[The effect of different antitubercular agents on phagosome fusion with lysosomes, on the F-actin content in mouse peritoneal macrophages and on G-actin polymerization in vitro]. / Vliianie razlichnykh protivotuberkuleznykh sredstv na sliianie fagosom s lizosomami, na soderzhanie F-aktina v peritoneal'nykh makrofagakh myshi i na polimerizatsiiu G-aktina in vitro]

Effects of various antituberculosis remedies (ATR)--isoniazid (INA), saluzid (SA), streptosaluzid (SS), ethambutol (EB), sodium para-aminosalicylate (SPAS) on phagosome-lysosome (PL) fusion, on F-actin content in mouse macrophages and on G-actin polymerization in vitro were studied. The ATR of choice have been shown to stimulate the PL fusion. INA (0.2 mM), SA (0.02 mM), SS (0.05 mM), EB (0.08 mM) and SPAS (0.5 mM) increased the F-actin content and changed its localization within macrophages. ATR changed the character of G-actin polymerization in vitro. Possible mechanisms of interrelation between cytoskeleton (actin part) changes and PL fusion are discussed. The results obtained suggest to use the ATR-induced changes in PL fusion and F-actin content in the cells for estimating therapeutic effects of respective antituberculosis remedies.

[The effect of chemical agents on lysosome fusion with phagosomes and on the F-actin content in murine peritoneal macrophages]. / Vliianie nekotorykh agentov na sliianie lizosom s fagosomami i na soderzhanie F-aktina v peritoneal'nykh makrofagakh myshei.

The influence of natural and synthetic polyamines, phalloidin, cytochalasin D, vinblastine, colchicine, puromycin, chlorpromazine, urea, glutaraldehyde, and ethanol on the phagosome-lysosome fusion and the content of F-actin in murine peritoneal macrophages has been studied. Fluorescent phallotoxin FITC-phalloidin was used to stain F-actin. Natural polyamines (spermine, spermidine, putrescine), phalloidin, ethanol (0.1 M) stimulated the phagosome-lysosome fusion and increased the mid-content of F-actin in macrophages. Cytochalasin D, vinblastine, colchicine, puromycin, chlorpromazine, urea, glutaraldehyde, ethanol (0.15 and 0.2 M) inhibited this process and decreased the mid-content of F-actin. Possible mechanisms of the interconnection of cytoskeleton and the phagosome-lysosome fusion are discussed.

[The effect of diamines on the process of lysosome fusion with phagosomes in mouse peritoneal macrophages]. / Vliianie diaminov na protsess sliianiia lizosom s fagosomami v peritoneal'nykh makrofagakh myshei.

Diamines (DA), characterized by a general formula H2N-(CH2) n-NH2 in which n varies from 2 to 10, inhibit the phagosome-lysosome fusion in murine peritoneal macrophages. The DA concentration was 0.2, 0.5 and 1.0 mM. The inhibitory effect increased with increasing the number of CH2-groups in the DA molecule. It was suggested that DA could influence the lysosomal membrane state. An additional proof of such changes was obtained with 1,6-diphenyl-1,3,5-hexatrien (DPH) as a fluorescent probe. Lysosomes isolated from murine peritoneal macrophages by differential centrifugation were used. It was found that DPH fluorescence intensity in lysosomal membrane increased under the influence of DA.

[The effect of polyanion, cationic and anionic dyes and nonelectrolytes on phagosome fusion with lysosomes in mouse peritoneal macrophages]. / Vliianie polianiona, kationnykh i anionnykh krasitelei i neélektrolitov na sliianie fagosom s lizosomami v peritoneal'nykh makrofagakh myshei.

Polyanion (mannansulphate), cation dyes (Toluidine blue, Nile blue), anion-dye (Trypan blue) also non-electrolytes (urea, glutaraldehyde) were found to inhibit the phagosome-lysosome fusion in murine macrophages. The mechanism of this effect is discussed.

[The effect of polyamines on lysosome fusion with phagosomes in mouse peritoneal macrophages]. / Vliianie poliaminov na sliianie lizosom s fagosomami v peritoneal'nykh makrofagakh myshei.

The influence of polyamines on the phagosome-lysosome fusion in murine peritoneal macrophages and on polymerization of G-actin from the rabbit muscle in vitro has been studied. Both natural polyamines (spermin, spermidin, putrescin) and synthetic phenyl derivates of polyamines (3,3'-diaminobensidin, 1,5-naphtalin diamine, 4,4'-diaminodiphenilmetan, dancylcadaverin) were used. Unlike the phenyl derivates of polyamines and putrescin, spermin and spermidin stimulate the phagosome-lysosome fusion to induce G-actin polymerization. Possible mechanisms of action of the above polyamines are discussed.

[Decreased G-actin content in cells at injury]. / Snizhenie soderzhaniia G-aktina v kletkakh pri ikh povrezhdenii.

The content of G-actin was measured in aqueous extracts of rat macrophages and human lymphocytes at hyperthermia and hypoxia. The G-actin content in altered cells decreases the greater the stronger the injury. This decrease correlates well with the data on visual estimation of cell light diffusing. A possible contribution of actin polymerization and its interaction with proteins in the colloidal reactions of altered protoplasm is discussed.

[Characteristics of the granules formed in nucleated erythrocytes as affected by cardiotrast]. / Kharakteristika granul, obrazuiushchikhsia v iadernykh éritrotsitakh pod vliianiem kardiotrasta.

Numerous granules are formed in frog erythrocytes under the influence of cardiotrast (C) (diethanolamine-3,5-diiodo-4-pyridone-1-acetic acid). However, as revealed by cytospectrophotometric investigation and X-ray microanalysis, no C was accumulated in these granules. It is known that C can dissociate into diethanolamine and 3,5-diiodo-4-pyridone-1-acetic acid. It was assumed that under the influence of C granule formation may occur at the expense of an uncharged diethanolamine form penetrating into lysosome-like structures to be accumulated by protonation process. Diethanolamine was found to provoke granule formation in frog erythrocytes. It is impossible to reveal substance in granules because it is colourless and has no ultraviolet absorbtion band. Under the influence of some inhibitors of energy metabolism on granule formation and the granules formed, their inhibitory effect is exerted on the process of granule formation. In granules isolated by differential centrifugation activity of some marker lysosomal enzymes was found which enabled us to attribute these granules to lysosome-like structures.

[Epsilon-aminocaproic acid inhibits the propagation of damage along the nerve fiber]. / Epsilon-aminokapronovaia kislota tormozit rasprostranenie povrezhdeniia v myshechnom volokne.

An inhibitor of proteinases--epsilon-aminocaproic acid--inhibits the propagation of destruction along the muscle fiber isolated from frog m. iliofibularis. It is assumed that generalization of the injury is hampered by a protein precipitate formed at the site of the injury, which is preserved due to inhibition of proteinase activity.

[Nature of the changes in the activity of water-soluble enzymes in exposure of muscles to harmful agents. IV. The study of hexokinase extractability from intact and altered muscles]. / K voprosu o prirode izmenenii aktivnosti vodorastvorimykh fermentov pri deistvii na myshtsy povrezhdaiushchikh agentov. IV. Izuchenie izvlekaemosti geksokinazy iz intaktnykh i al'terirovannykh myshts.

A possibility of hexokinase binding with actomyosin in skeletal muscles of Rana temporaria L., and the effect of thermal alteration (15 min at 36, 37, 38, 40 and 42 degrees C) on the binding were studied. Solutions of KCl (0.075 M and 0.15 M) extract more hexokinase from intact and altered muscles than does an non-electrolyte medium. Hexokinase freely dissolved in hyaloplasm is extracted in non-electrolyte medium. Hexokinase bound with structural components of the muscle cell is extracted upon the increase in ionic force of the extractant. The solubilizing effect of electrolytes on hexokinase is higher in alterated muscles than in the intact muscles indicating the increase in hexokinase binding under thermal alteration. Actomysin isolated from muscles reveals hexokinase activity. In reprecipitated actomyosin, the larger part of its hexokinase remains in actomyosin gel, the level of hexokinase activity not depending on the number of reprecipitation procedures or on the volume of washing solution. Hexokinase in actomyosin gel is less stable to the thermal action than in water supernatant of muscle extract. This may be due to the increase in hexokinase binding with actomiosin whose sorption activity increases under the thermal denaturation.

[Effect of metabolic inhibitors on formed novocaine and neutral red segregation zones in frog erythrocytes]. / Vliianie ingibitorov metabolizma na sformirovannye zony segregatsii novokaina i neitral'nogo krasnogo v éritrotsitakh liagushki.

Effects of some metabolic inhibitors, as well as of biologically active compounds (diakarb, ethidium bromide and a phenanthridine alkaloid sanguinarine) on the formed novocaine and neutral red segregation zones were studied. The volume of granules diminished under the influence of a glycolytic inhibitor iodoacetate, uncouplers of oxidative phosphorylation (2,4-dinitrophenol and carbonyl cyanide trifluoromethoxyphenylhydrozone), and respiratory inhibitors (antimycin A and rotenone), as well as under the influence of cycloheximide - an inhibitor of protein synthesis. Diakarb, ethidium bromide or sanguinarine also provoked a regression of the segregation zones. It has been found that all these compounds are inhibitors of ATPase activity of the isolated segregation zones. A possible mechanism of volume decreasing in segregation zones under the influence of both the metabolic inhibitors and diakarb, ethidium bromide and sanguinarine is discussed.

[ATPase activity of the novocaine segregation zones isolated from the erythrocytes of the common frog]. / ATFaznaia aktivnost' zon segregatsii novokaina, izolirovannykh iz éritrotsitov travianoi liagushki.

Novocaine segregation zones in frog's erythrocytes, isolated by differential centrifugation, were shown to be ATPase active. The enzyme displays half of its maximum activity at 0.18 Mm ATP concentration to be inhibited by high concentrations of ATP. ATPase is activated by both Mg2+ and Ca2+ (in a lesser degree), with the maximum activity being at pH 7.5. A 5 minutes heating without the substrate results in decreasing the enzyme activity at 30 degrees, and in the total inhibition at 50 degrees C. Along with ATP, the enzyme can hydrolyse GTP and, in a lesser degree, ADP and sodium pyrophosphate. The ATPase activity is not effected with oligomycin (0.5-1.5 mkg/ml) or ouabaine (0.1 mM). Oligomycin in concentration 5 micrograms/ml induced non-specific inhibition of ATPase. Uncouplers, like 2,4-dinitrophenol and carbonyl cyanid p-trifluorometoxyphenylhydrazone, stimulate the enzyme activity. The lack in the ATP-ase sensitivity to oligomycin (specific inhibitor of mitochondrial F1-ATPase) and ouabaine (specific inhibitor of Na+, K+-ATPase) may suggest that the ATPase activity of novocaine segregation zones in frog's erythrocytes is not associated with a random contamination with mitochondria or cytoplasmic membranes. The ATPase under study has much in common with the lysosomal +H-ATPase. The results obtained support a hypothesis that +H-ATPase may function as a course of protones for maintaining acidic medium in segregation zones and promote accumulation of weak bases by means of their protonation.

[Changes in hexokinase activity during muscle alteration]. / Izmeneniia aktivnosti geksokinazy pri al'teratsii myshtsy.

Changes in extractability and activity of hexokinase (HK) were studied under the action of heating and of urea on skeletal muscles of Rana temporaria L., and besides the stability of this enzyme in muscle extract to those agents in vitro was examined. Under a 15 minutes heating of muscle, a decrease in extractability (the activity calculated for 1 g of tissue) and activity (the activity calculated for 1 mg of protein) of hexokinase is first revealed at 37 degrees C. Then the enzyme extractability decreases gradually in accordance to the decrease in extractability of the total water-soluble protein; the level of hexokinase activity attained at 37 degrees does not change up to 40 degrees. At 42 degrees the activity of the enzyme is completely inhibited. Under the heating of the muscle extract, the decrease of enzyme activity takes place at 36 degrees, the level achieved being stable up to 42 degrees C. Under the action of urea on the muscle at the reversible phase of alteration (1 M urea from 5 minutes to 2 hours at room temperature, 1 M urea for 9 hours at + 4 degrees C), hexokinase activity increases, calculated for 1 g of tissue and for 1 mg of protein. Under the irreversible disappearance of muscle excitability (1 M urea during 9 hours, 2 M urea during 2 hours at room temperature) no hexokinase activity was revealed. The activation of the enzyme is discussed in connection with the data on the increase of ATP content in muscle under the urea alteration. The treatment of the enzyme in muscle extract with 1 M urea decreases its activity in 30 minutes down to 67%; the level achieved does not change during 20 hours.

[Effect of nonelectrolytes on the polymerization of G-actin]. / Vliianie neélektolitov na polimerizatsiiu G-aktina.

The non-electrolytes--urea, thiourea, sucrose, glycerin, polyethylendioxid, glutaraldehyde--inhibit polymerization of G-actin in vitro. The results are discussed in association with the capability of some non-electrolytes preventing colloid reactions of alterated protoplasm and increasing the stability of cells to injuring agents.

[Nature of the changes in water-soluble enzyme activity in the exposure of muscles to harmful agents. III. A study of creatine kinase extractability from intact and altered muscles]. / K voprosu o prirode izmenenii aktivnosti vodorastvorimykh fermentov pri deistvii na myshtsy povrezhdaiushchikh agentov. III. Izuchenie izvlekaemosti kreatinkinazy iz intaktnykh i al'terirovannykh myshts.

Binding of creatine kinase (CrK) in intact and thermally altered (15 minutes at 37 and 38 degrees C) skeletal muscles of frogs (Rana temporaria L.) was studied according to CrK extractability from muscles. The CrK activity in actomyosin isolated from muscles and its changes in thermally altered actomyosin were detected. Solutions of KCl (0.075, 0.15, 0.3 and 0.6 M) and of NaF (0.15 M) extract additional amounts of CrK from intact and altered muscles. Inhibiting effects of KCl, NaF and K2HPO4 on CrK in muscle extract were found. The latter exerts the most pronounced inhibiting effect, which prevents it from being used for CrK extraction purposes. Among the structural components of muscle that bind CrK actomyosin was detected. The actomyosin isolated from muscles revealed CrK activity. CrK in actomyosin gel is much more stable to thermal effect than that in a water extract of muscle. Such a thermostabity of CrK within the system of the whole muscle cell may apparently result from its being bound to actomyosin.

[NADPase and NADase in the peritoneal macrophages of mice]. / NADFaza i NADaza peritoneal'nykh makrofagov myshei.

Murine peritoneal macrophages are able to hydrolyse NAD+ and NADP+. The NADPase activity exceeds that of NADase by 22-24%. The pH optima for both the enzymes are, respectively, 6.0 and 7.0. NAD hydrolysis is considerably activated by Mg2+, whereas NADP hydrolysis remains not affected. NAD+ does not change NADPase activity, while NADase activity is inhibited by NADP by 25-30%. A diazonium salt of sulfanilic acid, known to be an inhibitor of cell plasma membranes, does not affect NADP+ hydrolysis and causes a 20-30% retardation of NAD+ hydrolysis. The data obtained suggest that murine peritoneal macrophages contain two hydrolytic enzymes: NADase and NADPase.

[Nature of the changes in water-soluble enzyme activity in the action of injurious agents on the muscles. II. The change in the extractability of aldolase from muscles exposed to urea]. / K voprosu o prirode izmenenii aktivnosti vodorastvorimykh fermentov pri deistvii na myshtsy povrezhdaiushchikh agentov. II. Izmenenie izvlekaemosti iz myshts al'dolazy pri deistvii mocheviny.

A study was made of solubilization of aldolase isolated from homogenates of skeletal muscles, both intact and being in the state of contracture due to urea action. Compared to water, electrolytes extract more aldolase from homogenates of intact and altered muscles. Almost the same amounts of aldolase were extracted with electrolytes from homogenates of muscles, which lost irreversibly their excitability, and of intact muscles. The actomyosin isolated from muscles displayed aldolase activity not removed under reprecipitation. The aldolase activity of actomyosin, the increase in sorption activity of proteins due to their conformational changes, and the decrease in excitability of aldolase isolated from homogenates of altered muscles by urea doses inducing denaturation of actomyosin and aldolase, all this may suggest that the action of injured agents on muscle stimulates the ability of aldolase and actomyosin to interact. The ratio of free and bound forms of aldolase differs in the intact and in the altered muscle cell.

[Nature of the changes in the activity of water-soluble enzymes as affected by noxious agents acting on the muscles. I. A study of the extractability of phosphofructokinase from intact and altered muscles]. / K voprosu o prirode izmenenii aktivnosti vodorastvorimyk fermentov pri deistvii na myshtsy povrezhdaiushchikh agentov. I. Uzuchenie izvlekaemosti fosfofruktokinazy iz intaktnykh i alt'terirovannykh myshts.

Binding of phosphofructokinase (PPK) in intact and thermally altered (15 minutes at 38 degrees C) skeletal muscles of frogs (Rana temporaria) the the extractability of PPK from muscles was studied. PPK activity in actomyosin was also studied. Inhibiting effect of electrolytes (KCl, NaCl, CaCl2, MgCl2) on PPK in muscle extract does not allow to use them for the decision of the question of the interprotein interactions of PPK. 5 mM Na2-EDTA extracts additional PPK from homogenates of intact and altered muscles in comparison with PPK extracted in the media without Na2-EDTA (for intact muscles and altered muscles--509 and 729%). Under alteration of muscle, the binding of PPK increases. Among the structural components of muscle which bind PPK, proteins of actomyosin complex have been found.