Synthesis and degradation of eicosanoids in primary rat hepatocyte cultures.

Arachidonic acid metabolites may play an important role in liver physiology, yet hepatocyte prostaglandin synthesis has not been characterized extensively. We used RIA to study production and clearance of several eicosanoids in confluent primary cultures of rat hepatocytes in serum-free, hormonally-defined medium. Under basal, unstimulated conditions 6-keto-PGF1 alpha (spontaneous breakdown product of prostacyclin) and 13,14-dihydro-15-keto-PGE (DHK-PGE, a metabolite of PGE) accumulated in the culture medium.

Inhibition of human neutrophil and Pseudomonas elastases by the amyloid P-component: a constituent of elastic fibers and amyloid deposits.

The amyloid P-component (AP), a ubiquitous component of amyloid fibrils, is also a plasma protein and a connective tissue constituent. Its proximity to elastin, in particular, suggested that AP might serve to protect elastic tissue from hydrolytic enzymes. The inhibition of pancreatic elastase by AP has been reported.

Approaches to radiolabeling of antibodies for diagnosis and therapy of cancer.

The development of monoclonal antibodies of high affinity and selectivity for tumor antigens has supported the development of radiolabeled antibodies for diagnostic localization and targeted delivery of therapeutic radionuclides. Several radionuclide chelating agent systems have been developed for indium-111 and technetium-99m that have shown good sensitivity and specificity for tumor detection in patients. Feasibility for therapy has been shown in animal models and a few patient studies with iodine-131 and yttrium-90.

The use of reversed phase cartridges (C18) in processing plasma for analysis of indomethacin by high performance liquid chromatography.

The purpose of these studies was to develop a high performance liquid chromatographic (HPLC) assay for plasma levels of indomethacin. A reversed phase (C18 Sep-Pak) cartridge was used to process plasma for absorption of indomethacin, the internal standard, and impurities. The recovery of plasma indomethacin and the added internal standard was quantitative from 0.

Investigation of the heterogeneity of rhesus monkey alpha 1-antitrypsin.

Rhesus monkey alpha 1-antitrypsin (n = 144) was examined for heterogeneity by acid starch gel electrophoresis, isoelectric focusing in agarose and agarose gel electrophoresis. In contrast to other studies, no heterogeneity of Rhesus monkey alpha 1-antitrypsin could be documented using specific antisera. Rhesus monkey alpha 1-antitrypsin contained a reactive thiol.

Castanospermine inhibits glucosidase I and glycoprotein secretion in human hepatoma cells.

We studied the effect of the plant alkaloid castanospermine on the biosynthesis and secretion of human hepatoma glycoproteins. The HepG-2 cells, grown in the presence or absence of the alkaloid, were labelled with [2-3H]mannose and then the labelled glycopeptides were prepared by Pronase digestion. This material was analysed by gel filtration on Bio-Gel P-4 before and after treatment with endo-beta-N-acetylglucosaminidase H.

Alpha 1-antitrypsin: the effect of anticoagulants on the trypsin inhibitory capacity, concentration and phenotype.

The effects of anticoagulants on the determination of both trypsin inhibitory capacity and the concentration of alpha 1-antitrypsin measured by radial immunodiffusion, and on the alpha 1-antitrypsin phenotype were investigated. These results were compared with those obtained for serum. The following anticoagulants were investigated: sodium citrate; sodium oxalate; buffered citrate; potassium oxalate/sodium fluoride; sodium heparin; and potassium EDTA.

Liver disease and the PI ElembergM phenotype of alpha 1-antitrypsin.

The purpose of this article is to document clinical and pathologic observations concerning liver disease associated with the PI ElembergM phenotype of alpha 1-antitrypsin. Deposits of alpha 1-antitrypsin that were periodic acid-Schiff positive and stained with an antiserum to alpha 1-antitrypsin were found in the liver of a markedly jaundiced, terminally ill patient with Stage IV primary biliary cirrhosis. A biopsy performed three years earlier failed to reveal alpha 1-antitrypsin deposits.

Serum prealbumin levels in alpha 1-antitrypsin deficiency (PiZ).

The relationship between low serum prealbumin levels and alpha 1-antitrypsin deficiency (PiZ) was investigated. Pi typing was done by acid starch gel electrophoresis followed by crossed antigen-antibody electrophoresis and/or immunofixation. Serum prealbumin levels were determined by radial immunodiffusion.

The fixation of C3b to pneumococcal cell wall polymers as a result of activation of the alternative complement pathway.

The present study was performed in order to determine the identity of the pneumococcal cell wall polymer(s) to which C3b becomes fixed after activation of the alternative pathway. Purified pneumococcal autolysin was used to solubilize pneumococcal cell walls to which C3b had been fixed via activation of the alternative pathway. The resulting soluble cell wall polymers were then examined for the presence of C3b.

Mucus-stimulating activity in the sera of patients with cystic fibrosis: demonstration and preliminary fractionation.

The ciliated, mucus-secreting urn cell complex (UCC) is found swimming in the coelomic cavity of the marine invertebrate Sipunculus nudus. This cell complex, which can be maintained in suspension cultures, responds to various stimuli by hypersecreting mucus in the form of a cohesive mucus "tail." This tail can be measured and expressed as a multiple of "urn cell diameters.

Anaphylactic release of a basophil kallikrein-like activity. I. Purification and characterization.

These studies describe the IgE-mediated relase of a basophil kallikrein-like enzyme that is an arginine esterase and is inhibited by plasma, diisopropylphosphofluoridate, and Trasylol. The substrate specificity for the synthetic amino acid ester substrates p-toluenesulfonyl-L-arginien methyl ester, benzoyl-arginine methyl ester, and acetyl-tyrosine methyl ester is similar for the basophil enzyme and plasma kallikrein. The interaction of arginine esterase-active fractions from ion-exchange (DEAE-Sephacel) and gel filtration (Sepharose 6B) chromatography, with human plasma kininogen, generates immunoreactive kinin.

An unusual type of alpha1-antitrypsin deficiency in a child.

An unusual variant of serum alpha1-antitrypsin is described in a 15 5/6-year-old white male with a history of chronic pulmonary disease. The patient had a very low level of this protease inhibitor as demonstrated by tryptic inhibitory capacity and electroimmunoassay. Even though the patient's serum alpha1-antitrypsin was partially purified and concentrated, no phenotypic pattern was seen using conventional Pityping procedures (acid starch gel with crossed antigen-antibody electrophoresis or isoelectric focusing).

Basophil kallikrein of anaphylaxis (BK-A)--purification and characterization.

These studies describe the IgE-mediated release of a basophil kallikrein of anaphylaxis (BK-A) that has arginine esterase activity and is inhibited by plasma, DFP, and Trasylol. The interaction of BK-A active fractions from ion exchange (DEAE-Sephacel) and gel filtration (Sepharose 6B) chromatography, with human plasma kininogen generates immunoreactive kinin. The BK-A and kinin-generating activities co-chromatograph on DEAE-Sephacel and Sepharose 6B columns, and the quantity of kinin generated is, in general, proportional to the BK-A activity of the column fractions, suggesting that these two activities are subserved by the same protease.

Presence of alpha-1-antitrypsin on mitogen-stimulated human lymphocytes.

Human lymphocytes that have undergone concanavalin A-induced blastogenetic transformation demonstrate surface alpha-1-antitrypsin by immunofluorescence when examined after 72 hr of culture, whereas unstimulated cells do not. These results may indicate a role for alpha-1-antitrypsin in lymphocyte blastogenesis.

Isolation and characterization of alpha-1-antitrypsin from rhesus-monkey serum.

A simple, relatively gentle, procedure for isolation of rhesus-monkey alpha-1-antitrypsis from serum is described. The method consists of chromatographic separation of the fraction precipitated by 50-75%-satd. (NH4)2SO4 from pooled monkey serum on DEAE-cellulose followed by affinity chromatography on Sepharose-bound concanavalin A.