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Biomedical subjects

J R Smith

Publications and source records attributed to J R Smith.

At least 253 records · Page 14Linked to original sources
PubMed1991

Senescent cells fail to express cdc2, cycA, and cycB in response to mitogen stimulation.

Senescent human diploid fibroblasts (HDF) contain no detectable cdc2 mRNA or p34cdc2 protein. Similarly, young quiescent HDF have only low levels of cdc2 mRNA and protein. After serum stimulation, quiescent HDF accumulate increasing amounts of cdc2 mRNA and protein and go through DNA synthesis and mitosis. In contrast, serum-stimulated senescent HDF fail to accumulate detectable amounts of cdc2 mRNA and protein and fail to enter S phase. Mitosis is likewise deficient in senescent cells even when they have been induced to synthesize DNA by simian virus 40 large tumor antigen. Since p34cdc2 or its homologues appear to be required for DNA synthesis and mitosis in eukaryotes, a lack of these molecules in serum-stimulated senescent HDF could be an important reason for their inability to enter S phase or mitosis. Nuclear microinjection of cdc2 DNA into senescent HDF causes rounding up of the cells but no induction of DNA synthesis. Since cyclins A and B are important cofactors of the protein kinase activity of p34cdc2 or its homologues, we analyzed expression of these genes in serum-stimulated senescent HDF and determined that they contain little or no cycA or cycB mRNA. These deficiencies may be relevant to the lack of DNA synthesis and mitosis in senescent HDF.

Amino Acid Sequence↗
PubMed1991

Inhibition of bovine brain calmodulin-dependent cGMP phosphodiesterase by peptide and non-peptide angiotensin receptor ligands.

Inhibition of a purified 60 KDa bovine brain calmodulin-dependent cGMP phosphodiesterase (PDE) was investigated for a number of peptides and non-peptides which are known to bind to angiotensin (ANG) receptors. The peptide antagonists sarilesin and sarmesin had KI = 120 and greater than 200 microM respectively, and the peptide agonists ANG II and ANG III had KI = greater than 200 and 45 microM respectively. Non-peptide ANG receptor antagonists related to DuP 753 exhibited KI values in the same range. For both peptide and non-peptide antagonists, inhibitory activities in the PDE assay reflected the order of antagonist potencies at ANG receptors in the rat isolated uterus assay and binding affinities at ANG receptors in rat uterine membranes, suggesting that molecular recognition factors are similar for both ANG receptors and cGMP PDE. The vasodilatory and blood pressure lowering effects of compounds related to DuP 753 may be due in part to inhibition of cGMP PDE. The differential effects of ANG II and ANG III at target tissues may relate in part to the marked differences in cGMP PDE inhibition associated with these two peptides hormones.

1-Sarcosine-8-Isoleucine Angiotensin II↗
PubMed1991

Genetic analysis of indefinite division in human cells: evidence for a cell senescence-related gene(s) on human chromosome 4.

Earlier studies had demonstrated that fusion of normal with immortal human cells yielded hybrids having limited division potential. This indicated that the phenotype of limited proliferation (cellular senescence) is dominant and that immortal cells result from recessive changes in normal growth-regulatory genes. In additional studies, we exploited the fact that the immortal phenotype is recessive and, by fusing various immortal human cell lines with each other, identified four complementation groups for indefinite division. Assignment of cell lines to specific groups allowed us to take a focused approach to identify the chromosomes and genes involved in growth regulation that have been modified in immortal cells. We report here that introduction of a normal human chromosome 4 into three immortal cell lines (HeLa, J82, T98G) assigned to complementation group B resulted in loss of proliferation and reversal of the immortal phenotype. No effect on the proliferation potential of cell lines representative of the other complementation groups was observed. This result suggests that a gene(s) involved in cellular senescence and normal growth regulation resides on chromosome 4.

Base Sequence↗
PubMed1991

Expression of a foreign gene in cats reconstituted with retroviral vector infected autologous bone marrow.

A Moloney murine leukemia virus based retroviral vector was used to transfer the bacterial neomycin resistance gene (neoR) into feline hematopoietic cells. We reconstituted four cats that had been lethally irradiated with autologous bone marrow that had been infected with the N2 or SAX retroviral vector. Bone marrow cells from all four cats expressed the neoR gene 30 days posttransplant and three of four cats still had the neoR gene and a low level of drug resistant colony-forming unit granulocyte-macrophage after more than 200 days. Two of the four cats unexpectedly developed diabetes mellitus 90 days posttransplantation. The expression of a foreign gene in cats, albeit at a low level, demonstrates that retroviral vectors can be used for gene transfer in noninbred large animal species and may be useful for gene therapy of humans. The development of diabetes mellitus in two of the subjects emphasizes the value of animal models for the study of possible deleterious effects of retroviral vector-mediated gene transfer.

Animals↗
PubMed1991

Prevalence of Chlamydia trachomatis infection in women having cervical smear tests.

OBJECTIVE: To determine the prevalence of sexually transmitted diseases in patients with normal and abnormal cervical smears. DESIGN: A prospective study of asymptomatic women with normal cervical smears attending their general practitioner and newly referred patients with abnormal smears attending a colposcopy clinic. SETTING: A hospital based colposcopy clinic and an urban general practice (list size 5500) in north west Glasgow. SUBJECTS: 197 asymptomatic women attending their general practitioner for cervical smear tests and 101 randomly selected patients attending the colposcopy clinic for investigation of abnormal smears. MAIN OUTCOME MEASURES: Presence of various sexually transmitted infections as determined by culture and serological tests. RESULTS: Of the 101 women with cytological abnormalities, six had current chlamydial infection proved by culture and none had gonococcal infection; of the 197 women with normal smears, 24 (12%) had a chlamydial infection and two had gonorrhoea. Serological studies for Chlamydia trachomatis specific antibody also indicated that a large proportion of patients had been exposed to this agent in both groups. There was no significant difference between the groups in the prevalence of any sexually transmitted disease studied. CONCLUSION: A high prevalence of chlamydial infection is present in women in north west Glasgow irrespective of their cervical cytological state.

Adult↗
PubMed1991

Detection of metabolites of toxic alkylmethylphosphonates in biological samples.

The major metabolites and breakdown products of some toxic organophosphonates are their respective alkymethylphosphonic acids. These acids ionize at physiological pH and are not amenable to gas chromatographic analysis in their underivatized forms. Their detection in biological samples has been difficult because of their presence at only trace levels. Existing analytical methods were developed mainly for measuring these phosphonic acids in environmental samples and at higher concentrations. In this study, we devised a gas chromatographic/mass spectrometric method to provide confirmation and quantification of the organophosphonic acids of soman (GD), sarin (GB) and GF in blood and urine. This report describes the various derivatization conditions that we have studied and demonstrates the characteristic mass spectra by different ionization techniques.

Animals↗
PubMed1991

Computerized seizure detection of complex partial seizures.

In this study, we describe a computerized method that uses 3 quantified EEG features and discriminant analysis to automatically detect seizure EEG. The quantified EEG features were relative amplitude, dominant frequency and rhythmicity. Using EEGs recorded from intracranial electrodes, the seizure detection method was applied to consecutive non-overlapping 2-channel EEG epochs. A seizure detection sensitivity, ranging from 90% to 100%, was associated with a false positive detection rate of 1.5-2.5/h. The performance of the seizure detection method remained stable for EEG recorded over variable time periods.

Discriminant Analysis↗
PubMed1991

Correlation between complementation group for immortality and DNA synthesis inhibitors.

Previous studies had demonstrated that a DNA synthesis inhibitor(s) was produced by senescent but not young human diploid fibroblasts (HDF). Analysis of immortal human cell lines led to the finding that SUSM-1, carcinogen-treated immortal human liver fibroblast cells, expressed a potent inhibitor of DNA synthesis that was active in proliferation-competent young HDF but did not affect the SUSM-1 cell line itself. To determine whether one mechanism of escape from senescence to the immortal phenotype involved the loss of response to such DNA synthesis inhibitors, we initiated the present study analyzing a larger number of immortal human cell lines representative of the four complementation groups for indefinite division identified to date. We have found a correlation between the assignment of a cell line to Complementation Group D and the production of DNA synthesis inhibitors coupled with inability to respond to the inhibitory factors. We have also observed a correlation between the ability of immortal cell lines to respond to such DNA synthesis inhibitory factors and assignment to Complementation Group B. These data suggest DNA synthesis inhibitors are involved in the limited lifespan of normal cells and that the immortalization process may involve alterations in the activity of or response to such inhibitors.

Biological Factors↗
PubMed1991

Fibronectin expression increases during in vitro cellular senescence: correlation with increased cell area.

Several changes in the functional characteristics of fibronectin have been noted as cells become senescent in culture. In this report we show that steady state levels of both fibronectin mRNA and protein increase significantly during the process of cellular aging. The greatest change in the proportion of cells expressing high levels of fibronectin occurs near the end of a culture's proliferative potential. The proportion of cells unable to synthesize DNA has previously been shown to follow a similar pattern. We also found that increasing cell size correlates closely with higher levels of fibronectin expression. Thus, there is a clear correlation between increased fibronectin mRNA content and in vitro cellular senescence. It remains to be determined whether the change in fibronectin production is a contributing cause or a result of in vitro cellular senescence.

Blotting, Northern↗
PubMed1991

Hippocampal contribution to verbal recent memory following dominant-hemisphere temporal lobectomy.

The effects of hippocampal encroachment in the language dominant-hemisphere were studied in 41 patients who underwent previous temporal lobectomy (TL). Patients undergoing dominant-hemisphere TL including anterior hippocampus (n = 13) performed significantly worse than nondominant TL patients (n = 16) on a verbal learning test (Selective Reminding; p less than or equal to .00001), thereby confirming the sensitivity of this procedure to lateralized temporal-lobe dysfunction. However, no significant difference was present on this or other primary measures of material-specific memory when contrasting dominant TL patients in whom the anterior hippocampus was spared (n = 12) to those in whom anterior hippocampus was resected. These data suggest that more extensive and posterior mesial temporal-lobe resection is not necessarily associated with a greater verbal material-specific memory deficit following dominant-hemisphere temporal lobectomy.

Adult↗
PubMed1991

Prohibitin, an evolutionarily conserved intracellular protein that blocks DNA synthesis in normal fibroblasts and HeLa cells.

Genes that act inside the cell to negatively regulate proliferation are of great interest because of their implications for such processes as development and cancer, but these genes have been difficult to clone. This report details the cloning and analysis of cDNA for prohibitin, a novel mammalian antiproliferative protein. Microinjection of synthetic prohibitin mRNA blocks entry into S phase in both normal fibroblasts and HeLa cells. Microinjection of an antisense oligonucleotide stimulates entry into S phase. By sequence comparison, the prohibitin gene appears to be the mammalian analog of Cc, a Drosophila gene that is vital for normal development.

Amino Acid Sequence↗
PubMed1991

Behavioral state-specific changes in human hippocampal theta activity.

Although there has been extensive examination of the behavioral and physiologic correlates of hippocampal theta activity in animals, the human literature consists of a single case study. We investigated the differential effects of four behavioral states on human hippocampal theta activity in 16 epilepsy surgery patients. Behavioral conditions included resting eyes closed (RC), resting eyes open (RO), eyes open with auditory word activation (AW), and eyes open with visuospatial activation (VS). Hippocampal theta activity decreased during both RO and VS compared to both RC and AW. There were reciprocal changes in delta activity. Comparisons of RO to VS and of RC to AW were nonsignificant. The results demonstrate state-specific changes in human hippocampal theta and are consistent with the animal literature that relates hippocampal theta to sensorimotor integration and forebrain volitional mechanisms.

Adolescent↗
PubMed1991

Effects of perfluorodecalin and perfluorooctylbromide emulsions on peak contractile force and ATP levels in the guinea pig left atrium.

The isolated, electrically-driven, guinea pig left atrium was used to study the ability of two perfluorocarbon emulsions to prevent anaerobic hypofunction in the myocardium. A 20% perfluorodecalin (PFD) emulsion maintained peak tension at 80% of aerated levels for more than 20 minutes. Emulsions of perfluorooctylbromide (PFOB) ranging from 25 to 100% produced a similar result, except that the 25% emulsion could not maintain contractions for the entire time period. Peak tension of atria bathed in K-H solution decreased to less than 50% over the same time period. Both 20% PFD and 100% PFOB maintained myocardial ATP levels at pre-hypoxic levels for at least twenty minutes after aeration was terminated. Unaerated atria, bathed in Krebs-Henseleit solution only, exhibited a significant decline in tissue ATP levels at this time. It appears that perfluorocarbon emulsions may delay oxygen desaturation and thereby protect cardiac tissue from ATP depletion and impaired cardiac function associated with hypoxia. This tissue preparation was found to be very useful for determining the efficacy of potential oxygen carriers.

Adenosine Triphosphate↗