[Mexidol-induced modification of lipid metabolism in pancreatitis]. / Modifikatsiia obmena lipidov pri pankreatite pod vliianiem meksidola.

The chronic experiments on mongrel dogs with a model pancreatitis showed that mexidol decreases manifestations of the inflammatory process. The treatment with mexidol led to a decrease in the degree of lipid transformations in the initial stage of pancreatitis development, with normaliation of the lipid metabolism according to the liver and blood plasma characteristics. The membranoprotector effect of mexidol, manifested in normalization of the lipid spectrum, is probably related to inhibition of the lipid peroxidation (LPO) process and to a decrease in the activity of phospholipase A2. The correlation between lipid metabolism, LPO, and phospholipase A2 activity in the tissues studied indicates that the therapeutic effect of mexidol in animals with pancreatitis is based on the cytoprotector activity of the drug.

[Dimephosphon in complex therapy of acute edematous pancreatitis]. / Dimefosfon v kompleksnoi terapii ostrogo otechnogo pankreatita.

Clinical investigations of 120 patients have shown that Dimephosphon included in complex treatment of acute edematous pancreatitis facilitates more rapid arrest of the symptoms. An important effect of the drug is its ability to decrease the level of endogenous intoxication by both the hydrophilic and hydrophobic components. It was established that one of the mechanisms of favorable action of the drug is its ability to inhibit LPO, to decrease activity of phospholipase A2. The drug possessing the cytoprotecting effect promotes rapid restoration of the functional state of the liver, in particular its detoxicating, albumin synthesizing ability that is not least of the factors of optimization of the complex therapy.

Histological studies of atrazine toxicity on the thyroid gland in rats.

Little is known about the toxic activity of the atrazine (a herbicide, commonly used in agricultural production) on the thyroid gland. In this study the compound was administered orally in female albino rats at sublethal exposure equivalent to 0.2 LD50 doses for 6 and 12 days. At termination of dosing the anesthetized animals were killed and blood was drawn for the determination of serum triiodothyronine (T3) and thyroxin (T4). A dose-dependent decrease of serum T3 concentration was observed in all the groups (control: 0.57 nmol-L-1; 6 days: 0.35 nmol-L-1; 12 days: 0.21 nmol-L-1). The thyroid gland was examined light-microscopically. Bouin's solution-fixed thyroids were embedded in paraffin and sections cut at 6 microns, stained separately with toluidine blue according to Slinchenko's method. Histologically in experimental groups epithelium featured small cuboidal cells and occasional structures of the follicles confluence within epitheliomers. A dose-dependent changes of the following parameters were observed: (a) increasing of number of follicle-building thyroid cells; (b) increasing of follicular volume; (c) decreasing of nucleus volume. Investigation of the whole population of thyroid mast cells disclosed no change in degranulation intensity. By contrast, degranulation intensity was decreased in perifollicular mast cells from groups treated with atrazine in dose-dependent manner. There are no changes observed in degranulation of stromal mast cells. These results suggesting that differences in response to the atrazine might account for an aspect of the functional heterogeneity within the rat thyroid mast cell population.

Neural cell adhesion molecule (N-CAM) distribution may predict the effect of neurotoxins on the brain.

The neural cell adhesion molecule (N-CAM) is a convenient neurospecific marker for investigating the effects of neurotoxins on cell migration, cell recognition and differentiation of neurons during development. In this report, we discuss the developmental toxicity of valproic acid studied by two different approaches (the immunochemical detection of N-CAM content and polypeptide composition, and immunohistochemical analysis of N-CAM topography). Immunohistochemical analysis of distribution of N-CAM as a surface marker on the neural cells predicted the effect of the neurotoxin.

A novel method for evaluation of carbohydrate-binding activity: enzyme-linked carbohydrate-binding assay (ELCBA).

A highly sensitive method for detection of the carbohydrate-binding activity of proteins is described. The method is based on interactions of carbohydrate-binding proteins, immobilized on a solid phase, with an enzyme-labeled soluble polysaccharide (peroxidase conjugated glycosaminoglycans-heparin, chondroitin sulfate or hyaluronic acid. Binding capacity was measured spectrofotometrically after enzymatic reaction with chromogenic substrate. The reliability of the assay was tested by use of two heparin-binding proteins-i) fibronectin (soluble) and ii) heparin-binding protein purified from the human brain (water-insoluble). Binding of heparin was dependent on metal ions, detergents and urea. The assay is believed to be applicable for the identification and characterization of a variety of carbohydrate(glycosaminoglycan)-binding proteins, especially, when traditional methods can not be applied (e.g., when proteins are water-insoluble).

[Blood proteinase inhibitors and cathepsin D in diffuse peritonitis]. / Ingibitory proteinaz i katepsin-D syvorotki krovi pri razlitom peritonite.

There were 52 patients with a diffuse peritonitis examined. The level of a proteinases alpha 1-inhibitor, alpha 2-macroglobulin, trypsin inhibitor, cathepsin-D was studied. Results witnesses the high activity of proteinases inhibitors cathepsin-D in the early, postoperative period. The level of alpha 2-macroglobulin and cathepsin-D in an elderly and senile patients approximated to normal values up to the 7-9th day after the operation. The dependence of proteinases inhibitors and cathepsin-D contents from the severity of peritonitis course, the number of interventions done to the patient, and the use of autologous blood photomodifization in the complex of treatment were not revealed.

Lateral patterns of the neural cell adhesion molecule on the surface of hippocampal cells developing in vitro.

In monolayer cultures of hippocampal neurons from newborn rats, an immunocytochemical quantitative study was carried out to investigate age-dependent arrangement of the neural cell adhesion molecules in different parts of cell membranes. On the fifth and 12th day in vitro, neural cell adhesion molecules were labelled with specific antibodies and protein A conjugated to colloidal gold particles. Samples of randomly selected electron micrographs that displayed labelled membrane fragments of cell bodies, growth cones, and axons were numerically analysed for the five- and 12-day in vitro neurons. Neural cell adhesion molecules surface topography was quantitatively described and compared, using a statistical stereological approach. The mean surface density of labelled neural cell adhesion molecules was found to be approximately 2.5 times higher in growth cone membranes relative to somatic and axonal membranes in five-day in vitro neurons. By the 12th day in vitro, this density decreases in somatic membranes (approximately 18%) and increases in axonal membranes (approximately 60%). Representative spectra of lateral intervals between labels as well as images that show typical topography of label on membrane surfaces were simulated. The results revealed regular patterns of neural cell adhesion molecules on the somatic surface and allowed consideration of neural cell adhesion molecules arrangement in a view of membrane adhesion properties. Participation of cytoskeleton in neural cell adhesion molecules rearrangement is discussed.

Changes in the neural cell adhesion molecule patterns on the rat glial cell surfaces with development and contact formation in vitro.

In monolayer cultures of newborn rat hippocampal cells, immunogold-labelling at the electron microscope level was used to study quantitatively the neural cell adhesion molecule (N-CAM) arrangement on the surface of glial soma and processes on 5 and 12 days in vitro (DIV). Four corresponding samples of micrographs were formed. To quantify the labelling, a stochastic geometry approach was used. Spectra of lateral distances between labels as well as simulated images of the surface label arrangement (invisible in micrographs) were derived and compared. The data show that, on both 5 and 12 DIV, N-CAM density on the surface of processes is approximately 2 times higher than that in somata; 12-DIV cells showing a lower (approximately 25%) N-CAM surface density as compared with the 5-DIV cells. This suggests that N-CAM expression in glia surfaces decreases while the cells form contacts, and N-CAM sorting between soma and processes remains stable. The simulated topographies of the lateral N-CAM arrangement might highlight fundamental mechanisms that underlie formation of the neural network.

[Immunoglobin-binding proteins in human placenta]. / Immunoglobulin-sviazyvaiushchie belki platsenty cheloveka.

Phenomena of the binding of poor-soluble placenta proteins (PSPP) with pregnant women sera IgG as well as placenta blood IgG were studied. PSPP were extracted from the placenta tissue, washed out from soluble proteins, by the use of 3M KCl solution containing 0.005 M PMSF. PSPP were separated by the use of two-dimensional isoelectrofocusing and SDS-PAG electrophoresis and more than 30 different polypeptides were visualized. Having used various ELISA procedures with pregnant women sera IgG, placenta blood IgG as well as its Fab and Fc-fragments we have shown that both the receptor-type and the antigen-antibody-like interaction of PSPP took place. Both the polypeptide compositions and the isoelectrofocusing points ranges of the antigen-antibody-like interacting IgG-binding PSPP were determined by the use of the peroxidase conjugated Fab-fragments of the placenta blood IgG.

[Chemiluminescence fiber optic immunosensor for detecting antibodies to the influenza virus]. / Khemiliuminestsentnyi optovolokonnyi immunosensor dlia opredeleniia antitel k virusu grippa.

A new variant of an optoimmunosensor for determination of antibodies to the influenza virus has been elaborated. Its advantages as compared to the traditional solid phase immunoenzyme analysis in respect to sensitivity and expressiveness are demonstrated. Time of the sensor response is below 17 min. When analyzing the 1:320 diluted serum, about 80% of response value is implemented after 6 min. Optimum conditions of the optoimmunosensor transducer regeneration are chosen. They permit reusing it for 20-30 cycles of measurements. A conclusion is made on the prospects of the developed variant of the optosensor for the immunoanalysis of antigens and antibodies under the equilibrium and kinetic conditions.

[Expression of glial fibrillary acidic protein in the developing human brain]. / Ekspressiia glial'nogo fibrilliarnogo kislogo belka v razvivaiushchemsiia mozge cheloveka.

It was shown that the glial fibrillary acidic protein (GFAP) content in developing (fetal) human brain is sharply increased. The expression of GFAP was observed already on the 7th-8th week after gestation, the GFAP concentration being less than 0.05% in comparison with adult brain. GFAP can be immunohistochemically detected in radial glial cells. At early stages of development the presence of antigenic determinants of 68 kDa and 100 kDa polypeptides interacting with monoclonal antibodies alongside with native GFAP (51 kDa) and its low molecular weight forms was demonstrated. These antigenic determinants cannot be detected at later stages of development and are absent in adult brain. The data obtained testify to changes in the gene expression of intermediate filament proteins at early stages of human brain ontogenesis.

[The effect of low doses of ionizing radiation on the intermediate filaments and the Ca2+-activated proteolysis system in the rat brain]. / Vliianie malykh doz ioniziruiushchei radiatsii na promezhutochnye filamenty i Ca2+-aktiviruemuiu sistemy proteoliza golovnogo mozga krysy.

The immunochemical methods were used to study the effect of low-level radiation (0.00645 C/kg and 0.0129 C/kg) on the content and polypeptide composition of glial intermediate filament proteins (GIFP) in different rat brain areas. Changes in glial fibrillar acidic protein (GFAP) concentration were more significant with the dose of 0.0129 C/kg than 0.00645 C/kg. It is suggested that soluble GIFP is more susceptible to the effect of Ca(2+)-dependent proteinases, calpains, than the filament one is, and degrades as early as the first few hours following irradiation. However, low radiation doses were ineffective with respect to calpains activity in the animal brain. The increased Ca2+ concentration enhances considerably GFAP degradation under the effect of calpains I and II. It is suggested that with low radiation doses the rearrangements of glial intermediate filaments may occur due to activation of calpains by releasing Ca ions.

[The distribution of glial fibrillary acidic protein and protein S-100 in experimental brain tumors in rats]. / Raspredelenie glial'nogo fibrilliarnogo kislogo belka i belka S-100 v éksperimental'nykh opukholiakh mozga krys.

The methods of quantitative immunoelectrophoresis and indirect immunofluorescence were used study the content of glial fibrillary acid protein in 10 serially reinoculated rat gliomas induced primarily by ethylnitrosourea (a total of 135 tumors). It was found that the GFAP content reduced with increase of malignancy. However, wide scattering of the GFAP content in some of the tumors was characteristic of all strains. In the group of slowly growing glial tumors (2 malignant astrocytomas and one malignant oligoastrocytoma) the GFAP content ranged from 50 to 600% and exceeded the normal content two-to threefold on the average. In the group of highly malignant gliomas (4 malignant ependymomas, 2 malignant gliomas, and one glioblastoma) the GFAP content was within the limits of 65-120%. In most cases the GFAP level was below normal or could not be determined at all. At the same time, tumors with a high GFAP content were encountered. The GFAP-positive cells were unevenly distributed in the gliomas: separately, in foci, and around the vessels. Their number increased in the direction of the periphery of the tumor. Intensive fluorescence was noted on the tumor--brain borderline. The content of protein S-100 in the experimental gliomas was always below normal.

[Characteristics of brain gastrin/cholecystokinin-binding proteins]. / Kharakteristika gastrin/kholetsistokininsviazyvaiushchikh belkov golovnogo mozga.

Gastrin/cholecystokinin-binding proteins were purified using the column affinity chromatography on immobilized pig tetragastrin and cholecystokinin. Immunoblotting analysis of different human tissue extracts with specific antisera obtained against gastrin-binding proteins was performed. It was found that high molecular weight polypeptide zones of 120 kDa and 35 kDa were characteristic of the brain only. Autoantisera of patients with type A gastric disease reacted with some gastrin/cholecystokinin-binding proteins in human brain and mucosa including human brain polypeptide of 120 kDa. It is supposed that there are neurospecific gastrin-binding proteins (possibly gastrin/cholecystokinin receptors in the brain).

[The CNS syndrome. The characteristics of the intermediate filaments of the rat brain]. / TsNS-sindrom. Kharakteristika promezhutochnykh filamentov golovnogo mozga krysy.

A study was made of the effect of ionizing radiation on the content and polypeptide composition of filamentous and soluble glial fibrillary acidic protein (GFAP) in different regions of rat brain. Ionizing radiation was shown to decrease considerably the level of soluble GFAP in cerebral cortex, cerebellum, middle brain and hippocampus. Polypeptide composition of soluble GFAP detected by the immunoblot method was found to be changed considerably in different brain areas of irradiated animals.

[Characteristics of concanavalin A-binding neurospecific glycoproteins from various regions of the rat brain during experimental neurosis]. / Kharakteristika konkanavalin-A-sviazyvaiushchikh neirospetsificheskikh glikoproteinov v razlichnykh otdelakh golovnogo mozga krys pri éksperimental'nom nevroze.

The polypeptide composition of neurospecific glycoproteins in different areas of the rat brain under experimental neurosis is characterized using SDS-PAG-electrophoresis followed by electroblot and immunofixation on nitrocellulose membranes. The soluble and membrane-bound glycoproteins are purified by Con A-Sepharose column chromatography. Changes in the glycoprotein polypeptide composition in different areas of the rat brain under experimental neurosis are qualitative. Soluble glycopolypeptide 27 kDa and membrane glycopolypeptide 32 kDa are not revealed in the midbrain and corpus striatum. Soluble polypeptide 47 kDa is absent in the cerebral cortex and hippocampus. It is suggested that the above mentioned glycopolypeptides are important for the CNS physiological functioning.

[A method of solid phase immunoenzyme analysis using antigen molecules immobilized on membranes]. / Metod tverdofaznogo immunofermentnogo analiza s ispol'zovaniem immobilizovannykh na membranakh molekul antigena.

The enzyme-linked immunoassay modification has been worked out. The method combines advantages of membrane technology of antigen immobilization which is used in the enzyme immunosensory technique and of conventional enzyme-linked immunosorbent assay. The nitrocellulose and polypropylene membranes are used as a solid-phase. The purified rabbit immunoglobulin G is immobilized on the surface of membranes as the first layer. The competitive immunoassay is employed. The immunoglobulin G concentration range is 1-1000 ng/ml. The membranes with the immobilized antigen can be repeatedly used after incubation in 0.1 M glycine buffer, pH 2.5. The dry membrane with the immobilized antigen can be used after keeping for 6 months in refrigerator at 4 degrees C without changing the concentration range measured.

[An analysis of blood serum proteins in chronic alcoholism patients by a cross immunoelectrophoresis method]. / Analiz belkov syvootki krovi bolnykh khronicheskim alkogolizmom metodom perekrestnogo immunoélektroforeza.

Prealbumin, albumin, alpha 1-antitrypsin, alpha 1-antichymotrypsin, ceruloplasmin, haptoglobin, transferrin, IgG, IgM and IgA were studied in blood serum of healthy donors and of patients with chronic alcoholism by means of cross immuno-electrophoresis and immunodiffusion. Only content of alpha 1-antitrypsin was distinctly altered in blood serum of the patients with alcoholism as compared with normal state, while individual variations in content of the proteins studied were considerably higher in blood serum of the patients. At the same time, distinct dissimilarity of the patterns studied was found between healthy donors and patients with chronic alcoholism when concentration ratios of some positively and negatively charged acute phase proteins were calculated (alpha 1-antitrypsin/albumin, haptoglobin/albumin, haptoglobin/transferrin).