Progressive inhibition of human glandular (urinary) kallikrein by human serum and identification of the progressive antikallikrein as alpha 1-antitrypsin (alpha 1-protease inhibitor).

Human urinary kallikrein was inhibited by human alpha 1-antitrypsin (alpha 1-protease inhibitor) in a similar way as by equivalent amounts of human serum. The inhibitor present in the kallikrein-inhibitor complex formed was identified as alpha 1-antitrypsin by two-dimensional immunoelectrophoresis. Under the experimental conditions applied, 90 mIU (= 185 microgram) alpha 1-antitrypsin inhibits about 9 microgram of human urinary kallikrein in 24 h at 37 degrees C, 1 ml of human serum, containing 2-4 mg alpha 1-antitrypsin, inhibits about 70 microgram kallikrein.

Inhibitors of acrosomal proteinase as antifertility agents. A problem of acrosomal membrane permeability.

In vitro studies were performed to investigate the accessibility of acrosin to various proteinase inhibitors inside the intact acrosome of testicular, ejaculated, and uterine human spermatozoa. As test system the gelatin plate assay was used. For this assay it was shown formerly that a correlation exists between the size of the digested lysis areas (halo formation) and acrosin activity estimated with synthetic substrates.

Effect of human granulocytic elastase on isolated human antithrombin III.

The interaction of elastase isolated from human granulocytes with purified human antithrombin III was investigated. Antithrombin III did not display any inhibitory effect on granulocytic elastase. Dependent on enzyme concentration, however, granulocytic elastase induced progressive inactivation of antithrombin III leading to an almost complete loss of the thrombin inhibitory activity at a molar ratio of elastase: antithrombin III = 0.

The suitability of carboxymethylcellulose as a vehicle in reproductive studies.

The 2% aqueous solution of a sodium carboxymethylcellulose (CMC) was administered by gavage to 20 male and 40 female albino rats at the daily volume of 1 ml/100 g body weight. The males were treated for at least 60 days and the females for at least 14 days prior to mating and during a 6-day mating period. In one half of the females treatment was continued until sacrifice on day 14 of pregnancy; in the other half of the female treatment was continued until weaning of the progeny.

Clotting and other plasma factors in experimental endotoxemia: inhibition of degradation by exogenous proteinase inhibitors.

Endotoxemia in dogs was induced by a slow intravenous infusion of E. coli endotoxin for 2 h. Thereby, a significant decrease was observed in the plasma levels of several clotting, fibrinolysis and complement factors.

Integration specificity of an artificial kanamycin transposon constructed by the in vitro insertion of an internal Tn5 fragment into IS2.

IS2 has been marked genetically by the in vitro insertion into its HindIII site of a 3.3 Kb HindIII fragment of Tn5 conferring resistance to kanamycin. The transposition of the IS2::Km, thus obtained, to lambda has been found and insertion sites were characterised.

The sequence of IS4.

IS-elements are devoid of easily recognizable transacting functions and exert their visible effects in the position cis only (recent reviews Calos and Miller 1980; Starlinger 1980). It has been a matter of debate, whether these elements encode functions for their own transposition. In the case of the E.

N-Terminal amino acid sequence of boar sperm acrosin. Homology with other serine proteinases.

Boar acrosin with a molecular weight of 38,000 had been isolated. An N-terminal amino acid sequence of 52 residues was determined on the S-carboxymethylated material. The protein contains a single peptide chain.

Purification and properties of boar acrosin.

An isolation procedure in the presence of non-ionic detergents has been developed for the large-scale preparation of boar acrosin. Five steps including hydrophobic interaction chromatography on phenyl-Sepharose resulted in a 161-fold purification of the enzyme with an accumulation yield of 41%. The resultant acrosin preparation had a molecular weight of 38,000, an isoelectric point of 10.

Structure of the elastase-cathepsin G inhibitor of the leech Hirudo medicinalis.

The leech Hirudo medicinalis contains three different groups of proteinase inhibitor proteins, the thrombin-specific hirudin, the bdellins directed against trypsin, plasmin and acrosin, and the eglins which were discovered only recently. We are interested in the eglins mainly for two reasons: (i) They form strong complexes with the granulocytic elastase and cathepsin G with Ki values close to 1 x 10(-10) mol/l. Due to this property they are potential candidates for the therapeutic treatment of various diseases.

Isolation of an enzymatically active glandular kallikrein from human plasma by immunoaffinity chromatography.

A glandular kallikrein from human plasma was isolated by immunoaffinity chromatography and characterized. The molecular weight was determined to 40,000 by gel filtration. The enzyme preparation liberates kinins from human HMW kininogen (specific activity: 0.

Interactions of boar acrosin with detergents.

The activity of boar acrosin was found to be stimulated up to 260% by non-ionic detergents if present in concentrations above their critical micelle concentration. In addition, the stability of acrosin was remarkably enhanced in the presence of 0.1% (w/v) detergents.

Effects of hydroxyurea on postnatal growth and behaviour of rats.

Hydroxyurea was administered intraperitoneally to pregnant albino rats in a single dose of 2000 mg/kg on day 14 of gestation. Both the perinatal and postnatal mortality rates of the progeny were increased and their weight gain reduced. Exploratory locomotion of 32-day-old rats, calculated by means of an 'activity index', was significantly reduced.

Papulacandins, a new family of antibiotics with antifungal activity. Structures of papulacandins A, B, C and D.

The structures of the papulacandins A, B, C and D, new antibiotics of Papularia sphaerosperma have been established by means of spectral analysis and degradation reactions. Base catalysed hydrolysis of the main product papulacandin B (1) gave two new hydroxylated long-chain unsaturated fatty acids 5 and 6 along with a hitherto unknown spirocyclic diglycoside 7. The structure of 7 was determined by further degradation reactions.

Oligomerisation of boar acrosin.

Boar acrosin, a glycoprotein present in the acrosome of spermatozoa, tends to aggregate in the absence of detergents and lipids. Self-association products were analyzed electrophoretically by the method of Ferguson. Molecular weights ranging from 44 000 up to 237 000 were found, corresponding to acrosin monomer up to hexamer.

Isolation and characterization of human urinary kallikrein.

Human urinary kallikrein was purified by gel filtration on Sephacryl S-200 and affinity chromatography on aprotinin-Sepharose, followed by ion exchange chromatography on DEAE-Sepharose. Thus an enzyme preparation with a specific activity (using AcPheArgOEt as substrate) of 1 100 U/mg protein was obtained. A specific bioligical activity of 2 300 KE/mg was measured in the dog blood pressure assay and of 0.