Search PubMed⌕ Search

Biomedical subjects

J R Smith

Publications and source records attributed to J R Smith.

At least 181 records · Page 10Linked to original sources
PubMed1995

Exit from G0 and entry into the cell cycle of cells expressing p21Sdi1 antisense RNA.

p21Sdi1 (also known as Cip1 and Waf1), an inhibitor of DNA synthesis cloned from senescent human fibroblasts, is an inhibitor of G1 cyclin-dependent kinases (Cdks) in vitro and is transcriptionally regulated by wild-type p53. In addition, p21Sdi1 has been found to inhibit DNA replication by direct interaction with proliferating cell nuclear antigen. In this study we analyzed normal human fibroblast cells arrested in G0 and determined that an excess of p21Sdi1 was present after immunodepletion of various cyclins and Cdks, in contrast to mitogen-stimulated cells in early S phase. Expression of antisense p21Sdi1 RNA in G0-arrested cells resulted in induction of DNA synthesis as well as entry into mitosis. These results suggest that p21Sdi1 functions in G0 and early G1 and that decreased expression of the gene is necessary for cell cycle progression.

Adenovirus E1A Proteins↗
PubMed1995

Identification of the active region of the DNA synthesis inhibitory gene p21Sdi1/CIP1/WAF1.

The cloning of the negative growth regulatory gene, p21Sdi1, has led to the convergence of the fields of cellular senescence, cell cycle regulation and tumor suppression. This gene was first cloned as an inhibitor of DNA synthesis that was overexpressed in terminally non-dividing senescent human fibroblasts (SD11) and later as a p53 transactivated gene (WAF1) and a Cdk-interacting protein (CIP1, p21) that inhibited cyclin-dependent kinase activity. To identify the active region(s) of p21Sdi1, cDNA constructs encoding various deleted forms of the protein were analyzed. Amino acids 22-71 were found to be the minimal region required for DNA synthesis inhibition. Amino acids 49-71 were involved in binding to Cdk2, and constructs deleted in this region expressed proteins that were unable to inhibit Cdk2 kinase activity in vitro. The latter stretch of amino acids shared sequence similarity with amino acids 60-76 of the p27Kip1 protein, another Cdk inhibitor. Point mutations made in p21Sdi1 in this region confirmed that amino acids common to both proteins were involved in DNA synthesis inhibition. Additionally, a chimeric protein, in which amino acids 49-65 of p21Sdi1 were substituted with amino acids 60-76 of p27Kip1, had almost the same DNA synthesis inhibitory activity as the wild-type protein. The results indicate that the region of sequence similarity between p21Sdi1 and p27Kip1 encodes an inhibitory motif characteristic of this family of Cdk inhibitors.

Amino Acid Sequence↗
PubMed1995

Intraoperative hippocampal cooling and Wada memory testing in the evaluation of amnesia risk following anterior temporal lobectomy.

OBJECTIVE: To determine the efficacy of memory testing following localized thermal inactivation (cooling) of the hippocampus during epilepsy surgery to assess risk for postoperative memory decline. DESIGN: Memory was assessed after intraoperative thermal inactivation of the hippocampus and during the Wada procedure to help determine the clinical utility of each procedure. SETTING: A university hospital, comprehensive epilepsy surgery program. PATIENTS: Individuals undergoing unilateral temporal lobectomy for relief of intractable seizures. PROCEDURE: After temporal tip resection, iced liquid was irrigated into the temporal horn of the lateral ventricle until the hippocampus was "cooled." MAIN OUTCOME MEASURES: Multiple neuropsychological measures of learning and memory. RESULTS: Wada memory testing suggested risk for memory impairment in 15 patients while cooling indicated risk in only five patients. Predictions arising from the two procedures were concordant in 13 patients and discordant in 12 patients. Among the 12 inconsistent predictions, hippocampal cooling suggested adequate contralateral memory support in 11 (92%) of 12 patients, and none of these 11 patients had a postoperative anterograde amnesia. CONCLUSION: Intraoperative hippocampal cooling may be useful in determining the risk of postoperative memory disorder among selected patients undergoing epilepsy surgery.

Adult↗
PubMed1995

Diminished responsiveness of senescent normal human fibroblasts to TNF-dependent proliferation and interleukin production is not due to its effect on the receptors or on the activation of a nuclear factor NF-kappa B.

The limited life span in culture of normal human diploid fibroblasts (HDF) has provided a model of cellular senescence. The short-term growth of these cells in culture is regulated by a number of different cytokines, including tumor necrosis factor (TNF), interleukin-1 (IL-1), and fibroblast growth factor (FGF). However, the effect of senescence on the responsiveness of HDF to these cytokines is not known. In the present report, we examined the effects of TNF on foreskin-derived HDF at different passage levels. We compared the response of HDF cells at population doubling (PD) 23 (young) with that of cells at PD 70 (senescent). Young cells proliferated in response to TNF in a dose-dependent manner. Under these conditions TNF had no effect on senescent HDF. The decrease in TNF responsiveness was found to be dependent on PD. The lack of response of senescent HDF was not unique to TNF, since FGF and IL-1 were also ineffective. In contrast to senescent HDF, TNF-dependent proliferation of young HDF could be further potentiated by IL-1 and FGF, suggesting an independent signaling mechanism. On exposure to TNF, senescent HDF produced IL-6 and IL-8, but to a much lower degree than that produced by young HDF. The diminished responsiveness of senescent HDF to TNF does not appear to be due to the difference in either receptor number or affinity, since senescent cells had two- to threefold higher number of TNF receptors than young HDF but the same affinity. TNF induced the activation of a nuclear transcriptional factor, NF-kappa B, equally in both young and senescent cells, which indicates the lack of a defect in the early events of TNF signal transduction in senescent fibroblasts. Overall, our results indicate that there is an age-dependent decline in TNF-induced proliferation and in the production of interleukins by fibroblasts; this unresponsiveness appears not to be due to TNF receptors or NF-kappa B activation. These results may have importance in understanding the diminished immune response, inflammation, and wound healing associated with aging.

Base Sequence↗
PubMed1995

Increased expression of p21Sdi1 in adrenocortical cells when they are placed in culture.

Expression of the cell cycle inhibitor p21Sdi1/WAF1/Cip1 was investigated in a differentiated cell type, the adrenocortical cell, at different stages of culture, from the preparation of cells from the adrenal gland to senescence after long-term growth. In bovine adrenocortical cells, expression of SDI1 was much higher in culture than in vivo. Elevation of SDI1 mRNA, accompanied by elevation of the level of a protein reacting with anti-p21Sdi1 antibodies, was observed as early as 3 h after the start of the tissue dissociation procedure used to prepare cells for culture. This level of expression was then maintained during plating and subsequent long-term growth in culture. Growth and quiescence in bovine adrenocortical cells can be modulated by inclusion or removal of FGF from the culture medium. In these cells SDI1 mRNA was not increased by long-term mitotic quiescence resulting from FGF deprivation. In cultured fetal human adrenocortical cells, SDI1 mRNA was also detected at all stages of the culture life span, including 2 days after isolation of cells from the adrenal gland and plating in culture. Mid-life-span cells had higher SDI1 mRNA than senescent human fibroblasts. Clones of human adrenocortical cells nearing senescence in culture had somewhat higher SDI1 mRNA than early passage cells. Thus, SDI1 expression in adrenocortical cells is not associated with mitotic quiescence either in vivo or in vitro, yet isolation of the cells and culturing them exerts a powerful inductive influence on its expression.

Adrenal Cortex↗
PubMed1995

Precision of dipole localization in a spherical volume conductor: a comparison of referential EEG, magnetoencephalography and scalp current density methods.

In this study, we determined the influence of dipole orientation, dipole location, and number of recording sites on the precision of dipole localization in a spherical volume conductor. We compared localization from referential EEG (R-EEG), scalp current density EEG (SCD-EEG) and magnetoencephalography (MEG). Dipole orientation had a small influence on the precision of dipole localization for R-EEG and SCD-EEG. Dipole location relative to the recording sites, dipole depth, and number of recording channels strongly influenced the precision of dipole localization. Assuming equal signal to noise conditions for each recording method, MEG and SCD-EEG had a similar precision for dipole localization of a single tangential dipole source and both methods were more precise than R-EEG. However, SCD-EEG was inferior to MEG for distinguishing a single tangential current source from a pair of deeper radial current sources. In summary, these results suggest that the MEG will be most useful for localization of multiple simultaneous dipole sources.

Brain↗
PubMed1995

Trisomy 22: prenatal diagnosis--a case report.

Multiple abnormalities of a fetus were detected on a routine antenatal anomaly scan at 19 weeks' gestation. Amniocentesis and karyotype analysis showed trisomy 22. The ultrasound and postmortem features are presented.

Abnormalities, Multiple↗
PubMed1995

Approach to genotyping errors caused by nontemplated nucleotide addition by Taq DNA polymerase.

Thermostable DNA polymerases can catalyze nontemplated addition of a nucleotide to the 3' end of amplification products. This presents a potential source of error in genotyping studies employing Taq DNA polymerase to amplify microsatellite loci. Although the activity is marker specific, experimental variation is often seen in the degree of modification. Consequently, for a given microsatellite marker, an allele may be inconsistently identified as either the unmodified or modified amplification product. Full automation of high-throughput genotyping has been hampered by the need for manual editing of data because of this source of allele misidentification. In this study we estimate a 1% to 3% error rate attributable to nontemplated nucleotide addition in the ABI PRISM genotyping system. We present a PCR-based strategy to minimize this source of error.

Alleles↗
PubMed1995

Intraoperative thermal inactivation of the hippocampus in an effort to prevent global amnesia after temporal lobectomy.

In an effort to assess the risk of amnesia after anterior temporal lobectomy (ATL), we conducted localized thermal inactivation (cooling) of the hippocampus with memory testing. Thirty-three ATL patients whose preoperative evaluation suggested risk for postoperative amnesia underwent hippocampal cooling. Cooling consisted of inserting a catheter in the temporal horn and irrigating it with an iced solution until a stable hippocampal temperature of approximately 20 degrees C was reached. Memory was assessed before and after cooling. In 12 of the 33 patients, memory testing was either aborted or suggested poor contralateral support, and the hippocampus was resected in 2 of these patients. The remaining 21 patients showed evidence of contralateral memory support, and the hippocampus was resected in 18. No patient became amnestic. These results suggest that intraoperative hippocampal cooling may be useful in selected cases. However, even among many patients who could cooperate with testing, discomfort, sedation, attentional deficits, confusion, and anxiety made test interpretation difficult.

Adolescent↗
PubMed1995

Multichannel magnetoencephalography in ablative seizure surgery outside the anteromesial temporal lobe.

Magnetoencephalography (MEG) was used to evaluate 40 candidates for seizure surgery thought to have foci outside the anteromesial temporal lobe. Of 29 cases with electrographic data suggesting a convexity focus, MEG spikes were recorded from 28. In 21, MEG and electrographic data were localized to the same area. Invasive studies were, or could have been, avoided in 13 cases based on MEG and other noninvasive data. MEG was not localizing value in 4 orbitofrontal or 7-depth-nonlocalized cases. Seventeen patients with MEG epileptiform data have had postoperative follow-up. Eight of 13 with electrographic and MEG data localized to the same area are seizure free. None of 4 with spatial discordance of MEG electrographic data are seizure free.

Electroencephalography↗
PubMed1995

Entamoeba histolytica trophozoites induce an inflammatory cytokine response by cultured human cells through the paracrine action of cytolytically released interleukin-1 alpha.

Infection with the protozoan parasite Entamoeba histolytica results in a high mortality worldwide. To initiate infection, E. histolytica trophozoites in the bowel lumen penetrate the epithelium, and cause extensive lysis of host cells. The acute amebic lesions in animal models are characterized by infiltration with inflammatory cells, particularly neutrophils. The acute host response is likely important for determining whether the infection will spread systemically, but little is known regarding the signals which initiate an acute inflammatory response to E. histolytica. In the studies reported herein, we used an in vitro model system to define the proinflammatory signals produced by epithelial and other host cells in response to infection with E. histolytica trophozoites. Coculture of human epithelial and stromal cells and cell lines with trophozoites is shown to increase expression and secretion of an array of chemoattractant and proinflammatory cytokines, including IL-8, GRO alpha, GM-CSF, IL-1 alpha, and IL-6. Moreover, high-level secretion of those cytokines is regulated by the paracrine action of cytolytically released IL-1 alpha. A second mechanism for trophozoite-induced IL-8 production involves trophozoite-target cell contact via a galactose-inhibitable amebic adherence protein, and appears to be mediated through increased intracellular calcium levels. These studies define novel mechanisms through which acute inflammation can be initiated in the host in response to a cytolytic pathogen, such as E. histolytica.

Animals↗
PubMed1995

Wada memory asymmetries predict verbal memory decline after anterior temporal lobectomy.

We examined Wada memory and neuropsychological memory function in 34 nonlesional patients who underwent anterior temporal lobectomy (ATL) and who were seizure free at 1-year follow-up. Patients who displayed a decline on verbal memory measures that exceeded 1 SD after left ATL had significantly smaller left/right Wada memory asymmetries than left ATL patients without a significant verbal memory decline. When Wada memory asymmetries were used to predict verbal memory decline after left ATL in individual patients, similar statistically significant effects were present. No significant relationship between Wada memory and postoperative memory was present in right ATL patients, and postoperative memory function was not related to Wada memory performance after either left hemisphere or right hemisphere injection alone. We conclude that Wada memory asymmetries provide one measure of the risk to material-specific decline in verbal memory after left ATL.

Analysis of Variance↗
PubMed1995

Effects of a 100% perfluorooctylbromide emulsion on ischemia/reperfusion injury following cardioplegia.

Protective effects of a perfluorooctylbromide emulsion on myocardial ischemia and reperfusion (MI/R) injury were evaluated in a modified Langendorff rat heart preparation. Isolated rat hearts were equilibrated in Krebs-Henseleit solution (KH) for 35 minutes and perfused with either cardioplegic solution (CPS) or a 100% perfluorooctylbromide (PFOB) emulsion in CPS for 3 minutes. Hearts were then bathed in the emulsion or CPS. Both groups were subjected to 30 minutes of ischemia. Following 30 minutes of ischemia and 30 minutes of reperfusion with KH solution, hearts subjected to the 100% PFOB emulsion showed improved recovery of left ventricular function. Tissue activities of the antioxidant enzymes glutathione peroxidase, superoxide dismutase, and catalase were not affected by the emulsion in this model. Activity of lactate dehydrogenase (LDH) in the bathing medium was elevated at the end of the experimental period in both control and PFOB-treated hearts. The PFOB emulsion reduced the decline in ATP and GSH levels produced by cardioplegia and subsequent reperfusion. No differences were noted in oxidized glutathione (GSSG) levels. These data suggest that the PFOB emulsion provides some protection for the myocardium against injury associated with cardioplegia.

Adenosine Triphosphate↗
PubMed1995

Induction of senescent cell-derived inhibitor of DNA synthesis gene, SDI1, in hepatoblastoma (HepG2) cells arrested in the G2-phase of the cell cycle by 9-nitrocamptothecin.

BACKGROUND: Recent studies have demonstrated that the plant-derived alkaloid camptothecin (CPT) and its derivative, 9-nitro-CPT (9NC), are cytotoxic in tumorigenic cells but cytostatic in nontumorigenic cells in vitro and in vivo. Also, CPT induces differentiation of human leukemia cells in vitro along specific lineages. In this study, we have investigated the effects of 9NC on nontumorigenic HepG2 cells derived from human hepatoblastoma. A newly discovered senescent cell-derived inhibitor (SDI1) plays a critical role in the cell cycle, so we evaluated the effect of 9NC on the expression of the SDI1 gene. EXPERIMENTAL DESIGN: The effects of 9NC on HepG2 cells were evaluated by monitoring DNA synthesis, morphologic and ultrastructural changes of cells, and perturbation in the cell cycle and by assessing the levels of p53 protein and SDI1 mRNA. RESULTS: Treatment of HepG2 cells with 9NC results in a dose-dependent inhibition of cell proliferation and DNA synthesis. Flow cytometric analysis of DNA content showed that 9NC-treated HepG2 cells are arrested in the G2-phase of the cell cycle. Light and electron microscopic examination revealed that 9NC at low concentrations induces morphologic and growth features that resemble properties highly differentiated or senescent cells, i.e., increased cell size and decreased nuclear/cytoplasmic ratio, as well as enlarged numbers of lysosomes, mitochondria, and lipid in the cytoplasm. No significant alteration in the p53 protein level was noted in 9NC-treated cells. In contrast to untreated, logarithmically grown HepG2 cells, 9NC-treated cells arrested at the G2-phase of the cell cycle and contained increased levels of SDI1 mRNA. Kinetic studies revealed gradual increases in SDI1 mRNA synthesis. CONCLUSIONS: Induction of SDI1 mRNA by 9NC represents the first documentation that the SDI1 gene can be overexpressed in the G2-phase of the cell cycle and provides a valuable cell culture system to dissect the events controlling the G2 checkpoint. In addition, this finding corroborates the hypothesis that genes up-regulated in senescent cells can also be induced in tumor-derived immortalized cells.

Antineoplastic Agents↗
PubMed1995

Trichotillomania: ophthalmic presentation.

PURPOSE: A case of trichotillomania, or compulsive hair-pulling, involving the eyelids is presented to alert ophthalmologists to this common, but frequently overlooked cause of eyelash and eyebrow alopecia. METHODS AND RESULTS: Clinical records of a 33-year-old woman suffering from trichotillomania were reviewed. Compulsive hair-pulling began in childhood and had become chronic. Psychiatric intervention was unsuccessful. CONCLUSIONS: The diagnosis of trichotillomania is made on history and slit-lamp examination findings. A skin biopsy may be necessary to exclude alopecia areata. Adults should be referred to a psychiatrist. Although childhood disease is usually benign, often reflecting a disturbed parent-child relationship, in adults hair-pulling is generally chronic and associated with psychiatric illness.

Adult↗
PubMed1995

Applications of magnetic source imaging in evaluation of candidates for epilepsy surgery.

Epilepsy was the first clinical entity to which magnetic source imaging was applied; however, the technology available severely limited early studies. The introduction of large-array magnetic source imaging systems has enabled striking advances in the application of magnetic source imaging in epilepsy surgery programs. The superimposition of functional localization of epileptic sources on anatomic images from MR imaging or computed tomography allows precise definition of the area and extent of epileptogenic tissue for possible resection. Concordance of magnetic source imaging indications with other noninvasive studies may reduce that need for invasive studies in certain patient populations.

Diagnostic Imaging↗