Patologicheskaia fiziologiia i eksperimental'naia terapiia最新文献

The purpose - comprehensive study of the purine metabolic enzymes in serum and immune cells in patients with pulmonary tuberculosis for the understanding of the pathogenesis of a specific lung disease.

Methods: The enzymes of purine metabolism (adenosine deaminase (ADA) and its isoenzymes (ADA-1 and ADA-2), dipeptidylpeptidase IV (DPPIV - CD26), ecto-5’-nucleotidase (5’-NC) in the blood and immune cells was studied in 29 and 76 patients with fibro-cavernous (FCPT) and infiltrative (IPT) pulmonary tuberculosis correspondingly.

Results: In patients found changes in purine metabolism, the severity and pathophysiological significance of which depend of clinical forms of tuberculosis, that is, from the gravity specific of the process. Reduced activity of ADA mononuclear cells was accompanied by a decrease in the expression of CD26 in patients with FCPT and the growth of the IPT ectopeptidase patients, that is, the concentration of CD26 mononuclear cells and neutrophils are associated with form of pulmonary tuberculosis. The increased levels of another enzyme purine metabolism - 5’-NC registered in both forms of pulmonary tuberculosis.

Conclusion: In the context of the ADA and CD26 association with the IPT can assume increased participation of each of them in the activation of cell proliferation and cytokine production. Low levels of CD26 immune cells in the absence of their connection with the activity of ADA is typical for patients with FCPT and reflects their inherent failure of cellular immunity. We can assume that the formation of complexes with the ADA ectopeptidases (CD26 and 5’-NC) for newly diagnosed IPT provides a balance CD26_ADA extracellular / intracellular adenosine and 5’-NC / adenosine and thereby adequate metabolism of immunocompetent cells.

Summary: A comparative study of the physical development and the rate of formation of sensory-motor reflexes offspring of rats with experimental preeclampsia (EP) was carried out. In the first experimental group EP was modeled intraperitoneal conduct of L-NAME at a dose of 25 mg/kg from 14 to 21 days of gestation, the second experimental group - the replacement of drinking water by 1.8% sodium chloride solution for the entire period of gestation. In the offspring of both groups, there was a delay of physical development, which was reflected in the later timing of the hair coat development, incisor eruption, pinna detachment as compared to the pups in the control group. It also noted the gap in the formation of sensory-motor reflexes and vestibular reactions. This was manifested in the delayed appearance of the olfactory response, auditory sensitivity, later performing tests «righting reflex», «negative geotaxis», «aerial righting reflex», «cliff avoidance», «horizontal wire test», «raising the head and forelegs», «supporting their bodies on hind legs» as compared to the indices of the pups of the female rats with an uncomplicated pregnancy. The most pronounced lag in postnatal development was observed in the offspring of rats with EP, which instead of drinking water was prepared 1.8% sodium chloride during the entire period of gestation.

The purpose: To make a comparative study of the impact of ADMA-like preeclampsia (PE) and preeclampsia modeled by the replacement of drinking water consumed by female rats during gestation with 1.8% NaCl solution on the physical development and the rate of the maturation of sensory motor reflexes of their offspring.

Methods: The study was performed on three groups of pregnant female rats aged 3-4 months whose original weight was 210-250 g and their pups. They were divided into three groups: 1: Control group including female rats with an uncomplicated pregnancy (n = 6) and their pups (n = 49); 2. Experimental group 1 - pregnant female rats with PE (n = 6) induced by intraabdominal introduction of L-NAME at a dose of 25 mg/kg from 14 to 21 day of gestation (ADMA-like preeclampsia) and their offspring (n = 35); 3. Experimental group 2 - pregnant female rats with PE (n = 6) modeled by the replacement of drinking throughout gestation with 1.8% NaCl solution and their pups. When studying the physical development of the pups we considered the terms of pinna detachment, hair coat development, incisor eruption and the time when they began to open their eyes. To estimate the rate of the maturation of sensory motor reflexes and motor coordination of the pups of the rats with PE we analyzed the time when they started to support their bodies on hind legs, lift their bodies off the floor, crawl, raise their head and forelegs, show the aerial righting reflex and the righting reflex, negative geotaxis, reactions to auditory and olfactory stimuli as well as the tim

The purpose of the study was to determine the relative number and degree of lymphocyte-platelet adhesion in patients with frostbites of III-IV degree of extremities with malnutrition and eutrophia.

Methods: Non-randomized prospective controlled study was performed in 49 patients of both sexes, aged from 16 to 60 years, with frostbites of III-IV degree of extremities. The object of the study was the blood plasma. Test subjects were divided into two groups according to the trophic status on the scale of Luft V.M and Kostyuchenko A.L. By the original method of the Professor Vitkovsky Yu.A. (1999) number of lymphocyte-platelet coaggregations was counted in 100 cells (percentage), which is relative number of the lymphocyte-platelet adhesion (LTA). In this case lymphocyte which adhered to its surface one or more platelets was named the coaggregation. Lymphocyte-platelet index (LTI) or the degree of LTA was determined as the arithmetic mean of the number of platelets which have adhered to a single lymphocyte.

Results: The relative number of LTA greater in patients with eutrophia (n = 19) vs. (p<0.001) the control group (n = 20) and vs. (p = 0.008) patients with malnutrition (n = 20). The relative number of LTA is not different in patients with malnutrition vs. (p = 0.085) the control group. LTI greater in patients with eutrophia vs. the control group (p<0.001), and vs. patients with malnutrition (p = 0.020). LTI greater in patients with malnutrition vs. the control group (p = 0.006).

Conclusion: It was established that LTI increased irrespective of premorbid trophic status, and LTA percentage increased only in patients with eutrophia. In patients with malnutrition LTA degree and LTA percentage were detected less than in patients with normal nutritional status.

The purpose of study was evaluation of extra and intracellular cytokine production in blood cultures and cytokine-pain relationship at different stages od addiction.

Methods: Classic and multiplex enzyme immunoassay, flow, cytometry, algometry, visual analogue scale were used in 34 abusers, 12 users and 20 control subjects.

Results: Cytokine profile correlated with clinical parameters and pain sensitivity in abusers (mostly IL-6 and IL-1β). IL-10 production is increased in episodic and systematic users. Activation of intracellular IL-6 synthesis was found in users. Both parameters are expected to affect the emotional evaluation of pain.

Conclusion: To assess the predisposition to substance dependence most informative are balance of extra and intracellular production of IL-6 and IL-10 and algometry.

The purpose: This study investigated the role of alpha-synuclein in the development of dopaminergic neurons.

Methods: In this study a new SNCA knockout mouse line has been used to model the deficiency of alpha-synuclein function. In the knockout and control mice the dynamics of the formation of two distinct populations of dopaminergic neurons differently affected in patients with PD was studied by the comparative morphometric analysis.

Results: Here, we revealed a prominent modulating effect of alpha-synuclein on the developing DA neurons in substantia nigra (SN) which is the most affected region in PD patients. Yet, alpha-synuclein had no effect on the formation of DA neurons in ventral tegmental area which is much less susceptible to degeneration in PD patients.

Conclusion: The new line of knockout mice is a convenient model for studying pathophysiologic aspects of selective impairment of DA neurons.

The purpose. Identifying the capacity of the medicinal leech novel original recombinant thrombolytic preparation Destabilase-Lysozyme-2 to inhibit the blood platelet aggregation.

Methods: Gene of destabilase-lysozyme. ds2 (mlDL-Ds2 ), was cloned in E.coli cells. Recombinant protein was isolated in denaturing conditions using metal-chelate chromatography followed by denaturation of the polypeptide by rapid dilution in exact accordance with the procedure described by Kurdyumov A.S. et al. ( 2016, Russian Journal of Bioorganic Chemistry, v.42, s. 42-52). Blood was collected from the jugular vein of 18 horses. The functional status of platelets in the presence of different destabilase-lysozyme concentrations were evaluated for their aggregation in Platelet Rich Plasma ( PRP) and in Washed Platelet suspension (WP) using aggregometers Chrono-Log-700 and Сhrono-Log-560, USA560, США. As used aggregation inducers of ADP, collagen type III and human thrombin.

Results: First demonstrated the ability of newly synthesized (Kurdyumov A.S. et al. 2016, Russian Journal of Bioorganic Chemistry, v42, s. 42-52) thrombolytic recombinant enzyme destabilase-lyzosyme to inhibit more than 40% of ADP-stimulated PRP aggregation and ADP- stimulated aggregation of horse blood washed platelets.

Conclusion: The ability of destabilase-lyzosyme -2 to inhibit platelets aggregation extends biological properties of recombinant thrombolytic enzyme, pre-clinical trials which resulted in the end of 2015.

The purpose of the research: To study the water balance of lung and NO level in blood in experimental autoimmune encephalomyelitis combined with capsaicin blockade of vagus nerve.

Methods: Experiments were conducted on 47 adult (16-week-old) male rats weighing 220-280 g. To simulate the experimental autoimmune encephalomyelitis (EAE) rats were subcutaneously injected with encephalitogenic mixture in complete Freund's adjuvant (0.2 ml; the content of inactivated Mycobacterium tuberculosis was 5 mg/ml) at the rate of 100 mg of homologous spinal cord homogenate per animal. Сapsaicin blockade was performed by bilateral application of 50 uM capsaicin («Sigma») on the neck portions of vagus nerves. The animals were divided into 4 groups: intact rats - control group1; rats with EAE; rats with capsaicin application on vagus nerve + EAE; sham operated rats subjected to vagus nerves allocation without the subsequent capsaicin application + EAE - control group 2. The next parameters were detected: the content of nitric oxide in blood plasma; protein content in broncho-alveolar lavage fluid; lung water balance indices including the amount of total, extra- and intravascular fluid and blood supply of lungs, which were calculated based on wet and dry lung mass and the hemoglobin content in blood and lung tissue determined by hemiglobincyanide method.

Results: It was found that EAE is accompanied by an increase of total fluid, extravascular fluid (EVF) and blood supply of lungs on the background of increasing content of nitric oxide in arterial (art) and venous (ven) blood. In EAE and its combination with bilateral capsaicin blockade of vagus nerve a strong negative correlation between the NOart / NOven coefficient and EVF amount was found out. The blockade of capsaicin-sensitive vagal afferents normalized lung water balance impaired in EAE and restored the levels of nitric oxide in blood plasma.

Conclusion: The obtained results suggest that capsaicin-sensitive vagal afferents with NO-ergic mechanisms involvment take part in the development of pulmonary hyperhydration during experimental autoimmune encephalomyelitis.

The comparative analysis of surfactant metabolism and water balance of the lungs in modeling the formation and neurodegeneration focus of pathological activity in the substantia nigra of the brain.

Methods: Experiments were performed on male rats - nonlinear, including the control, sham operated with unilateral stereotaxic administration of the neurotoxin 6-hydroxydopamine (Sigma) in the substantia nigra compact part and the implantation of cobalt metal nanopowder (Berlin) in the reticular part of the structure. Complex investigations included determination of surfactant phospholipid fractions by thin layer chromatography, total phospholipids and cholesterol in the bronchoalveolar lavage and surface-active properties by the Wilhelmy method, phospholipase activity, lipid peroxidation intensity on the content of TBA-active products in the lung tissue, evaluation of water balance using the gravimetric method.

Results: It has been established that the introduction of intranigral neurotoxin and implantation of cobalt in the structure of the brain called the deterioration of the surface activity of the alveolar lining set against the backdrop of an imbalance of fractional composition of surfactant lipids with multidirectional changes in the amount of phospholipids and the degree of hydration of the lung. Induction focus of pathological activity in the substantia nigra was accompanied by an increase in alveolar phospholipids by lysophosphatidylcholine, sphingomyelin, phosphatidylethanolamine, intensification of lipid peroxidation of the lung tissue and organ hyperhydration. Modeling neurodegeneration - reduction of total phospholipids, phosphatidylcholine and lysophosphatidylcholine accumulation in terms of activation of phospholipase hydrolysis, increasing blood supply to the lungs.

Conclusion: Metabolism of surfactant lipids and water balance in the lung, along with changes of rhythmogenesis and mode of ventilation, may lead to the development of disregulation of pneumopathy with dysfunction of the substantia nigra of the brain.

The purpose of the research consisted in the measurement of nonequilibrium radiation in superhigh frequency (SHF) range from aqueous solution of albumin upon its mechanical stimulation.

Methods: the monitoring of change in the ratio between brightness temperatures TSHF and TIR values after the mechanical stimulation of aqueous solution of albumin in the measuring cell at 35-39°С. The measurements of brightness temperatures were carried out with use of radiothermometer. SHF frequency range corresponded to 3.4-4.2 GHz, SHF frequency range corresponded to 8-13 mm.

Results: It was found that mechanical stimulation of aqueous solution of albumin at the temperature ~39°С, the change in the ratio between TSHF and TIR occurs. This corresponds to emergence of nonequilibrium SHF radiation from the solution near the phase transition with ~39°С temperature.

Conclusion: The effect of emergence of nonequilibrium SHF radiation from protein solution near its temperature 39°С was found. This temperature corresponds to the temperature of human organism upon a number of pathological states connected with inflammatory processes. The discovered effect can be used in the development of novel non-invasive methods of disease diagnostics.

The purpose of the research - studying the features of the production of pro- and anti-inflammatory cytokines, as well as indicators of thyroid status in patients with nephrotic variant of glomerulonephritis (GN). Research methods.

Methods: The examination involved 78 patients with primary GN, including 30 patients with nephrotic syndrome (NS) and 48 GN patients who had no NS symptoms. Laboratory researches included the determination of the concentration of the main cytokines circulating in the blood - IL-1b, IL-2, IL-4, IL-10, IFN-g and the receptor antagonist of IL-1b - Rа-IL-1b by the method of solid-phase enzyme linked immunosorbent assay enzyme immunoassay (ELISA) in the system of the bideterminant definition of antigen with the use of peroxidase as indicator enzyme using standard sets (“Cytokine”, St.-Petersburg) according to the technique attached to a set. The investigation of the basic indicators of thyroid status - free thyroxine (FT4), free triiodothyronine (FT3), thyroid-stimulating hormone (TSH), anti-thyroid peroxydase antibodies (TPOAb) is carried out by the ELISA using standard sets and NGO techniques «Diagnostic systems» (N-Novgorod). The researches were conducted twice - before the hospitalization (1-2 days) and after the end of a stationary stage of treatment (12-14 days).

Results: In 90% of patients with nephrotic option of GN there have been identified laboratory signs of hypothyroidism of different degrees of severity accompanied by increasing of production levels of proinflammatory cytokine IL-1b and IL-4, related to the activity of a humoral link of adaptive immunity. The reduction of glomerular, erythropoietic, concentration kidney functions, as well as proteinuria in patients with nephrotic option GN are associated with the decrease of T4 levels in the blood and increased levels of the cytokines circulating in the blood - IL-1b and IL-4.

Conclusion: The obtained data demonstrate that the high level of production of IL-1b and IL-4 in GN patients causes hypothyroidism resulting in the formation of NS.