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Biomedical subjects

W E Hopkins

Publications and source records attributed to W E Hopkins.

36 records · Page 2Linked to original sources
PubMed1993

Comparison of color flow imaging and peripheral venous saline contrast during transesophageal echocardiography to evaluate right-to-left shunt at the atrial level.

Intravenous injections of agitated saline (contrast) are used to identify right-to-left atrial shunt during transesophageal echocardiography (TEE). Color flow imaging (CFI) with TEE can be used to detect left-to-right atrial shunt but its application to detect right-to-left shunt is not known. We performed CFI and contrast during TEE in 72 patients for detection of right-to-left shunt. Frame-by-frame review of CFI demonstrated discrete laminar jets of the left side of the atrial septum at the lower or upper margin of the fossa ovalis (FO) in 41 (57%) of 72; only 22 had positive contrast. Timing of shunts was late diastolic or early systolic and not related to pulmonary venous inflow. The length of the FO at end-systole was greater in those with positive CFI compared to negative CFI (13.5 +/- 5 vs 11 +/- 4 mm, P = 0.02). CFI was positive for right-to-left shunt in ten of 14 with atrial septal aneurysms. Contrast was positive for right-to-left shunt in 29 (40%) of 72; all but six had positive CFI. The degree of left atrial opacification was minimal in 19 and moderate to severe in ten. FO size was greater in positive versus negative contrast (14 +/- 4 vs 11 +/- 5 mm, P = 0.02). Contrast was positive for shunt in ten of 14 with septal aneurysms. Thus, right-to-left atrial shunt more often occurs with increased FO size and septal aneurysms.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1993

Two-dimensional Doppler color flow imaging in adults with L-transposition of the great arteries.

Abnormalities of the left atrioventricular (AV) (tricuspid) valve and pulmonic stenosis are part of the spectrum of manifestations in patients with corrected transposition of the great arteries (L-TGA). However, the utility of Doppler color flow imaging combined with two-dimensional echocardiography in L-TGA has not been established. Thus, we evaluated 14 adults (mean age 33 +/- 8 {SD} years) with L-TGA. Two-dimensional echocardiography demonstrated Ebstein's anomaly in five patients. Color flow imaging detected left AV valve regurgitation in 13 (moderate to severe in six); the other patient had a prosthetic valve. Right AV valve regurgitation was detected in two (mild in both). Aortic regurgitation was present in five (moderate in three). Pulmonic regurgitation was present in nine (moderate to severe in three). Pulmonic stenosis was detected in three (subvalvular in two, valvular in one) and two patients had pulmonary atresia. One patient had a bicuspid aortic valve with mild stenosis. Mean right ventricular ejection fraction (apical, area-length) was 44%; only 1 of 9 patients had a value greater than 55%. Thus, two-dimensional Doppler echocardiography with color flow imaging detects a high incidence of systemic AV valvular regurgitation and reduced right (systemic) ventricular systolic function in adults with L-TGA.

Adult↗
PubMed1992

Frequency and significance of intrapulmonary right-to-left shunting in end-stage hepatic disease.

Intrapulmonary vascular abnormalities consisting of arteriovenous malformations and capillary dilatations have been described in patients with severe liver disease. These intrapulmonary vascular abnormalities can result in intrapulmonary right-to-left shunting and hypoxemia. Twenty-five of 53 patients (47%) with end-stage hepatic disease were found to have contrast echocardiographic evidence of intrapulmonary right-to-left shunting. There was no difference in mean age, gender distribution, or severity of hepatic disease in those with and without evidence of such shunting. Although there was no difference in mean partial arterial oxygen pressure (PaO2) values in the 2 groups (82 +/- 11 vs 76 +/- 11 mm Hg), the mean PaO2 value of those with at least 2+ left ventricular opacification (2 to 4+) was significantly lower (66 +/- 3 mm Hg, n = 8; p less than 0.01). Unexpectedly, patients with evidence of intrapulmonary shunting had a lower mortality rate before transplantation (3 of 25, 12%) than those without evidence of shunting (10 of 28, 36%) resulting in a significant difference in actuarial survival (p less than 0.05) by the end of the follow-up period. It is concluded that intrapulmonary right-to-left shunting occurs frequently in patients with end-stage liver disease and may be a marker of a positive biologic process that, in some way, leads to improved short-term survival.

Adult↗
PubMed1992

Synergistic induction of plasminogen activator inhibitor type-1 in HEP G2 cells by thrombin and transforming growth factor-beta.

Plasminogen activator inhibitor type-1 (PAI-1) is a physiologic modulator of the fibrinolytic system. Its activity in plasma increases in diverse thrombotic states. The large synthetic capacity of the liver make it a source of potentially large amounts of PAI-1. Because thrombin activity increases in association with thrombotic disorders and because specific binding sites for thrombin have been identified on hepatocytes, we characterized the effect of thrombin on hepatocyte PAI-1 production. Incubation of Hep G2 cells with human alpha-thrombin resulted in a dose- and time-dependent increase in the concentration of PAI-1 in conditioned media. This effect was inhibited completely by hirudin and by antithrombin III. Steady-state levels of both the 3.2-kb and 2.2-kb forms of PAI-1 mRNA increased after stimulation of the cells with thrombin, indicating that thrombin influences PAI-1 expression in Hep G2 cells at the pretranslational level. Incubation of Hep G2 cells with alpha-thrombin and either platelet lysates or purified transforming growth factor-beta (TGF-beta), both previously shown to augment hepatocyte PAI-1 expression, resulted in a synergistic increase in the concentration of PAI-1 in conditioned media. PAI-1 mRNA appeared to be synergistically increased as well. Thus, thrombin increases expression of both PAI-1 protein and mRNA in Hep G2 cells and exerts synergistic effects with TGF-beta. These results underscore the potential importance of inhibition of thrombin under conditions in which thrombolysis is induced pharmacologically.

Antithrombin III↗
PubMed1991

Mediators of induction of augmented expression of plasminogen activator inhibitor type-1 in Hep G2 cells by platelets.

Plasminogen activator inhibitor type-1 (PAI-1) is a physiologic modulator of the fibrinolytic system. We have shown previously that PAI-1 biosynthesis in cultured cells depends on several factors in serum. Because platelets are richly endowed with specific growth factors and because the release reaction is an integral part of thrombosis, the present study was performed to determine whether platelets augment PAI-1 production and if so, to define mediators responsible. Hep G2 cells were used to determine whether platelet lysates increased PAI-1 synthesis in a dose and time-dependent manner. In cells labeled metabolically with 35S-methionine for 6 h, an increase in labeled PAI-1 was elicited indicative of de novo synthesis as well as increased secretion of PAI-1 mediated by platelet lysates. Steady state levels of both the 3.2 and 2.2 kb forms of PAI-1 mRNA increased after 2 h and peaked in 3-5 h in a dose-dependent fashion as well. Incubation of Hep G2 cells with collagen activated platelets resulted in a similar induction of PAI-1 mRNA. The increase in PAI-1 mRNA occurred with exposure of the cells to platelet lysates for intervals as brief as 15 min and was not inhibited by cycloheximide indicating its independence of new protein synthesis. In order to identify the factors in platelets responsible for the induction of PAI-1 synthesis in the Hep G2 cell model system, neutralizing antibodies were used to inhibit specific platelet associated growth factors. Antibodies to transforming growth factor-beta (TGF-beta) and to the epidermal growth factor (EGF)/transforming growth factor alpha (TGF-alpha) receptor inhibited the platelet lysate-mediated increase in PAI-1 protein by 77%.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Platelets↗
PubMed1991

Multiple transforming growth factor-beta-inducible elements regulate expression of the plasminogen activator inhibitor type-1 gene in Hep G2 cells.

Regulation of plasminogen activation by plasminogen activator inhibitor type-1 (PAI-1) is a critical feature of many biological processes. Transforming growth factor-beta (TGF-beta) induces PAI-1 mRNA and protein in several types of cultured cells, including Hep G2 cells. The present study was performed to define mechanisms by which PAI-1 gene expression is regulated by TGF-beta. Nuclear run-on assays performed on Hep G2 cells stimulated with TGF-beta for 6 h showed a 3.8-fold increase in PAI-1 gene transcription. TGF-beta increased the half-life of PAI-1 mRNA in Hep G2 cells 2.5-fold over control values. To characterize transcriptional regulatory mechanisms, we constructed chimeric genes containing PAI-1 5'-flanking DNA fused upstream of the bacterial chloramphenicol acetyltransferase gene in the vector pSVOCAT and transfected Hep G2 cells. Promoter deletion analysis demonstrated that sequences between -791 and -328 and -328 to -186 base pairs upstream of the PAI-1 gene cap site contain TGF-beta responsive elements that conferred TGF-beta inducibility in an orientation and position-independent manner. Further characterization of the larger TGF-beta-inducible enhancer (-791 to -328) located a TGF-beta-inducible element at nucleotides -791 to -546 upstream of the PAI-1 gene cap site. These results demonstrate that PAI-1 gene regulation by TGF-beta in Hep G2 cells is mediated both at a transcriptional level by two specific inducible elements, as well as by post-transcriptional mechanisms.

Blotting, Northern↗
PubMed1991

Transcriptional regulation of plasminogen activator inhibitor type-1 mRNA in Hep G2 cells by epidermal growth factor.

Secretion of plasminogen activator inhibitor type-1 (PAI-1) by cultured cells is increased after exposure to specific cytokines and growth factors. We have shown previously that incubation of Hep G2 cells with epidermal growth factor (EGF) results in a marked increase in steady state levels of PAI-1 mRNA (Lucore, C.L., et al. (1988) J. Biol. Chem. 263, 15845-15848). The present study was undertaken to determine whether the regulation of expression of PAI-1 mRNA by EGF is mediated at the level of transcription and/or by post-transcriptional mechanisms. The rate of transcription of the PAI-1 gene measured by nuclear run-on assays was found to be increased within 2 h after stimulation of the cells with EGF (5 ng/ml) (3.2 fold increase relative to control, n = 2, range 3.0-3.4). It reached a maximum in 3 h, (9.2 fold increase relative to control, n = 2, range 8.8-9.6) and returned to baseline in 5 h. Exposure of the cells to EGF did not increase the rate of transcription of the glyceraldehyde-3-phosphate dehydrogenase gene. The half life of PAI-1 mRNA in Hep G2 cells was 120 min as determined by RNA blot analysis after exposure of the cells to actinomycin D to inhibit transcription. Stimulation of the cells with EGF did not result in significant change in the half life of PAI-1 mRNA. The results demonstrate that exposure of Hep G2 cells to EGF increases PAI-1 gene transcription.

Cycloheximide↗
PubMed1991

Metabolism of glyphosate in Sprague-Dawley rats: tissue distribution, identification, and quantitation of glyphosate-derived materials following a single oral dose.

Five groups of male Sprague-Dawley rats were orally administered a mixture of [14C]- and [12C]-glyphosate (N-phosphonomethylglycine) at a dose level of 10 mg/kg body weight. The majority of radioactivity 2 hr after administration was associated with the gastrointestinal contents and small intestinal tissue. Approximately 35-40% of the administered dose was absorbed from the gastrointestinal tract, and urine and feces were equally important routes of elimination. The total body burden 7 days after administration was approximately 1% of the administered dose and was primarily associated with the bone. Total recovery for this study ranged from 95 to 102% of the administered dose. Metabolic profiles of tissues containing greater than 1% of the administered dose at various times after administration indicated that nearly 100% of the body burden of radioactivity was present as unmetabolized parent glyphosate. A minor component constituting less than 0.1% of the administered dose (less than 0.4 ppm) was observed in colon tissue from animals 2 hr after the administration of glyphosate and was also present in the GI contents of one animal 28 hr after administration of the radiolabel. The retention time for this metabolite was similar, but not identical, to the retention time for AMPA (aminomethylphosphonic acid), the major bacterial metabolite of glyphosate found in soil. Tissue extraction efficiency was always greater than 90% and stability assays indicated no significant effect of storage on either parent glyphosate or AMPA. The results from this study indicate that virtually no toxic metabolites of glyphosate were produced since there was little evidence of metabolism and essentially 100% of the body burden was parent compound with no significant persistence of material.

Animals↗
PubMed1991

Mechanisms contributing to increased synthesis of plasminogen activator inhibitor type 1 in endothelial cells by constituents of platelets and their implications for thrombolysis.

We recently hypothesized that after pharmacologically induced coronary thrombolysis, increased activity of plasminogen activator inhibitor type 1 (PAI-1) retards recanalization, contributes to early reocclusion, or both. This hypothesis was based on the increased elaboration of PAI-1 that we observed in cultured liver cells exposed to growth factors releasable from platelets activated at sites of thrombosis in vivo. PAI-1 released locally is particularly likely to attenuate lysis of thrombi that are targets of thrombolytic drugs. Accordingly, the present study was performed to determine whether synthesis of PAI-1 by endothelial cells is augmented by products of platelets. Lysates from platelets (0.5-8.0 x 10(4)/mm3 media, i.e. less than 10% of the concentration of platelets in blood) increased synthesis and release of PAI-1 into both the extracellular matrix and conditioned media (by 2.8-fold and 3.3-fold within 6 and 24 hours, respectively). Synthesis of neither tissue-type plasminogen activator nor overall protein increased. Increased synthesis of PAI-1 was confirmed by immunoprecipitation of [35S]PAI-1 after metabolic labeling of cells. The increased elaboration of PAI-1 was consistent with increased transcription as reflected by the observed increase in PAI-1 mRNA of 2.2-fold in 4 hours. Effects of platelet lysates were simulated by transforming growth factor beta (TGF-beta), known to be present in platelet alpha-granules and released with platelet activation. Antibody to TGF-beta reduced the stimulation of PAI-1 synthesis by TGF-beta, as expected, by 82%.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Platelets↗
PubMed1990

Induction of synthesis of plasminogen activator inhibitor type-1 by tissue-type plasminogen activator in human hepatic and endothelial cells.

Plasminogen activator inhibitor type-1 (PAI-1) can modify fibrinolytic activity in vitro and in vivo. The present study was performed to determine whether pharmacologic concentrations of tissue-type plasminogen activator (t-PA) can initiate negative feedback by stimulating PAI-1 synthesis. In both human hepatoma cells (Hep G2) and human umbilical vein endothelial cells (HUVEC), t-PA increased the total concentrations and appearance of newly synthesized protein in conditioned media of free PAI-1 and PAI-1 complexed with t-PA in a dose and time dependent fashion judging from results after immunoprecipitation of metabolically labeled PAI-1. The t-PA effect was not attributable simply to release of stored or matrix-bound PAI-1. In HUVEC, Northern blot analyses indicated that t-PA increased steady-state levels of PAI-1 mRNA two-fold. In contrast PAI-1 mRNA expression was not increased in Hep G2 cells. Thus, mechanisms of stimulation appeared to differ in the two cell lines. The results obtained are consistent with the hypothesis that increased PAI-1 synthesis and secretion in response to t-PA may limit or attenuate fibrinolysis locally or systemically in vivo.

Cells, Cultured↗
PubMed1989

Potential attenuation of fibrinolysis by growth factors released from platelets and their pharmacologic implications.

Increased concentrations of the fast-acting tissue-type plasminogen activator (t-PA) inhibitor attenuate the fibrinolytic activity of pharmacologically administered activators of the fibrinolytic system such as t-PA. Accordingly, it was hypothesized that augmentation of synthesis and elaboration of inhibitor from the liver, leading to increased concentrations of inhibitor in plasma, or from endothelial cells in the vicinity of thrombi undergoing lysis, leading to increased concentrations locally, may contribute to failure of pharmacologically induced thrombolysis or to early reocclusion. Because platelets are rich in transforming growth factor beta and epidermal growth factor-like activity, it was thought that release of growth factors from platelets activated in vivo could mediate increases of the inhibitor in plasma by stimulating its formation in the liver and its local release from endothelial cells in the vicinity of thrombi. If so, fibrinolysis might be rendered more effective by concomitant prevention of platelet growth factor release. Transforming growth factor beta, a major constituent of platelets, increased concentrations of the t-PA inhibitor messenger ribonucleic acid (mRNA) in human hepatoma cells in a specific and dose-dependent manner. A peak effect was seen with 5 ng/ml and a 10-fold increase in 6 hours. Release of inhibitor protein into conditioned media increased as well. Induction of the inhibitor mRNA increase was elicited by exposure as brief as 30 minutes. Cycloheximide, an inhibitor of protein synthesis, was not inhibitory. The mechanisms responsible differed from those seen with epidermal growth factor, shown previously in the laboratory to increase inhibitor mRNA. In addition, the 2 factors were synergistic. Platelet lysates elicited effects simulating those of the purified growth factors.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1983

Hypothalamo-adenohypophyseal-thyroid interrelationships in the chick embryo. IV. Immunocytochemical demonstration of TSH in the hypophyseal pars distalis.

The cells that synthesize thyroid-stimulating hormone (TSH) in the pars distalis of the chick embryo were identified immunocytochemically (immunoperoxidase and immunofluorescence) using anti-bovine TSH-beta and anti-human TSH-beta sera. TSH cells are first demonstrable on Day 6.5 of incubation. By Day 11.5, when the two lobes (rostral and caudal) of the pars distalis are easily recognized, TSH cells are confined exclusively to the rostral lobe. TSH cells identified by means of immunofluorescence were stained with the periodic acid-Schiff component of the performic acid-Alcian blue periodic acid-Schiff's Orange G stain. Immunoreactive TSH cells in the pares distales of Day 13.5 chick embryos, injected at 5.5 days of incubation with thiourea, were more intensively stained than their normal counterparts. The marked change in immunocytochemically demonstrable TSH on Day 11.5 corresponds with physiological and morphological events occurring within the hypothalamus, adenohypophysis, and the thyroid gland of the developing chick during this midincubational (midgestational) period. The data suggest that not only is hypophyseal TSH present in greater quantities after Day 10.5, but that adenohypophyseal synthesis and secretion of TSH may be stimulated by another factor (hypothalamic TRH) at this time, signaling functional maturation of the hypothalamo-adenohypophyseal-thyroid axis.

Animals↗
PubMedDate not supplied

Succinylcholine sensitivity and plasma cholinesterase deficiency.

Cholinesterase deficiency is a relatively rare condition. However, if unrecognized, this condition can be potentially fatal. The authors present a case report of cholinesterase deficiency and a review of the literature. Discussion of the preoperative evaluation and preventive measures is also included.

Adolescent↗