A Perfluorochemical Oxygen Carrier (Fluosol‐43) in a Synthetic Medium Used for Perfusion of Isolated Rat Liver

Nováková, V.; G, Birke; Plantin, Lars; Wretlind, A

doi:10.1111/j.1748-1716.1976.tb10320.x

Cited by 12 publications

(7 citation statements)

References 14 publications

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“…Rat livers from Sprague-Dawley rats (250-400 g) were perfused at 370C through the portal vein with a recycling perfusate (80 ml) of perfluorotributylamine/pluronic F-68 emulsion (Fluosol 43; Green Cross Corporation, Osaka, Japan), free of plasma proteins and erythrocytes, heparinized with 3000 units of heparin, and maintained in an atmosphere of 95% 02/5% CO2 by means of a membrane oxygenator. A similar nonhemoglobin oxygenation system has been shown to maintain viability of the liver for at least 5 hr (16). Synthesis of albumin and transferrin and secretion of bile were similar to those previously found with a recycling erythrocyte/plasma perfusate (17).…”

supporting

confidence: 73%

Production and release of plasminogen by isolated perfused rat liver.

Saito¹,

Hamilton²,

Tavill³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

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In view of conflicting evidence for a major hepatic role in the synthesis of circulating plasminogen, the precursor of the fibrinolytic enzyme plasmin, we carried out the present study with a sensitive assay in order to measure the accumulation of small quantities of plasminogen in a recycling rat liver perfusion system. We have purified plasminogen from Sprague-Dawley rat plasma and have raised a monospecific antiserum against it in rabbits. Isolated rat liver perfusions were performed with an oxygenated recycling perfusate consisting of a perfluorotributylamine/Pluronic F-68 emulsion (Fluosol 43) These data indicate that theliver is a major site of plasminogen production.Plasminogen, a protein of Mr t85,000, is present in all mammalian plasmas and is a precursor of the proteolytic (fibrinolytic) enzyme plasmin. Activation of the precursor initiates the process of dissolution of intravascular fibrin deposits (1).The site of synthesis of plasminogen has not been established with certainty, and several organs and tissues have been proposed as possible sources. Earlier immunofluorescence studies demonstrated plasminogen in eosinophilic granulocytes and led to the suggestion that the protein was produced in developing eosinophils of the bone marrow (2). Although the kidney has been implicated as a site of synthesis (3), plasma concentrations and de novo biosynthesis of plasminogen are maintained in anephric rats (4). In contrast, the decreased titer of plasma plasminogen in patients with advanced hepatic cirrhosis (5) suggests that the liver may be a major site of production in man. This conclusion is supported by the recent observation that, after hepatic homotransplantation in man, the plasminogen phenotype of the host converted to that of the donor (6).Although a previous study failed to identify release of plasminogen by the isolated perfused rat liver (7), the search for small quantities of the protein in the perfusate may have been hindered by the lack of a sensitive assay technique at that time.In the present paper we describe a specific and sensitive radioimmunoassay for rat plasminogen that is capable of detecting increasing titers of plasminogen in the plasma-free perfluorochemical perfusing the isolated rat liver. Inhibition of the increase in titer and of incorporation of a labeled leucine by cycloheximide confirmed that the liver is a site of de novo biosynthesis of plasminogen. Furthermore, the rate of release of plasminogen is compatible with the conclusion that the liver is the predominant site of synthesis in vivo. The findings of our study have been presented in abstract form (8). Since this work was completed, evidence has appeared for the capacity of rat hepatocytes in culture to synthesize plasminogen during a period of 48 hr (9).MATERIALS AND METHODS Materials. Rat plasminogen was isolated from pooled, citrated plasma of Sprague-Dawley rats (Pel-Freez) by lysineSepharose affinity chromatography (10) and Sephadex G-150 column chromatography. The purified preparation showed a sharp si...

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supporting

confidence: 73%

Production and release of plasminogen by isolated perfused rat liver.

Saito¹,

Hamilton²,

Tavill³

et al. 1980

Proc. Natl. Acad. Sci. U.S.A.

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“…Livers perfused with FC-43 emulsion secreted significantly less bile during 4-hr perfusions in comparison with livers perfused with RBC medium. A similar, lower and gradually decreasing bile secretion was reported for livers perfused with FC-43 emulsion for 4 hr (52) and in shorter perfusions (1 to 2.5 hr) by other investigators (14,17,20,28,29,49). Van Dyke, Gollan and Scharschmidt (49) also observed a lack of correlation between oxygen consumption and bile secretion without the addition of bile salts.…”

Section: Discussionsupporting

confidence: 81%

“…FC-43 emulsion was used to flush blood and trapped lipoproteins from hepatic sinusoids. Thereafter, 40 ml of FC-43 emulsion was recirculated in the perfusion system through the livers at a constant flow rate of 24 mVmin to achieve sufficient oxygen transport (13,(14)(15)(16)(17)(18)(19)(20)(21)(22)(23). The actual flow rate was calculated using the wet weight of livers at the end of perfusions.…”

mentioning

confidence: 99%

A comparison of lipoprotein secretion, bile production and hepatic morphology in isolated rat livers perfused with a perfluorocarbon emulsion or rat erythrocytes

et al. 1991

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Isolated rat livers were perfused with an oxygenated perfluorocarbon emulsion, FC-43 emulsion for 1 to 4 hr. FC-43 emulsion contained 20% FC-43 (wt/vol) perfluorotributylamine (the fluorocarbon component for the transport of oxygen and carbon dioxide) emulsified with 2.56% Pluronic F-68 (a nonionic surfactant) in Krebs-Ringer bicarbonate buffer. FC-43 emulsion also contained 3% hydroxyethyl starch as an oncotic agent and 1.8 mg/ml glucose. The viability (oxygen consumption), bile secretion, structural integrity and secretion of nascent lipoproteins by FC-43-perfused rat livers was compared with livers perfused with Krebs-Henseleit bicarbonate buffer that contained rat erythrocytes (25% hematocrit) and 1.5 mg/ml glucose (red blood cell medium). Oxygen consumption was somewhat higher in livers perfused with FC-43 emulsion. Bile secretion of livers perfused with FC-43 emulsion for 4 hr was reduced significantly to 40% of that by red blood cell medium. The structural integrity of livers perfused with FC-43 emulsion varied from normal to marked cellular damage. Light-microscopical examination of rat livers perfused with FC-43 emulsion showed ballooning of sinusoids, presence of vacuoles in sinusoidal lining cells in some hepatocytes and detachment of endothelium in sinusoids. The number of vacuoles progressively increased in longer perfusions. Electron-microscopical studies showed the presence of small (60 to 100 nm) vesicles of varying electron density, presumably fluorocarbon particles inside the vacuoles in sinusoidal lining cells (Kupffer and endothelial) and hepatocytes. After 4 hr of perfusion with FC-43 emulsion, most of the sinusoidal endothelia were denuded, and the microvilli of the hepatocytes all but disappeared. In contrast, the ultrastructure of rat livers perfused with red blood cell medium for 4 hr was unaltered. The accumulation of nascent lipoproteins in perfusates of FC-43-perfused livers was markedly reduced, and no normal very-low-density lipoprotein, low-density lipoprotein or high-density lipoprotein were isolated. Chemical analysis showed the presence of Pluronic F-68 in all lipoprotein fractions. Our data strongly suggest that, during recirculating liver perfusions with FC-43 emulsion (between 1 and 4 hr), the nonionic surfactant detergent Pluronic F-68 dissociated from the emulsion and markedly affected hepatic structure, lipoprotein secretion and the composition of lipoproteins isolated from perfusate. Therefore FC-43 emulsion is not a suitable liver-perfusion medium for studies of lipoprotein metabolism.

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“…Thus, the possibility that HF is generated by contaminating blood cells (21) is unlikely. A similar preparation has been shown to maintain viability of the perfused liver for at least 5 h (22), and previous studies in this laboratory have demonstrated that the liver in this system synthesizes plasminogen at physiologic rates (16). Furthermore, similar rates of production of albumin were observed in the present study, as were found in an earlier study that used a more physiological perfusate of plasma and erythrocytes (23).…”

Section: Discussionsupporting

confidence: 80%

Synthesis and release of Hageman factor (Factor XII) by the isolated perfused rat liver.

Saito¹,

Hamilton²,

Goodnough³

et al. 1983

J. Clin. Invest.

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A B S T R A C T The site of synthesis of Hageman factor (HF, Factor XII) has not been previously demonstrated with certainty. We have studied the production and release of HF in the isolated perfused rat liver and have compared rates of synthesis in this system with absolute rates of degradation measured in vivo. Rat livers, perfused for 5 h with a recycling fluid consisting of a perfluorochemical emulsion (Fluosol 43), were used to demonstrate a cumulative increase of HF in the perfusate as measured by a specific and sensitive radioimmunoassay. The rate of increase in the perfusate pool of HF during the final 4 h of perfusion yielded a mean synthetic rate of 3.5 Ag/h per 100 g body wt, which was -0.2% of the synthetic rate of albumin in the same system. The cumulative appearance of albumin and transferrin was linear after 1 h and calculated rates of synthesis were 2,012 tg/h per 100 g and 263 tsg/h per 100 g body wt, respectively.De novo synthesis of HF was confirmed by demonstrating incorporation of [14C]lysine into specific immunoprecipitates of HF, and by the observations that both specific incorporation of labeled amino acid and net release of immunoassayable HF were inhibited by the administration of cycloheximide. Finally, it was evident that the rates of synthesis observed in the isolated perfused liver agreed closely with absolute rates of degradation of HF measured in vivo with '25I-rat

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A Perfluorochemical Oxygen Carrier (Fluosol‐43) in a Synthetic Medium Used for Perfusion of Isolated Rat Liver

Cited by 12 publications

References 14 publications

Production and release of plasminogen by isolated perfused rat liver.

Production and release of plasminogen by isolated perfused rat liver.

A comparison of lipoprotein secretion, bile production and hepatic morphology in isolated rat livers perfused with a perfluorocarbon emulsion or rat erythrocytes

Synthesis and release of Hageman factor (Factor XII) by the isolated perfused rat liver.

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