Biomedical subjects

S Chien

Publications and source records attributed to S Chien.

At least 73 records · Page 4Linked to original sources

PubMed1997

Antiapoptotic compound to enhance hypothermic liver preservation.

BACKGROUND: Apoptosis (programmed cell death) occurs as a consequence of global organ ischemia during isolation and storage prior to transplantation. If apoptosis is inhibited during ischemia, organ preservation should be improved, and the length of time for permissible storage may be increased. The objective of this study was to test the effect of a newly developed antiapoptotic compound, LXR-015, during extended hypothermic liver preservation. METHODS: Three groups of 12 rats each were studied. In the normal group, liver function was studied immediately after harvesting. In the study group, harvested livers were flushed with Euro-Collins solution (30 ml/kg body weight) containing LXR-015 at a concentration equivalent to 9 mg/kg animal body weight (300 microg/ml). The livers were then stored at 4 degrees C for 24 hr before liver function was studied. In the control group, harvested livers were flushed with Euro-Collins solution without LXR-015 and then stored at 4 degrees C for 24 hr before liver function was studied. RESULTS: Portal venous flow was higher (P<0.05) in the normal and study groups compared with the control group. Portal venous resistance was lower (P<0.05) in the normal and study groups compared with the control group. Liver tissue oxygen consumption in the study group was significantly higher than in both the normal and control groups (P<0.05). Liver enzyme production (aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, creatine kinase) was higher in the control group than in either the study or normal group (P<0.05). Bile production in both the normal and study groups was higher than in the control group (P<0.05). The liver tissue wet to dry weight ratio in both the normal and study groups was lower than in the control group (P<0.05). Histopathology studies revealed fewer apoptotic bodies (P<0.05) in both the normal (1.70+/-0.15 per high-power field) and study groups (2.08+/-0.10 per high-power field) than in the control group (7.92+/-.33 per high-power field). CONCLUSIONS: Adding an antiapoptotic compound, LXR-015, to Euro-Collins solution significantly improves hypothermic preservation of the rat liver compared with Euro-Collins solution alone.

Alanine Transaminase↗

PubMed1997

Shear stress induction of the tissue factor gene.

Using flow channel, we report that the application of a laminar shear stress induced a transient increase of tissue factor (TF) procoagulant activity in human umbilical vein endothelial cells (HUVEC), which was accompanied by a rapid and transient induction of the TF mRNA in the HUVEC. Functional analysis of the 2.2 kb TF 5' promoter indicated that a GC-rich region containing three copies each of the EGR-1 and Sp1 sites was required for induction. Mutation of the Sp1 sites, but not the EGR-1 sites, attenuated the response of TF promoter to shear stress. Thus, Sp1 is a newly defined shear stress responsive element. Electrophoretic mobility shift assays showed there was no increase in binding of nuclear extracts from sheared cells to an Sp1 consensus site. In contrast, immunoblotting of these nuclear extracts with antibody against transcription factor Sp1 demonstrated that shear stress increased the phosphorylation of Sp1. We also showed that shear stress, like the phosphatase inhibitor okadaic acid, increased the transcriptional activity of Sp1. These findings suggest that the shear stress induction of TF gene expression is mediated through an increased Sp1 transcriptional activity with a concomitant hyperphosphorylation of Sp1.

Endothelium, Vascular↗

PubMed1997

Micropipette aspiration of human erythrocytes induces echinocytes via membrane phospholipid translocation.

When a discocytic erythrocyte (RBC) was partially aspirated into a 1.5-microns glass pipette with a high negative aspiration pressure (delta P = -3.9 kPa), held in the pipette for 30 s (holding time, th), and then released, it underwent a discocyte-echinocyte shape transformation. The degree of shape transformation increased with an increase in th. The echinocytes recovered spontaneously to discocytes in approximately 10 min, and there was no significant difference in recovery time at 20.9 degrees C, 29.5 degrees C, and 37.4 degrees C, respectively. At 11 degrees C the recovery time was significantly elevated to 40.1 +/- 6.7 min. At 20.9 degrees C the shape recovery time varied directly with the isotropic RBC tension induced by the pipetting. Sodium orthovanadate (vanadate, 200 microM), which inhibits the phospholipid translocase, blocks the shape recovery. Chlorpromazine (CP, 25 microM) reversed the pipette-induced echinocytic shape to discocytic in < 2 min, and the RBC became a spherostomatocyte-II after another 30 min. It was hypothesized that the increase in cytosolic pressure during the pipette aspiration induced an isotropic tension in the RBC membrane followed by a net inside-to-outside membrane lipid translocation. After a sudden release of the aspiration pressure the cytosolic pressure and the membrane tension normalized immediately, but the translocated phospholipids remained temporarily "trapped" in the outer layer, causing an area excess and hence the echinocytic shape. The phospholipid translocase activity, when not inhibited by vanadate, caused a gradual return of the translocated phospholipids to the inner layer, and the RBC shape recovered with time.

Blood Specimen Collection↗

PubMed1997

Influence of network topology on the elasticity of the red blood cell membrane skeleton.

A finite-element network model is used to investigate the influence of the topology of the red blood cell membrane skeleton on its macroscopic mechanical properties. Network topology is characterized by the number of spectrin oligomers per actin junction (phi a) and the number of spectrin dimers per self-association junction (phi s). If it is assumed that all associated spectrin is in tetrameric form, with six tetramers per actin junction (i.e., phi a = 6.0 and phi s = 2.0), then the topology of the skeleton may be modeled by a random Delaunay triangular network. Recent images of the RBC membrane skeleton suggest that the values for these topological parameters are in the range of 4.2 < phi a < 5.5 and 2.1 < phi s < 2.3. Model networks that simulate these realistic topologies exhibit values of the shear modulus that vary by more than an order of magnitude relative to triangular networks. This indicates that networks with relatively sparse nontriangular topologies may be needed to model the RBC membrane skeleton accurately. The model is also used to simulate skeletal alterations associated with hereditary spherocytosis and Southeast Asian ovalocytosis.

Actins↗

PubMed1997

Relationship between endothelial cell turnover and permeability to horseradish peroxidase.

The quantitative relations between cell turnover (cell mitosis and death) and macromolecular leakage were studied at the level of individual endothelial cells (ECs) in the thoracic aortae of 32 adult male Sprague-Dawley rats. The experiments were performed on en face preparations of aortic specimens obtained 1, 3, 5 or 10 min after the intravenous administration of horseradish peroxidase (HRP). Mitotic ECs were identified by hematoxylin nuclear staining; dying or dead ECs containing cytoplasmic immunoglobulin G were detected by indirect immunocytochemistry and endothelial leakages to HRP were visualized by light microscopy. The number and size of HRP spots increased with time and the spots fused to form large brown areas in 10 min. Quantitative data on the contributions of EC mitosis and EC death to the transendothelial leakages of HRP were obtained in the same animals. Although mitotic ECs (0.01%) and dying ECs (0.1%) were infrequent in occurrence, the great majority (over 90%) of these ECs were associated with focal HRP uptake. These mitotic and dying ECs, however, accounted for only 17% of the total leakage sites indicating that significant leakage of the 4-5 nm HRP also occurs in normal ECs not morphologically identified as being in mitosis or death. The percentages of leaky spots attributable to mitosis or cell death were greater for the 6 nm albumin and the 22 nm low density lipoprotein (LDL) which probably cannot traverse the normal junctions and use the leaky junctions during cell turnover as the major pathway.

Animals↗

PubMed1997

Role of integrins in cellular responses to mechanical stress and adhesion.

Mechanical stresses are important environmental cues for both normal cellular functions and pathophysiological changes in conditions such as cardiac hypertrophy and atherosclerosis. There is increasing evidence that mechanotransduction processes in response to mechanical stresses share many common features with processes in cell adhesion, such as an increase in tyrosine phosphorylation of proteins in the focal adhesion sites. Recent findings suggest that integrins may function as mechanotransducers in cells.

Animals↗

PubMed1997

Dietary methods research in the third National Health and Nutrition Examination Survey: underreporting of energy intake.

Assessment of diet is a critical component of the third National Health and Nutrition Examination Survey (NHANES III), which was designed to describe the health and nutritional status of the US population. We analyzed data collected with the primary dietary assessment instrument used in NHANES III, the 24-h recall, for 7769 nonpregnant adults aged > or = 20 y to investigate underreporting of total energy intake. Underreporting was addressed by computing a ratio of energy intake (EI) to estimated basal metabolic rate (BMRest). EI:BMRest was 1.47 for men and 1.26 for nonpregnant women; a population level of 1.55 is expected for a sedentary population. About 18% of the men and 28% of the women were classified as underreporters. Underreporting of energy intake was highest in women and persons who were older, overweight, or trying to lose weight. Underreporting varied according to smoking status, level of education, physical activity, and the day of the week the 24-h recall covered. Additionally, underreporting was associated with diets lower in fat (P < 0.01) and alcohol (P < 0.01 in women) when expressed as a percentage of total energy intake.

Adult↗

PubMed1997

Morphometry and strain distribution in guinea pig duodenum with reference to the zero-stress state.

The aim of the present study is to determine the distribution of residual circumferential strains along the duodenum in anesthetized guinea pigs. A silicone elastomer was allowed to harden in the duodenal lumen under a pressure of 0.7 kPa. The duodenum was excised with the cast and photographed. The zero-stress state was obtained by cutting rings of duodenum radially. The geometric configuration at the zero-stress state is of fundamental importance, because it is the basic state with respect to which the physical stresses and strains are defined. A basic piece of information is the way the tangent vector rotates from one end of the circumference to the other. In the duodenum at zero-stress state, the total rotation of the tangent from one tip to the other is -500 to -850 , with the lowest absolute value in the proximal duodenum. In other words, the duodenum usually turns itself inside out on changing from a loaded state to the zero-stress state. The serosal circumference, the duodenal wall thickness, and the ratio of wall thickness to mucosal circumference decreased in the distal direction. In the pressurized state, the serosal Cauchy strain was tensile and increased in the distal direction; the mucosal Cauchy strain was compressive in the proximal half of the duodenum and tensile in the distal half. The large circumferential residual strains must be taken into account in a study of physiological problems in which the stresses and strains are important, e.g., the bolus transport function.

Animals↗

PubMed1997

A model for the initiation and growth of extracellular lipid liposomes in arterial intima.

There is considerable evidence that lipoprotein cholesterol, after crossing the arterial endothelium and entering the intima from the vascular lumen, lodges in extracellular lipid packets (labeled "liposomes") bound to the extracellular matrix. These liposomes appear to form by occasional attachment of a low-density lipoprotein (LDL) to the intimal matrix and to grow in place mainly by appending available free LDL. The liposome size distributions observed in chronically hypercholesteremic (WHHL) and in short-term cholesterol-fed rabbits are quite different. We propose a hierarchy of simple nucleation-polymerization models to describe liposome formation and growth. Even the simplest of these (with only one adjustable parameter) agrees extremely well with the WHHL data. In contrast, the cholesterol-fed rabbit data seem to result from the short-term nonuniform intimal history of LDL supply, which is a consequence of the focal nature of the transendothelial LDL flow through isolated transient leakyjunctions. The same models used for the WHHL data, together with this intimal nonuniformity, superimposed on a slow uniform transendothelial seepage also account very well for this cholesterol-fed rabbit data.

Animals↗

PubMed1997

A fiber matrix model for the filtration through fenestral pores in a compressible arterial intima.

We advance a new hypothesis to explain the changes in hydraulic conductivity of an intact artery wall with transmural pressure previously observed by Tedgui and Lever [Am. J. Physiol. 247 (Heart Circ. Physiol. 16): H784-H791, 1984] and Baldwin and Wilson [Am. J. Physiol. 264 (Heart Circ. Physiol. 33): H26-H32, 1993]. This hypothesis suggests that compaction due to pressure loading of the proteoglycan matrix in the arterial intima near fenestral pores of the internal elastic lamina (IEL) leads to a narrowing of the pore entrance area and a large decrease in local intrinsic Darcy permeability of the matrix. To quantitatively assess the feasibility of this mechanism, a local two-dimensional model is proposed to study filtration flow in the vicinity of fenestral pores in a compressible intima. Using a heterogenous fiber matrix theory, we first predict the change in Darcy permeability with intimal thickness (Li). The model then calculates local velocity profiles and pressure distributions in the intima and media. The results show a marked nonlinear steepening of intimal pressure profiles near fenestral pores when the intima thins at higher luminal pressures. The predicted relative change in resistances of the IEL (with intima, R(I)) and of the media (Rm) shows a steep increase in R(I)/Rm when Li is <20% of its unstressed value. Numerical results also suggest that intimal compression has a limiting behavior in which the much stiffer collagen fibrils inhibit further compaction at high pressures after the proteoglycan matrix is maximally compressed. Predictions are also presented to show how different transmural pressures alter growth of an intimal horseradish peroxidase spot that derives from a localized (a single cell's boundary) endothelial leakage. Such a prediction is amenable to experimental verification.

Animals↗

PubMed1997

Effects of shear stress on protein kinase C distribution in endothelial cells.

We studied the effects of shear stress on protein kinase C (PKC) in cultured human umbilical vein endothelial cells by use of a flow channel and a monoclonal antibody (MAb 1.3) that recognizes the PKC beta-isozyme. The fluorescence intensity (FI) of the secondary antibody, crystalline tetramethylrhodamine isothiocyanate, was determined by image analysis. The results on each of five shearing experiments were normalized by using the paired stationary control. After 30-min shearing at 2 N/m2, FI per cell increased to 1.6 times that of control, as did the mean FI per unit cell area. The FI per unit stained area and the stained area/cell area ratio were also increased significantly by shearing. The distribution of immunostaining in each cell was determined for its cortical, cytoplasmic, perinuclear, and nuclear regions. The normalized FI per unit area in all four regions and the stained area/cell area ratio in cortical and cytoplasmic regions were significantly higher in the sheared cells than in control; the increases were greatest in the cortical area. Double staining with rhodamine-phalloidin and MAb 1.3 showed the association of actin with the PKC isozyme in both stationary and sheared cells.

Actins↗

PubMed1997

Canine lung transplantation after more than twenty-four hours of normothermic preservation.

BACKGROUND: Consistent clinical results have not been achieved when lung preservation times exceed 6 hours. The aim of this study was to use an alternative normothermic autoperfusion technique for lung preservation and transplantation. METHODS: In six paired dogs, donor lungs were removed, along with the heart, liver, pancreas, duodenum, and both kidneys, and were preserved for 24 to 33 hours in a normothermic autoperfused multiple organ block. Orthotopic left lung transplantation was performed at the end of the preservation period. RESULTS: Lung function was good during the preservation period. With a gas mixture of 50% O2 + 3% CO2 + 47% N2 delivered to the multiorgan block, arterial oxygen tension ranged from 331 +/- 19 to 383 +/- 8 mm Hg; carbon dioxide tension ranged from 18 +/- 5 to 32 +/- 5 mm Hg; and pH ranged from 7.36 +/- 0.02 to 7.45 +/- 0.08. After transplantation, the dogs were kept anesthetized and ventilated for 24 hours with the same gas mixture. The opposite pulmonary artery was occluded 0 to 6 hours after transplantation. Arterial blood pressures were stable after surgery. Arterial oxygen tension was maintained between 205 +/- 39 and 320 +/- 57 mm Hg, and arterial carbon dioxide tension was maintained between 23 +/- 2 and 34 +/- 2 mm Hg. Lung tissue wet/dry weight ratio was 4.94 +/- 0.17 after preservation; this ratio did not differ from that found in normal controls (4.91 +/- 0.10). CONCLUSIONS: This study shows that the lungs were well preserved for more than 24 hours of preservation when the normothermic multiorgan block preparation was used. The transplanted left lung was able to support the anesthetized dog after the opposite pulmonary artery was occluded.

Animals↗

PubMed1996

An elastic network model based on the structure of the red blood cell membrane skeleton.

A finite element network model has been developed to predict the macroscopic elastic shear modulus and the area expansion modulus of the red blood cell (RBC) membrane skeleton on the basis of its microstructure. The topological organization of connections between spectrin molecules is represented by the edges of a random Delaunay triangulation, and the elasticity of an individual spectrin molecule is represented by the spring constant, K, for a linear spring element. The model network is subjected to deformations by prescribing nodal displacements on the boundary. The positions of internal nodes are computed by the finite element program. The average response of the network is used to compute the shear modulus (mu) and area expansion modulus (kappa) for the corresponding effective continuum. For networks with a moderate degree of randomness, this model predicts mu/K = 0.45 and kappa/K = 0.90 in small deformations. These results are consistent with previous computational models and experimental estimates of the ratio mu/kappa. This model also predicts that the elastic moduli vary by 20% or more in networks with varying degrees of randomness. In large deformations, mu increases as a cubic function of the extension ratio lambda 1, with mu/K = 0.62 when lambda 1 = 1.5.

Biophysical Phenomena↗

PubMed1996

delta Opioid extends hypothermic preservation time of the lung.

To test the hypothesis that a delta opioid, DADLE ([D-Ala2, D-Leu5]-enkephalin), could protect tissue from ischemic damage during hypothermic lung preservation, we studied three groups of rats. In group 1 (n = 8), lung function was studied immediately after harvesting. In group 2 (n = 8), the lung was flushed with 4 degrees C Euro-Collins solution and preserved for 24 hours. In group 3 (n = 8), the lung was flushed with 4 degrees C Euro-Collins solution plus DADLE (1 mg/kg) and preserved for 24 hours. Lung function was studied by using a living rat perfusion model. Venous blood from the host rat perfused the pulmonary artery of the isolated lung. Blood from the isolated lung was returned to the carotid artery of the host rat with a roller pump. Severe pulmonary edema, hemorrhage, and occlusive pulmonary artery resistance occurred in group 2 within 30 minutes of perfusion. Perfusion studies were carried out for more than 60 minutes in groups 1 and 3. Pulmonary blood flow was lower in group 2 than in either group 1 or group 3. Pulmonary vascular resistance was much higher in group 2 than in groups 1 and 3 (p < 0.05). Airway pressure and airway resistance were much higher in group 2 than in groups 1 and 3 (p < 0.05). Airway resistance was also higher in group 3 than in group 1 after 20 minutes of perfusion (p < 0.05). Oxygen tensions from the pulmonary vein of the isolated lung in group 2 were lower than those in groups 1 and 3 (p < 0.05). Alveolar-arterial oxygen difference was much higher in group 2 than in groups 1 and 3 (p < 0.05). Lung tissue wet/dry weight ratio after perfusion was much higher in group 2 than in groups 1 and 3. The results clearly show, for the first time, that DADLE can effectively enhance hypothermic lung preservation in rats.

Airway Resistance↗

PubMed1996

Effect of verapamil on renal haemodynamics in a portal hypertensive rat model.

In order to investigate the effects of verapamil on renal haemodynamics in rats with portal hypertension, verapamil was given at either a low (0.2 mg/kg) or high (2 mg/kg) dose to rats after portal vein ligation. An approximate 12% decrease in mean arterial pressure followed administration of low dose verapamil, with a significant decrease in cardiac output and renal blood flow, as well as reduced portal pressure, observed; these signs were all indicative of a rise in renal vascular resistance. In contrast, the marked fall in both mean arterial pressure and cardiac output with high dose verapamil, accompanied by a significant decrease in portal pressure and no change in renal blood flow, suggests a reduction in renal vascular resistance. This study shows that the acute effects of verapamil on renal haemodynamics may vary with the dose used. Also, acute verapamil administration tends to decrease renal blood flow to alter the autoregulation of the kidney; thus, caution should be taken in the clinical use of verapamil in the treatment of cirrhosis with portal hypertension.

Animals↗

PubMed1996

The Ras-JNK pathway is involved in shear-induced gene expression.

Hemodynamic forces play a key role in inducing atherosclerosis-implicated gene expression in vascular endothelial cells. To elucidate the signal transduction pathway leading to such gene expression, we studied the effects of fluid shearing on the activities of upstream signaling molecules. Fluid shearing (shear stress, 12 dynes/cm2 [1 dyne = 10(-5)N]) induced a transient and rapid activation of p21ras and preferentially activated c-Jun NH2 terminal kinases (JNK1 and JNK2) over extracellular signal-regulated kinases (ERK-1 and ERK-2). Cotransfection of RasN17, a dominant negative mutant of Ha-Ras, attenuated the shear-activated JNK and luciferase reporters driven by 12-O-tetradecanoylphorbol-13-acetate-responsive elements. JNK(K-R) and MEKK(K-M), the respective catalytically inactive mutants of JNK1 and MEKK, also partially inhibited the shear-induced luciferase reporters. In contrast, Raf301, ERK(K71R), and ERK(K52R), the dominant negative mutants of Raf-1, ERK-1, and ERK-2, respectively, had little effect on the activities of these reporters. The activation of JNK was also correlated with increased c-Jun transcriptional activity, which was attenuated by a negative mutant of Son of sevenless. Thus, mechanical stimulation exerted by fluid shearing activates primarily the Ras-MEKK-JNK pathway in inducing endothelial gene expression.

Animals↗

PubMed1996

Fluid shear stress induces the phosphorylation of small heat shock proteins in vascular endothelial cells.

The small molecular mass heat shock protein of 27 kDa (HSP27) has been shown to influence actin filament dynamics and endothelial cell behavior in ways similar to those observed during laminar flow. We have employed human umbilical vein endothelial cells to determine whether fluid shear stress affects HSP27 expression or phosphorylation. After a shear stress of 16 dyn/cm2, HSP27 became more highly phosphorylated, with maximum increase in phosphorylation levels (3-fold) attained by 30 min and sustained for at least 20 h. HSP27 antigen levels did not change; however, HSP27 mRNA levels decreased by 20% after 16 h. In bovine aortic endothelial cells stably transfected with the wild-type human HSP27 gene, shear stress induced the phosphorylation of both the exogenous human HSP27 and the endogenous bovine HSP25. The product of a transfected mutant HSP27 gene in which the putative phosphorylation sites Ser-15, Ser-78, and Ser-82 had been replaced with Gly was not phosphorylated. Thus the modulation of HSP27 and its activity by shear stress is mediated through a posttranslational mechanism and differs from the shear stress induction of immediate early genes at the level of transcription.

Animals↗

PubMed1996

A systematic study of hypothermic lung preservation solutions: Euro-Collins solution.

BACKGROUND: Testing lung function after preservation is difficult because a suitable model is still lacking; thus, the effectiveness of different solutions for lung preservation has not been confirmed. This study tested the effectiveness of Euro-Collins solution alone for hypothermic preservation of rat lungs. METHODS: A living rat perfusion model was used, which allowed more than 5 hours of continuous perfusion for isolated lung function studies. Group 1 lungs (control, n = 8) were tested without preservation. In groups 2 through 6 (n = 8 lungs each), the lungs were flushed with 4 degrees C Euro-Collins solution and preserved for 4, 6, 8, 12, and 24 hours, respectively. Lung function studies were carried out after preservation. RESULTS: In groups 1 and 2, pulmonary arterial blood flow and pulmonary venous oxygen tension were higher and pulmonary resistance was lower than in the other groups. Airway pressure and resistance were lowest in group 1. Lungs in groups 5 and 6 demonstrated the worst function, but the lung tissue wet to dry ratio was higher only in group 6. CONCLUSIONS: At 4 degrees C, Euro-Collins solution can effectively preserve rat lungs for 4 hours. Six to 8 hours of preservation resulted in depressed lung function. More than 12 hours of preservation resulted in uniformly deficient lung function, rendering the lungs unsuitable for transplantation.

Airway Resistance↗