Inhibition of MMPs Cat G and downregulates the signaling of TGF-ß/Smad in chronic photodamaged human fibroblasts.

OBJECTIVE: To understand the action of Cathepsin G (Cat G) and matrix metalloproteases (MMPs) on the ß/Smad pathway of transforming growth factor ß (TGF-ß) in chronically photodamaged human fibroblasts. Cat G plays a significant role in the process of skin photoaging and in collagen synthesis and degradation which is induced by UV irradiation it could interact with TGF-ß/Smad signaling. No available studies have thoroughly explored its molecular mechanisms of photoaging regulation. PATIENTS AND METHODS: Fibroblasts were divided into 4 groups: (1) control, (2) UVA irradiation of 25 J/cm2, (3) UVA irradiation of 25 J/cm2 + MMPs inhibitor, and (4) 25 J/cm2 UVA irradiation + Cat G inhibitor. All treatments were repeated daily for 21 days. Western blot and ELISA was employed to detect Protein levels for Cat G, MMPs, and several smads. RESULTS: Compared to UVA-irradiated cells, the addition of MMPs inhibitor downregulated the expression of smad2, smad3, and smad4 as well as TGF-ß. The addition of Cat G inhibitor downregulated the expression of smad2, smad3, and smad4 as well as TGF-ß. These data suggest that TGF-ß/Smad signaling was decreased by inhibition of MMPs and Cat G decreased in chronically human fibroblasts which are photo-damaged. CONCLUSIONS: These results may help expand our knowledge of mechanisms mediating photoaging and is possibly instrumental to the exploration of novel anti-photoaging treatments.

Effects of dietary phospholipid level in cobia (Rachycentron canadum) larvae: growth, survival, plasma lipids and enzymes of lipid metabolism.

A study was conducted to determine the effects of dietary phospholipid (PL) levels in cobia (Rachycentron canadum) larvae with regard to growth, survival, plasma lipids and enzymes of lipid metabolism. Fish with an average weight of 0.4 g were fed diets containing four levels of PL (0, 20, 40 and 80 g kg(-1)dry matter: purity 97%) for 42 days. Final body weight (FBW), weight gain (WG) and survival ratio were highest in the 8% PL diet group and mortality was highest in PL-free diet group. We examined the activities of lipoprotein lipase (LPL) and hepatic lipase (HL) in liver, lecithin-cholesterolacyltransferase (LCAT) in plasma as well as plasma lipids and lipoprotein. LCAT activity showed a decrease of more than two-fold in PL-supplemented diet groups compared with the PL-free diet group. HL activity was highest in the 8% PL diet group and the other three groups showed no difference. LPL activity was significantly higher in the PL-supplemented diet groups than in the PL-free diet group. The dietary intervention significantly increased plasma phospholipids and total cholesterol (TC) levels, and the higher free cholesterol (FC) level contributed to the TC level. However, the fish fed PL exhibited a significantly decreased plasma triglyceride (TG) level. The lipoprotein fractions were also affected significantly by the PL. The PL-supplemented diet groups had significantly higher high-density lipoprotein (HDL) compared with the PL-free diet group, but showed a marked decrease in very low-density lipoprotein (VLDL). The results suggested that PL could modify plasma lipoprotein metabolism and lipid profile, and that the optimal dietary PL level may well exceed 80 g kg(-1) for cobia larvae according to growth and survival.

A proposal for a theory and detection method of cavitation potential of mitral bileaflet heart valve.

We believe that cavitation damage to mitral bileaflet heart valve (MBHV) is caused by microcracks of hydrophobic pyrolite carbon. Our proposal is to detect cavitation potential of MBHV with an acoustic technique under various conditions of transvalvular pressures, ventricular dp/dt, leakage flow, and gas concentration in fluid, This modality, different from recent research in theory and technique, will lead to a standardized noninvasive and sufficiently sensitive technique for the detection of MBHV, to reject those valves believed to cause cavitation damage, and to develop cavitation-resistant MBHV.

Modified Wicomb's apparatus for preservation of heart for 24 hours.

To prolong the time of heart preservation, we modified the Wicomb's perfusion apparatus, in which oxygen flow acts as the source of power and provides oxygenation for the perfused myocardium. Ten adult porcine hearts which had been preserved for 24 hours were resuscitated successfully and continued to beat steadily for more than 1.5-3 hours after reperfusion. Myocardial ultrastructure was observed at the end of preservation and 15-120 minutes after reperfusion. The damages of the myocardial ultrastructure at the end of preservation were reversible.

Effect of pulsatile reperfusion on myocardial high energy phosphates following global ischemia.

To evaluate the effects of pulsatile reperfusion (PR) on the postischemic myocardial phosphometabolites, 17 sheep were put on cardiopulmonary bypass (CPB) and randomly divided into a pulsatile group (P group) and nonpulsatile group (NP group). The heart was arrested by global ischemia for 45 minutes, then defibrillated and reperfused for 2 hours while the circulation was supported by CPB. Myocardial needle biopsies were obtained, and ATP, ADP, and AMP were measured with high performance liquid chromatography. There were no significant differences between the two groups in myocardial ADP and AMP. However, after 120 minutes of reperfusion, the myocardial ATP was restored in the P group, but continued to decrease further in the NP group. Experimental results imply that PR might reduce reperfusion injury and promote recovery of the ischemic myocardium.

Reduction of granulocyte-mediated lung injury in canine cardiopulmonary bypass by anisodamine infusion.

The effect of infusing the M-cholinergic receptor blocker anisodamine during cardiopulmonary bypass (CPB) on granulocyte-mediated lung injury was evaluated in sixteen dogs undergoing CPB for 120 min with the aorta crossclamped for 90 min. The treated dogs (n = 8) received a total dose of 15 mg/kg anisodamine hydrochloride before and during CPB whereas the controls (n = 8) received saline only. A significantly reduced sequestration of granulocytes in the lungs was found in the treated dogs as indicated by a lower right-to-left atrium granulocyte gradient 15 minutes after pulmonary recirculation during CPB (p less than 0.05) and less granulocyte accumulation in the lung capillaries after CPB, as shown by histological examination (p less than 0.01). Moreover, oxygen free radical release as indicated by plasma malondialdehyde concentrations was significantly lower (p less than 0.05) in the treated dogs than in the controls towards the end of CPB. Finally, a significantly reduced lung-water content was found 30 min after CPB in the treated dogs as compared with the controls (p less than 0.01). These results suggest that anisodamine administration in this experimental model significantly inhibits pulmonary granulocyte sequestration in CPB and the consequent lung injury induced.

Membrane oxygenator prevents lung reperfusion injury in canine cardiopulmonary bypass.

The effect of blood activation on lung reperfusion injury during cardiopulmonary bypass was investigated in 20 dogs with the use of a bubble oxygenator (n = 10) or a membrane oxygenator (n = 10). In the bubble oxygenator group, significant leukocyte and platelet right to left atrium gradients were found 15 minutes after lung reperfusion (p less than 0.05, p less than 0.01) accompanied by a sharp increase in plasma malondialdehyde concentration 5 minutes after lung reperfusion, whereas no significant right to left atrium gradient of leukocytes or platelets nor significant increase in plasma malondialdehyde concentration was observed in the membrane oxygenator group. In both the bubble oxygenator and membrane oxygenator group, similar mild to moderate lung histological changes were found before lung reperfusion. After lung reperfusion, however, more endothelial cell swelling (p less than 0.05), leukocyte (p less than 0.01) and platelet (p less than 0.01) accumulation in lung capillaries, leakage of erythrocytes into the alveolar space (p less than 0.05), and type I cell damage (p less than 0.05) were found only in the bubble oxygenator group. Eventually, a significantly higher lung water content was found in the bubble oxygenator group than in the membrane oxygenator group (p less than 0.01) after cardiopulmonary bypass. This study indicated that lung injury during cardiopulmonary bypass starts mainly after lung reperfusion, which was correlated with lung leukocyte and platelet sequestration associated with different types of oxygenators.

In vitro and in vivo evaluations of the antithrombogenic properties of the spiral vortex pump for temporary left heart bypass.

From conception to realization, the design emphasis of the spiral vortex diaphragm pump has been on promoting efficient blood flow patterns in order to lower thrombogenicity. The tracer method was used to visualize flow patterns in this pump and the results were compared with those of a conventional diaphragm pump with paraxial inlet and outlet ports. During diastole, the flow through a 45 degrees angled inlet is tangential to the axis of the pump forming a continuous vortex central along the pump axis. During systole, the vortex converges to pass through the apically located outlet. No areas of turbulence or stasis could be found, whilst in the conventional pump only random flow with recirculation could be demonstrated. Dye washout tests confirmed good washout at the periphery of this pump with no signs of stasis. However, large areas of stagnation with incomplete washout at diaphragm-housing (D-H) junctions were observed in the conventional pump. The comparative in vitro haemolysis test revealed that the level of free plasma haemoglobin was doubled in a commercially available pump compared with that of the spiral vortex pump. No thrombus formed within the pump housing after up to 50 hours of pumping in five acute animal experiments without postoperative anticoagulants. However, thrombi were found at the D-H junction of this pump due to imperfect fabrication techniques in one of the four sheep that survived 14-21 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Limitation of myocardial infarct size by a right ventricular assist device.

To evaluate the effects of an RVAD on myocardial ischemic injury during right coronary artery (RCA) ligation, a pneumatically driven pusher plate pump was inserted between the right atrium and pulmonary artery, and the RCA was ligated at its origin for two hr. There were 18 sheep, of which nine each served as the control and RVAD groups. Hemodynamic parameters and blood samples were collected from the coronary sinus. Area at risk and area of infarct were detected by gentian violet injection and triphenyltetrazolium (TTC) staining. Hemodynamic parameters (left ventricular systolic pressure (LVSP), cardiac output (CO], and creatine kinase levels in coronary sinus blood tended to be beneficial, to some degree, in the RVAD group. Electron microscopic examination revealed chromatin clumping and mitochondrial destruction, however this ultrastructural damage was more severe in the control group. The ratio of area of infarct to area at risk in the RVAD group was significantly less than that in the control group, being 20.25 +/- 6.93% vs. 47.24 +/- 10.53% (p less than 0.05). The infarct size induced by RCA occlusion was significantly reduced by the RVAD, due to the right ventricular unloading via reduced myocardial oxygen demand.