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Biomedical subjects

A Koj

Publications and source records attributed to A Koj.

At least 91 records · Page 5Linked to original sources
PubMed1978

Distinction between binding and endocytosis of human asialo-transferrin by the rat liver.

The ability of the rat liver to bind and endocytose human asialo-transferrin was investigated in vivo. Asialo-transferrin was separated from incompletely desialylated transferrin and neuraminidase by chromatography before being labelled with (125)I. Plasma radioactivity curves and hepatic radioactivity contents measured over a 1270-fold dose range led to the following observation. At the lowest dose (0.4mug/100g body wt.), the distribution of asialo-transferrin between plasma and liver resembled a reversible reaction reaching equilibrium in approx. 20min. After 35min, 93% of the dose was recovered with the plasma and liver as protein-bound radioactivity. Most of the asialo-transferrin associated with the liver could be displaced by asialo-orosomucoid, indicating that binding of asialo-transferrin to the galactose-specific lectin on the plasma membrane of hepatocytes was not followed by a signal for endocytosis. A range of doses, up to an average of 509.2mug of asialo-transferrin per 100g body wt., resulted in progressive increments in asialo-transferrin catabolism, as evidenced by lower dose recoveries and increased concentrations of non-protein-associated radioactivity in the liver and plasma volume. These observations indicate that binding and endocytosis of human asialo-transferrin by the rat hepatocyte are distinct phenomena. Individual asialo-transferrin molecules, although readily bound by the hepatic lectin, lack either the quantity or spacing of terminal galactose residues necessary for triggering endocytosis. Although endocytosis is induced by several asialo-transferrin molecules acting synergistically, preliminary experiments with asialo-glycopeptides and other substances have so far failed to provide further insight into the chemical basis of the signal for endocytosis.

Animals↗
PubMed1978

Elimination of asialofetuin and asialoorosomucoid by the intact rat. Quantitative aspects of the hepatic clearance mechanism.

The capacity of the liver to eliminate asialofetuin and asialoorosomucoid was investigated in intact rats. From plasma radioactivity curve measurements and assays on tissue homogenates the liver is shown to be able to dispose of an average of 19.8 microgram of asialofetuin/min per 100 g body weight. No other major route is identified for the disappearance of asialofetuin from the plasma, although trace amounts of the protein were detectable in the urine. From analyses of the plasma radioactivity curves the elimination process for asialoorosomucoid appears to be comparatively complex because of the existence of extrahepatic disposal routes. Quantification of labelled asialoorosomucoid in liver homogenates indicates, however, that the hepatic clearance rate for asialoorosomucoid is similar to that for asialofetuin. Urinary excretion significantly contributes to the disappearance of asialoorosomucoid from the plasma but the hepatic and renal routes do not account for all the protein lost from this compartment. At plasma concentrations above the maximal eliminative capacity of the liver, the hepatic clearance of asialofetuin obeys zero-order kinetics and is remarkably constant. Elimination of a quantity of asialoglycoprotein which exceeds the calculated total number of binding sites in the liver does not reduce the efficiency of the pathway, and studies of [3H]leucine incorporation indicate that the lectin, unlike the bound asialoglycoprotein, is not destroyed in the elimination process. Cytochalasin B (80 microgram/100 g body wt.) had no measureable effect on the hepatic clearance of asialofetuin. Administration of colchicine (10 mg/100 g body wt.) resulted in transitory accumulations of asialoorosomucoid in the liver, presumably due to interference with the intracellular transport of the endocytised protein.

Animals↗
PubMed1978

Synthesis of antithrombin III and alpha-1-antitrypsin by the perfused rat liver.

Livers isolated from control or turpentine-injected rats were perfused for 3 h with human red cells suspended in Krebs-Henseleit solution containing bovine serum albumin, dextran, glucose, heparin, cortisol, insulin, a mixture of 20 amino acids and [3H]leucine. Changes in the concentrations of antithrombin III and alpha-1-antitrypsin were evaluated by rocket immunoelectrophoresis using specific antisera, and incorporation of the 3H radioactivity into the total protein, albumin, antitrhombin III and alpha-1-antitrypsin in the perfusate was measured. The results indicate that both antithrombin III and alpha-1-antitrypsin are synthesized in the liver. Local inflammation induced in the liver donors moderately stimulated the synthesis of alpha-1-antitrypsin but it affected only marginally that of antithrombin III.

Albumins↗
PubMed1978

Isolation and partial characterization of rabbit plasma alpha1-antitrypsin.

Alpha1-Antitrypsin was isolated from rabbit plasma by salting out with (NH4)2SO4 followed by ion-exchange chromatography either on DEAE-Sephadex or DEAE-cellulose (each at pH8.8 and 6.5), and affinity chromatography on Sepharose-Cibacron Blue and Sepharose-concanavalin A. The protein thus obtained was homogeneous during crossed immunoelectrophoresis by using an antiserum to whole rabbit plasma, but it migrated as two broad bands when electrophoresed in alkaline polyacrylamide gels. Under optimal loading conditions, two or three subcomponents could be distinguished in each band. The two major forms of rabbit alpha1-antitrypsin, designated components F and S, were separated by preparative polyacrylamide-gel electrophoresis, and some of their physico-chemical properties were established. Both forms reacted with trypsin at a molar ratio of 1:1. Their elution volumes from a Sephadex G-200 column were identical, corresponding to a mol.wt. of 58000; however, some heterogeneity was observed after sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Isoelectric focusing in polyacrylamide gel in a pH 4-6 gradient revealed a multiple-band pattern for each form in the range of pH4.4-4.9. The two forms of rabbit alpha1-antitrypsin possessed the same N-terminal amino acid (glutamic acid) and had very similar amino acid and carbohydrate compositions.

Amino Acids↗
PubMed1978

Effect of experimental inflammation on the synthesis and distribution of antithrombin III and alpha1-antitrypsin in rabbits.

Local inflammation, induced by s.c. injection of turpentine, evoked characteristic changes in the metabolism of antithrombin III, and alpha1-antitrypsin. For a period of approximately 36 h, the plasma half-lives of both protease inhibitors were shortened to 70--74% of the respective preinjection values. Similar changes were also observed in the slope of iodine-labelled albumin, suggesting that increased capillary permeability was primarily responsible for the losses of labelled proteins from the circulation. Incorporation of [3H]- or [14C]-leucine into albumin changed little during inflammation, but markedly increased values were measured for anti-thrombin III (3-fold), alpha1-antitrypsin (4-fold) and, above all, for fibrinogen (7-fold) 24 h and 48 h after the injection of turpentine. These changes in synthesis and elimination rates resulted in the following net balances: fibrinogen concentrations in plasma rose substantially during the early phase of inflammation; alpha1-antitrypsin concentrations increased gradually but to a significantly lesser extent, peak concentrations being reached after a reverse trend in fibrinogen concentrations had become apparent; antithrombin III concentrations remained steady throughout at levels which were only marginally above the pretreatment values.

Animals↗
PubMed1978

The effect of D-galactosamine on plasma protein synthesis by the perfused rat liver from turpentine-stimulated donors.

D-galactosamine (100 mg) was added to the reconstituted blood during 4h perfusion of livers isolated either from control rats or those injected with turpentine 20 h or 5 h earlier. This dose of galactosamine administered 30 min before [3H]lysine significantly inhibited the incorporation of the label into liver proteins, and even more into plasma proteins, but albumin and acute-phase reactants (fibrinogen, seromucoid fraction, Concanavalin A-adsorbed glycoproteins) were all similarly affected. When galactosamine was administered in vivo simultaneously with turpentine, and the liver was isolated 5 h later, trauma-induced fibrinogen synthesis was selectively inhibited. This can be explained either by a differential control of synthesis of various acute-phase reactants, or by augmentation of catabolism of fibrinogen in galactosamine-treated rats. Crossed immunoelectrophoresis of the full perfusate or Concanavalin A-adsorbed glycoproteins did not reveal any significant effect of galactosamine on the protein pattern obtained from control or turpentine-stimulated liver donors.

Animals↗
PubMed1976

Isolation and some molecular parameters of elastase-like normal proteinases from horse blood leucocytes.

Cytoplasmic granules were isolated from horse blood polymorphonuclear leucocytes by the heparin method and extracted with 0.9% NaCl by repeated freezing. Soluble proteins were separated on a column of Sephadex G-75 followed by chromatography on a column of CM-Sephadex with a NaCl gradient. Gel filtration, density-gradient centrifugation, isoelectric focusing and 0.1% sodium dodecyl sulphate/polyacrylamide-gel electrophoresis at pH 7.0 and at pH 4.5 were used to determine molecular parameters of proteinases. Three enzymes hydrolysing both casein and N-benzyloxycarbonyl-L-alanine nitrophenyl ester were found in the granule extract: proteinase 1, mol.wt. 38000, pI5.3; proteinase 2A, mol.wt. 24500, pI8.8; and proteinase 2B, mol.wt. 20500, pI above 10. The latter two elastase-like proteinases were purified to apparent homogeneity.

Animals↗
PubMed1976

Substrate specificity and modifications of the active centre of elastase-like neutral proteinases from horse blood leucocytes.

Two proteinases (2A and 2B) purified from the granular fraction of horse blood leucocytes degrade casein (Km values 12.8 and 6mg/ml respectively) with maximum activity at pH 7.4 and in the presence of 2m-urea. Urea-denatured haemoglobin, fibrinogen, albumin and resorcin/fuchsin-stained elastin are digested at a slower rate. The enzymes hydrolyse synthetic substrates of elastase, N-benzyloxycarbonyl-L-alanine 4-nitrophenyl ester (Km 0.114 and 0.178 mM) and N-acetyl-tri-L-alanine methyl ester (Km 5.55 and 0.98 mM), but they do not hydrolyse synthetic substrates of trypsin, chymotrypsin and thrombin. The examined proteinases are completely inhibited by 2 mM-di-isopropyl phosphorfluoridate and show a sensitivity to butyl and octyl isocyanates similar to that of pancreatic elastase. The pH-dependence of their photoinactivation in the presence of Rose Bengal indicates the presence of histidine in the active centre. Proteinase 2A rather insensitive to iodination by IC1 as is pancreatic elastase, whereas proteinase 2B is totally inactivated after incorporation of five iodine atoms per enzyme molecule.

Animals↗
PubMed1976

Inhibition of the liver and plasma protein acute-phase response in mice by D-galactosamine.

Local inflammation evoked in Swiss albino mice by subcutaneous injection of Celite resulted in a rise of liver tyrosine aminotransferase activity and plasma level of fibrinogen and seromucoid, while liver alanine aminotransferase activity and plasma level of fibrinogen and seromucoid, while liver alanine aminotransferase activity and the plasma level of albumin and total protein remained unaltered. By measuring the incorporation of [14C] leucine, stimulation of liver and plasms protein synthesis by Celite injection was demonstrated. Administration of D-galactosamine (2-5 mg/10 g body weight) inhibited the enhanced synthesis of liver proteins, and especially of trauma-induced synthesis of plasma fibrinogen and seromucoid. The inhibitory effect of galactosamine was most pronounced when the amino sugar was injected simultaneously with Celite and then protein synthesis was measured 6 h later. The results obtained support the idea that high doses of galactosamine inhibit transcription of trauma-inducible mRNA in the liver and thus block the acute-phase response.

Animals↗