Role of injury in the appearance of “activated” smooth muscle cells

In experiments on rats the ultrastructure of the muscle tissue of the inferior vena cava after disturbance of the outflow of blood, of the muscular coat of the stomach after resection of 50% of its fundus, and of the muscular coat of the cecum after constriction of its ascending portion was studied. Activation of smooth muscles was shown to reflect the phase of injury to the ultrastructure of the cells, followed by processes of intracellular regeneration. Analysis of the ratio between DNA-synthesizing and activated cells showed the local origin of the latter from differentiated myocytes.Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR. No. 4 Main Board, Ministry of Health of the USSR. Institute of Human Morphology, Academy of Medical Sciences of the USSR. Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 7, pp. 112–115, July, 1977.     

Analysis of the effect of morphine and trimeperidine on the uptake and liberation of noradrenalin by myocardial tissue

The effect of morphine and trimeperidine on the concentration, uptake, and liberation of noradrenalin (NA) in the rat myocardium was investigated. Trimeperidine lowers the NA level in the myocardium. Morphine does not affect the liberation of NA-¹⁴C from the isolated perfused heart, whereas trimeperidine significantly increases it, affecting both the slow and the rapid release of the mediator. Trimeperidine does not affect the uptake of NA-¹⁴C by the perfused heart but morphine significantly lowers it. Competition between morphine and NA is characterized by an incomplete inhibition effect: Morphine and NA mutually affect the affinity of each other for the receptor and their interaction depends on their relative concentrations.Laboratory of Pharmacology of the Cardiovascular System, Institute of Pharmacology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR V. V. Zakusov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 11, pp. 1345–1347, November, 1976.     

A highly sensitive electroimmunodiffusion method of antigen detection on cellulose acetate films

A highly sensitive electroimmunodiffusion method of detecting antigens on cellulose acetate films is suggested. There are three stages: concentration of the antigen in a discontinuous buffer system on cellulose acetate films; detection of the antigen on the same films by immunodiffusion using a standard test systems; staining the washed films with a protein stain to detect precipitation bands if the reaction takes place in the visible zone, or further treatment by growing precipitates with antiglobulin antibodies or by autoradiography. The method can detect nanogram amounts of -fetoprotein and can be used to discover antigens of different molecular weight and electrophoretic mobility.Laboratory of Immunochemistry and Diagnosis of Tumors, N. F. Gamaleya Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR G. V. Vygodchikov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 7, pp. 121–124, July, 1977.     

Transformation of substrate specificity of EcoRl restrictase by glycerol

In the presence of glycerol, EcoRl restriction endonuclease hydrolyzes DNA into a larger number of fragments than under ordinary conditions. On the addition of glycerol to 50% concentration, this enzyme begins to act by what is called the EcoRl' type of restriction, which was producible experimentally only by reducing the ionic strength and increasing the pH of the solution. However, under these extremal conditions, the enzyme is quickly inactivated and does not give reproducible results, especially during the hydrolysis of high-molecular-weight DNA. The conditions found for manifestation of EcoRl' activity yield reproductible results, which is essentially equivalent to the discovery of a new restriction endonuclease.The nomenclature of the restriction enzymes follows that of Smith and Nathans [6].Laboratory of Biochemistry, D. I. Inanovskii Institute of Virology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR S. S. Debov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 7, pp. 46–48, July, 1977.     

Properdin and the protein composition of the lymph and blood during resuscitation

Experiments on dogs showed that terminal blood loss followed by resuscitation by injection of autologous blood into the bone marrow causes a regular redistribution of proteins between the blood, limbs, and tissues. Retention of properdin and -globulins is observed in the interstitial tissue and is not abolished by the resuscitation measures; a stress discharge of -globulins is also found from the lymph nodes.Department of Pathological Physiology, N. P. Ogarev Mordovanian University, Saransk. (Presented by Academician of the Academy of Medical Sciences of the USSR P. D. Gorizontov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 7, pp. 24–26, July, 1977.     

Ultrastructure of conjugates of cytolytic T lymphocytes and target cells

Conjugates of target cells and cytolytic T lymphocytes, isolated on the 11th day after alloimmunization, were investigated. Conjugates were formed by small and medium-sized lymphocytes, in the cytoplasm of which mature secretory granules, crystalloid structures, and lipids were found. The lymphocyte was spherical in shape and its area of contact with the target cell did not exceed 5–15%. Cytolysis of the target cells was observed after incubation for 30–60 min. The lymphocyte became flatter, its nucleus became oval in shape, and the area of its contact with the target cell increased. Meanwhile, hypertrophy and a change in the orientation of the Golgi complex were found in the zone of contact with the target cell, fusion of the secretory granules with the lipids and crystalloid structures took place, and immature secretory granules and vacuolar degeneration of the mitochondria appeared. Peeling of the lymphocyte membrane was observed, and structures connected with it and called membranosomes are described. It is suggested that secretory processes are activated in the cytoplasm of cytolytic T lymphocytes during their interaction with target cells.Laboratory of Systemic Blood Diseases, Oncologic Scientific Center, Academy of Medical Sciences of the USSR. Department of Electron Microscopy, N. F. Gamaleya Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR V. M. Zhdanov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 5, pp. 560–565, May, 1978.     

Electron-microscopic study of interaction between cytolytic T lymphocytes and target cells

Thymocytes stimulatedin vitro in mixed culture were adsorbed by centrifugation on to the surface of target cells for an electron-microscopic study of the cytology of immune T lymphocytes and of the early stages of cytolysis. A well-developed Golgi apparatus and clusters of tubular structures 50–60 nm in diameter, communicating with the cisternae of the granular endoplasmic reticulum, with emptied vesicles, and with the plasma membrane of the lymphocyte, were found in the cytoplasm of the lymphocytes. Over a wide area the plasma membrane formed numerous contacts with the membrane of the target cells, so that closed slit-like spaces were formed. With these data and also modern views regarding interconversion of membranes and intracellular transport in mind, a hypothetical scheme for the mechanism of cytolysis of the target cell by the immune T lymphocyte is suggested.Oncologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. N. F. Gamaleya Institute of Epidemiology and Microbiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR O. V. Baroyan.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 10, pp. 443–447, October, 1977.     

Peroxidation of lipids and degeneration of photoreceptors in the retina of rats with avitaminosis E

The formation of peroxidation products of lipids (hydroperoxides, dialkyl peroxides, intermolecular cross linkages) is sharply intensified in the retina of rats with alimentary avitaminosis Ein vivo and degeneration of the photoreceptors (mainly the layer of outer segments of the rods) develops. The experimental results are discussed on the basis of views regarding the antioxidant mechanism of action of -tocopherolin vivo.Laboratory of Physical chemistry of Biological Membranes, Departament of Biochemistry, Biological Faculty, M. V. Lomonosov Moscow State University. Institute of Chemical Physics, Academy of Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR S. E. Severin.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 4, pp. 411–413, April, 1977.     

Heterogenous antigens in vaccine preparations ofVibrio cholerae

Serological similarity was found between antigens of the human small intestine, stomach, and liver and antigens of various fractions of cholera vibrios. An antigenic similarity was found on testing the heart and kidney. Heterogenous antigen was found not only in somatic antigen ofV. cholerae strain 569 (B), but also in the cholerogen, the toxoid which is the most widely used prophylactic preparation in use at the present time, obtained from it.Mikrob Plague Research Institute, Ministry of Health of the USSR. Research Laboratory of Experimental Immunobiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 8, pp. 961–962, August, 1976.     

Action of phytohemagglutinin on transplantable human cells

Agglutinating and growth-stimulating properties of phytohemagglutinin (PHA) preparations were studied in experiments on transplantable human cells (clonal line HeLa k-41 and Cave). PHA and its -globulin fraction were found to possess weak hemagglutinating properties for human erythrocytes of groups A, B, and O, but strong cytoagglutinating properties with respect to HeLa k-41 and Cave cells. With large doses of PHA (100 and 500 g/ml) proliferation and the mitotic index of the cells of the cultures were lower but the percentage of dead cells and the agglutinin titer in the preparations were higher (1:256). With smaller doses of PHA (5 and 25 g/ml) growth was much more intensive and the percentage of dead cells was smaller. The agglutinin titer in the preparation fell to 1:16–1:32. The -globulin fraction of PHA had the strongest growth-stimulating action and gave the smallest number of dead cells. However, the agglutinin titer in the preparations was high (1:128). It is concluded that the inhibitory and growth-stimulating action of PHA preparations on transplantable human cells is directly linked with the agglutinin content in the preparations, for the -globulin fraction of PHA had the strongest cytoagglutinating and growth-stimulating action.Research Laboratory of Experimental Immunobiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. N. Zhukov-Verezhnikov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 8, pp. 970–972, August, 1976.     

The rhombencephalon and vasomotor reflexes to impulses in somatic A-fibers

Cats were decerebrated so that the medulla preserved its connections with a greater or lesser part of the pons, after which they were immobilized with flaxedil or succinylcholine and the response of their arterial blood pressure (BP) to volleys of impulses in various subgroups of fibers of the tibial nerve (frequency of volleys 10 sec^–1) was investigated. Impulses in A + A₁-afferents and the most excitable fraction of A₂-afferents lowered BP in all animals. Application of impulses in all A₂-afferents potentiated the fall in BP in 79% of the experiments and led to a rise in BP in only 21% of the experiments. The addition of impulses in A₃-fibers to the afferent flow either did not change the mean decrease in BP or led to a small or sometimes more considerable increase in BP. The critical factor producing a sharp reduction in the pressor reflexes or their replacement by depressor reflexes was the integrity of the structures of the pontobulbar junction and the most rostral part of the medulla. This region of the hindbrain thus contains formations causing tonic depression of the excitatory action of impulses in somatic A-afferents on vasoconstrictor neurons and which thereby unmask the existence of an inhibitory component of their action on these neurons. In unanesthetized cats this action of hindbrain structures is depressed tonically by the mesencephalon.Laboratory of Biophysics and Pathophysiology of the Circulation, Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. M. Chernukh.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 11, pp. 519–522, November, 1978.     

Decrease in tissue-specific resistance of the gastric epithelium to acute cell death induced by inhibition of DNA synthesis

Administration of hydroxyurea to mice caused acute death of a few cells synthesizing DNA in the epithelium of the glandular stomach. If actinomycin D was given 8 h before the hydroxyurea, cell death was sharply intensified and about 80% of cells synthesizing DNA died during administration of the hydroxyurea. Administration of actinomycin D simultaneously with hydroxyurea had no potentiating effect on cell death. It is postulated that actinomycin D stimulates protein synthesis in stomach cells (the superinduction effect), thus increasing their sensitivity to inhibition of DNA synthesis.Sector of Kinetics of Chemical and Biological Processes, Institute of Chemical Physics, Academy of Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR L. M. Shabad.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 4, pp. 491–492, April, 1977.     

Esterase spectrum in hepatocytes and Kupffer cells of the regenerating liver

The esterase spectrum in hepatocytes and Kuppfer cells of the regenerating rat liver was determined by electrophoresis in starch gel. The relative number of Kupffer cells, the percentage of phagocytic Kupffer cells, and the frequency of mitosis in the hepatocytes were determined. The esterase spectrum of hepatocytes isolated from the intact liver consisted of six zones, and that of the Kupffer cells of five zones of enzyme activity. The spectrum of the hepatocytes was simplified 2.5 h after partial hepatectomy, additional bands of activity appeared toward 9 h, and these persisted until 24 h after the operation. The original esterase spectrum was restored 48 h after the operation. A similar reorganization of the esterase spectrum was observed in the Kupffer cells but this was not fully restored 72 h after partial hepatectomy.Laboratory of Pathophysiology, Institute of Clinical and Experimental Medicine, Siberian Branch, Academy of Medical Sciences of the USSR, Novosibirsk. (Presented by Academician of the Academy of Medical Sciences of the USSR V. V. Kaznacheev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 7, pp. 44–46, July, 1977.     

Relations between musuclar and connective tissue components of the rat heart in experimental myocardial infarction

After the first day of experimental myocardial infarction in rats the volume and mean number of connective tissue stromal cells are increased in the intact zones of the left ventricle; on the second day of the experiment these indices reach their maximum, and by the 20th day they are equal to the values for the myocardium of control animals. On the third day of the experiment the number of muscle nuclei per standard area of cross section is increased, evidently as the result of amitotic division of the myocyte nuclei, for the number of paired nuclei is increased at the same time.Central Pathological Anatomical Laboratory, Institute of Human Morphology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 4, pp. 486–488, April, 1977.     

Posthypoxic changes in the cerebral cortex of dogs in the late recovery period after hypovolemic hypotension for 4 hours

On the 14th–21st day of the recovery period after hypovolemic hypotension lasting 4 h the total RNA level in the tissue of the gray matter of the brain fell by 20.9% and the DNA level by 13%. In the postmitochondrial supernatant the concentration of prealbumins fell by 26.5%, of -globulins by 19.2%, and of -globulins by 59.8%; the concentration of albumins and -globulins increased by 12.6 and 50% respectively. Activity of acid cathepsins rose by 50% and of acid phosphatase by 44%. Activity of total lactate dehydrogenase (LD) and glutamate dehydrogenase did not differ significantly from the control values. However, the LD isozyme spectrum showed a decrease in LD₃₊₄₊₅ from 31.9 to 14.2%. Analysis of densitograms of electrophoresis in polyacrylamide gel showed changes in the physiocochemical properties of the protein molecules similar to denaturation in nature. The number of Purkinje cells in the cerebellar cortex was 41.3% below the control level.Laboratory of Experimental Physiology of Resuscitation, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR V. A. Negovskii.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 8, pp. 933–936, August, 1976.     

Action of hemolysate of polycythemic animals on proliferations of bonne marrow cells

Erythrocytic inhibitor from polycythemic rats depressed mitotic activity of cells of of the erythroid series in mice by 40%. The inhibitory effect lasted about 12 h. The inhibitor acted on the G₂ period. The points of application of the inhibitor were not only blast forms of the erythron but also hematopoietic stem cells.Department of Pathological Physiology, Moscow Medical Stomatological Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR N. A. Fedorov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 3, pp. 337–339, March, 1977.     

Pattern of antinuclear autoantibodies in diseases with systemic connective tissue lesions

Several types of antinuclear antibodies (ANA) were discovered by the fluorescent antibody method in diseases accompanied by systemic lesions of connective tissue and also in certain other diseases and in clinically healthy blood donors, depending on the character of fluorescence in the nuclei. Diffuse fluorescence of nuclei, diffuse fluorescence without fluorescence of the nucleoli, annular fluorescence, fluorescence in the form of granules, selective fluorescence of nucleoli, and fluorescnece in the form of long, thin, interweaving bands with simultaneous fluorescence in the region of the nuclear membrane were distinguished. The last type of ANA was observed only in various forms of lupus erythematosus, and the character of fluorescence in the nuclei differed from the reticular and filamentous types of fluorescence described previously.Department of Pathological Anatomy and Department of Skin Diseases, N. I. Pirogov Second Moscow Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 7, pp. 75–78, July, 1977.     

Quantitative analysis of stimulation frequency transformation in the neuromuscular apparatus

The dependence of the parameters of neuromuscular transmission on the frequency of stimulation was determined. This dependence was analyzed in relation to fatigue of the synapse arising during prolonged repetitive stimulation of muscle. The proposed mathematical model and the method of statistical analysis of the records of evoked responses of the muscle derived from it permit approximate estimates to be made from the experimental data of parameters quantitatively reflecting frequency (transmission) properties of the neuromuscular apparatus.Laboratory of Mathematical Modeling and Processing of Experimental Data, Institute of General Pathology and Pathophysiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. M. Chernukh.) Translated from Byullten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 8, pp. 1016–1018, August, 1976.     

Transformation ofBacillus subtilis in mice after immunological suppression of exogenous deoxyribonuclease 1 activity

Transformation ofBacillus subtilis was carried out intraperitoneally in mice. The frequency of transformation was considerably reduced by intraperitoneal injection of bovine deoxyribonuclease 1 (DNase 1) into the animals in a dose of 3–5 g. Immune rabbit -globulins, containing antibodies against bovine DNase 1, inhibit the activity of DNase 1 in vivo, thus protecting the transforming DNA against the hydrolytic action of that enzyme. The model suggested can be used to search for ways of preserving a nucleic acid introduced into an animal for the purposes of genetic engineering.Research Laboratory of Experimental Immunobiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 7, pp. 68–70, July, 1977.     

Down-regulation of tumor necrosis factor α activity by acute ethanol treatment in human peripheral blood monocytes

As the most commonly used drug that can modulate both metabolic and immune pathways, ethanol is evaluated in this report as a regulator of tumor necrosis factor (TNF) production in human peripheral blood monocytes (M) in combination with a variety of stimuli. While acute ethanol treatment did not induce TNF in M, it was a potent down-regulator of M TNF production whether induced by the combination of interferon- plus muramyl dipeptide (MDP) (P<0.001), lipopolysaccharide (LPS) alone (P<0.01), or interferon- plus LPS. Down-regulation of M TNF by ethanol was dose dependent and statistically significant in the biologically relevant, 25–150 mM, ethanol concentration range. We also demonstrate that these ethanol concentrations did not affect M viability. TNF down-regulation by ethanol was most effective when ethanol was administered 4 hr prior to MDP stimulation; however, it was also effective—though to a lesser extent—if it was added at the time of MDP stimulation. Furthermore, ethanol also down-regulated TNF production of thein vivo preactivated M of trauma patients, which produce hyperelevated levels of TNF. We have previously shown that the majority of posttrauma elevated M TNF is produced by the M subpopulation expressing high-affinity type I Fc receptors (FcRI). When the FcRI cross-linking-stimulated M subpopulation was treated with acute ethanol, TNF production was suppressed again both inin vivo preactivated M of trauma patients and in M of normal controls. In experiments utilizing cyclooxygenase inhibitor, we also demonstrate that ethanol has a direct, prostaglandin E₂-independent, effect on M TNF production. These results demonstrate that acute ethanol exposure has the potential to down-regulate M production of TNF significantly regardless of the TNF-inducing stimulus. Decreased capacity of M to produce TNF might, therefore, contribute to the immunological and metabolic abnormalities described after ethanol uptake.