Polyvinylchloride (PVC) particles implantation in mouse liver. A technique for experimental study of schistosome eggs-induced liver pathology

Summary An injection of suspended PVC particles in the caecal vein of mice induces a foreign-body portal granuloma reaction in the liver. Plastic casts of the portal system, after PVC particles implantation, show modifications in the portal bed and are compared with plastic casts obtained in mice infested bySchistosoma mansoni. This technique can be useful to study the cellular dynamics of the portal granuloma and can be a model for schistosomal eggs induced liver pathology.This work was supported by an ATP (INSERM) 18.75.41 and a scholarship from the Société d'Hépatologie Expérimentale.     

Eine Methode zur fortlaufenden Blutentnahme aus der Vena portae von Hunden

Summary A method is described for the introduction of a PVC.-Catheter into the portal vein of dogs. The catheter is introduced into one of the mesenteric veins of the ileum and pushed through the vena mesenterica communis until the portal vein is reached. The catheter, the end of which is fixed to the skin on the outside, can be removed without any injury to the animal by just drawing it out through the skin. The method can be applied for taking blood at various intervals during experiments on absorption, measurement of blood pressure in the portal vein and its branches, for the injection of substances directly into the liver, etc., on unanaesthetized animals.     

Properties of glutamine aminohydrolases in subcellular fractions of liver of tumour bearing mice.

Glutamine aminohydrolase is found to be present in microsomal and soluble supernatant in liver of EAC-bearing mice. Enzymes obtained from these two sources were characterized and found to behave differently from the mitochondrial glutaminase of both normal and tumour-bearing mice.     

Effect of orotic acid on liver glycogen of different animal species.

The effect of orotic acid on the liver glycogen content in the mice, frogs and catfish was studied. It was observed that the orotic acid significantly increases the glycogen content in the liver of mice and catfish as it does in rats. On the other hand it causes a fall of the glycogen level in frogs in experiments made both in autumn and spring. This effect was modified by amino acids administered together with orotic acid.     

The implantation of sepharose beads in mouse livers as an aid in the study of hepatic schistosomal fibrosis

Summary An experimental model of schistosomal portal fibrosis is described. Sepharose beads the size of schistosome eggs, loaded or not with soluble egg antigen (SEA) fromSchistosoma mansoni, are injected into the coecal vein of C3H/Sn mice and become embolized in the liver. Only SEA-coated beads evoke a granulomatous reaction; this is enhanced by simultaneous priming of the mice with spleen cells fromSchistosoma mansoni-infected syngeneic animals. The fibrosis, which ensues around the beads, is stable and is much more evident after priming. Preliminary collagen tissue immunotyping reveals the presence of collagen deposits of types I and III collagen. Type IV collagen remains unchanged in the portal tracts. The model appears to be well suited for studies of the pathogenesis of portal fibrosis.This work was supported by a contract from INSERM (Action Spéciale No 3).     

Comparative studies on the covalent binding of the carcinogen benzo(a)pyrene to DNA in various model systems.

The covalent binding of tritiated benzo(a)pyrene (BP) to DNA has been determined in rat liver in vivo, in rat liver perfused in situ, after incubation of BP with liver single cells, with liver homogenate, with liver microsomes and DNA, with fibroblasts from a rat granuloma pouch, and with 2 cell lines. Liver single cells were found to be a valuable compromise between the most sensitive system (microsomal incubation of BP and DNA) and the biologically most relevant system (in vivo).     

Ambiquitous behavior of rabbit liver lactate dehydrogenase

Rabbit liver mitochondrial fraction shows lactate dehydrogenase activity. The enzyme can be released from particles by increasing the pH and the ionic strength of the medium. There is a narrow range of pH (6.8-7.4) and ionic strength (20-50 mM NaCl) in which the solubilization sharply increases. It has been shown that divalent anions (SO4(2-) and cations (Mg2+, Ca2+) are highly effective specific solubilizing agents. NADH (1.5 mM) and ATP (1.0 mM) were effective in solubilizing 50% of the enzyme bound, whereas the same concentrations of the analogs NAD+ and ADP had little effect. Cytosolic lactate dehydrogenase bound to the mitochondrial fraction and a saturation of particles by enzyme was observed in all experiments performed. The in vitro binding requires a short period of incubation between the enzyme and particles and the binding is independent of the temperature in the 0-37 degrees C range. Binding was prevented by 0.15 M NaCl. The bound enzyme is approximately 20% less active than the soluble one. The results described give support to the proposal that rabbit liver lactate dehydrogenase has an ambiquitous behavior, like other glycolytic enzymes, which have not a fixed intracellular localization.     

Pathological changes in the liver of mice given 2,3,7,8-tetrachlorodibenzo-p-dioxin.

In C57BL/6 mice a single oral dose of 2,3,7,8-tetrachlorodibenzodioxin (LD50 126 mug/kg) results in loss of body weight and death with an enlarged fatty liver after ca. 21 days. A progressive necrotic centrilobular liver lesion is also seen.     

Pre- and postnatal distribution of lipids in the liver of genetic diabetic mice.

Triglycerides, phospholipids, cholesterol and cholesterol esters were determined by thin layer chromatography in the fetal and neonate livers of normal (Swiss albino) and genetic diabetic (KK) mice. In general, the lipids were elevated in the fetal liver of the KK mice. Despite this elevation in liver lipids, no increase in the weight of the newborn was observed.     

Unchanged r-RNA-gene dose in mice liver cells of different developmental stages.

The content of ribosomal DNA in mice liver at the beginning as well as near the end of the hematopoietic period was measured by RNA/DAN-hybridization in solution. At both stages the amount of ribosomal DNA was the same and comparable to that of postnatal liver.     

Ambiquitous behavior of rabbit liver lactate dehydrogenase

Summary Rabbit liver mitochondrial fraction shows lactate dehydrogenase activity. The enzyme can be released from particles by increasing the pH and the ionic strength of the medium. There is a narrow range of pH (6.8–7.4) and ionic strength (20–50 mM NaCl) in which the solubilization sharply increases. It has been shown that divalent anions (SO ₄ ^2– ) and cations (Mg²⁺, Ca²⁺) are highly effective specific solubilizing agents. NADH (1.5 mM) and ATP (1.0 mM) were effective in solubilizing 50% of the enzyme bound, whereas the same concentrations of the analogs NAD⁺ and ADP had little effect. Cytosolic lactate dehydrogenase bound to the mitochondrial fraction and a saturation of particles by enzyme was observed in all experiments performed. The in vitro binding requires a short period of incubation between the enzyme and particles and the binding is independent of the temperature in the 0–37°C range. Binding was prevented by 0.15 M NaCl. The bound enzyme is approximately 20% less active than the soluble one. The results described give support to the proposal that rabbit liver lactate dehydrogenase has an ambiquitous behavior, like other glycolytic enzymes, which have not a fixed intracellular localization.     

The effect of urethane and pentobarbital anaesthesia and hepatic portal vein catheterization on liver blood flow in the rat

Summary The effect of urethane and pentobarbital anaesthesia and hepatic portal vein catheterization on liver blood flow was investigated in the rat. Liver blood flow with pentobarbital anaesthesia was 40% greater than with urethane. Hepatic portal vein catheterization had no effect under pentobarbital anaesthesia whereas it produced an 18% fall in liver blood flow with urethane.Acknowledgments. This work was supported by grants from ICI Pharmaceuticals and Mersey Regional Health Authority (Research Scheme No. 338). We wish to thank Mr P.J. Roberts for much skilled assistance and Evans Medical Ltd for the kind gift of drugs.     

Comparative studies on the covalent binding of the carcinogen benzo(a)pyrene to DNA in various model systems

Summary The covalent binding of tritiated benzo(a) pyrene (BP) to DNA has been determined in rat liver in vivo, in rat liver perfused in situ, after incubation of BP with liver single cells, with liver homogenate, with liver microsomes and DNA, with fibroblasts from a rat granuloma pouch, and with 2 cell lines. Liver single cells were found to be a valuable compromise between the most sensitive system (microsomal incubation of BP with DNA) and the biologically most relevant system (in vivo).Presented in part at the 10th Annual Meeting of the Union of Swiss Societies of Experimental Biology, Experientia34, 925 (1978), abstract.Acknowledgment. We thank Dr G. Kistler, Institute of Anatomy, University of Zurich, for generously supplying the SIRC and VERO cell lines, and Mr P. Manser for the preparation of the granuloma pouch fibroblasts.     

An improved noninfections murine skin model of organized granulomatous inflammation

Summary An improved model of granulomatous inflammation in skin was developed by second passage skin grafting of isolated, lyophilized skin granulomas, originally elicited in naive mice by inoculations of lyophilized hepatic schistosome egg granulomas. The tissue reaction is caused by a single exposure to a noninfectious, acellular granulomagenic stimulus and occurs in healthy mice free of systemic disease. The model should prove useful for isolation of granuloma initiation factor(s). Furthermore, because there is a time lag before new granuloma formation begins, a window exists for analytical dissection of the initiation process. In this study we described the responses of host cells by autoradiography, and light and electron microscopy. The activity of angiotensin-converting enzyme and proline-specific endopeptidase showed a modulation during granuloma formation. In addition we found that severe immunosuppression with high dose cyclosporine therapy did not alter granuloma formation, supporting the idea that initiation of organized granulomas is T-cell independent.     

An improved noninfectious murine skin model of organized granulomatous inflammation

An improved model of granulomatous inflammation in skin was developed by second passage skin grafting of isolated, lyophilized skin granulomas, originally elicited in naive mice by inoculations of lyophilized hepatic schistosome egg granulomas. The tissue reaction is caused by a single exposure to a noninfectious, acellular granulomagenic stimulus and occurs in healthy mice free of systemic disease. The model should prove useful for isolation of granuloma initiation factor(s). Furthermore, because there is a time lag before new granuloma formation begins, a window exists for analytical dissection of the initiation process. In this study we described the responses of host cells by autoradiography, and light and electron microscopy. The activity of angiotensin-converting enzyme and proline-specific endopeptidase showed a modulation during granuloma formation. In addition we found that severe immunosuppression with high dose cyclosporine therapy did not alter granuloma formation, supporting the idea that initiation of organized granulomas is T-cell independent.     

A simple method of obtaining multiple blood samples from the portal vein and the hepatic vein in the rat in vivo

A very simple and rapid technique for inserting a catheter in the portal vein and the hepatic vein in the anesthesized rat in vivo is described. The pointed, saline-containing PE tubing is frozen in liquid nitrogen, whereupon it is used as a 'needle' to insert the catheter into the blood vessel. Multiple blood samples can be obtained from the portal and the hepatic vein at the same time, so that in situ extraction of drugs by the liver can be measured in vivo, since hepatic blood flow is uninterrupted.     

Über die Entwicklung des Fremdkörpergranuloms in verschiedenen Organsystemen

Summary The weight has been measured of the granuloma produced after implantation of the same foreign body in the kidneys, the testes, the liver and the subcutaneous tissue of male rats. It was found that granulomae in the testes and liver are nearly twice as heavy as in the kidney or in the subcutaneous tissue. Enhancement or inhibition of this reaction by treating the animals with cortexone shows organ-dependent effects on granuloma formation.     

Intracellular distribution of two lysosomal enzymes in newborn rat liver]

The distribution of two lysosomal markers (beta-acetylglucosaminidase and acid phosphatase) between liver fractions was studied in the newborn Rat. The results indicate that after birth the hydrolase-bearing particles increased in size and had a lower density than the primary lysosomes. These modifications may be related to the autophagic-vacuole formation known to occur during this period.     

Effect of orotic acid on liver glycogen of different animal species

Summary The effect of orotic acid on the liver glycogen content in the mice, frogs and catfish was studied. It was observed that the orotic acid significantly increases the glycogen content in the liver of mice and catfish as it does in rats. On the other hand it causes a fall of the glycogen level in frogs in experiments made both in autumn and spring. This effect was modified by amino acids administrated together with orotic acid.