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Biomedical subjects

K Tran

Publications and source records attributed to K Tran.

At least 55 records · Page 3Linked to original sources
PubMed1995

The enhancement of phosphatidylcholine biosynthesis by angiotensin II in H9c2 cells.

The effect of angiotensin II on the biosynthesis of phosphatidylcholine in rat heart myoblastic (H9c2) cells was investigated. Cells were incubated with [methyl-3H]choline, and the labelling of phosphatidylcholine at different time intervals was examined. When cells were pretreated with angiotensin II, a significant increase in the labelling of phosphatidylcholine was observed. Analysis of the labelled phosphatidylcholine precursors indicated that the conversion of phosphocholine to CDP-choline was enhanced by angiotensin II treatment. Determination of enzyme activities in the CDP-choline pathway revealed that the activities of choline kinase or CDP-choline: diacylglycerol cholinephosphotransferase were not changed, but the activities of CTP:phosphocholine cytidylyltransferase were stimulated in both the particulate and soluble fractions. The stimulation of the cytidylyltransferase by angiotensin II was not abolished by okadaic acid, indicating that the activation of the enzyme was not mediated via the okadaic-sensitive dephosphorylation mechanism. Alternatively, the stimulation of the cytidylyltransferase activity was completely abolished by protein kinase C inhibitors. Immunoblotting studies revealed that levels of the cytidylyltransferase in the soluble and particulate fractions were not affected by angiotensin II treatment. We conclude that the increase in phosphatidylcholine biosynthesis by angiotensin II was a direct result of the enhancement of the cytidylyltransferase activity. The enhancement of enzyme activity was not mediated via enzyme translocation, but by a mechanism which was intimately associated with the protein kinase C cascade.

Angiotensin II↗
PubMed1995

Enhancement of phospholipid hydrolysis in vasopressin-stimulated BHK-21 and H9c2 cells.

The hydrolysis of phospholipids in vasopressin-stimulated baby hamster kidney (BHK)-21 and H9c2 myoblastic cells was investigated. Phosphatidylcholine and phosphatidylethanolamine in these cells were pulse labelled with [3H]glycerol, [3H]myristate, [3H]choline or [3H]ethanolamine, and chased with the non-labelled precursor until linear turnover rates were obtained. When cells labelled with [3H]glycerol or [3H]myristate were stimulated by vasopressin, no significant decrease in the labelling of phosphatidylcholine was detected, but the labelling of phosphatidic acid was elevated. However, the labellings of phosphatidylethanolamine and its hydrolytic product were not affected by vasopressin stimulation. When the cells were pulse labelled with [3H]-choline, vasopressin stimulation caused a decrease in the labelled phosphatidylcholine with a corresponding increase in the labelled choline. The apparent discrepancy between the two types of labelling might be explained by the recycling of labelled phosphatidic acid back into phosphatidylcholine, thus masking the reduction in the labelled phospholipid during vasopressin stimulation. Alternatively, the labelled choline produced by vasopressin stimulation was released into the medium, thus reducing the recycling of label precursor back into the phospholipid and making the decrease in the labelling of phosphatidylcholine readily detectable. Further studies revealed that vasopressin treatment caused an enhancement of phospholipase D activity in these cells. The presence of substrate-specific phospholipase D isoforms in mammalian tissues led us to postulate that the differential stimulation of phospholipid hydrolysis by vasopressin was caused by the enhancement of a phosphatidylcholine-specific phospholipase D in both BHK-21 and the H9c2 cells.

Animals↗
PubMed1994

Pain management for elderly patients with cancer at home.

BACKGROUND: Pain is an important problem for patients with cancer and is particularly important for elderly patients with cancer and their family care givers. Increasingly, cancer is managed on an outpatient basis with pain management responsibility assumed by the family at home. This study evaluated a structured pain education program that included three components: basic pain management principles and assessment, pharmacologic interventions, and nondrug treatments. METHODS: The pain education intervention was implemented across three home visits with two points of follow-up evaluation. Outcomes of the 66 elderly patients with cancer completing the educational program included measures of quality of life, patient knowledge and attitudes regarding pain, and use of a self-care log to document drug and nondrug interventions and their effectiveness. RESULTS: Repeated measurement analysis was used to evaluate the outcomes of the three-part education intervention. Results indicate an improvement in knowledge and attitudes regarding pain as well as the use of drug and nondrug interventions. Outcomes of the quality of life instrument suggest significant effect of pain on all aspects of quality of life, including physical well being, psychological well being, social concerns, and spiritual well being. CONCLUSIONS: The investigators concluded that the pain education intervention provided important support to elderly patients with cancer and family members at home. Structured pain education based on an evolving science of pain relief should become a part of the standard health care for pain management. Improved pain management includes quality of life for the elderly patient with cancer as well as for family care givers.

Aged↗
PubMed1994

Vitamin E suppresses diacylglycerol (DAG) level in thrombin-stimulated endothelial cells through an increase of DAG kinase activity.

The present study has examined the role of vitamin E, a natural lipid antioxidant, in the production of diacylglycerol (DAG) and phosphatidic acid (PA) in thrombin-stimulated human endothelial cells. Cells were labelled with [3H]myristate and the incorporation and distribution of [3H]myristate into cellular lipids was not affected by vitamin E. However, in response to thrombin stimulation, considerably more PA and less DAG were formed in cells enriched with vitamin E. The time-course of thrombin stimulation indicated that vitamin E attenuated the accumulation of sustained DAG levels with a concomitant increase in PA. Direct determination of DAG mass further confirmed that vitamin E suppresses the accumulation of DAG induced by thrombin. In the presence of ethanol, the formation of [3H]phosphatidylethanol (PEt) in [3H]myristate-labelled cells stimulated by thrombin was unaffected by vitamin E enrichment. DL-Propranolol, a PA phosphohydrolase inhibitor, caused an accumulation of PA, without affecting DAG formation in either vitamin E-treated and untreated cells. This indicated that the increase in PA and decrease in DAG in vitamin E-treated cells was not due to a stimulation of phospholipase D or an inhibition of PA phosphohydrolase. Determination of inositol phosphates formation in response to thrombin showed that the change of DAG levels elicited by vitamin E was independent of phospholipase C-induced hydrolysis of inositol phospholipids. In contrast, analysis of DAG kinase activity revealed that vitamin E enrichment enhanced the activity of the enzyme in both basal and thrombin-stimulated cells. Taken together, these data indicated that vitamin E caused an increased conversion of DAG to PA by activating DAG kinase activity without causing any change in the activities of phospholipase D, PA phosphohydrolase or phospholipase C.

Cells, Cultured↗
PubMed1994

Vitamin E enhances the acylation of 1-O-alkyl-sn-glycero-3-phosphocholine in human endothelial cells.

1-O-Alkyl-2-acyl-sn-glycero-3-phosphocholine (alkylacyl-GPC) is the precursor of platelet-activating factor. It is formed via the CoA-independent transacylase reaction, which transfers the polyenoyl acyl group from the sn-2 position of a diacyl phospholipid to the sn-2 position of 1-O-alkyl-sn-glycero-3-phosphocholine (alkyl-GPC). We have reported previously that vitamin E alters phospholipid turnover in the endothelial cells by increasing arachidonic acid release and prostacyclin synthesis. In the present study, the role of vitamin E in the formation of alkylacyl-GPC was investigated. Incubation of endothelial cells with vitamin E resulted in an increase in the formation of [3H]alkylacyl-GPC from [3H]alkyl-GPC. The effect of vitamin E was dose-dependent at concentrations below 23 microM. However, vitamin E did not have a direct effect on the transacylase activity. When endothelial cells were incubated with vitamin E, the CoA-independent transacylase activity in the cell homogenate was found to be enhanced. Kinetic analysis of the transacylase activity in the pre-incubated cells showed that the enhancement of enzyme activity was at the enzyme-substrate level. When endothelial cells were incubated with vitamin E analogues (Trolox, tocol and tocopherol acetate), only limited enhancement of the transacylation process was detected. It is clear that vitamin E enhanced the synthesis of alkylacyl-GPC from alkyl-GPC in a very specific manner by an indirect stimulation of the CoA-independent transacylase activity. The regulation by vitamin E of the formation of alkylacyl-GPC may mediate the transfer of arachidonate from the diacyl phospholipid pool into the ether-linked phospholipid pool.

Acetyl-CoA C-Acyltransferase↗
PubMed1994

Regulation of lens beta-adrenergic receptors by receptor occupancy and dexamethasone.

Beta-adrenergic binding sites in primary cultures of chick lens annular pad (CLAP) cells were characterized with dihydroalprenolol (DHAP). Binding site affinities and densities were similar to beta-adrenergic receptors (BARs) previously characterized on crude membranes from freshly isolated cells. In competitive displacement studies, the beta-blocker propranolol was shown to increase the number of available binding sites in a concentration dependent manner. Acute exposure of CLAP cells to propranolol prior to DHAP binding also resulted in an increase in the number of available binding sites. Finally, lens beta-adrenergic binding site levels could be modulated by dexamethasone treatment. These results indicate that lens BARs are subject to common regulatory mechanisms and further implicate ophthalmic pharmaceuticals as possible cataractogenic agents.

Animals↗
PubMed1994

Loss of red blood cell viability associated with limited thermal inactivation of extracellular HIV-1.

The effects of incubation at mildly elevated temperatures on HIV-1 inactivation and in vitro red blood cell properties were investigated. Red cells (55% Hct) were leukodepleted (3 log10) by filtration, maintained at 45 or 47 degrees C for 4 or 8 h, and then stored at 4 degrees C. Hemolysis was twice that of controls after 42-day storage for samples treated for 4 h at 45 degrees C, and five times larger for samples heated at 47 degrees C. There was also a significant increase in the rate of potassium loss, an early decrease in ATP levels, and an initial drop in pH for samples treated at either temperature. Larger differences were observed for samples exposed to these elevated temperatures for 8 h. Osmotic deformability curves obtained by ektacytometry showed dramatic decreases in red cell deformability at both temperatures and for both time periods. HIV-1 inactivation in red cells treated at 45 degrees C (approximately 0.25 log10/h) was considerably less than that obtained in tissue culture medium (1-2 log10/h). Since the decrease in red cell deformability is likely to indicate reduced red cell function and survival, and the rate of HIV-1 inactivation is low, mild heat treatment is not an adequate process for viral inactivation of red cell products.

Blood↗
PubMed1994

Long-term reconstitution of mice after ex vivo expansion of bone marrow cells: differential activity of cultured bone marrow and enriched stem cell populations.

In this report, we evaluated the short-term expansion of murine bone marrow mononuclear cells (BMMNC) and enriched stem cell populations to determine the capacity of these cells for long-term rescue and engraftment to lethally irradiated recipients. In our study, nonadherent bone marrow mononuclear cell (NBM-MNC) and Thy1+Lin- stem cells populations were cultured with interleukin-3 (IL-3) or IL-3 plus stem cell factor (SCF) for periods up to 6 days. By day 6 of culture, the mononuclear cells (MNC) decreased to 6% of input cell number, whereas Thy1+Lin- cells increased by 2310%. Doses of 95,000; 100,000; 50,000; and 250,000 NBM-MNCs at 0, 1, 2, and 6 days of culture, respectively, rescued 50% of lethally irradiated mice. When 250,000 MNCs were cultured for 0, 1, 2, and 6 days, 71, 61, 100, and 50% of the animals survived lethal irradiation for greater than 24 weeks. In contrast, doses of 8,000 and 21,000 Thy1+Lin- cells cultured 0 and 1 day, respectively, yielded 50% survival rates. These same cells cultured for 6 days failed to rescue recipients even at high doses. Twenty thousand Thy1+Lin- cells cultured for 0, 1, 2, and 6 days, even in the presence of SCF, produced decreasing survival rates of 86, 43, 26, and 0%, respectively. The proliferative responses of these different populations in combination with their long-term rescue abilities indicated that the absolute number of long-term rescue units (LD50, 24 weeks) in the cultured Thy1+Lin- population decreased faster than in similarly cultured NBM-MNCs. Studies evaluating donor cell engraftment demonstrated that animals rescued with cultured Thy1+Lin- and NBM-MNCs maintained high levels of donor reconstitution [7]. The percent donor T cell engraftment did not significantly change between 2 and 17 months post-bone marrow transplantation (post-BMT). Therefore, those animals who received sufficient cells to survive lethal irradiation generally established and maintained high levels of donor engraftment. The data suggest a role for accessory cells and/or factors in the preservation of stem cell activity.

Animals↗
PubMed1993

Beta-adrenergic mechanisms affect cell division and differentiation in cultured chick lens epithelial cells.

Primary cultures of chick lens annular pad (CLAP) cells were exposed to beta-adrenergic drugs and lipid soluble cAMP analogues. In dose-dependent manners, these agents inhibited re-entry into the cell cycle when these normally post-mitotic cells were cultured in the presence of serum. The inhibition of cell division was evidenced by a lack of DNA accumulation and greatly reduced tritiated thymidine incorporation into DNA. Treated cells were also largely inhibited from spreading onto the substrate. Instead, the epithelial monolayers originally placed into culture formed nearly spherical aggregates. Treated cultures continued to synthesize crystallins and exhibited increased levels of a 49-kDa cytoskeletal protein which accumulates in fiber cells. These results indicate that beta-adrenergic mechanisms may be involved in regulating cell division and terminal differentiation of lens fiber cells.

Animals↗
PubMed1992

Comparative uptake of alpha- and gamma-tocopherol by human endothelial cells.

The intake of gamma-tocopherol by North Americans is generally higher than that of alpha-tocopherol. However, the levels of alpha-tocopherol in human blood have consistently been shown to be higher than those of gamma-tocopherol suggesting differential cellular retention of the two tocopherol forms. We sought to resolve this question by studying tocopherol metabolism by human endothelial cells in culture. The time- and dose-dependent uptake of gamma-tocopherol by endothelial cells was similar to that of alpha-tocopherol. To determine the comparative uptake between alpha- and gamma-tocopherol, we adopted two approaches in which cells were enriched with either increasing concentrations of an equimolar mixture of alpha- and gamma-tocopherol; or cells were enriched with a fixed concentration of tocopherols in which the alpha to gamma ratio was varied. Our results indicated that there was a preferential uptake of gamma-tocopherol by the cells. When cells were enriched with either alpha- or gamma-tocopherol and the disappearance of individual tocopherols was monitored over time, gamma-tocopherol exhibited a faster rate of disappearance. The faster turnover of gamma-tocopherol can explain the discrepancy between high intake and low retention of gamma-tocopherol in man.

Cells, Cultured↗
PubMed1991

Capsaicin effects on non-neuronal plasma membranes.

Capsaicin has been touted as a pharmacological tool specific for sensory afferent neurons and is widely used in neurophysiological studies. However, we have recently demonstrated that in concentrations commonly employed within the gastrointestinal tract, capsaicin inhibits platelet aggregation to at least three different stimuli. Since this was observed in a nerve free system it raised the question of how specific capsaicin is. In this communication we report that capsaicin has profound effects on physical properties of non-neuronal cell plasma membranes. These effects were observed while measuring the effect of capsaicin upon the fluidity of both intact cell membranes and a variety of purified membrane preparations. Membrane fluidity was assessed with the fluorescent probes diphenylhexatriene (DPH) and its trimethylamino derivative TMA-DPH and demonstrated concentration-dependent capsaicin effects. Furthermore, the effects were cell specific and for full expression required both intact cells and a non-lipid extractable component of the plasma membrane. These non-neuronal effects must be carefully considered when contemplating the explanation for capsaicin-induced effects.

Animals↗
PubMed1991

Regeneration of vitamin E in human platelets.

Human platelets possess active lipoxygenase and cyclooxygenase which convert arachidonic acid to (12S)-12-hydroperoxy-5,8,10,14-eicosatetraenoic acid (12-HPETE) plus (12S)-12-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE) and thromboxane B2 plus 12-hydroxy-5,8,10-heptadecatrienoic acid (HHT), respectively. When platelet homogenates were incubated with arachidonate, there was a rapid consumption of platelet tocopherol. Time course analysis revealed that within 0.5 min, over half of arachidonate and tocopherol were metabolized. Mass formation of 12-HPETE and 12-HETE or thromboxane B2 and HHT exceeded that of the mass of tocopherol oxidized. Preincubation with the lipoxygenase inhibitor 5,8,11,14-eicosatetraynoic acid (ETYA) completely abolished this arachidonate-induced tocopherol oxidation whereas cyclooxygenase inhibitors (indomethacin and aspirin) further potentiated tocopherol oxidation, indicating that this oxidation is closely linked with platelet 12-lipoxygenase activity. Incubation with lipoxygenase metabolites of arachidonic acid showed that only 12-HPETE caused a rapid tocopherol oxidation which was followed by a gradual tocopherol regeneration. By using nordihydroguaiaretic acid (NDGA), a lipoxygenase inhibitor which is also a strong reductant, over 60% of the arachidonate-induced oxidized tocopherol was regenerated. Tocopherol regeneration declined with increasing oxidation time induced by arachidonate, and after 30-60 min virtually no regeneration could be observed, suggesting that the precursor molecule was unstable. We postulate that the precursor molecule is the tocopheroxyl radical. In the presence of ETYA, a lipoxygenase inhibitor without antioxidant properties, either ascorbate or GSH provided significant tocopherol regeneration. Kinetic studies showed that tocopherol regeneration after the addition of ascorbate was essentially completed by 1 min. By contrast, GSH addition caused a steady increase in tocopherol which peaked after 10 min of its addition. To determine whether this rapid regeneration is chemical or enzymic, regeneration was studied in the presence of chloroform and methanol. Comparison of various reductants in this denaturing condition for enzymes showed that ascorbate and NDGA afforded significant regeneration whereas GSH was ineffective, indicating that there are distinct enzymic and non-enzymic mechanisms for tocopherol regeneration. This study provides direct evidence from mass analysis that tocopherol can be regenerated in human cell homogenates. This finding implies that maintenance of membrane tocopherol status may be an essential function of ascorbate and GSH which operate in concert to ensure maximum membrane protection against oxidative damage.

Arachidonate 12-Lipoxygenase↗
PubMed1990

R,R,R-alpha-tocopherol potentiates prostacyclin release in human endothelial cells. Evidence for structural specificity of the tocopherol molecule.

Human umbilical vein endothelial cells (HUVEC) in culture synthesize prostacyclin (PGI2) as the predominant metabolite of arachidonic acid which is derived from the deacylation of phospholipids. Under basal-unstimulated condition, PGI2 release from HUVEC is extremely low; however, when endothelial monolayers were preincubated with the natural vitamin E (R,R,R-alpha-tocopherol), we found a dose-dependent potentiation of basal PGI2 release. When HUVEC were stimulated with arachidonate or ionophore A23187, there was a dose-dependent increase of PGI2 release in response to tocopherol enrichment. When HUVEC were labelled with [Me-3H]choline followed by A23187 stimulation, a significantly higher lysophosphatidylcholine was found in the tocopherol-enriched cells, suggesting a change in enzymes involved in phosphatidylcholine metabolism. Analysis of these enzymes revealed that phospholipase A2 activity was enhanced by tocopherol enrichment, whereas lysophospholipase and acyl-CoA acyltransferase were unaffected. To determine the specificity of the tocopherol molecule, different analogues were tested for their PGI2 potentiating activity. Results showed that the free hydroxyl group on the chromanol ring as well as the phytyl side-chain are absolutely required to stimulate PGI2 release, whereas, different methyl locations and substituents on the chromanol ring had no effect. These studies demonstrated that tocopherol potentiates basal PGI2 release in HUVEC and in contrast to its reported inhibitory role in rat platelets, myocardium and neutrophils, tocopherol stimulates phospholipase activity in HUVEC.

Acetyl-CoA C-Acyltransferase↗
PubMed1990

The uptake of (R,R,R)alpha-tocopherol by human endothelial cells in culture.

Endothelial cells from human umbilical cord vein in culture incorporate physiological and pharmacological amounts of (R,R,R)alpha-tocopherol in a time-dependent and dose-dependent manner. Incorporated tocopherol was found to associate predominantly with membrane fractions of the cell. When expressed on the basis of organelle protein, the highest amount of tocopherol was found in plasma membrane, and decreasing amounts in mitochondria, endoplasmic reticulum and cytosol. With the relatively wide range of tocopherol concentrations used in these studies (23.2-92.8 microM), there was no apparent toxicity on the cells as judged by unaltered cell numbers and cell viability. When the cells were enriched with tocopherol and cell tocopherol levels were monitored in tocopherol-free medium, there was a rapid phase of tocopherol disappearance, which was followed by a slower phase. The half-time for the disappearance of incorporated tocopherol was found to be approximately 65 +/- 8.6 hr (mean +/- SD, n = 3). The results of this study clearly show that human endothelial cells in culture are a feasible model for the study of vitamin E uptake. The cell culture model could potentially be used to study other fat-soluble vitamins and essential nutrients.

Cell Membrane↗
PubMed1990

Beneficial long-term effects of hydralazine in aortic regurgitation.

The long-term effects of vasodilators in asymptomatic patients with aortic regurgitation have not been studied extensively. We retrospectively reviewed the echocardiograms of 19 asymptomatic patients with significant aortic regurgitation followed up annually for up to 4 years (average +/- SD, 3.1 +/- 0.7 years). Of these 19 patients, 12 were not receiving vasodilators and 7 were receiving hydralazine hydrochloride, 40 to 200 mg daily. In the patients not receiving vasodilators, left ventricular diastolic and systolic dimensions increased progressively in all patients by an average of 8% and 13%, respectively, after 3 years. In the patients receiving hydralazine, left ventricular dimensions increased by 9% and 5% in the year or more before hydralazine use and decreased by 7% and 7%, respectively, during the first year after using hydralazine. The reduction was observed in all patients during the first year, but an increase was detected in 3 patients followed up beyond that period. The results suggest that the progression of left ventricular dilatation in asymptomatic patients with aortic regurgitation can be delayed by long-term therapy with vasodilators. Pending further confirmation, such therapy may possibly influence the natural history of the disease and delay the timing of operation.

Adult↗
PubMed1989

Effects of dietary vitamin E on the biosynthesis of 5-lipoxygenase products by rat polymorphonuclear leukocytes (PMNL).

Activation of polymorphonuclear neutrophils (PMNL) leads to the release of arachidonate from cellular phospholipids via a phospholipase A2, and conversion of products of the 5-lipoxygenase pathway. Evidence to date indicates the dietary vitamin E ((R,R,R)-alpha-tocopherol) can influence both cyclooxygenase and phospholipase A2 activities and that the effect of this vitamin is cell/tissue specific. The present study was undertaken in order to examine the effects of varying dietary tocopherol on PMNL tocopherol content and 5-lipoxygenase product profile using the ionophore A23187 as stimulant in the presence and absence of exogenous arachidonate. Feeding semi-purified diets containing 0, 30 or 3000 ppm of (R,R,R)-alpha-tocopherol acetate to weanling rats for 17 weeks resulted in a dose-related enrichment of PMNL tocopherol. Stimulation of PMNL elicited a significant and rapid loss of tocopherol. When PMNL were stimulated with A23187 alone, the synthesis of 5-HETE, LTB4 and 19-hydroxy-LTB4 was decreased in proportion to increasing dietary tocopherol concentrations. However, when exogenous arachidonate was provided with A23187, intermediate amounts of dietary tocopherol (30 ppm) still suppressed the formation of 5-lipoxygenase products, but high doses (3000 ppm) did not have any additional inhibitory effect. This differential response to high concentrations of vitamin E in the presence and absence of exogenous arachidonate highly suggest that at these concentrations, tocopherol may act principally at the level of substrate release whereas at lower concentrations, 5-lipoxygenase is inhibited. Data from this study demonstrated that attenuation of the formation of 5-lipoxygenase products in PMNL can be achieved by dietary vitamin E enrichment.

Animals↗
PubMed1989

Immunoassay for estrogen receptor does not detect inactivated receptor.

Accurate quantification of estrogen receptor (ER) is essential for optimal clinical characterization of individual cases of breast cancer. If breast tumors are mishandled, the relatively labile ER protein may lose its steroid-binding capacity (become inactivated) and not be measurable by the routine steroid-binding assay. We tested whether the commercial enzyme immunoassay of Abbott Laboratories could quantify inactivated ER. Samples of powdered breast tumors from humans were exposed to various temperature and homogenization conditions known to inactivate ER, and any remaining ER was quantified by both the immunoassay and the steroid-binding assay. For all inactivation conditions tested, the two assay methods detected the same proportions of remaining ER. We conclude that the inactivation reaction for ER also alters one or both of the antigenic site(s) necessary for the immunoassay. Hence, for breast tumors mishandled to the extent of inactivating ER, the immunoassay offers no advantage over the more conventional steroid-binding assay for quantifying any remaining ER.

Breast Neoplasms↗