Immunoassay of serum galactosyltransferase isoenzyme II in cancer patients and control subjects.

Serum galactosyltransferase isoenzyme II (GT-II) was assayed in 409 coded serum samples obtained from the National Cancer Institute Tumor Serum Bank using a monoclonal antibody-based immunoassay. The serum panel consisted of samples from patients with confirmed, metastatic ovarian, breast, stomach, esophageal, pancreatic, lung, colorectal, bladder, prostate, and cervical cancer, as well as benign disease controls corresponding to each cancer type, and confirmed healthy normal controls. The serum panel was matched for age and sex; 176 of 179 cancer patients had metastatic disease, and many had undergone previous therapy. GT-II was significantly elevated (P less than 0.01) in all pairwise tests (Wilcoxon) comparing cancer cases with normals and cancer cases with benign disease cases of the same site. A cutpoint of 200 milliunits of GT-II activity/ml of serum was selected, and only one of 50 normal control sera was elevated above this value, yielding a specificity of 98%. The overall sensitivity of the GT-II assay was 55.3%, with higher sensitivity shown by pancreatic (77%), prostate (65%), esophageal (64%), cervical (59%), and bladder cancer (58%).

Clinical value of serum glycoprotein galactosyltransferase levels in different histological types of ovarian carcinoma.

Serum glycoprotein galactosyltransferase levels were determined in 28 healthy women and 113 patients with ovarian carcinoma with various histological types, at different clinical stages. Ovomucoid, which possesses terminal N-acetylglucosaminyl residues, was used as glycoprotein acceptor. Clinical correlations between galactosyltransferase levels and tumor burden were examined, as well as the variations due to histology. Follow-up studies could be done for 60 patients, and correlations with clinical evolution, established. Galactosyltransferase might be a promising marker for the diagnosis and follow-up of ovarian carcinomas.

Uridine 5'-diphosphate-galactose:glycoprotein galactosyltransferase activity in the ovarian cancer patient.

Uridine 5'-diphosphate-galactose:glycoprotein galactosyltransferase activity was demonstrated in homogenates of normal ovary and ovarian epithelial adenocarcinomas. The specific activity of the enzyme in ovarian tumors was 3 to 5 times higher than in normal ovaries when the enzyme was assayed under identical conditions. The glycoprotein fetuin, from which terminal sialic acid and penultimate galactose were removed (fetuin minus N-acetylneuraminis acid and galactose), acted as an excellent exogenous acceptor. Galactosyltransferase from normal ovary and ovarian tumor cells had similar properties. Both required Mn2+ and Triton X-100 and had broad pH optima between 5.5 and 7. Galactosyltransferase activity was also measured in serum samples from ovarian cancer patients and normal healthy individuals in the presence of fetuin minus N-acetylneuraminic acid and galactose as exogenous acceptor. The enzyme levels were significantly elevated in the sera of ovarian cancer patients as compared to normal controls. The differences in the levels of this enzyme in the tissues and sera of normal individuals and ovarian cancer patients were not due to differential levels of the degrading enzymes such as uridine 5'-diphosphate-galactose pyrophosphatase or beta-D-galactosidase. Serial determinations were carried out on the sera of 5 ovarian cancer patients over a long period of time. The serum level of galactosyltransferase activity appeared to correlate with tumor volume as well as with the clinical status of the patient, which suggests possible leakage of the tumor enzyme into the host sera. Serial determination of this enzyme level in ovarian cancer patients seems promising in measuring tumor progression or success of therapeutic approaches.

Serum UDP-galactose: glycoprotein galactosyltransferase in diabetics with microangiopathy.

1. Levels of serum UDP-galactose:glycoprotein galactosyltransferase in 117 unselected diabetics were compared with those in 60 non-diabetic healthy controls. 2. Enzyme activity (mean +/- 2 S.D.) of control sera was found to be 90.2 +/- 21.5 etamoles/ml/hr at 37 degrees. In 30 of the 117 diabetic sera (26%) enzyme activity was elevated (greater than mean + 2 S.D. of the controls). Sixteen of 19 (84%) patients with retinopathy, 16 of 26 (62%) patients with peripheral vasculopathy and 13 of 26 (50%) patients with neuropathy had higher levels of serum enzyme. When serum enzyme levels of groups of diabetics with retinopathy, peripheral vasculopathy and neuropathy were compared with the enzyme level in all diabetics, there was a significant difference with p values of 0.001, 0.05 and 0.05 respectively.

Glycoprotein sialyl- and galactosyl transferase activities in erythrocyte membranes in alcoholic patients and healthy controls.

In investigating possible mechanisms underlying carbohydrate deficiencies in serum transferrin and erythrocyte membranes in alcoholics, a total of 27 alcoholic patients and 27 healthy controls were examined for the activities of sialytransferase in serum and erythrocyte membranes and galactosyltransferase in erythrocyte membranes. The enzymes were assayed with endogenous and different exogenous glycoprotein acceptors and with [14C] CMP-sialic acid and [14C] UDP-galactose as substrates. No decrease in enzyme activities were found in alcoholic patients compared to controls, indicating that chronic ethanol abuse does not exert any direct inhibitory effect on these glycosyltransferases in isolated erythrocyte membranes or on sialytransferase in serum. Further studies of the effect of ethanol on the metabolism of complex carbohydrates are clearly necessary.

Human plasma uridine diphosphate galactose-glycoprotein galactosyltransfertase. Purification, properties and kinetics of the enzyme-catalysed reaction.

The soluble galactosyltransferase of human plasma catalysed the transfer of galactose from UDP-galactose to high- and low-molecular-weight derivatives of N-acetylglucosamine, forming a beta-1-4 linkage. The enzyme was purified by using (NH4)2SO4 precipitation and affinity chromatography on an alpha-lactalbumin-Sepharose column. The galactosyltransferase was maximally bound to this column in the presence of N-acetylglucosamine, and the enzyme was eluted by omitting the amino sugar from the developing buffer. The molecular weight of the enzyme was estimated to be 85000 by gel filtration. The assay conditions for optimum enzymic activity was 30 degrees C and pH7.5. Mn2+ ion was found to be an absolute requirement for transferase activity. The Km for Mn2+ was 0.4 mM and that for the substrate, UDP-galactose, was 0.024 mM. The Km for the acceptors was 0.21 mM for alpha1-acid glycoprotein and 3.9 mM for N-acetylglucosamine. In the presence of alpha-lactalbumin, glucose became a good acceptor for the enzyme and had a Km value of 2.9 mM. Results of the kinetic study indicated that the free enzyme reacts with Mn2+ under conditions of thermodynamic equilibrium, and the other substrates are added sequentially.

[Separation of isoenzymatic forms of human serum galactosyltransferase by chromatofocusing and isoelectric focusing. Application to cancerology]. / Séparation des formes isoenzymatiques de la galactosyltransférase du sérum humain par chromatofocalisation et focalisation isoélectrique. Application à la cancérologie.

Chromatofocusing is used to separate the multiple isoenzyme forms of human serum galactosyltransferase. At least 11 major peaks of activity are observed in normal sera, which are eluted between pH 4.3 and 6.9; a fraction of activity is eluted above pH 7.0. The normal patterns are compared with those obtained with sera from cancer patients and with an ascitic fluid. Chromatofocusing appears as resolutive as agarose isoelectric focusing.

Serum beta-(1----4)-galactosyltransferase activity with synthetic low molecular weight acceptor in human ovarian cancer.

A modified procedure was developed for the determination of UDP-galactose: 2-acetamido-2-deoxy-glucopyranoside beta-(1----4)-galactosyltransferase (GT) in human serum which employed the synthetic substrates p-nitrophenyl 6-0-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)-beta-D-galactopyranoside and p-nitrophenyl 6-0-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)-alpha-D- mannopyranoside as acceptors. The enzyme products were identified by thin layer chromatography with authentic reference compounds, and the galactosyl linkage was characterized by hydrolysis with beta-D-galactosidase from jack beans. The diagnostic value of this GT for ovarian cancer was tested by measuring the serum enzyme activity in 28 ovarian cancer patients with disease, 20 ovarian cancer patients with no clinical evidence of disease, and 22 healthy females. Although the level of the enzyme activity was significantly higher (P less than 0.002) in the serum of patients with active disease when compared to healthy controls, an appreciable overlap of enzyme activity was found between them. Also, no correlation was found between enzyme activity and tumor size. Differences in methodology and selection of patients makes it difficult to compare results from other reports. However, based on our improved assay procedure, we suggest caution should be exercised in evaluating the merits of GT as a diagnostic marker for ovarian cancer.

Glycoprotein metabolism in human renal disease: serum glycoproteins and glycoprotein: glycosyl transferase levels in chronic renal failure.

Glycoprotein: galactosyl and glycoprotein: sialic acid transferase activity were measured in the serum of patients with renal insufficiency. Significant elevations in enzyme activity were found in all patients including those on regular hemodialysis. Galactosyl transferase activity was elevated in a group of renal transplant recipients with normal serum creatinine. Serum glycoproteins from renal failure serum have an altered carbohydrate composition, the major finding being a reduced galactose content. A smaller reduction in sialic acid content was also observed. The findings are discussed in terms of glycoprotein metabolism and the clinical setting of chronic renal failure.