[Determination of esterase activity of human and animal serine proteinases using fluorogenic esters of amino acids as substrates]. / Opredelenie ésteraznoi aktivnosti serinovykh proteinaz cheloveka i zhivotnykh s ispol'zovaniem v kachestve substratov fliuorogennykh éfirov aminokislot.

Two fluorogenic derivatives of amino acids are proposed as substrates for the purpose of enzymatic assay: N-benzyloxycarbonyl-phenylalanine-4-methyl umbelliferyl ester (substrate-1) and tert-butyloxycarbonyl-alanine-4-methyl-umbelliferyl ester (substrate-II). Chymotrypsin-like (hydrolysis of substrate-1), elastase-like (hydrolysis of substrate-II) esterase activity of bovine pancreatic chymotrypsin, activities of cathepsin G and elastase from human, porcine and rat neutrophils and esterase activity of human, porcine and rat serum were assayed. Differences in the level of chymotrypsin-like and elastase-like activities of human, porcine and rat serum were established. Activities of purified elastase and cathepsin G from human and animal neutrophils were shown to have no significant distinctions.

[Determination of the esterase activity of serine proteinases using synthetic substrates]. / Opredelenie ésteraznoi aktivnosti serinovykh proteinaz s ispol'zovaniem sinteticheskikh substratov.

The authors suggest a highly sensitive rapid and simple method for measuring esterase activities of bovine pancreatic chymotrypsin, human neutrophilic cathepsin D, and elastase, and of human blood serum chymotrypsin-like esterase and elastase-like esterase activities, with fluorogenic synthetic ethers, amino acid derivatives, employed as substrates: N-benzoxycarbonylphenylalanine 4-methylumbelliferyl ester (Z-Phe-OMC) and tret-butyloxycarbonyl-1-alanine 4-methylumbelliferyl ester (BOC-Ala-OMC).

[Alpha-2 macroglobulin from cerebrospinal fluid in neurosurgical diseases]. / Alpha-2-makroglobulin v spinnomozgovoi zhidkosti pri neirokhirurgicheskikh zabolevaniiakh.

Content of alpha 2-macroglobulin (alpha 2-MG) was estimated in cerebrospinal fluid (CSF) of patients with neurosurgical impairments. Minimal content of the globulin was found in patients with brain concussion (0.011 +/- 0.001 g/L, control group), maximal concentration--in severe craniocerebral trauma with brain contraction (0.056 +/- 0.007 g/L) and moderately increased content of alpha 2-MG was detected in intracranial tumors and drug-resistant epilepsy, 0.028 +/- 0.004 g/L and 0.025 +/- 0.004 g/L, respectively. Alteration in content of alpha 2-MG during postoperational period corresponded to clinical state of patients. Estimation of alpha 2-MG in CSF might be used as a criterion of brain impairment severity as well as for monitoring the treatment course.

[Various properties of cathepsin G and elastase from swine peripheral blood neutrophils]. / Nekotorye svoistva katepsina G i élastazy iz neitrofilov perifericheskoi krovi svin'i.

Using gel filtration through Sephadex G-100 and bioaffinity chromatography on contrical-Sepharose, cathepsin G and elastase were isolated from pig peripheral blood neutrophil granules and purified to homogeneity. Both enzymes hydrolyzed the total histone from calf thymus as well as synthetic substrates--tert-butoxy-L-alanine p-nitrophenyl ester (elastase) and benzoyltyrosine ethyl ester (cathepsin G). The use of natural and synthetic protease inhibitors showed that both enzymes were related to the group of serine proteases. The molecular mass of the cathepsin G subunit as determined by SDS polyacrylamide gel electrophoresis is 28-29 kD, that of elastase--30-31 kD. The pH optima for the hydrolysis of proteinaceous and synthetic substrates for cathepsin G and elastase are 8.0-8.5 and 7.0-7.5, respectively. The isoelectric points for elastase and cathepsin G are 9.7-10.0 and greater than 10, respectively; the temperature optima--30-40 degrees C and 50-60 degrees C, respectively. The amino acid composition of the two enzymes from pig granulocytes revealed a high content of arginine and was similar to that of human granulocytes.

[Structural and functional modifications of phospholamban and regulation of calcium transport in the sarcoplasmic reticulum under conditions of metabolic cardiac insufficiency]. / Strukturno-funktsional'nye modifikatsii fosfolambana i reguliatsiia transporta kal'tsiia v sarkoplazmaticheskom retikulume pri metabolichesk oi nedostatochnosti serdtsa.

The phosphorylation of phospholamban from dog myocardium is shown to correlate with the protein sensitivity to tryptic attack both in undamaged myocardium and under conditions of circulatory hypoxia and experimental infarction. In the absence of "exogenous" protein kinase a decreased phosphorylation of phospholamban is observed in the incubation mixture containing 10(-6) M cAMP both for circulatory hypoxia and myocardial infarction. In the latter case the phosphorylation remains diminished in the presence of "exogenous" protein kinase as well. The sarcoplasmic reticulum from damaged myocardium exhibited changes in the velocity of the oxalate-dependent transport of calcium which correlated with the corresponding degree of the phospholamban phosphorylation.

[Separation of proteolytic enzymes of polymorphonuclear leukocytes of pig peripheral blood]. / Razdelenie proteoliticheskikh fermentov polimorfnoiadernykh leikotsitov perifericheskoi krovi svin'i.

Application of gel filtration through Sephadex G-100 (pH 7,5) and ion-exchange chromatography on KM-Sephadex G-50 (pH 7.2) made it possible to detect high-active proteolytic enzymes: elastase-like esterases and serine proteinases in the granular fraction of polymorphonuclear leucocytes of pig peripheral blood. Their pH optimum is for hydrolysis of p-butyltetroxy-L-alanine n-nitrophenyl ester in the weak alkaline medium, for the splitting of histones-in the neutral and weakly alkaline media. Ion-exchange chromatography revealed a sharp increase in the total activity of elastase-like esterase, which may evidence for a separation of the inhibitor at the given stage of the enzyme purification. This is also confirmed by a considerable decrease in the activity of the serine proteinase fraction when adding proteins possessing the activity of elastase-like esterase.