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Biomedical subjects

J D Smith

Publications and source records attributed to J D Smith.

At least 235 records · Page 13Linked to original sources
PubMed1989

Ca2+ induces charybdotoxin-sensitive membrane potential changes in rat lymphocytes.

There is disagreement regarding the existence of Ca2+-activated K+ channels in lymphocytes. Depolarization, hyperpolarization, or little change in membrane potential (Em) has been reported following elevation of free cytosolic Ca2+ concentration ([Ca2+]i). Patch-clamping studies have demonstrated inhibition of voltage-gated K+ channels, but Ca2+-activated K+ channels have not been detected. We used charybdotoxin (CTX), a potent inhibitor of Ca2+-activated K+ channels, to assess their presence in rat thymic lymphocytes. Fluorescent probes were used to measure Em and [Ca2+]i in cell suspensions treated with ionomycin. At basal [Ca2+]i, CTX had no effect on Em, suggesting that Ca2+-activated K+ channels do not contribute importantly to the resting potential. Elevation of [Ca2+]i in the submicromolar range induced a hyperpolarization that was dependent on the outward K+ gradient. The shape and duration of the Em change closely followed the elevation of [Ca2+]i. This hyperpolarization was inhibited by nanomolar concentrations of CTX. When [Ca2+]i approached or exceeded 1 microM, a biphasic Em change was recorded. A transient, CTX-sensitive hyperpolarization was followed by a sustained depolarization. The latter was greatly reduced when external Na+ was omitted. The data suggest that thymic lymphocytes possess Ca2+-sensitive K+ channels, which are activated by moderate increases in [Ca2+]i, resulting in hyperpolarization. At higher [Ca2+]i, the effect of K+ channels on Em is superseded by opening of nonselective cation channels, producing depolarization. Variations in the level of [Ca2+]i attained in earlier studies can explain existing discrepancies.

Animals↗
PubMed1989

G syndrome and its otolaryngologic manifestations.

G syndrome is a familial constellation of congenital anomalies that include a distinctive facies, ocular hypertelorism, prominent occiput and forehead, short lingual frenulum, stridor, hoarse cry, and laryngotracheoesophageal (LTE) clefts, as well as hypospadias and cryptorchidism. Approximately one third of the reported cases have also involved cleft lip and/or palate. Thirty cases of this syndrome have been reported in the pediatric and genetic literature, with variable expression of the syndrome characteristics. The purpose of this article is to report two additional cases of G syndrome with LTE cleft. A technique for repair of LTE cleft via a lateral pharyngeal approach is described. Although G syndrome is rare, otolaryngologists should be aware of its association with LTE cleft and its potential life-threatening problems.

Abnormalities, Multiple↗
PubMed1989

Clinical responses with active specific intralymphatic immunotherapy for cancer--a phase I-II trial.

We evaluated the method of active specific intralymphatic immunization to treat cancer in 32 patients with various tumor types as part of a broad-based phase I-II evaluation and describe the results of 3 sequential series. In series 1, the patients (n = 13) received 2 or more injections of autologous, cryopreserved, irradiated tumor cells directly into the lymphatic system through the cannulation of a dorsal pedal lymphatic channel. In series 2, the patients (n = 7) received low-dose cyclophosphamide, 300 mg per m2, 3 days before the autologous cell vaccine was administered. Series 3 (12 patients) was similar to series 2 except that the tumor cells were treated with cholesteryl hemisuccinate immediately before irradiation. Patients received from 2 to 6 injections of cells, depending on availability, at 2-week intervals. In all, 91 treatments are evaluated in this study. Clinical responses occurred in 7 of the 32 patients and were seen in all 3 series with about the same frequency. These responses occurred in cases of melanoma, lung cancer, colon cancer, and sarcoma. Regressions occurred in both visceral and subcutaneous sites. There was little toxicity, the chief side effect being local discomfort or inflammation. This experience indicates that active specific intralymphatic immunotherapy is safe, produces antitumor effects, and requires more investigation to increase the frequency and duration of observable tumor regression.

Adult↗
PubMed1989

Surgical correction of temporomandibular joint ankylosis.

A better understanding of facial growth processes and pathophysiology of temporomandibular joint ankylosis should lead to an improved surgical outcome. Surgical anatomy, pathophysiology, and treatment options are reviewed. Suggestions for varying the procedure in patients in whom facial growth is not complete are discussed.

Ankylosis↗
PubMed1989

The health and economic burden of cigarette smoking in Georgia in 1985.

Using the estimated prevalence of current and former cigarette smoking among Georgians, death certificate data in 1985, and the available literature on smoking-related health risks, we estimated the health and economic impact of cigarette smoking in Georgia in a single year. We estimated, using relatively conservative assumptions, the mortality and years of life lost prematurely, as well as the costs of medical care for ill individuals, lost wages due to death, and lost wages due to illness attributable to smoking. In 1985, past or present smoking was the cause of death for more than 7,700 Georgians, accounting for over 120,000 years of expected life lost prematurely, and with an overall societal cost exceeding $1.5 billion dollars. Cardiovascular diseases were the major cause of all estimated deaths associated with smoking. The prevalence of current smokers is higher in Georgians than the national average; this approach to examining the risks may be useful in lowering that prevalence by influencing patient education in Georgia.

Adult↗
PubMed1988

Increased turnover of the messenger RNA encoding tyrosine aminotransferase can account for the desensitization and de-induction of tyrosine aminotransferase by 8-bromo-cyclic AMP treatment and removal.

Treatment of H-4 rat hepatoma cells with 8-bromo-cyclic AMP (8-Br-cAMP) resulted in a transient induction of the gluconeogenic enzyme tyrosine aminotransferase. Synthesis of tyrosine aminotransferase and the level of its corresponding mRNA peaked 2 h after the addition of the cyclic nucleotide and declined thereafter. Tyrosine aminotransferase synthesis and mRNA failed to respond to the readdition of fresh 8-Br-cAMP, a process which we defined as desensitization. Removal of 8-Br-cAMP resulted in a decrease in tyrosine aminotransferase synthesis and mRNA, a process defined as de-induction. The relative transcription rate of the tyrosine aminotransferase gene and the turnover of its mRNA were determined by labeling intact cells with [3H]uridine. 8-Br-cAMP led to an increase in the rate of tyrosine aminotransferase transcription which was sustained for at least 4 h. The transcription rate declined upon de-induction. In addition, 8-Br-cAMP increased the turnover rate of tyrosine aminotransferase mRNA, but only after a 1.5-3 h time lag. This increased degradation rate persisted for at least 1.5 h after the removal of 8-Br-cAMP. These two contrasting and temporally distinct processes could account for the observed changes in tyrosine aminotransferase mRNA levels in response to 8-Br-cAMP treatment and removal.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1988

Use of 3-aminopropanol as an ethanolamine analogue in the study of phospholipid metabolism in Tetrahymena.

About 50% of the ethanolamine in phosphatidylethanolamine in Tetrahymena is replaced by 3-aminopropan-1-ol when the compound is added to the growth medium. The phosphatidylpropanolamine which is formed is not converted into the corresponding phosphatidylcholine analogue by methylation. There is an increase in phosphatidylcholine formed by the phosphotransferase pathway from free [3H]choline and a decrease in the phosphatidylcholine formed by the methylation pathway from [14C]methionine. The nature of the observed phospholipid alterations suggests that the regulation of phosphatidylcholine biosynthesis in Tetrahymena may be different from that found in higher eukaryotes.

Aldehydes↗
PubMed1988

Activation of Na+/H+ exchange and the expression of cellular proto-oncogenes in mitogen- and phorbol ester-treated lymphocytes.

It has been suggested that an intracellular alkalinization, resulting from stimulation of Na+/H+ exchange, is a necessary step and perhaps the signal leading to cellular proliferation in cells stimulated by mitogens. This hypothesis was tested by measuring the early stages of the proliferative cascade in cells where antiport activity was precluded by omission of Na+ or by the addition of potent amiloride analogs. To circumvent possible nonspecific effects due to long incubations under these conditions, an early response to mitogens, the increased level of c-fos mRNA, was monitored. In rat thymic lymphocytes, the increase in the level of c-fos RNA induced by the combination of 12-O-tetradecanoylphorbol 13-acetate and ionomycin was unaffected by inhibition of the antiport with 5-(N-ethyl-N-propyl)amiloride. Increased c-fos RNA was also observed in the absence of Na+ and when alkalinization was prevented by means of nigericin. Similar results were obtained with phytohemagglutinin-stimulated human T lymphocytes. Moreover, although the lectin stimulated the antiport in these cells, an alkalinization was not observed, due to the concomitant occurrence of an acidifying process. It was concluded that the stimulation of the Na+/H+ antiport that accompanies the addition of mitogens is neither sufficient nor necessary for the initiation of cellular proliferation.

Amiloride↗
PubMed1988

Expression of the human apolipoprotein E gene is regulated by multiple positive and negative elements.

Apolipoprotein E (apoE), unlike the other major lipoproteins, is synthesized in a variety of tissues. We examined which regions of the human apoE gene contributed to its tissue-specific expression using HepG2 and HeLa cells as examples of expressing and nonexpressing tissues, respectively. Regions between -360 bp and -80 bp and within the first intron were shown to be necessary for full expression activity in HepG2 cells by a nuclease protection assay which demonstrated correct transcriptional initiation of the transfected constructions. To fine map the regulatory regions, we constructed a series of deletions fused to the reporter gene chloramphenicol acetyltransferase. We discovered eight regions which had a positive effect on expression and three regions that had a negative effect on expression, in both HepG2 and HeLa cells. In addition we found three regions which had a tissue-specific negative effect on expression in HeLa cells and one region with a tissue-specific positive effect in HepG2 cells. A DNase I protection assay revealed eight footprints within the proximal 5'-flanking sequence and the first intron. Seven of these footprints fell within closely defined regions with positive expression activity. Sequence analysis of these footprint elements revealed the presence of previously identified elements and two novel elements related to each other, identified here as B1 and B2. We also defined another repeated sequence, the A element; all three of the tissue-nonspecific negative regions contained this element or sequences with homology to it. In the context of a heterologous promotor, a synthetic oligonucleotide containing the B1 and B2 elements behaved like a classical enhancer, having a positive effect on expression, even when placed at a distance. This effect was neutralized by a different synthetic oligonucleotide containing an A element repeat.

Animals↗
PubMed1988

The induction, desensitization and de-induction of tyrosine aminotransferase by 8-bromo-cyclic AMP in rat hepatoma cells.

Addition of 1-3 mM-8-bromo-cyclic AMP to monolayer cultures of H-4 rat hepatoma cells resulted in a rapid but short-lived increase in tyrosine aminotransferase (EC 2.6.1.5) activity. The transient nature of this induction is due to desensitization to 8-bromo-cyclic AMP. Throughout this time course of induction and desensitization, removal of 8-bromo-cyclic AMP resulted in a rapid and significant decrease in tyrosine aminotransferase activity, a process referred to as 'de-induction' in this study. We showed that the changes in tyrosine aminotransferase activity in its induction, desensitization and de-induction by 8-bromo-cyclic AMP were directly attributable to changes in the synthesis rate of the protein, and the amount of translatable and hybridizable mRNA encoding for tyrosine aminotransferase (mRNATAT). We further showed that this desensitization was specific to cyclic AMP. First, only active analogues of cyclic AMP and agents which increased cellular concentrations of cyclic AMP elicited this desensitization. Second, the desensitized cells were refractory only to the effects of 8-bromo-cyclic AMP; dexamethasone and insulin induced the tyrosine aminotransferase activity in the 8-bromo-cyclic AMP-desensitized cells in a manner similar to that of the controls. Studies on the metabolism of 8-bromo-cyclic AMP suggest that neither its degradation nor the accumulation of its primary metabolite, 8-bromoadenosine, played a significant role in modulating the expression of tyrosine aminotransferase during the time course of action of 8-bromo-cyclic AMP. These results provide evidence for a specific pretranslational mode of action of cyclic AMP in the control of tyrosine aminotransferase expression in its desensitization and de-induction, in addition to the early phase of induction.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1988

Malignant nerve sheath tumors of the head and neck.

Malignant nerve sheath tumors comprise approximately 5% of all soft tissue sarcomas; only 8% to 15% of these tumors arise in the head and neck. Most tumors appear as a rapidly expanding nonpainful mass in the face or lateral neck. Reported association with Von Recklinghausen's disease varies from 26% to 70%. Wide surgical excision is generally the recommended primary treatment. Recently, there has been a trend to include postoperative radiation therapy as a primary modality.

Adult↗
PubMed1987

Mechanism of activation of lymphocyte Na+/H+ exchange by concanavalin A. A calcium- and protein kinase C-independent pathway.

Treatment of thymic lymphocytes with the mitogenic lectin concanavalin A (ConA) increases the intracellular free Ca2+ concentration and stimulates phosphoinositide turnover. ConA also induced a rapid, amiloride-sensitive, Na+-dependent increase in cytosolic pH of 0.13 +/- 0.01, indicative of stimulation of the Na+/H+ antiport. To investigate the mechanism underlying activation of Na+/H+ exchange by ConA, the intracellular free Ca2+ concentration changes induced by this lectin were precluded by loading the cells with Ca2+-buffering agents and suspension in Ca2+-free media. Under these conditions, the ConA-induced cytoplasmic alkalinization proceeded normally. Two approaches were used to assess the role of protein kinase C. First, this enzyme was inhibited by the addition of 1-(5-isoquinolinysulfonyl)-2-methylpiperazine. In the presence of this potent antagonist, stimulation of the antiport by 12-O-tetradecanoylphorbol-13-acetate was greatly inhibited. In contrast, stimulation by ConA was unaffected. Second, protein kinase C was depleted by overnight incubation with phorbol esters. Following this treatment, Na+/H+ exchange was no longer activated by 12-O-tetradecanoyl-13-acetate, but was still stimulated by ConA. These data suggest that a Ca2+- and protein kinase C-independent mechanisms mediates the activation of Na+/H+ exchange by ConA. The possible role of GTP-binding proteins in the activation was also studied. The antiport was not stimulated by either fluoroaluminate or vanadate. Moreover, pretreatment with pertussis toxin failed to inhibit the ConA-induced cytoplasmic alkalinization. In contrast, preincubation with cholera toxin partially inhibited activation. Under these conditions, cholera toxin significantly elevated intracellular cAMP levels. Inhibition was also observed in cells treated with forskolin at concentrations that increased [cAMP]. The data suggest that a novel cAMP-sensitive signaling mechanism not involving Ca2+ and protein kinase C is involved in the stimulation of Na+/H+ exchange by mitogens in T lymphocytes.

Animals↗
PubMed1987

Asymmetry of the Na+/H+ antiport of dog red cell ghosts. Sidedness of inhibition by amiloride.

Amiloride is a potent inhibitor of the Na+/H+ antiport. Inhibition is generally competitive with extracellular Na+ and therefore believed to result from binding to the outward-facing transport site. It is not known whether amiloride can interact with the internal aspect of the antiport. This question was addressed by trapping the drug inside resealed dog red cell ghosts. The antiport, which is quiescent in resting ghosts, was activated by acid-loading the cytoplasm. This was accomplished by exchanging extracellular Cl- for internal HCO-3 through capnophorin, the endogenous anion exchanger. The activity of the Na+/H+ antiport was detected as an increase in cell volume, resulting from the net osmotic gain associated with coupled Na+/H+ and Cl-/HCO-3 exchange, or as the uptake of 22Na+. Intracellular amiloride, at concentrations in excess of 100 microM, failed to inhibit Na+/H+ exchange. This is approximately 10 times higher than the concentration required for half-maximal inhibition when amiloride is added externally. Independent experiments demonstrated that failure of internal amiloride to inhibit exchange was not due to leakage of the inhibitor, to differences in pH, or to binding or inactivation of amiloride by the soluble contents. It was concluded that the antiport is functionally asymmetric with respect to amiloride. This implies that the transport site undergoes a conformational change upon translocation across the membrane or, alternatively, that a second site required for amiloride binding is only accessible from the outside.

Amiloride↗
PubMed1987

Fluid balance and the adult respiratory distress syndrome.

This study examined the effect of fluid balance on survival in ARDS. Of the 213 patients entered into a prospective data collection study, we evaluated 113 patients who met strict criteria for ARDS. Multiple variables were analyzed for as long as 14 days after intubation including cardiac output, pulmonary capillary wedge pressure, mean blood pressure, intake minus output (I-O), cumulative intake minus output (cum I-O), and change in weight (delta wt). We found significant differences in cum I-O and delta wt between survivors and nonsurvivors on almost every day. Survivors lost weight and had a significantly lower cum I-O compared with nonsurvivors. Logistic regression was used to determine if delta wt and cum I-O could predict survival. Patients who lost 3 kg or more weight had a much higher survival than did those who gained 3 kg or more weight (67 and 0%, respectively, on Day 14). Similar results were obtained using comparably low and high values for cum I-O. The logistic regression equations demonstrated that weight loss and low cumulative I-O correlated with improved survival. Although cause and effect relationships are difficult to determine from these types of analyses, they can be used to formulate prospective studies and predict survival in patients.

Body Weight↗