Jeanne Feger

Effect of polymeric nanoparticle administration on the clearance activity of the mononuclear phagocyte system in mice

We investigated the consequences of acute and subacute administration of mice with polyisobutylcyanoacrylate (PIBCA), polyisohexylcyanoacrylate (PIHCA), poly(D,L-lactic) acid (PLA), and polystyrene (PS) nanoparticles on the mononuclear phagocyte system phagocytic function. This was done by measuring the clearance rate of colloidal carbon. Single administration of PIBCA and PIHCA (but not PLA and PS) nanoparticles reduced carbon clearance in both a time- and dose-dependent fashion. Since clearance of preopsonized carbon was normal, it was assumed that PIBCA and PIHCA nanoparticles deplete opsonins specific for carbon recognition. A decrease in plasma fibronectin levels resulting from nanoparticle administration suggested its implication in their removal from blood. However, fibronectin does not seem to be responsible for PIBCA and PIHCA blockade. Phagocytic function was preserved after repeated treatment with nanoparticles, probably as a result of increased Kupffer cell phagocytic activity and the contribution of spleen macrophages. Neither toxicity nor effects due to nanoparticle hepatic accumulation were observed.

Evaluation of hepatic antioxidant systems after intravenous administration of polymeric nanoparticles

We have investigated the modifications of the levels of intracellular markers of the oxidative stress in hepatocytes, after single or repeated injections of poly(isobutyl cyanoacrylate) (PIBCA) and polystyrene (PS) nanoparticles. Nanoparticles were administered intravenously at single doses of 20 and 100 mg kg 1 for 14 days. Levels of reduced (GSH) and oxidized (GSSG) glutathione, superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CT) and the peroxidation of membrane lipids were measured. Single and repeated administration of PIBCA and PS nanoparticles induced a transient depletion of GSH and GSSG levels, a transient inhibition of SOD activity and a slight increase in CT activity. However, GPx activity was not modified and lipid peroxidation was not observed, suggesting that hepatocytes are not strongly affected by these modifications. Since nanoparticles do not distribute in hepatocytes, oxidative species could proceed from hepatic macrophages, activated after nanoparticle phagocytosis. It is unlikely that poly(alkyl cyanoacrylate) degradation products might be responsible for the oxidative attack because non-biodegradable PS nanoparticles induced the same effect. Uptake of polymeric nanoparticles by Kupffer cells in the liver induce modifications in hepatocyte antioxidant systems, probably due to the production of radical oxygen species. However, the depletion in glutathione was not great enough to initiate hepatocyte damage, since no changes in lipid peroxidation and reversible alterations were observed. This is an important factor to be considered in the use of polymeric nanoparticles as drug carriers.

FIVE ISOENZYMES OF PROTEIN KINASE C ARE EXPRESSED IN NORMAL AND STZ-DIABETIC RAT HEPATOCYTES : EFFECT OF PHORBOL 12-MYRISTATE 13-ACETATE

Using isoenzyme-specific antisera, five Protein Kinase Cs (PKCs) were detected in cytosol and membrane hepatocytes from normal rats: PKC alpha (80 kDa), PKC beta II (40, 50, 55, 85 kDa), PKC delta (74, 76 kDa), PKC epsilon (95 kDa), PKC zeta (65, 70 kDa). STZ-diabetes induced a lower expression of the five PKCs, a higher localization in the cytosol, a preferential expression of PKC delta as the 76 kDa phosphorylated species and a decreased kinase activity towards Histone III-S. A 1 microM phorbol 12-myristate 13-acetate (PMA) incubation induced similar translocation to the membrane of PKCs alpha, native 85 kDa beta II and epsilon. The 74 kDa PKC delta was switched to the 76 kDa species, the normal form in STZ-diabetic cells. The truncated PKC beta II and PKC epsilon were unchanged.

Evaluation of the degree of desialylation of serum α1-acid glycoprotein and α1-antitrypsin

Abstract A method for evaluating the degree of desialylation of α 1 -AGP and α 1 -at has been developed. It consists in their simultaneous determination by radial immunodiffusion (RID) and electroimmunodiffusion (EID). When a desialylation exists, an underestimation by EID relative to RID is found. 1. (1) No significant desialylation of α 1 -AGP and α 1 -AT occurred in normal subjects. 2. (2) No correlation between desialylation of α 1 -AGP and α 1 -AT and their amounts existed. 3. (3) Desialylation was preferentially observed in patients with severe hepatic damage but also with inflammatory disorders.

Involvement of sialic acid residues in electroimmunodiffusion of α1-acid glycoprotein. A method for determining the degree of sialylation of serum glycoproteins

When the two immunological methods, radial immunodiffusion (RID) and electroimmunodiffusion (EID), were used for the determination of alpha 1-acid glycoprotein, a significant discrepancy in the results was encountered depending on the degree of sialylation. It appeared that with desialylated alpha 1-acid glycoprotein, amounts estimated by EID were much lower than those actually present as assayed by the RID method. Determination of glycoprotein samples treated with neuraminidase for varying periods of time revealed an increasing underestimation by the EID method with decreasing sialic acid content. Partial resialylation of asialo-alpha 1-acid glycoprotein by bovine colostrum beta-galactoside alpha (2 leads to 6) sialyltransferase on the other hand resulted in less underestimation. Differential determination of alpha 1-acid glycoprotein by the two immunological methods thus offers a method for the estimation of the degree of sialylation of alpha 1-acid glycoprotein and other sialoglycoproteins in serum and body fluids.

Effect of phenobarbital on the oligosaccharide structure of rat α-acid glycoprotein

Abstract We have recently shown that the administration of phenobarbital to rats leads t an increased serum α 1 -acid glycoprotein content with alterations in the relative proportion of the sugar moiety. Therefore, α 1 -acid glycoprotein was purified from normal ( α 1 -acid glycoprotein N ) and phenobarbital-treated rats ( α 1 -acid glycoprotein PB ). Glycans were separated by AX-10 chromatography and analysed by gas chromatography. It appears that, compared to α 1 -acid glycoprotein N , α 1 -acid glycoprotein PB had a higher carbohydrate content (31.7% compared to 26%) and a non-negligible amount of neutral oligosaccharide (12.2% compared to 1.3%). No tetrasialyl oligosaccharides in α 1 -acid glycoprotein PB were detected, whereas their relative proportion in α 1 -acid glycoprotein N was 27%.

Evaluation of the degree of desialylation of serum C1-inactivator and haemopexin

The calibration curves for evaluating the degree of desialylation of C1-inactivator (sialic acid content 12.5%) and haemopexin (sialic acid content 4%) have been plotted. No desialylation of either glycoprotein occurs in normal subjects. In the patients (liver damage) studied, C1-inactivator is often desialylated, whereas haemopexin is not. In a previous report, we had shown that alpha1-acid glycoprotein is more often desialylated than alpha1-antitrypsin. Thus, it appears that the degree of desialylation of the sialic acid-rich glycoproteins is a more sensitive index of the severity of hepatic injury than that of the sialic acid-poor glycoproteins. This could be due to a defect in the sialylation process during synthesis.

Isolement du sérum humain normal d'une protéine capable de diminuer l'activité biologique de l'histamine sur l'iléon isolé de Cobaye: Étude préliminaire du mode d'action

Summary We have isolated from human serum a chemically well-defined compound which is able to decrease the biological activity of histamin on Guinea-pig isolated ileon. Modern protein partition technique made it possible to isolate a highly purified protein which reproduces the total effect. The first part of the study describes the techniques used to isolate the active principle and states the physico-chemical characteristics of this molecule. The purified protein enabled us to study its mode of action. The pharmacological activity is partly linked to the large affinity of this factor for histamin. We give the first prove for the formation of a stable combination between histamin and the protein causing this effect.

in vivo Ageing of human erythrocytes and cell-surface labeling by metaperiodate and sodium borotritide.

Young and old human erythrocytes, separated in vitro according to age, were labeled at the surface-sialic acid residues by sodium periodate and borotritide treatment. No qualitative difference was observed between the sialic acid derivatives of young- and old-erythrocyte membranes. The number of labeled residues was significantly decreased in the in vivo aged erythrocytes (12 +/- 2.8 X 10(6); n = 8) when compared to the young ones (19.4 +/- 2.8 X 10(6); n = 8). Analysis of the labeled glycoproteins by sodium dodecyl sulfate gel-electrophoresis showed that this decrease affects the PAS-1, PAS-4, PAS-2, and PAS-3 bands (glycophorins A and B) of the old-erythrocyte membranes.

Utilisation du fructose par I'hépatocyte isolé de rat nourri Zucker obèse fa/fa.

The utilization of fructose by isolated hepatocytes was investigated in fed obese Zucker fa/fa rats compared with their lean littermates (Fa/?) and Sprague-Dawley rats. Hepatocytes were incubated duriThe utilization of fructose by isolated hepatocytes was investigated in fed obese Zucker fa/fa rats compared with their lean littermates (Fa/?) and Sprague-Dawley rats. Hepatocytes were incubated during 3 h using U14C fructose (20 mM). Our results show: a significant increase of fructose consumption, glucose, lactate and pyruvate production and faster turnover of glycogen by fa/fa rats. In these animals, synthesis of acylglycerol was also significantly enhanced. Our results suggest that fructose in fa/fa rats was used preferentially as precursor for lipid synthesis not only by the liver but also by the adipose tissue after a prior transformation into glucose by hepatocytes. All these abnormalities result in an accumulation of acylglycerols maintaining an obesity state in fa/fa rats.