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1.
Perit Dial Int ; 12(4): 359-64, 1992.
Article in English | MEDLINE | ID: mdl-1420493

ABSTRACT

Reports in the literature have linked a low phosphatidylcholine content in continuous ambulatory peritoneal dialysis (CAPD) effluent to ultrafiltration loss. Clinical evidence suggests that adding phosphatidylcholine to the dialysis solution enhances ultrafiltration. A clinical study has been designed to clarify the effect of phosphatidylcholine on ultrafiltration in CAPD patients with normal ultrafiltration. A weekly measurement of the peritoneal equilibration test was conducted per patient in the hospital. A comparison between the control dialysis solution (three-week period) and the phosphatidylcholine premixed solution (three-week period) was performed on a total of 12 patients. This study shows that a phosphatidylcholine premixed dialysis solution significantly enhances ultrafiltration. Since ultrafiltration per osmotic driving force (mL/g glucose) is enhanced, the patient's glucose load per day is reduced to achieve equal ultrafiltration. In the presence of phosphatidylcholine, peritoneal permeability remained unchanged, as indicated by membrane transport characteristics. No side effects were observed.


Subject(s)
Peritoneal Dialysis, Continuous Ambulatory , Phosphatidylcholines/administration & dosage , Biological Transport/physiology , Dialysis Solutions/chemistry , Female , Humans , Infusions, Parenteral , Kidney Failure, Chronic/therapy , Male , Middle Aged , Peritoneum/physiology , Phosphatidylcholines/therapeutic use , Ultrafiltration
2.
Toxicol Appl Pharmacol ; 97(3): 473-9, 1989 Mar 01.
Article in English | MEDLINE | ID: mdl-2609344

ABSTRACT

Tetracycline is known to cause hepatic dysfunction in humans by inducing steatosis. Accumulation of fat in the liver could result from biochemical effects at various levels in the sequence from protein and triglyceride synthesis to lipoprotein secretion. The effects of tetracycline on the synthesis and secretion of triglycerides and proteins were studied in isolated rat hepatocytes surviving in suspension for up to 2.5 hr. Interpretation of the results obtained for tetracycline was made by comparison with results obtained, under the same experimental conditions, for the well-known steatogenic compounds, cycloheximide and colchicine. The data indicate that tetracycline produces a concentration-dependent inhibition of 14C-triglyceride secretion without affecting triglyceride synthesis. This inhibition explains the intracellular triglyceride accumulation. However, tetracycline does not affect protein secretion. Furthermore, it was demonstrated that the effect of tetracycline on protein synthesis was not related to inhibition of triglyceride release. In conclusion, it is proposed that the effect of tetracycline could be at the level of the association between triglycerides and apoproteins to form lipoproteins.


Subject(s)
Lipid Metabolism , Liver/drug effects , Tetracycline/toxicity , Animals , Apoproteins/metabolism , Colchicine/pharmacology , Cycloheximide/pharmacology , In Vitro Techniques , Liver/metabolism , Male , Proteins/metabolism , Rats , Rats, Inbred Strains , Triglycerides/metabolism
3.
In Vitro Cell Dev Biol ; 22(12): 706-12, 1986 Dec.
Article in English | MEDLINE | ID: mdl-3782009

ABSTRACT

Adult rat liver slices were maintained for 20 h in a novel organ culture system with minimal loss of viability and function. Potassium and adenosine triphosphate levels were maintained at in vivo levels, following an initial recovery period (2 to 4 h), for up to 20 h. Protein synthesis and secretion were linear for 20 and 16 h, respectively. In addition, the liver slices synthesized glycogen between 4 and 12 h in culture. Finally, the liver slices were hormonally responsive during the 20 h culture period as exemplified by glucagon-stimulated glucose production. This system provides a simple and effective method for the culture and biochemical maintenance of adult rat liver for 20 h with minimal loss of biochemical function.


Subject(s)
Liver/physiology , Organ Culture Techniques , Adenosine Triphosphate/analysis , Animals , Glucose/biosynthesis , Liver/analysis , Liver/metabolism , Liver Glycogen/analysis , Male , Potassium/analysis , Protein Biosynthesis , Rats , Rats, Inbred Strains
4.
Drug Chem Toxicol ; 9(3-4): 223-37, 1986.
Article in English | MEDLINE | ID: mdl-3816613

ABSTRACT

The effect of volatile anesthetics on protein synthesis and secretion by isolated rat hepatocytes in suspension was investigated. Halothane and enflurane inhibited protein synthesis in a dose-dependent manner. Diethyl ether had little effect on protein synthesis while isoflurane caused a mild inhibition. This effect was more pronounced in hepatocytes from phenobarbital treated male rats when compared to hepatocytes from control rats. Protein synthesis in hepatocytes from phenobarbital treated female rats was inhibited similar to that seen with control male rat hepatocytes. Isoflurane, enflurane, and halothane also caused a dose-dependent inhibition of protein secretion, while diethyl ether was only mildly inhibitory. From these studies it appears that inhibition of protein synthesis and secretion might be an early and sensitive indicator of cellular injury by volatile anesthetics.


Subject(s)
Anesthetics/toxicity , Liver/metabolism , Protein Biosynthesis , Animals , Biotransformation , Cells, Cultured , Kinetics , Leucine/metabolism , Phenobarbital/pharmacology , Rats , Rats, Inbred Strains , Sex Factors
5.
Fundam Appl Toxicol ; 4(3 Pt 1): 441-50, 1984 Jun.
Article in English | MEDLINE | ID: mdl-6745533

ABSTRACT

Isolated hepatocytes from adult male Wistar rats are a suitable experimental model to study the cytotoxicity of chemicals. Indeed, the isolated cells incubated in suspension in a Waymouth medium supplemented with 10% newborn calf serum maintain critical biochemical functions such as cytochrome P-450-dependent monooxygenase activity, glycogen, and protein synthesis capacities. This cellular model is used to detect the early biochemical effects of various xenobiotics, i.e., chlorpromazine, promethazine, bromobenzene, paracetamol, and isoniazid. Both cellular lysis (measured by the LDH leakage) and metabolic competence of the hepatocytes (glycogen deposits and protein synthesis) are modified as a function of both the duration of exposure to, and the concentration of, the chemicals. These results point out that the evaluation of metabolic functions of isolated cells surviving in suspension might be a sensitive test to predict early cell injury. Indeed, changes in the cellular behavior may occur before or without cell death. Furthermore, since both the cytochrome P-450 content and its dependent monooxygenase activity together with critical biochemical functions of the isolated cells remain stable, this model is of significant interest in ascertaining the mechanisms of toxicity.


Subject(s)
Liver/drug effects , Acetaminophen/toxicity , Animals , Bromobenzenes/toxicity , Chlorpromazine/toxicity , Cytochrome P-450 Enzyme System/metabolism , In Vitro Techniques , L-Lactate Dehydrogenase/metabolism , Liver/cytology , Liver/metabolism , Liver Glycogen/metabolism , Male , Mixed Function Oxygenases/metabolism , Promethazine/toxicity , Protein Biosynthesis , Rats , Rats, Inbred Strains , Time Factors
6.
Toxicology ; 26(1): 47-54, 1983 Jan.
Article in English | MEDLINE | ID: mdl-6829029

ABSTRACT

Diethyl maleate is commonly used in toxicological and drug metabolism research using isolated adult rat hepatocytes. At the highest concentrations used the effect of diethyl maleate is, however, not limited to glutathione depletion. In these conditions it inhibits protein synthesis and it impairs the "L" system for amino acid transport. It has, however, no effect on the cytochrome P-450 content or its dependent aldrin monooxygenase. The present report shows that a concentration of diethyl maleate as low as 0.2 mM is sufficient to deplete glutathione without affecting glycogen and protein synthesis, transport of amino acid or monooxygenase activity in isolated adult rat hepatocytes.


Subject(s)
Liver/metabolism , Maleates/pharmacology , Protein Biosynthesis , Animals , Biological Transport/drug effects , Cell Survival/drug effects , Cytochrome P-450 Enzyme System/metabolism , Glutathione/biosynthesis , In Vitro Techniques , Male , Rats , Rats, Inbred Strains
8.
IARC Sci Publ ; (41): 561-72, 1982.
Article in English | MEDLINE | ID: mdl-7141561

ABSTRACT

The present communication reports the effects of N-nitroso-pyrrolidine (NPYR) on some essential metabolic pathways in isolated hepatocytes. Isolated cells prepared by the collagenase-perfusion technique are incubated for 4 hours in suspension in a Waymouth medium, either in the absence or in the presence of NPYR (20 to 40 mmol/l). Under these conditions, NPYR, even at 40 mmol/l, has no effect on the vital staining of the cells by erythrosin B and does not increase the leakage of LDH, indicating no lethal effect. However, the glycogen synthesis which occurs in control cells is inhibited in presence of NPYR and the intracellular glycogen is even degraded. This effect is accompanied by reactivation of phosphorylase 'a'. NPYR also affects the protein synthesis capacity of the isolated hepatocytes. Incubation in the presence of non-metabolizable labelled amino acids has demonstrated that such an effect could be explained by a specific action of NPYR on the L-transport system for amino acids through the hepatic plasma membrane.


Subject(s)
Liver/drug effects , N-Nitrosopyrrolidine/toxicity , Nitrosamines/toxicity , Animals , Cell Membrane/drug effects , In Vitro Techniques , Liver/metabolism , Liver Glycogen/biosynthesis , Male , Protein Biosynthesis , Rats , Rats, Inbred Strains
9.
Biochim Biophys Acta ; 632(4): 619-29, 1980 Nov 03.
Article in English | MEDLINE | ID: mdl-7437478

ABSTRACT

An improved method for the homogenization and the subsequent subcellular fractionation of hepatocytes isolated from adult rat liver is described. The homogenization procedure developed in the present study allows the preservation of the integrity of subcellular structures, as demonstrated by measurement of the activities of representative enzymes as well as by determination of their latency. The activities of representative marker enzymes, as calculated on subcellular fractions obtained by differential centrifugation of the homogenate, are identical whether the homogenate arises from isolated hepatocytes or from the whole liver. Moreover, there is a close similitude between the kinetic parameters (Km and V) of two microsomal cytochrome P450-dependent mixed-function oxidases, namely aniline hydroxylase and aminopyrine demethylase determined on microsomal preparations obtained either from isolated cells or from the whole liver.


Subject(s)
Liver/ultrastructure , Animals , Cell Fractionation/methods , Cell Nucleus/ultrastructure , Kinetics , Male , Microscopy, Electron , Microsomes, Liver/ultrastructure , Mitochondria, Liver/ultrastructure , Mixed Function Oxygenases/analysis , Rats , Ultracentrifugation/methods
10.
Toxicology ; 18(3): 213-23, 1980.
Article in English | MEDLINE | ID: mdl-7222052

ABSTRACT

Freshly isolated hepatocytes in suspension were used to evaluate the possible effects of certain chemicals. Conditions including the choice of the incubation medium have been defined for maintaining the cells competent for a sufficient length of time. Using paracetamol alone or in combination with diethylmaleate, we have been able to show that these chemicals markedly alter the metabolic state of the cells, as indicated by an inhibition of glycogen synthesis and even by an enhancement of glycogen degradation, without modifying membrane integrity. These effects are dose-dependent and probably mediated through modification of glycogen phosphorylase activity.


Subject(s)
Liver Glycogen/metabolism , Acetaminophen/metabolism , Animals , Carbon Tetrachloride/metabolism , Cell Membrane/drug effects , In Vitro Techniques , L-Lactate Dehydrogenase/metabolism , Liver/cytology , Liver/drug effects , Liver/enzymology , Male , Phosphorylase a/metabolism , Rats , Time Factors
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