treated with 10 microM NAAAF (3.1 adducts/14 kilobases); DNA adduct removal attains 70% for both sequences within 24 h. This result contrasts with that obtained for the repair of cyclobutane dipyrimidines in the DHFR gene, in which the repair efficiency is much higher in the coding region than in the 3' downstream noncoding region. Our results suggest that in CHO cells the repair pathway for aminofluorene DNA adducts is not the same as that for cyclobutane dipyrimidines. This new technique has the potential to detect a variety of chemical carcinogen induced DNA adducts at the gene level in cultured cells and in DNA isolated from animal tissues.

2-Acetylaminofluorene

Carbonyl sulfide: an alternate substrate for but not an activator of ribulose-1,5-bisphosphate carboxylase.

Carbonyl sulfide, a competitive inhibitor of ribulose-bisphosphate carboxylase with respect to CO2 (Laing, W. A., and Christeller, J. T. (1980) Arch. Biochem. Biophys. 202, 592-600), is an alternate substrate. Thiocarboxylation was monitored by mass spectrometry as the stoichiometric consumption of carbonyl sulfide. The product, 1-thio-3-phosphoglycerate, was identified by 13C NMR and UV absorption spectroscopy and measured by enzymic conversion to thiolactate, coupled to the oxidation of NADH. The expected stoichiometry of thiocarboxylation was confirmed. The maximal rates of thiocarboxylation for the spinach and Rhodospirillum rubrum enzymes were close to the maximal rates of carboxylation for these two enzymes. Both enzymes favored CO2 over carbonyl sulfide (with Mg2+ as metal ion) by a factor of about 110. Thiocarboxylation could only be demonstrated with enzymes carbamylated with CO2. Incubation of the carbamylated E.ACO2.Mg complex with excess carbonyl sulfide caused the displacement of the activator carbamate. The thiocarbamylated enzyme was catalytically incompetent and did not form a stable quaternary complex with 2'-carboxyarabinitol bisphosphate. Incubation of the thiocarbamylated enzyme with excess CO2 resulted in the displacement of the thiocarbamate, the re-formation of the carbamylated E.ACO2.Mg complex and the restoration of catalytic competence. Computergraphic simulation of the thiocarbamylated quaternary complex indicated unfavorable van der Waals interactions associated with the thiocarbamate.

Binding Sites

Isolation of a novel receptor cDNA establishes the existence of two PDGF receptor genes.

A genomic sequence and cloned complementary DNA has been identified for a novel receptor-like gene of the PDGF receptor/CSF1 receptor subfamily (platelet-derived growth factor receptor/colony-stimulating factor type 1 receptor). The gene recognized a 6.4-kilobase transcript that was coexpressed in normal human tissues with the 5.3-kilobase PDGF receptor messenger RNA. Introduction of complementary DNA of the novel gene into COS-1 cells led to expression of proteins that were specifically detected with antiserum directed against a predicted peptide. When the new gene was transfected into COS-1 cells, a characteristic pattern of binding of the PDGF isoforms was observed, which was different from the pattern observed with the known PDGF receptor. Tyrosine phosphorylation of the receptor in response to the PDGF isoforms was also different from the known receptor. The new PDGF receptor gene was localized to chromosome 4q11-4q12. The existence of genes encoding two PDGF receptors that interact in a distinct manner with three different PDGF isoforms likely confers considerable regulatory flexibility in the functional responses to PDGF.

Amino Acid Sequence

A cyanobacterial mutant requiring the expression of ribulose bisphosphate carboxylase from a photosynthetic anaerobe.

Ribulose bisphosphate carboxylase is essential for both photoautotrophic and photoheterotrophic growth of the cyanobacterium Synechocystis 6803. However, a mutant lacking cyanobacterial carboxylase could be obtained by replacing the natural carboxylase gene with the corresponding gene from Rhodospirillum rubrum, a photosynthetic anaerobe. This treatment produced an organism whose growth depended on the activity of the structurally and functionally dissimilar foreign carboxylase. As a further consequence of this mutagenic replacement, the mutant also lacked microscopically observable carboxysomes, the subcellular inclusion bodies in which the wild-type carboxylase naturally resides. The mutant, dependent on a carboxylase with an inferior relative specificity for CO2 versus O2 and apparently lacking carboxysomes, is extremely sensitive to the CO2/O2 ratio supplied during growth and is unable to grow at all in air. This response to the gas composition should prove useful for selection of various R. rubrum carboxylase mutants with altered specificities for CO2 and O2.

Anaerobiosis

Patterns of Doppler recordings and its relationship to varicocele in infertile men.

Incompetence of the testicular vein appears to be the basic pathology of testicular dysfunction in varicocele. Doppler recording is a very sensitive method for detecting this reflux even when varicocele is not evident clinically. One hundred and seventy-eight men with infertility were studied. The presence of reflux in the pampiniform plexus as demonstrated by Doppler recording was compared with clinical varicocele. Reflux patterns were recorded on graph paper and various grades of reflux were observed. The three grades of reflux identified varied between a momentary reflux during vigorous Valsalva manoeuvre to significant reflux on minimal increase in intra-abdominal pressure brought about by normal respiration and deep breathing. Ninety-four per cent of the patients with clinical signs of varicocele had refluxes of grade 2 and 3 on Doppler study. Forty per cent of the patients without clinical evidence of varicocele were found to have reflux of grade 1 and 2 in the testicular veins.

Humans

Coronary risk factor outcomes following coronary artery bypass surgery.

The coronary risk factor status of patients prior to and following coronary artery bypass surgery (CABG) has been poorly investigated. Two consecutive series of CABG patients were surveyed following CABG. One hundred and thirty patients were assessed immediately following CABG and 530 patients were assessed 12-30 months following CABG. For the long-term post-CABG group, over 80% of those who had ever smoked had ceased. Sixty-four per cent of these males and 50% of females were classified as being overweight. Twenty-five per cent of males and 34% of females reportedly had high serum cholesterol (i.e. greater than or equal to 6.5 mmol/L). Comparing these CABG data with age-adjusted National Heart Foundation Risk Factor Prevalence Survey data, there was a higher prevalence of ex-smokers, overweight, hypertension, and elevated cholesterol. It was concluded that on most coronary risk factors, except for smoking, these CABG patients had a worse profile than the general Australian community. This problem warrants further research and the development of appropriate treatment programs.

Adult