sepharose and Liver-Diseases

Topics: Artificial Organs; Biological Products; Charcoal; Chromatography, Affinity; Dialysis; Extracorporeal Circulation; Humans; Kidney; Kidney Failure, Chronic; Liver; Liver Diseases; Perfusion; Renal Dialysis; Resins, Synthetic; Sepharose; Serum Albumin

In recent years, with the widespread use of laparoscopic cholecystectomy and liver transplantation, complications involving the biliary system are increasing. All techniques available have a high risk of recurrence or high-morbidity problems. We developed a three-dimensional collagen duct modified with agarose hydrogel, to substitute the affected extrahepatic bile duct.. We used it in 40 guinea pigs and studied the histology and physiology at 4 wk, 3, and 6 mo after transplantation. Blood test, histologic techniques, and cholangiography were performed in all of them.. All experimental animals survived up to their sacrifices. Our graft showed highly potential applications to treat hepatobiliary diseases that require surgery.

Topics: Animals; Bile Duct Diseases; Bile Ducts, Extrahepatic; Biocompatible Materials; Cattle; Collagen; Gels; Guinea Pigs; Liver Diseases; Models, Animal; Sepharose; Tissue Engineering

Close monitoring of liver iron content is necessary to prevent iron overload in transfusion-dependent anemias. Liver biopsy remains the gold standard; however, MRI potentially offers a noninvasive alternative. Iron metabolism and storage is complicated and tissue/disease-specific. This report demonstrates that iron distribution may be more important than iron speciation with respect to MRI signal changes. Simple synthetic analogs of hepatic lysosomes were constructed from noncovalent attachment of horse-spleen ferritin to 0.4 microm diameter phospholipid liposomes suspended in agarose. Graded iron loading was achieved by varying ferritin burden per liposome as well as liposomal volume fraction. T1 and T2 relaxation times were measured on a 60 MHz NMR spectrometer and compared to simple ferritin-gel combinations. Liposomal-ferritin had 6-fold stronger T2 relaxivity than unaggregated ferritin but identical T1 relaxivity. Liposomal-ferritin T2 relaxivity also more closely matched published results from hemosiderotic marmoset liver, suggesting a potential role as an iron-calibration phantom.

Topics: Animals; Callithrix; Ferritins; Gels; Iron Overload; Liposomes; Liver Diseases; Lysosomes; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Phantoms, Imaging; Reproducibility of Results; Sepharose

The subforms of MM isozyme of creatine kinase (ATP:creatine N-phosphotransferase, EC 2.7.3.2, CK) in sera obtained from healthy adults and patients were determined by agarose gel isoelectric focusing (IEF). The patients were classified into six groups according to serum CK-MM activities and IEF patterns. The IEF spectra offered useful information on cell hyperplasia, augmented cell membrane permeability, cell destruction and release time of CK-MM in the circulation from the cells for diagnosis, progress observation and prognosis, especially in the cases of chronic hepatic diseases, acute myocardial infarction and muscular dystrophy. Macro CKs were also determined by IEF. Macro CKs could be completely distinguished from each other, and CK isozymes consisting of macro CK type 1 could be presumed by isoelectric points.

Topics: Chromatography, Gel; Creatine Kinase; Health Status; Humans; Immunoenzyme Techniques; Isoelectric Focusing; Isoenzymes; Liver Diseases; Muscular Dystrophies; Myocardial Infarction; Neoplasms; Prognosis; Sepharose

Topics: Aminohydrolases; Electrophoresis; Electrophoresis, Agar Gel; Guanine Deaminase; Humans; Liver Diseases; Sepharose

A new method is described for the separation of hydrophobic and hydrophilic forms of gamma-glutamyltransferase (GGT) present in human serum using hydrophobic chromatography on phenyl-Sepharose CL-4B columns. The analysis of sera of 30 normal healthy people showed that 71.49 +/- 8.73% (mean +/- SD) of the GGT was in the hydrophilic form. In sera from 52 patients with various liver diseases the major part of the total enzyme was hydrophobic GGT, contributing 67.38 +/- 13.8% of the total GGT activity. Elevated total serum GGT and a hydrophobic to hydrophilic ratio of more than one helped in diagnosing hepatobiliary disease in 51 of 52 patients. Papain digestion of serum converted hydrophobic to hydrophilic GGT, which was similar to soluble form of liver GGT. The results are discussed in relation to the proposed mechanisms of release of GGT from the liver to the circulation.

Topics: Adult; Chemical Phenomena; Chemistry; Chromatography, Affinity; Electrophoresis, Polyacrylamide Gel; Female; gamma-Glutamyltransferase; Humans; Hydrolysis; Liver Diseases; Male; Papain; Sepharose; Solubility; Water

Topics: Albumins; Animals; Blood; Dogs; Liver Diseases; Polysaccharides; Sepharose; Ultrafiltration

UDP-glucuronlytransferase, E.C. 2.4.1.17, has been solubilised from the microsomal fraction of liver homogenate from phenobarbital pretreated rabbits by lipase or detergent treatments. A 110-fold purification of the enzyme with respect to the crude homogenate was achieved by precipitation and column separations. The cholate-detergent solubilised enzyme was far more stable than that prepared by the lipase method. The partially purified UDP-glucuronyltransferase has been covalently bound to cyanogen bromide-activated agarose in the form of large haemocompatible beads to the extent of 0.22 mg protein per mg agarose dryweight, equivalent to about 25 mg of swollen gel. The acceptors for glucuronidation employed were the non-physiological phenolic compounds p-nitrophenol and 1-naphthol, and an exogenous and endogenous substance of physiological importance, namely paracetamol and phenol respectively. The immobilised enzyme exhibited at least 80% of the original activity of the solubilised enzyme, and the catalytic function was preserved for a much longer period of time in the carrier-bound form. The system described in this publication could well be applied in an extracorporeal liver assist device for the replacement of glucuronidation function.

Topics: Animals; Artificial Organs; Enzymes, Immobilized; Female; Glucuronates; Glucuronosyltransferase; Hemoperfusion; Hepatic Encephalopathy; Liver; Liver Diseases; Microsomes, Liver; Polysaccharides; Proteins; Rabbits; Sepharose; Uridine Diphosphate Glucuronic Acid

Polyclonal rheumatoid factor (pRF) was isolated from sera seropositive for RF and it was radioiodinated with lactoperoxidase method. Using the 125I-pRF and IgG-p-azobenzamidoethyl Sepharose 6B, a competitive inhibition radioassay for detecting immune complexes is described. The assay can be performed in 90 min utilizing 10 microliter of serum with good reproducibility and endogenous RF in test serum did not interfere the reaction. The test can detect complexes nearly to 8s and as low as 0.1 microgram/ml of aggregated human IgG. Decomplementation by heating test sera is unnecessary. The assay was performed at 4 degrees C but less inhibition activity was obtained when it was carried out at 24 degrees C.

Topics: Antigen-Antibody Complex; Binding, Competitive; Centrifugation, Density Gradient; Humans; Immunoenzyme Techniques; Immunoglobulin G; Liver Diseases; Radioimmunoassay; Rheumatoid Factor; Sepharose; Temperature

Topics: Artificial Organs; Carrier Proteins; Exchange Transfusion, Whole Blood; Glucuronidase; Humans; Liver; Liver Diseases; Perfusion; Renal Dialysis; Sepharose

In this investigation a new possibility of isoenzyme-differentiation of the gamma-glutamyl-transferase (GGT) (EC Nr.2.3.2.2.) was demonstrated by Concanavalin A and Con A-Sepharose. Because of the different sugar content of the glycoproteins distinction between liver- and kidney-GGT is possible. Furthermore it was possible for the first time to show a different precipitation behaviour of one glycoprotein to Concanavalin A in certain diseases. In cases of alcoholic hepatitis GGT looses its Concanavalin A-affinity because of increased neuraminic acid concentration. The possible reasons of the different behaviour to the binding affinity of Con A and Con A-Sepharose and GGT as well as additional use for enzyme-differentiation by Con A-Sepharose affinity chromatography are discussed.

Topics: Azathioprine; Chemical and Drug Induced Liver Injury; Chemical Precipitation; Chromatography, Affinity; Concanavalin A; Ethanol; gamma-Glutamyltransferase; Glycoproteins; Humans; Isoenzymes; Kidney Diseases; Liver Diseases; Neuraminic Acids; Neuraminidase; Polysaccharides; Sepharose

Topics: Albumins; Animals; Charcoal; Dogs; Humans; In Vitro Techniques; Liver; Liver Diseases; Rabbits; Rats; Renal Dialysis; Resins, Synthetic; Sepharose