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At least 469 records · Page 26Linked to original sources
PubMed2005

The third gas: H2S regulates perfusion pressure in both the isolated and perfused normal rat liver and in cirrhosis.

The regulation of sinusoidal resistance is dependent on the contraction of hepatic stellate cells (HSC) around sinusoidal endothelial cell (SEC) through paracrine cross-talk of vasoconstrictor and vasodilator agents. Hydrogen sulfide (H2S), a recently discovered gas neurotransmitter, is a putative vasodilator whose role in hepatic vascular regulation and portal hypertension is unexplored. Four-week bile duct-ligated (BDL) rats with cirrhosis and control rats were treated daily with NaHS (56 micromol/kg) for 5 days. Isolated livers were perfused first with NaHS for 20 minutes and then with norepinephrine (NE) and the intrahepatic resistance studied. In normal rats and animals with cirrhosis, administration of NE resulted in a dose-dependent increase of portal pressure. This effect was attenuated by H2S treatment (P < .05). The H2S-induced relaxation of hepatic microcirculation was attenuated by glibenclamide, an adenosine triphosphate (ATP)-sensitive K+ channel inhibitor. L-Cysteine, a substrate of cystathionine-gamma-lyase (CSE), decreased vasoconstriction in normal rat livers (P < .05) but failed to do so in livers with cirrhosis. BDL resulted in a downregulation of CSE mRNA/protein levels and activity (P < .05). Our in vitro data demonstrate that CSE is expressed in hepatocytes, HSCs, but not in sinusoidal endothelial cells (SEC). HSC activation downregulates CSE mRNA expression, resulting in a defective production of H2S and abrogation of relaxation induced by L-cysteine. In conclusion, CSE-derived H2S is involved in the maintenance of portal venous pressure. The reduction of CSE expression in the liver with cirrhosis contributes to the development of increased intrahepatic resistance and portal hypertension.

Animals↗
PubMed1986

Metabolism of bradykinin in isolated perfused rat kidney. Measurement of kininase activity in perfusate and urine.

The effect of bradykinin and kininase on renal function and the metabolism of bradykinin in the kidney were examined in isolated perfused rat kidney. In this experimental system, exogenous bradykinin did not affect the water and electrolyte handling by the kidney. Most of bradykinin and kininase in urine were derived from kidney but not from circulating medium. Kininase II may not have a major role in bradykinin destruction by the kidney.

Animals↗
PubMed1988

Immunohistochemical detection of antibody in tissue sections of non-perfused and ex vivo-perfused organs using a tetrazolium alkaline phosphatase substrate.

We have used nitroblue tetrazolium (NBT) as a color reagent to localize antibody-bound alkaline phosphatase in frozen tissue sections. In the method described, NBT is reduced to a stable black diformazan reaction product that contrasts well with nuclear counterstains such as hematoxylin and stands out strongly in black and white photographs. We have found NBT to be a suitable color reagent for the alkaline phosphatase: anti-alkaline immunohistochemical technique. The reaction product also contrasts well with fast red and can therefore be used as second reagent for two color immunoenzyme studies. In this report, we describe a novel two color immunoenzyme method to assess the ex vivo binding of antibodies against Class II histocompatibility antigens in whole organs connected to a perfusion circuit.

Alkaline Phosphatase↗
PubMed1988

Oxyntomodulin (glicentin-(33-69)): pharmacokinetics, binding to liver cell membranes, effects on isolated perfused pig pancreas, and secretion from isolated perfused lower small intestine of pigs.

The pharmacokinetics of purified synthetic oxyntomodulin were studied after infusing it into euglycaemic pigs at two rates. The elimination of the peptide from plasma was characterized by two components, a fast one (t1/2 7.2 +/- 0.6 min) and a slow one (t1/2 20.4 +/- 3.8 min) (mean +/- S.E.M., n = 7). The metabolic clearance rate was independent of infusion rate (6.96 +/- 0.99 vs 7.44 +/- 0.98 ml/kg . min (mean +/- S.E.M., n = 7). The synthetic peptide bound to pig hepatic glucagon receptors, but with approximately 2% of the affinity of glucagon, and showed insulinotropic and somatostatinotropic effects when infused into isolated perfused pig pancreases at concentrations higher than 10(-10) M. A dose-dependent increase was also shown for pancreatic glucagon output. A naturally occurring peptide, identified as oxyntomodulin by gel filtration and HPLC, was released into the circulation from the pig lower small intestinal mucosa upon intraluminal administration of glucose, and represented 25 +/- 3.8% of the secreted glucagon-like immunoreactivity. 11 +/- 2.3% of the secreted glucagon-like immunoreactivity was indistinguishable from glucagon itself upon gel filtration; thus at least 36% of the glucagon-like immunoreactivity secreted from the intestinal mucosa is already in an active form.

Animals↗
PubMed1991

Cholyl-lysylfluorescein: synthesis, biliary excretion in vivo and during single-pass perfusion of isolated perfused rat liver.

A fluorescent bile salt, cholyl-lysylfluorescein (cholyl-lys-F), was synthesised so that it retained both an intact steroid ring and a side chain structure with an unblocked carboxyl group. Its biliary kinetics and hepatic extraction were studied in Wistar rats and in the isolated perfused rat liver, respectively. The synthetic method used excess N-epsilon-CBZ-l-lysine methyl ester hydrochloride (7 mmol) and cholic acid (5 mmol) via EEDQ with a yield of 94% for cholyl-lys. Cholyl-lys-F was synthesized employing equimolar amounts of cholyl-lys (sodium salt) and fluorescein isothiocyanate (FITC) in bicarbonate buffer (pH 9.5) over 16 h at room temperature (21 degrees C) with a yield of 70%. The fluorescent property of cholyl-lys-F was similar to fluorescein with a strong apple-green fluorescence. In bile-fistula rats under pentobarbital anaesthesia, the cumulative 20 min biliary excretion as a percentage of injected dose were as follows: cholyl-lys-F, 94.4 +/- 0.3%, [14C]cholylglycine (CG), 93.1 +/- 1.2% and fluorescein (F), 34.8 +/- 0.5. Furthermore the single-pass hepatic extraction of cholyl-lys-F was 64.1 +/- 3.9%, [14C]CG was 66.1 +/- 1.2% and F was 16.5 +/- 2%. The similarity in biliary output and hepatic extraction of cholyl-lys-F to that of the natural bile acid cholylglycine suggest that both compounds are handled in a similar fashion. The greater biliary excretion and hepatic extraction of cholyl-lys-F relative to free fluorescein further suggest that conjugation with a bile salt may be an efficient way of targeting compounds to the liver.

Animals↗
PubMed2006

Phospholamban phosphorylation sites enhance the recovery of intracellular Ca2+ after perfusion arrest in isolated, perfused mouse heart.

OBJECTIVE: To investigate the importance of the phosphorylation of Ser16 and Thr17 sites of phospholamban (PLN) on intracellular Ca2+ (Cai2+) handling and contractile recovery of the stunned myocardium. METHODS: Cai2+ (Rhod-2, pulsed local-field fluorescence microscopy) and contractility (isovolumic left ventricular developed pressure, LVDP) were simultaneously measured in Langendorff perfused hearts from transgenic mice expressing either intact PLN (PLN-WT) or PLN with both phosphorylation sites mutated to Ala (PLN-DM), subjected to 12 min of global ischemia followed by a reperfusion period of 30 min. RESULTS: Pre-ischemic values of Cai2+ and LVDP were similar in both groups. In PLN-WT, a transient increase in Thr17 phosphorylation at early reperfusion preceded a recovery of Ca2+ transient amplitude, virtually completed by the end of reperfusion. LVDP at 30 min reperfusion was 67.9+/-7.6% of pre-ischemic values, n=14. In contrast, in PLN-DM, there was a poor recovery of Cai2+ transient amplitude and LVDP was significantly lower (28.3+/-6.7%, n=11, 30 min reperfusion) than in PLN-WT hearts. Although myofilament Ca2+ responsiveness and troponin I (TnI) degradation did not differ between groups, the episodes of mechanical alternans, typical of Cai2+ overload, were significantly prolonged in PLN-DM vs. PLN-WT hearts. CONCLUSIONS: PLN phosphorylation appears to be crucial for the mechanical and Cai2+ recovery during stunning and protective against the mechanical abnormalities typical of Cai2+ overload. The importance of PLN phosphorylation would primarily reside in the Thr17 residue, which is phosphorylated during the critical early phase of reperfusion. Our results emphasize that, although ablation of PLN phosphorylation does not affect basal contractility, it does alter Ca2+ handling and mechanical performance under stress situations.

Animals↗
PubMed2006

Repeat isolated limb perfusion with TNFalpha and melphalan for recurrent limb melanoma after failure of previous perfusion.

AIM: To assess the effectiveness of isolated limb perfusion (ILP) with tumour necrosis factor-alpha (TNFalpha) and melphalan for recurrent or persistent melanoma lesions after previous ILP. METHODS: Between 1978 and 2001, 21 patients (mean age 65, range 29-83 years) underwent repeat ILP for recurrent or persistent melanoma after a previous ILP. First ILPs had been performed with melphalan alone in 13 patients and with addition of TNFalpha in eight, for a median of nine lesions (interquartile (IQ) range 2-23 lesions). Repeat ILP was performed with TNFalpha and melphalan in all 21 patients for a median of nine lesions (IQ range 5-25 lesions). Median follow-up after repeat ILP was 18 months (IQ range 6-36 months). RESULTS: Thirteen patients attained a complete response (CR) after repeat ILP compared to 11 of 17 with measurable lesions after the first ILP. Nine patients relapsed after CR. Median limb recurrence-free survival was 13 months. Fourteen patients had mild acute regional toxicity after repeat ILP compared to 18 after the first ILP (n.s.). One patient underwent amputation for critical limb ischemia 10 months following repeat ILP. The limb salvage rate was 95%. Overall median survival was 62 months after CR compared to 13 months for those without CR (P=0.05). CONCLUSION: Repeat ILP with TNFalpha and melphalan is feasible after previous ILP with mild regional toxicity. The CR rate is relatively high and comparable to the first procedure with good limb recurrence-free survival and high limb salvage rate.

Adult↗
PubMed1989

Influence of unbound fraction and perfusate flow rate on taurocholate elimination by perfused rat liver: applicability of three distributed models.

The applicability of three different distributed models to the kinetics of elimination of taurocholate by isolated perfused rat liver was examined by fitting each model to literature data. Each of the models was able to predict the effect of changing hepatic blood flow on elimination, but only the model which incorporates separate density functions describing the degree of sinusoidal heterogeneity of blood flow and intrinsic clearance was able to predict the effect of changing unbound fraction on elimination.

Animals↗
PubMed1979

The effect of reduction of perfusion rate on lactate and oxygen uptake, glucose output and energy supply in the isolated perfused liver of starved rats.

1. Lactate and O2 uptake and glucose output were studied in isolated livers from starved rats at perfusate flow rates varying from 100 to 7% of "normal" (11.25-0.75 ml/min per 100 g body wt.). 2. With moderate diminution of flow rate, lactate and oxygen uptake fell more slowly than would be expected if uptake purely depended on substrate supply. 3. Use of a mathematical model suggests that the intrinsic capacity of the liver for lactate uptake is unaffected until the flow rate falls below 25% of "normal". 4. Some lactate uptake was always observed even at 7% of the "normal" flow rate. 5. At flow rates below 33% of the "normal", lactate was increasingly metabolized by pathways other than gluconeogenesis, which became a progressively less important consumer of available O2. 6. ATP content decreased with diminution of flow rate, but substantially less markedly than did lactate uptake and glucose output. 7. Intracellular pH fell from a mean value of 7.25 at "normal" flow rate to 7.03 at 7% of the "normal" flow rate.

Adenine Nucleotides↗
PubMed1983

Use of a transcutaneous PO2 regional perfusion index to quantify tissue perfusion in peripheral vascular disease.

In order to develop transcutaneous oxygen tension (PtcO2) measurements into a practical method for assessing peripheral vascular disease, the relationships between extremity and chest wall PtcO2 were examined in subjects with and without systemic atherosclerotic disease. The ratio of extremity to chest PtcO2, or transcutaneous regional perfusion index (RPI) assessed limb oxygenation more reliably than did direct PtcO2 measurement by obviating the effects of changes in systemic oxygen delivery upon local PtcO2. The authors find that transcutaneous oximetry can be used during treadmill exercise testing and that the RPI is unchanged by exercise in all normal subjects. PtcO2 and RPI were then measured during rest, position change, and exercise testing in patients with intermittent claudication. Whereas normal subjects maintain a constant thigh and calf RPI during exercise, patients with intermittent claudication consistently manifested large decreases in RPI in these areas when they were exercised until symptomatic. The authors find no overlap between the responses of normal subjects and patients with claudication; positive findings are, therefore, highly specific for exercise-induced limb ischemia. Since transcutaneous RPI exercise testing is easily performed and highly reproducible, it is well suited to clinical use in the diagnosis and documentation of intermittent claudication. Furthermore, since limb ischemia can be quantified, this method lends itself both to grading the severity of disease and to evaluating clinical progression of disease. It is suggested that such a quantitative approach to evaluation of intermittent claudication may allow refinement and extension of the indications for operative intervention in patients with intermittent claudication.

Adult↗
PubMed1985

Venovenous perfusion in ECMO for newborn respiratory insufficiency. A clinical comparison with venoarterial perfusion.

Venoarterial (VA) extracorporeal membrane oxygenation (ECMO) has been successful in the treatment of newborns less than 1 week of age and greater than 2000 gm birthweight with respiratory failure resistant to current medical and surgical management. While VA ECMO supports the heart as well as the lungs, it has the disadvantage of requiring carotid artery ligation and the possibility of perfusing air bubbles or particles into the arterial tree. We have treated 11 newborns with respiratory failure with venovenous (VV) ECMO returning the oxygenated blood to a cannula in the distal iliac vein. We compared these patients with 16 patients treated during the same period of time with VA ECMO. Three of the 11 VV patients required conversion to VA ECMO because of inadequate oxygenation and unstable hemodynamic situations. Ten of the 11 VV patients survived. Eleven of the 16 VA patients survived. The better survival in these patients treated with VV ECMO is attributed to their more favorable initial condition compared to patients treated with VA ECMO. The disadvantages of VV ECMO include a longer operative time to place the cannulas, groin wound problems, and persistent leg swelling along with the necessity to convert some patients to VA ECMO. Although this experience demonstrates that newborns with severe respiratory failure can be supported with VV ECMO, the complications and lack of practical advantages over VA lead us to recommend VA ECMO for routine clinical use at present.

Female↗
PubMed1985

Xeno- and auto-perfusion of rabbit kidney. Machine perfusion with blood at 37 degrees C.

Five rabbit kidneys were perfused with human blood and another five with their own blood in a re-circulating oxygenated system at 37 degrees C. The flow decreased to 2 ml/min. within 30 min. in all xenoperfusions, while none of the autoperfused had decreased to this level by 60 min. Endothelial damage, exudation, and IgG deposits along the basement membrane of the glomerular capillaries were the discriminative features of the xenoperfusion. In these experiments, we were unable to demonstrate any major role of platelets in the process leading to decreased blood flow.

Animals↗
PubMed1992

Aortic perfusion pressure as early determinant of beta-isomyosin expression in perfused hearts.

Pressure overload in vivo induces an increase in cardiac protooncogene and stress protein expression that may initiate the long-term genetic changes observed in hypertrophy. To known whether mechanical stimulus is linked to specific gene transcription, expression of immediate early genes and synthesis of total proteins and myosin heavy chains (MHCs) were studied in beating and KCl-arrested isolated rat hearts perfused for 2 h under various coronary pressures. The main result of this study is that in the beating heart an augmentation of aortic pressure from 60 to 120 mmHg results in a pronounced enhancement of the synthesis of MHC (+59%) and of the expression of the beta-MHC isomyosin mRNA (iso-mRNA; +104%). Also, total protein synthesis and the amounts of poly-(A)+, c-fos, c-myc, and heat-shock protein HSP68 mRNAs were increased. To arrest the heart at 60 mmHg has no effect on total protein synthesis and on the amounts of poly(A)+, alpha-MHC and beta-MHC iso-mRNAs, and mRNAs coding for oncoproteins, but the synthesis of MHC decreased by 24%. By contrast with what we have observed in the beating heart, the augmentation of the coronary pressure in the arrested heart stimulates total protein synthesis and increases the amount of poly(A)+, c-fos, c-myc, and HSP68 mRNAs but has no effect on the expression of both MHC iso-mRNAs. In conclusion, the activation of myosin synthesis by high coronary pressure in this model has mainly a pretranslational origin when the heart is beating.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1998

Prophylactic isolated limb perfusion for localized, high-risk limb melanoma: results of a multicenter randomized phase III trial. European Organization for Research and Treatment of Cancer Malignant Melanoma Cooperative Group Protocol 18832, the World Health Organization Melanoma Program Trial 15, and the North American Perfusion Group Southwest Oncology Group-8593.

PURPOSE: Patients with primary cutaneous melanoma > or = 1.5 mm in thickness are at high risk of having regional micrometastases at the time of initial surgical treatment. A phase III international study was designed to evaluate whether prophylactic isolated limb perfusion (ILP) could prevent regional recurrence and influence survival. PATIENTS AND METHODS: A total of 832 assessable patients from 16 centers entered the study; 412 were randomized to wide excision (WE) only and 420 to WE plus ILP with melphalan and mild hyperthermia. Median age was 50 years, 68% of patients were female, 79% of melanomas were located on a lower limb, and 47% had a thickness > or = 3 mm. RESULTS: Median follow-up duration is 6.4 years. There was a trend for a longer disease-free interval (DFI) after ILP. The difference was significant for patients who did not undergo elective lymph node dissection (ELND). The impact of ILP was clearly on the occurrence-as first site of progression - of in-transit metastases (ITM), which were reduced from 6.6% to 3.3%, and of regional lymph node (RLN) metastases, with a reduction from 16.7% to 12.6%. There was no benefit from ILP in terms of time to distant metastasis or survival. Side effects were higher after ILP, but transient in most patients. There were two amputations for limb toxicity after ILP. CONCLUSION: Prophylactic ILP with melphalan cannot be recommended as an adjunct to standard surgery in high-risk primary limb melanoma.

Adolescent↗
PubMed1991

Increase in glucose and lactate output and perfusion resistance by stimulation of hepatic nerves in isolated perfused rat liver: role of alpha 1-, alpha 2-, beta 1- and beta 2-receptors.

Rat liver was perfused in situ via the portal vein without recirculation: 1) Electrical stimulation of the nerve bundles around hepatic artery and portal vein increased glucose and lactate output, reduced flow and caused an overflow of noradrenaline into the hepatic vein. The alpha-agonist phenylephrine also augmented glucose and lactate output and lowered flow with an ED50 of about 1 microM, while the beta-agonist isoproterenol increased glucose output but reduced lactate output with an ED50 of about 0.2 microM and left flow unaltered. 2) The alpha 1-receptor antagonist prazosin (KI at alpha 1-sites approximately 1 nM, at alpha 2-sites approximately 100 nM) inhibited the nerve stimulation-dependent increase in glucose and lactate output and reduction of flow with an ID50 of about 1 nM, while the alpha 2-receptor antagonist yohimbine (KI at alpha 2-sites approximately 10 nM, at alpha 1-sites approximately 1500 nM) was inhibitory only with an ID50 of about 400 nM. 10 nM prazosin clearly reduced the nerve actions, completely blocked the effects of 1 microM phenylephrine and left the effects of 0.2 microM isoproterenol unaltered. 10 nM yohimbine did not affect the nerve actions nor the effects of phenylephrine or isoproterenol. 3) The beta 1-receptor antagonist metoprolol (KI at beta 1-sites approximately 100 nM, at beta 2-sites approximately 1.2 microM) at 10 microM concentrations did not interfere with the nerve stimulation-dependent increase in glucose and lactate output or the decrease in flow. It did not have any specific alpha-antagonistic influence either on the changes brought about by 1 microM phenylephrine; however, it blocked the beta 2-mediated increase in glucose output by isoproterenol.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1976

[Auto-perfused multi-organ preparations. III. Interpretation of histologic changes in perfused organs].

The functional instability of "multi-organ" preparations has been studied by morphological and histochemical assessment of 46 autoperfused heart-lung-kidney-liver-duodenopancreas preparations. The organs of the autoperfused preparations were damaged to different degrees, but the damage was always a function of perfusion time (1 to 3 hours). The liver suffered the most serious lesions (stagnation, oedema, hepatocytolysis, hydropic degeneration) but lesions such as interstitial haemorrhage and oedema though in a milder form appeared also in the kidney. The heart structure was comparatively well preserved.

Animals↗
PubMedDate not supplied

[Phagocytic activity of isolated perfused liver of rats of different ages. Biochemical and structural studies following perfusion with Cu-ceruloplasmin].

The phagocytic activity of young and old isolated perfused rat livers was measured referred to a glycoprotein (Cu-coeruloplasmin). It was shown that the phagocytic activity of the old rat liver is significantly higher than that of the young rat liver. There was no age-dependent difference of the parameters of liver function GOT, GPT, Kt and the quotients beta-OH-Butyric acid/Acetoacetate and lactate/butyrate.

3-Hydroxybutyric Acid↗
PubMed1981

[Phagocytosis activity of the isolated perfused liver in rats of different ages. Biochemical and ultrastructural examination after perfusion with colloidal ink].

Liver contains about 85% of reticulohistiocytic system. There are no problems to perfuse the isolated organ. The phagocytic activity was measured of young and old rat livers referred to phagocytosis of colloidal carbon. There is no difference between the phagocytic activity of young and old rat livers. The consumption of oxygen of rat livers increases with age.

Aging↗