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Biomedical subjects

D R Cohen

Publications and source records attributed to D R Cohen.

53 records · Page 3Linked to original sources

A zinc finger-encoding gene coregulated with c-fos during growth and differentiation, and after cellular depolarization.

Egr-1 is an early growth response gene that displays fos-like induction kinetics in fibroblasts, epithelial cells, and lymphocytes following mitogenic stimulation. Sequence analysis of murine Egr-1 cDNA predicts a protein with three DNA binding zinc fingers. The human EGR1 gene maps to chromosome 5 (bands 5q23-31). Egr-1 mRNA increases dramatically during cardiac and neural cell differentiation, and following membrane depolarization both in vitro and in vivo. Thus, Egr-1 and c-fos are often coregulated with strikingly similar kinetics. These results, in conjunction with the Egr-1 primary structure, suggest that Egr-1 may function as a transcriptional regulator in diverse biological processes.

Amino Acid Sequence↗

fra-1: a serum-inducible, cellular immediate-early gene that encodes a fos-related antigen.

A set of proteins antigenically related to the c-fos protein (Fos) are induced by serum in fibroblasts. To isolate cDNA clones of genes encoding such proteins, a lambda gt11 expression cDNA library constructed from serum-stimulated rat fibroblasts was screened with antibodies raised against a hydrophilic region (amino acids 127 to 152) of Fos. One of the positive clones identified, termed fra-1 (Fos-related antigen) was characterized. It encoded a protein that shared several regions of extensive amino acid homology with Fos (including the region that showed similarity to both the yeast GCN4 regulatory protein and the protein encoded by the jun oncogene), although its nucleotide sequence was considerably diverged from that of the c-fos gene. Only a subset of the agents and conditions that activated c-fos also induced fra-1. Induction of fra-1 expression following serum stimulation was delayed compared with that of c-fos. However, like c-fos, fra-1 was induced rapidly by serum in the presence of protein synthesis inhibitors. Thus, a family of Fos-related, inducible genes are involved in the cellular immediate-early transcriptional response to extracellular stimuli.

Amino Acid Sequence↗

Mapping patterns of c-fos expression in the central nervous system after seizure.

A dramatic and specific induction of c-fos was observed in identifiable neuronal populations in vivo after administration of the convulsant Metrazole. This effect was time- and dose-dependent and was abolished by prior treatment with the anticonvulsant drugs diazepam or pentobarbital. About 60 minutes after administration of Metrazole, c-fos messenger RNA reached a maximum and declined to basal levels after 180 minutes. A further decrease below that in normal brain was observed before a return to basal levels after 16 hours. While Metrazole still elicited seizures during this period, reinduction of c-fos was largely refractory. At 90 minutes, c-fos protein was observed in the nuclei of neurons in the dentate gyrus, and in the pyriform and cingulate cortices. Subsequently, c-fos protein appeared throughout the cortex, hippocampus, and limbic system. Thus, seizure activity results in increased c-fos gene expression in particular subsets of neurons.

Animals↗

Analysis of Fos protein complexes and Fos-related antigens by high-resolution two-dimensional gel electrophoresis.

Protein complexes containing the c-fos protein (Fos) and (Fos)-related antigens were isolated from serum-stimulated fibroblasts and from nerve growth factor plus benzodiazepine-treated pheochromocytoma (PC12) cells, and investigated by high-resolution two-dimensional gel electrophoresis. The results show that Fos is complexed with a basic 39-kDa protein (p39) in fibroblasts, and primarily with an acidic 40-kDa protein (p40) in PC12 cells. Whole cells lysates from both cell types contain p40, suggesting that the interaction of Fos and other cellular proteins is dependent on the differentiated state of the cell. In addition to p39 and p40, a heterogeneous population of polypeptides of approximately 48 kDa are present in Fos complexes isolated from non-denatured extracts of either cell type. These proteins may represent a minor class of Fos-binding proteins. Analysis of extracts prepared under denaturing conditions using antisera raised against a Fos peptide (amino acids 127-152) reveals a series of Fos-related antigens. These antigens are induced, some with a slower kinetics than Fos, in fibroblasts and PC12 cells. Thus, c-fos may represent a marker for a family of genes, some of which are antigenically related, that are part of an early cellular transcriptional response to diverse extracellular stimuli.

Animals↗

Cloning and expression of the rat interleukin-3 gene.

Genomic clones carrying the rat interleukin-3 (IL-3) gene have been isolated and the nucleotide sequence of the gene determined. Alignment of this sequence with that of the mouse IL-3 gene has allowed the structure of the rat IL-3 gene to be deduced. The intron-exon boundaries are conserved and extensive nucleotide homology (approx 90%) is present in the 5' flanking region and the portion of the gene coding for the signal peptide. Several proposed regulatory sequences are conserved and an analogous element to the tandem repeat in intron 2 of the mouse gene is also present. The predicted amino acid sequence for mature rat IL-3 shows surprisingly low homology (54%) with its murine counterpart, although all four cysteine residues are conserved. The rat IL-3 gene was expressed in monkey COS-1 cells and colony assays established that rat IL-3 is a multi-lineage haemopoietic growth regulator. There was little cross-reactivity of the respective IL-3 species on mouse and rat bone marrow cells suggesting that rat IL-3, in concert with its receptor, has evolved significantly away from the mouse IL-3/receptor system.

Amino Acid Sequence↗

pIN32: a cointegrate plasmid with IncHI2 and IncFII components.

An Enterobacter cloacae strain isolated from the faeces of a child with diarrhoea in Indonesia contained a transferable 216 MDa plasmid, pIN32, exhibiting IncHI2 phenotypic characters, including temperature sensitivity of transfer and the expression of H serotype pili at a repressed level. A derivative plasmid (pIN32-1), which had lost the IncHI2 phenotype, and contained only 60 MDa of the original replicon, was obtained after mating at 37 degrees C. It was IncFII, showed regions of homology with plasmid R100, determined IncFII serotype conjugative pili constitutively and was transfer-derepressed. After overnight growth at 37 degrees C in non-selective medium, pIN32 gave rise to another derivative, pIN32-2 (size 184.3 MDa), which retained the IncHI2 phenotype and several other pIN32 characters.

Child↗

Regulation of the production of granulocyte-macrophage colony-stimulating factor by macrophage-like tumour cell lines.

Macrophage tumour cell lines (PU5-1.8, P388D1) produced detectable granulocyte macrophage colony-stimulating (CSF-2) activity measured using a factor-dependent cell line FDC-P1. The production of CSF-2 was enhanced by endotoxin and inhibited by serum, and correlated inversely with [3H]TdR incorporation. mRNA isolated from PU5-1.8 or P388D1 cells initiated CSF-2 production when injected into Xenopus laevis oocytes. The specific activity in this assay was unaltered in mRNA isolated from endotoxin-treated cells. The results suggest that endotoxin acts at a post-transcriptional level.

Animals↗

Utility model of preventive behaviour.

The utility model of preventive behaviour is presented as an alternative to psychosociological analysis. All preventive actions are viewed as the consumption of those "goods" that affect the risk of illness or injury. Levels of consumption depend on, inter alia, utility (perceived benefit) and cost. Total utility yielded by risk affecting goods is the sum of that derived from the use value of the good, if any, and that from reduced anxiety which results from reduced risk. Many risk affecting goods are consumed mainly for their use value. An examination of the nature of risk affecting goods is presented, followed by an analysis of the factors that determine the anxiety associated with unwanted outcomes. The utility model places preventive behaviour within the framework of the economic theory of demand. The economic principles used in this analysis are explained and their relevance to the consumption of risk affecting goods is determined.

Economics↗

Co-ordinating prevention.

A major reason why the balance between pronouncements on prevention and action is tipped so much towards the former is that, in most countries, responsibility for prevention is fragmented and vaguely defined. If an effective and efficient national prevention policy is to come about, then it is important that overall responsibility be placed in the hands of a single agency, and for that agency to recognise that priorities in prevention must be a function of costs as well as benefits. The first task of such an agency will be to sort out the fundamental objectives of public policy by clarifying what is meant by prevention and specifying the rationale for government intervention and hence the sorts of prevention which should be the responsibility of the public sector. Given this, national expenditure on prevention from any source which contributes to government prevention strategy can be identified and expressed in the form of a programme budget showing the proportions of total prevention expenditure going to each of the defined programme areas (environment, occupational health and safety, screening, health promotion, etc.). This framework gives a broad overview of the existing situation and by forcing consideration of benefit valuation in any decision to alter the balance of expenditure between programmes, ought to lead to a more rational prevention strategy than would result from a focus on individual programmes in isolation. Moreover, through an emphasis on costs and benefits, the overseers of prevention policy will be better placed to consider the relative efficiency of existing policies--since all share the common objective of reducing future morbidity and mortality. Finally, consideration will also need to be given to issues of equity. The approach described, illustrated by the case of the United Kingdom, will, if adopted, increase the likelihood that a rational and sensible national prevention policy will emerge.

Budgets↗

A cost benefit study of milk-borne salmonellosis.

An estimate of the benefits which would result from a ban on the sale of non-pasteurized milk in Scotland has been assessed by costing a recent outbreak of milk-borne salmonellosis in the Grampian Region. The cost of such a ban would not exceed the benefits under any but the most severe assumptions about the values attached to intangible benefits.

Animals↗

The Howie code: is the price of safety too high?

The code of practice for the prevention of infection in clinical laboratories and postmortem rooms (the "Howie code") was produced in order to standardise laboratory safety procedures at a level which would minimise risks to laboratory workers. The costs of implementing this code, which were not given proper consideration when the code was drawn up, are seen to be very high. This paper questions whether the benefits of risk reduction resulting from the code can justify the high cost of its implementation. Beginning with an examination of the incidence in recent years of laboratory-acquired infections, the paper looks at the perceptions of risk by laboratory workers and the feasibility of low cost alternatives to those of the Howie code. The potential benefits of the code are then viewed against the cost of upgrading the laboratories of two Scottish hospitals to Howie standards. The fact of high costs relative to benefits reinforces the paper's call for the proper consideration of economic issues to be given at the development rather than the implementation stage of any regulatory codes.

Cost-Benefit Analysis↗

Specification of surface mating systems among conjugative drug resistance plasmids in Escherichia coli K-12.

Representative plasmids for most incompatibility groups in Escherichia coli K-12 were transferred to a "bald" strain to compare transfer frequencies for liquid and solid media. Standard broth matings were used for a liquid environment, but for solid surface mating, conjugation was allowed to take place on nutrient plates before washing off the cells for transconjugant selection on plates containing appropriate drugs. Plasmids that determine rigid pili transferred at least 2,000x better on plates than in broth. Some plasmids that determine thick flexible pili transferred 45 to 470x better, whereas others transferred equally well in both environments, as did plasmids of the I complex, which determine thin flexible pili. These results clearly distinguished a number of surface mating systems where most plasmids were derepressed for transfer and determined conjugative pili constitutively. The temperature-independent IncH2 plasmid R831b transferred best on plates, but other IncH plasmids transferred equally well in broth. This inconsistency led to the reclassification of R831b as IncM.

Conjugation, Genetic↗

Basic characterization of W-pili.

The bacterial drug-resistance plasmids Sa, R388 and R7K that comprise the W compatability group were transferred by conjugation to species in the genera Escherichia, Salmonella, Shigella and Pseudomonas, chosen for their lack of common pili. On receiving the W plasmids, all strains produced pili, which were similar morphologically, at average frequencies of up to 3-0 pila/cell. The pili determined by the W plasmids were also related serologically, but unrelated to those of other plasmids. R- segregant strains which had lost their plasmids showed a simultaneous loss of both pili and drug-resistance characteristics. W-pili are pointed flexible filaments 10 to 12 nm thick, with an average length of 450 nm. The best host found was Salmonella typhimurium.

Bacteria↗

Molecular cloning of cDNA for murine interleukin-3.

The cDNA sequence for murine interleukin-3, one of the colony stimulating factors that regulate haematopoiesis, codes for a polypeptide of 166 amino acids including a putative signal peptide. The predicted amino acid sequence indicates that formation of mature interleukin-3 involves proteolytic removal of not only the signal peptide but additional amino-terminal amino acids.

Animals↗

Evaluation of clitoral blood flow by color Doppler ultrasonography.

The feasibility of reliably measuring clitoral blood flow using standard color Doppler ultrasonography was evaluated by two independent assessors in a sample of 40 pre- and post-menopausal women. High positive correlations with no significant mean differences between examiners were found for three major standard blood flow measures including maximum velocity, resistance, and pulsatility indices. With further methodological refinements, ultrasonographic evaluation of clitoral blood flow will be adaptable for clinical and research use and is likely to become the standard physiological measure of female sexual arousal.

Adult↗