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J R Smith

Publications and source records attributed to J R Smith.

At least 163 records · Page 9Linked to original sources
PubMed1996

Effect of aging on regulation of sdi-1 in rat hepatocytes.

We examined basal and EGF stimulated DNA synthesis as well as sdi-1 mRNA and protein in primary hepatocyte cultures, and basal levels of sdi-1 mRNA and protein in whole liver homogenates from 6 and 24 month old rats. Since EGF stimulated DNA synthesis decreases with age, it was hypothesized that basal and EGF stimulated levels of sdi-1 mRNA and protein, an inhibitor of DNA synthesis, might increase. Surprisingly, however both sdi-1 mRNA and protein actually decreased both in cells and homogenates of old rats. These results indicate that the age-related impairment in EGF stimulated DNA synthesis in hepatocytes appears to occur prior to or parallel with sdi-1 expression and cannot be explained on the basis of increased inhibition due to elevated levels of this protein.

Aging↗
PubMed1996

Replicative senescence: implications for in vivo aging and tumor suppression.

Normal cells have limited proliferative potential in culture, a fact that has been the basis of their use as a model for replicative senescence for many years. Recent molecular analyses have identified numerous changes in gene expression that occur as cells become senescent, and the results indicate that multiple levels of control contribute to the irreversible growth arrest. These include repression of growth stimulatory genes, overexpression of growth inhibitory genes, and interference with downstream pathways. Studies with cell types other than fibroblasts will better define the role of cell senescence in the aging process and in tumorigenesis.

Aging↗
PubMed1996

CCAAT/enhancer-binding protein alpha (C/EBP alpha) inhibits cell proliferation through the p21 (WAF-1/CIP-1/SDI-1) protein.

C/EBPalpha has a role in growth arrest and differentiation of mouse preadipocytes. To study the mechanism of C/EBPalpha-induced growth arrest, we developed a cell line, HT1, that contained the human C/EBPalpha gene under Lac repressor control. IPTG-induced C/EBPalpha caused inhibition of cell proliferation and DNA synthesis as measured by colony growth assays, cell counting, and BrdU uptake. A number of proteins that are known to be involved in the regulation of the cell cycle, such as cyclin-dependent kinase (CDK)2 and CDK4, proliferating cell nuclear antigen (PCNA), p53, c-fos, and the CDK inhibitor p16 and p27 were investigated by Western analysis. No change in their expression was observed. However, the p21 (WAF-1/CIP-1/SDI-1) protein was significantly elevated in growth-arrested HT1 cells. Elevation of p21/SDI-1 mRNA (threefold) and activation of the p21/SDI-1 promoter by C/EBPalpha did not account for the 12- to 20-fold increase in p21/SDI-1 protein. Protein synthesis inhibition by cycloheximide (CHX) treatment indicated that the half-life of p21/SDI-1 in dividing HT1 cells was approximately 30 min. However, in C/EBPalpha growth-arrested cells, the level of the p21/SDI-1 did not change for > 80 min after CHX addition. Our studies demonstrate that C/EBPalpha activates p21/SDI-1 by increasing p21/SDI-1 gene expression and by post-translational stabilization of p21/SDI-1 protein. Furthermore, induction of p21/SDI-1 is responsible for the ability of C/EBPalpha to inhibit proliferation because transcription of antisense p21/SDI-1 mRNA eliminated growth inhibition by C/EBPalpha.

Base Sequence↗
PubMed1996

Myocardial ischemia/reperfusion protection using monophosphoryl lipid A is abrogated by the ATP-sensitive potassium channel blocker, glibenclamide.

OBJECTIVES: Monophosphoryl lipid A (MLA), a detoxified derivative of the lipid A portion of the endotoxin molecule, given as a pretreatment 24 h prior to cardiac ischemia/reperfusion reduces myocardial stunning and infarction in dogs. This study was undertaken to evaluate the ability of MLA pretreatment to reduce infarct size in a rabbit model of in situ regional myocardial ischemia and reperfusion. Secondly, the potential role of modulation of ATP-sensitive potassium (KATP) channel in MLA's cardioprotection was evaluated using in vivo pharmacologic antagonism with a KATP channel blocker, as was the role of tumor necrosis factor using an enzyme-linked immunosorbent assay method of serum cytokine analysis. METHODS: Rabbits were pretreated intravenously with MLA or vehicle injection 24 h prior to initiation of 30 min in situ left anterior descending coronary artery occlusion followed by 3 h reperfusion. In animals receiving glibenclamide, the potassium channel antagonist was administered 30 min prior to inducing ischemia. Animals receiving glibenclamide, which possesses hypoglycemic effects, underwent serial blood glucose evaluation prior to drug and throughout the ischemia and reperfusion periods. Hemodynamics were monitored; infarct size and area at risk were assessed by contrast dye staining (triphenyltetrazolium chloride). Serum tumor necrosis factor was measured by enzyme-linked immunosorbent method in animals administered cardioprotective doses of MLA as well as pyrogenic doses of MLA and endotoxin (positive control) to determine if elaboration of this cytokine could be associated with the cardioprotective effect of MLA. RESULTS: MLA administered as a single intravenous dose 24 h prior to ischemia reduced infarct size, expressed as a percent of the area at risk, 64 and 71% at doses of 35 and 10 micrograms/kg, respectively. Lower doses of MLA (2.5 and 5 micrograms/kg) did not significantly reduce infarct size. Administration of glibenclamide (300 micrograms/kg) 30 min prior to ischemia completely blocked the ability of MLA pretreatment to limit infarct size, while MLA vehicle-glibenclamide-treated control rabbits displayed infarcts not significantly different from MLA-vehicle-treated control rabbits. A cardioprotective dose of MLA (35 micrograms/kg) did not induce the elaboration of tumor necrosis factor into rabbit serum (within the limits of assay sensitivity). CONCLUSIONS: Single-dose pretreatment with MLA administered intravenously to rabbits substantially reduces infarct size when administered 24 h prior to ischemia. Pharmacologic preconditioning with MLA appears to be mediated through KATP channels as the channel blocker, glibenclamide, reversed the cardioprotective activity of MLA when administered 1 day following MLA pretreatment, yet 30 min prior to ischemia. In this model the cardioprotective does not appear to be associated with increases in serum tumor necrosis factor.

Adenosine Triphosphate↗
PubMed1996

The significance of ictal depth EEG patterns in patients with temporal lobe epilepsy.

We reviewed 187 depth recorded seizures in 33 patients with non-lesional temporal lobe complex partial seizures. All patients had a minimum of 1 year follow-up following temporal lobectomy. We classified seizure onset pattern as rhythmic activity, attenuation, or repetitive spikes or spike wave complexes. The most common pattern of seizure onset was rhythmic activity and the next most common pattern was repetitive spikes. Seventy-five seizures (49%) had only one seizure onset pattern, and 79 seizures (51%) had a combination of seizure onset patterns. The degree of hippocampal gliosis strongly predicted the type of seizure onset pattern (Chi square = 24.07, 2 d.f., P < 0.01). The rhythmic activity pattern was associated with mild gliosis, and the repetitive spike pattern was associated with severe gliosis. We classified seizure onset as focal or regional based on the number of electrode contacts that were involved by the ictal EEG. A focal seizure onset was associated with an excellent outcome following temporal lobectomy.

Brain↗
PubMed1996

Local electrogram changes in response to a high-voltage intracardiac shock in humans.

INTRODUCTION: Transvenous defibrillators may have difficulty sensing ventricular fibrillation following an unsuccessful shock. This study was undertaken to characterize the changes that occur in intracardiac electrograms following a defibrillator shock that may contribute to the failure to redetect arrhythmias. METHODS AND RESULTS: Unipolar and bipolar electrogram recordings were made during sinus rhythm before and following monophasic and biphasic test shocks delivered in random order through a single lead defibrillator system in 15 patients. An additional 14 patients received only a biphasic shock. Electrogram amplitude, dV/dt, and activation recovery time were determined. Following biphasic shocks, unipolar electrogram amplitude decreased by a mean of 19.6% and the bipolar amplitude by a mean of 20.8% (P = NS). The mean dV/dt in the unipolar recordings decreased by 30.7% and in the bipolar recordings decreased by 33.0% (P = NS). Time constants for recovery were approximately 60 seconds. Reduction in dV/dt was greater when the shocking coil was closer to the distal sensing electrode. There was no significant influence of shock waveform (monophasic vs biphasic), recording type (unipolar vs bipolar), or clinical variables on the change in electrogram characteristics. CONCLUSION: Changes in intracardiac electrograms following defibrillator shocks through transvenous leads are seen with shocks delivered in sinus rhythm and may be related to the proximity of the shocking coil to the distal sensing electrode.

Aged↗
PubMed1996

Inhibition of E2F activity by the cyclin-dependent protein kinase inhibitor p21 in cells expressing or lacking a functional retinoblastoma protein.

p21Sdi1/WAF1/Cip1 inhibits cyclin-dependent protein kinases and cell proliferation. p21 is presumed to inhibit growth by preventing the phosphorylation of growth-regulatory proteins, including the retinoblastoma tumor suppressor protein (pRb). The ultimate effector(s) of p21 growth inhibition, however, is largely a matter of conjecture. We show that p21 inhibits the activity of E2F, an essential growth-stimulatory transcription factor that is negatively regulated by unphosphorylated pRb. p21 suppressed the activity of E2F-responsive promoters (dihydrofolate reductase and cdc2), but E2F-unresponsive promoters (c-fos and simian virus 40 early) were unaffected. Moreover, the simian virus 40 early promoter was rendered p21 suppressible by introducing wild-type, but not mutant, E2F binding sites; p21 deletion mutants showed good agreement in their abilities to inhibit E2F transactivation and DNA synthesis; and E2F-1 (which binds pRb), but not E2F-4 (which does not), reversed both inhibitory effects of p21. Despite the central role for pRb in regulating E2F, p21 suppressed growth and E2F activity in cells lacking a functional pRb. Moreover, p21 protein (wild type but not mutant) specifically disrupted an E2F-cyclin-dependent protein kinase 2-p107 DNA binding complex in nuclear extracts of proliferating cells, whether or not they expressed normal pRb. Thus, E2F is a critical target and ultimate effector of p21 action, and pRb is not essential for the inhibition of growth or E2F-dependent transcription.

3T3 Cells↗
PubMed1996

A comparison of 2 analgesic regimens for the control of postoperative periodontal discomfort.

A single blind study of 24 patients compared the postoperative periodontal pain relief and adverse effects associated with a pretreatment regimen with etodolac, a nonsteroidal anti-inflammatory drug (NSAID), to a typical pro re nada (prn) regimen with a combination of acetaminophen with hydrocodone. Patients selected required one or more periodontal osseous surgeries that were judged to involve relatively similar degrees of surgical manipulation. Patients in the etodolac group received two 300 mg capsules 30 minutes prior to surgery and then redosed themselves prn. Patients who received the combination drug were not premedicated and followed a prn regimen. The subjects used a verbal analogue scale to report levels of pain hourly for the first 8 hours (starting 30 minutes prior to surgery) and also indicated any side effects experienced during the first week after surgery. Specific parameters monitored were the mean sum of hourly pain scores, mean hourly pain scores, time to first medication, number of postoperative doses, and adverse effects. Of the parameters studied, the only one that showed a statistically significant difference was the time to first medication. The time span from 30 minutes prior to the beginning of surgery to the first postsurgical dose was greater for etodolac than for the combination drug. However, the total number of medications taken under both regimens was similar. The side effects were minimal for both of the drugs studied. It was concluded that the analgesic regimens tested under clinical practice conditions were comparable in providing analgesia with minimum side effects in uncomplicated periodontal osseous surgery. Studies with larger numbers of patients are needed to definitively address whether these regimens are truly equivalent.

Acetaminophen↗
PubMed1996

Modulation of non-templated nucleotide addition by Taq DNA polymerase: primer modifications that facilitate genotyping.

Taq DNA polymerase can catalyze non-templated addition of a nucleotide (principally adenosine) to the 3' end of PCR-amplified products. Recently, we showed that this activity, which is primer-specific, presents a potential source of error in genotyping studies based on the use of short tandem repeat (STR) markers. Furthermore, in reviewing our data, we found that non-templated nucleotide addition adjacent to a 3' terminal C is favored and that addition adjacent to a 3' terminal A is not. It was clear, however, that features of the template in addition to the 3' terminal base also affect the fraction of product adenylated. To define consensus sequences that promote or inhibit product adenylation, we transplanted sequences between the 5' ends of the reverse primers of markers that are adenylated and those of markers that are not adenylated. It proved difficult to identify a single sequence capable of protecting the products of all markers from non-templated addition of nucleotide. On the other hand, placing the sequence GTTTCTT on the 5' end of reverse primers resulted in nearly 100% adenylation of the 3' end of the forward strand. This modification or related ones (called "PIG-tailing") should facilitate accurate genotyping and efficient T/A cloning.

Base Sequence↗
PubMed1996

Supplementary sensorimotor area epilepsy in adults.

Based on stimulation studies, observation of seizure phenomena in patients with epilepsy, and diagnostic studies documenting SSMA anatomic and functional abnormalities, there is evidence that epileptic involvement of the SSMA results in a set of clinical phenomena that include asymmetric tonic posturing of the extremities, abduction and elevation of the contralateral upper extremity, and speech arrest with preserved consciousness. Auras and vocalization occur less frequently. These seizures tend to be brief and to have an abrupt onset and cessation. They are more common during sleep than during wakefulness, and they tend to occur multiple times per night. The abolition of seizures following a resection limited to the SSMA provides conclusive evidence that the SSMA is the epileptogenic zone in some patients with this type of seizure. On the other hand, there is variation in seizure phenomena among patients whose seizures originate in the SSMA. In addition, there are patients with seizures suggestive of SSMA involvement whose seizures originate in a broad epileptogenic zone that includes the SSMA, other patients whose seizures originate outside the SSMA and spread to the SSMA, and still other patients whose seizures originate outside the SSMA and remain outside the SSMA. To better understand the phenomena associated with SSMA seizure onset and to determine the specificity of certain phenomena to SSMA involvement, additional data are needed from structural and functional imaging studies, video and EEG monitoring studies with noninvasive and invasive electrodes, and pathologic and surgical outcome studies. Since one cannot be certain at this time of the specificity of seizure phenomena to SSMA involvement, and because localization for possible surgery is the primary rationale for detailed study of these patients, we recommend that the term SSMA seizure refer only to those seizures that originate in the SSMA and that SSMA epilepsy refer only to epilepsies in which the patient's seizures originate in the SSMA. Since the present classification of epileptic seizures is not of value in describing the major phenomena of a partial seizure or in localization of the site of onset, a descriptive classification is needed to improve communication concerning seizure phenomena and to provide clues to localization.

Adult↗
PubMed1996

Human immunodeficiency virus and pregnancy.

There are now proven ways to reduce the transmission of human immunodeficiency virus from infected mothers to their offspring, including the use of zidovudine during pregnancy and labour, and avoidance of breast-feeding. Therefore, screening programmes should be extended to enable at-risk mothers to benefit from these advances.

Female↗
PubMed1995

Effect of increased drive-train stimulus intensity on dispersion of ventricular refractoriness.

BACKGROUND: Most studies evaluating the effects of high-intensity drive-train (S1) stimulation on the measurement of the ventricular effective refractory period (VERP) demonstrated a shortening of the VERP. Because this effect may be due to the local release of catecholamines, VERP shortening would be expected to occur only near the site of stimulation. Local shortening in the VERP should then result in an increased dispersion of refractoriness during high-intensity drive-train stimulation. Thus, this study evaluated the spatial distribution of the VERP shortening resulting from high-intensity S1 stimulation and its effect on dispersion of refractoriness. METHODS AND RESULTS: Three groups of patients were studied. In group 1, 10 subjects without structural heart disease had VERP determinations performed at the right ventricular apex (RVA) and outflow tract (RVOT) while the S1 site was changed to evaluate the effects of low-intensity S1 stimulation on the measured VERP. In group 2, the effect of high-intensity S1 stimulation on the VERP was studied 0, 7, 14, and 21 mm away from the S1 site to measure the spatial distribution of VERP shortening and the effect on dispersion of refractoriness; 10 additional subjects without structural heart disease made up group 2. Because increased dispersion of refractoriness may be deleterious in certain clinical situations, the effect of high-intensity S1 stimulation was studied in group 3, which comprised 10 subjects with chronically implanted transvenous defibrillators; noninvasive measurements of the VERP through the chronic lead were made while the S1 stimulus intensity was varied from low to high intensity. All VERP determinations were performed during continuous pacing by use of an incremental method and a low stimulus intensity for the extrastimulus. In group 1, the RVA VERPs were 218 +/- 9 and 214 +/- 10 ms when the S1 site was the RVA and RVOT, respectively (P = NS). The RVOT VERPs were also unchanged when the S1 site was changed from the RVOT to the RVA. In group 2, high-intensity S1 changed the VERP from 224 +/- 8 (at twice the threshold) to 203 +/- 10 ms (P < .01), 220 +/- 11 to 209 +/- 12 ms (P < .01), 222 +/- 12 to 221 +/- 12 ms, and 220 +/- 11 to 221 +/- 11 ms at 0, 7, 14, and 21 mm away from the S1 site, respectively. High-intensity S1 stimulation led to an increase in the dispersion of refractoriness from 13 +/- 4 to 22 +/- 9 ms (P = .006). In group 3, high-intensity S1 stimulation shortened the VERP from 309 +/- 23 to 285 +/- 30 ms (P = .0003). CONCLUSIONS: Low-intensity S1 stimulation has no significant effect on the VERP. High-intensity S1 stimulation shortens the refractory period maximally at the site of stimulation; the VERP shortening dissipates between 7 and 14 mm away from the site of S1 stimulation, resulting in an increased dispersion of refractoriness. The local VERP shortening with high-intensity stimulation is noted in patients with chronically implanted defibrillator leads, which may have implications for the mechanism of proarrhythmia during high-intensity stimulation.

Adult↗
PubMed1995

The C-terminal region of p21SDI1/WAF1/CIP1 is involved in proliferating cell nuclear antigen binding but does not appear to be required for growth inhibition.

The cyclin-dependent kinase (Cdk) inhibitor p21SDI1/WAF1/CIP1 has been found to be involved in cell senescence, cell cycle arrest, and differentiation. p21SDI1 inhibits the activity of several Cdks, in contrast to other inhibitors such as p15INK4B and p16INK4A, which act on specific cyclin-Cdk complexes. Of interest were reports that p21SDI1 also bound proliferating cell nuclear antigen (PCNA), an auxiliary protein for DNA polymerase delta, and inhibited DNA replication but not DNA repair in vitro. To better understand the function of this interaction in vivo, we first determined the region of p21SDI1 that was needed for PCNA binding. Analysis of deletion mutants of p21SDI1, which covered the majority of the protein, revealed that deletion of either amino acids 142-147 or 149-154 resulted in loss of ability to bind a glutathione S-transferase-PCNA fusion protein. Site-directed mutagenesis in this region led to the identification of the PCNA binding motif RQXXMTXFYXXXR and demonstrated that mutation of either amino acid Met-147 or Phe-150 resulted in almost complete ablation of PCNA binding. Interestingly, when we determined DNA synthesis inhibitory activity of deletion mutants or point mutants that were unable to bind Cdk2 and/or PCNA, we found that loss of binding to PCNA did not affect inhibitory activity, whereas lack of Cdk2 binding greatly reduced the same. This result suggests that the primary mechanism for inhibition of DNA synthesis by p21SDI1 occurs via inhibition of Cdk activity.

Amino Acid Sequence↗
PubMed1995

Mutant p53 rescues human diploid cells from senescence without inhibiting the induction of SDI1/WAF1.

Although the cyclin-dependent kinase inhibitor p21SDI1 (WAF1/CIP1) has been proposed as the mediator of p53-induced cell cycle arrest following DNA damage, several stimuli now appear to induce SDI1 independent of p53 function. We have examined the behavior of p53 and SDI1 in an isogeneic model by manipulating p53 status in normal diploid human fibroblasts using an amphotropic retroviral vector. Following DNA strand break damage induced by bleomycin, both SDI1 induction and G1-S cell cycle arrest are p53 dependent, consistent with SDI1 being the key mediator. In contrast, in cellular senescence (and following UV irradiation), induction of SDI1 occurs independent of p53 function yet growth arrest is still p53 dependent. We conclude (a) that redundant pathways exist for induction of SDI1, but that (b) SDI1, while perhaps necessary, is not sufficient for inhibition of cell cycle progression, requiring the cooperation of an additional factor (possibly another cyclin-dependent kinase inhibitor) whose expression, at least in the case of senescence, is strictly p53 dependent.

Bleomycin↗