[Basal membrane proteins]. / Belki bazal'nykh membran.

The review summarizes recent achievements in biochemistry of basal lamina which is highly specialized structural element of extracellular matrix. Structural and functional characteristics of main basal lamina proteins are reviewed including collagen type IV, laminin, nidogen, and heparan sulfate-containing proteoglycans. Special attention is paid to characteristics of structure and biochemical composition of renal glomerular basal lamina which is one of the main components of renal filtration barrier. Possible methods of investigation and characterization of new basal lamina proteins are discussed which help in understanding of biochemical composition of this structure.

[Carnosine as a stimulator of cytotoxic and phagocytic function of peritoneal macrophages]. / Karnozin kak stimuliator tsitostaticheskoi i fagotsitarnoi funktsii peritoneal'nykh makrofagov.

Biochemical changes in peritoneal macrophages and their relatedness to the cytostatic and phagocytotic function in C3HA mice injected with a single intraperitoneal dose of 0.45 mM carnosine and 4-methyluracil or stimulated with peptone have been studied. During the first 24 hours after injection both carnosine and 4-methyluracil increase the activity of adenosine deaminase and purine nucleoside phosphorylase, the key enzymes of purine catabolism which is the main source of O2-. radicals in macrophages. In carnosine-stimulated macrophages the activity of membrane 5'-AMP nucleotidase decreases on days 1-3 after injection which points to alleviation of adenosine-induced inhibition as well as to macrophage activation. Carnosine increases the cytostatic and phagocytotic activities of macrophage coupled to O2-. production. The mechanism of the stimulating effect of carnosine on macrophages seems to consist in the dipeptide interaction with specific receptors localized on the plasma membrane of macrophagal cells.

[Mapping proteins from various structural regions of human kidney by two-dimensional electrophoresis]. / Kartirovanie belkov razlichnykh strukturnykh otdelov pochki cheloveka metodom dvumernogo élektroforeza.

The protein composition of various structural divisions of human kidney was studied using two-dimensional electrophoresis. Two-dimensional electrophoregrams of the cortical substance of human kidney revealed 165 polypeptide fractions within the pH range of 4.5-7.5, having molecular masses of 10 to 330 kDa. Electrophoresis of glomerular proteins gave 155 fractions with M(r) = 15-300 kDa, whereas fractionation of glomerular basement membrane proteins gave 40 fractions with M(r) = 30-330 kDa within the same range of pH. The M(r) values for all fractions and the relative electrophoretic mobility in the forward direction were determined. A comparative analysis of the electrophoregrams was conducted. The data obtained were used to construct two-dimensional maps of the cortical substance and glomerular proteins of human kidney.

[Effects of 4-methyluracil and carnosine on healing of skin wounds in rats]. / Vliianie 4-metiluratsila i karnozina na zazhivlenie kozhnykh ran u krys.

In experiments in vivo and in vitro the authors studied antioxidative properties of 4-methyluracil and carnosine, their capacity to inhibit sex and accelerate healing of skin wounds. 4-methyluracil and carnosine discover almost the same capacity to decrease in the tissues of the wound and the blood serum in the formation of various intermediate products of free radical oxidation. Data are given on the study of the dynamics of wound healing after a 5-day treatment with equimolar quantities.

[Synthesis of nucleic acids and dynamics of morphological indices of the granulation tissue of skin wounds in rats treated with 4-methyluracil]. / Sintez nukleinovykh kislot i dinamika morfologicheskikh pokazatelei granuliatsionnoi tkani kozhnykh ran u krys, lechennykh 4-metiluratsilom.

Effect of 4-methyl uracil on dynamics of morphological patterns and the rate of nucleic acids synthesis was studied in granulation tissue developed during healing of full-layer skin wounds. Application of ointment containing 4-methyl uracil onto the wounds exhibited an antiinflammatory efficiency and decreased accumulation of leukocytes in the wound. The drug shortened the mitotic cycle in dividing fibroblasts, as a result of which periods of DNA synthesis became more frequent exhibiting two days duration. The rate of RNA synthesis was increased 1.5-2-fold. Content of acid glycosaminoglycans and collagen was increased in intercellular space surrounding fibroblasts.

[Heparin and its antagonist--a quaternary ammonium salt of oligomer-25-conidine: distribution in subcellular organelles, effect on DNA synthesis in the regenerating rat liver]. / Geparin i ego antagonist--chetvertichnaia ammoniinaia sol' oligomera-25-konidina: raspredelenie po subkletochnym organellam, vliianie na sintez DNK v regeneriruiushchei pecheni krys.

Dynamics of distribution in subcellular organelles as well as effects on DNA synthesis of heparin, polycanidine (ChAS oligomer of 25 canidine) and their complexes were studied in regenerating rat liver tissue after partial hepatectomy. Polycanidine and heparin penetrated from circulation into hepatocyte cells, nuclei and mitochondria. After consecutive administration of polyelectrolytes polycanidine increased 2-2.5-fold the amount of heparin entering into cells. Polycanidine formed stable complexes with DNP in nuclei. Heparin and its complexes with polycanidine decreased the DNA synthesis within first day. Heparin appears to be responsible for the inhibitory effect, whereas administration of polycanidine only into animals caused a slight increase in 3H-thymidine incorporation into DNA as compared with controls.

[Dynamics of nucleic acid synthesis in the granulation tissue of skin wounds in the rat]. / Dinamika sinteza nukleinovykh kislot v granuliatsionnoi tkani kozhnykh ran krys.

Dynamics of nucleic acid synthesis was studied in granulation tissue, harvested within 2-11 days after an injury of rats with full-layer skin wounds, simultaneously with evaluation of the morphometric patterns of the tissue. Synthesis of nuclear DNA exhibited a wave-shaped dynamics. The intervals between the maximal values of the DNA synthesis constituted 3 days. Within the early steps of wound healing the intensive incorporation of 3H-thymidine into mitochondrial DNA was found, which appears to occur due to active mitochondriogenesis. In dynamics of total and nuclear RNA contents two peaks were observed, which did not coincide in time, thus suggesting the wave-shaped increase in biosynthetic processes in cells.

[Effect of the synthetic heparin antagonist, conidine oligomer 25, on the synthesis of nucleic acids in regenerating liver cells]. / Vliianie sinteticheskogo antagonista geparina oligomera-25 konidina na sintez nukleinovykh kislot v kletkakh regeneriruiushchei pecheni.

Localization of 14C-conidine oligomer (the synthetic antagonist of heparin) and of polyelectrolyte complex heparin-conidine oligomer 25 was studied in cells of regenerating liver tissue within 20 hrs after partial hepatectomy in rats. Effects of heparin, conidine oligomer 25 and of their polyelectrolyte complex on synthesis of nucleic acids were studied. Conidine oligomer 25 penetrated from circulation into hepatocytes and subcellular organelles of regenerating liver cells and was localized mainly in cytosol (75-88%). In nuclei and mitochondria of the hepatocytes 14C-conidine oligomer 25 was mostly found in the fraction of low density of the organelles. Polyanion heparin, polycation conidine oligomer 25 and their polymeric complex impaired replication and transcription processes.

[Evaluation of the hepatotoxic activity of several chlor-nitro derivatives of benzoic acid]. / Otsenka gepatotoksicheskogo deistviia nekotorykh khlor- nitroproizvodnykh benzoinoi kisloty.

Hepatotoxic effects of 4-chloro-3-nitrobenzoic acid (x-NBA), 3-nitrobenzoic acid (NBA) and 4-chlorobenzoic acid (CBA) were studied at the doses corresponding to LD50, 1/10 LD50 and 1/50 LD50. The toxic effects were estimated by monitoring alterations in activity of protein-synthesizing system in liver tissue and by the analyses for the presence in blood serum of two tissue-specific cytoplasmic enzymes of liver cells--urokaninase (EC 4.2.1.49) and histidase (EC 4.3.1.3). Single peroral administration of the toxic substances at a dose equal to LD50 led to dissimilar alterations in synthesis of liver proteins: activation within 0.5-5 hrs, distinct increase up to the maximal value within 60 hrs, with a gradual decrease approaching the control values within 8 days. Urokaninase and histidase activities were found in blood serum within 8 hrs after the intoxication, reaching the maximum within 15 hrs and exhibited the constant level during 25 hrs. Then the enzymatic activity was gradually decreased but it did not reach the control values within 70 hrs. After daily peroral administration of these toxic substances at the doses corresponding to 1/10 LD50 and 1/50 LD50, only the first dose inhibited the protein synthesis within 2 weeks. Within this period the enzymes studied were not found in blood serum. Distinct inhibition of the protein synthesis was observed within 4 weeks after administration of both these doses. In this case enzymatic activity was present in blood serum (up to 3 mumole/I L blood serum). Within 2 months after administration of these preparations the alterations studied were the most distinct.

[Stability and viability of spermatogenic cells after their separation and fractionation]. / Stabil'nost' i zhiznesposobnost' spermatogennykh kletok posle ikh vydeleniia i fraktsionirovaniia.

Spermatogenic cells isolated from seminiferous tubules by enzymatic treatment are very sensitive to the action of Triton X-100. The cells isolated by mechanical dissociation of spermatogenic epithelium are destroyed during storage. On the contrary, the cell separation by sedimentation velocity technique in human serum protein gradient leads to cell stabilization. As the result, the cells in collected fractions kept their living capacity and, what is more, their specific features.

[Modulation of the rate of mRNA poly(A)-segment shortening during embryonal and postnatal development]. / Moduliatsiia skorosti ukorachivaniia poli (A)-segmenta mRNK v embrional'nom i postnatal'nom razvitii.

A procedure for determination of the rate of poly(A) degradation based on a comparative analysis of poly(A)-segments of impulse-labelled mRNA under conditions of subsequent blocking of poly(A) synthesis by cordycepin was developed. The method described was used to analyze the rate of shorting of the poly(A)-segment of mRNA in the liver and brain cortex of the rat during the embryonic and postnatal development. It was shown that throughout ontogenesis the rate of poly(A) degradation in liver cells is significantly higher than in brain cortex cells for both types of polyribosomal mRNAs, the rate of poly(A) degradation within free polyribosomal mRNA being higher as compared to the membrane-bound mRNA for both tissues. On the 17th embryonic day the rate of poly(A) degradation is high in both polyribosomal types; it is decreased by the moment of birth and is either increased in the postnatal life or remains unchanged (free polyribosomes of the liver and membrane-bound polyribosomes of brain cortex).

[Purification, properties and regulation of urocaninase from rat liver]. / Ochistka, svoistva i reguliatsiia urokaninazy pecheni krys.

Urocaninase (EC 4.2.1.4.9) from rat liver homogenate has been purified, using protein precipitation at pH 4,8, ammonium sulfate fractionation, gel-filtration through Sephadex G-200 and chromatography on DEAE-cellulose. Upon DEAE-cellulose chromatography urocaninase is separated from the proteins possessing the activity of 3',5'-AMP-dependent protein kinase. The purified enzyme becomes activated after addition of ATP and exogenous protein kinase or one of the fractions resulting from DEAE-cellulose chromatography. Using [gamma-32P]ATP, it has been shown that such activation is accompanied by incorporation of at least one phosphate residue into the enzyme molecule. The mol. weight of urocaninase as determined by gel-filtration is about 110 000. The Km value for urocanate is 15 . 10(-6) M, the isoelectric point lies at 5,6. The mechanism of regulation of the urocaninase activity in rat liver is discussed.

[Substrate specificity, inhibitors and kinetics of deamidase AG (asparaginase-glutaminase) from Pseudomonas fluorescens AG]. / Nekotorye dannye o substratnoi spetsifichnosti, ingibitorakh i kinetike dezamidazy AG (asparaginazy-glutaminazy) iz Pseudomonas fluorescens AG.

Deamidase AG (asparaginase-glutaminase) from Pseudomonas fluorescens AG was shown to hydrolyze 1-glutamine and 1-asparagine highly effectively. Besides, the enzyme exhibited the rather high rate of deamidation of D-asparagine and D-glutamine (70% and 100%, respectively), Nalpha-butyl asparagine (63%) and among peptides -- of glycyl-L-asparagine (40%). L-glutamic acid gamma-methyl ester was hydrolyzed only slightly (5%). Effect of several substrate analogues on the deamidase AG activity was studied as well. Albiciine (alpha-amino-beta-ureide propionic acid) proved to be the strongest inhibitor (100%). Beta-Methyl aspartic acid, S-carbamoyl cysteine, alpha-ketoglutaric acid showed the slight inhibitory effect (20%). Amount of active centres per enzyme molecule was estimated by means of 14C-albiciine. Deamidase AG had apparently only one active centre. In estimation of relationship between the rate of reaction and substrate (L-asparagine) concentration, the reaction was found to follow Michaelis-Menten kinetics, K(m) = 4.5 with 10-4 M.

[Some properties of cytoplasmic thymidine kinase and nucleoside phosphotransferase from rat liver]. / Nekotorye svoistva tsitoplazmaticheskikh timidinkinazy i nukleozidfosfotoransferazy iz pecheni krys.

The activities of two deoxythymidine-phosphorylating enzymes--thymidine kinase and nucleoside phosphotransferase--were found in the cytoplasmic fraction of normal and regenerating rat liver. The specific activity of nucleoside phosphotransferase appeared to be by 50% higher than that of thymidine kinase. Nucleoside phosphotransferase has a broad specificity for the phosphate donor. This enzyme is more stable to heating and prolonged dialysis as compared to thymidine kinase. The enzymes respond differently to the addition of d-TTP, d-CTP and sturins A and B: thymidine kinase is strongly inhibited by these agents whereas nucleoside phosphotransferase is insensitive to d-TTP and d-CTP and is only slightly inhibited by sturins. On the other hand the activity of nucleoside phosphotransferase is considerably decreased after addition of ATP. Changes in the activities of both enzymes during 50 hrs following partial hepatectomy were studied. Two activity maxima were observed at 20-22 and 40-46 hrs of regeneration. Using polyacrylamide gel electrophoresis, three isoforms of both enzymes were found. The ratio between the isoenzyme content of the two enzymes from the cytoplasmic fraction of regenerating liver varied as compared to normal.

[The role of 3',5'-AMP in regulation of activity of histidase from rat liver and skin]. / Rol' 3':5'- AMP v reguliatsii aktivnosti gistidazy kozhi i pecheni krys.

A dependence of activity of histidase from rat liver and skin on the agents affecting the activity of the adenylate cyclase systeme was studied in vitro. Under conditions optimal for the activity of liver phosphorylase protein kinase the skin extract histidase was activated 2-3-fold. This is indicative of a possibility of regulation of the skin histidase activity via the adenylate cyclase system by modification of enzyme by phosphorylation-dephosphorylation, which is performed by 3':5'-AMP-dependent protein kinase. Theophylline at concentrations of 10(-4) M and 10(-3) M activates partially purified histidase (both liver and skin forms), probably in the course of direct interaction with the enzyme.