[Risk factors and mechanisms of acute pyelonephritis development after contact ureterolithotripsy].

AIM: of the study is to identify risk factors for the development of acute pyelonephritis after contact urethrolithotripsy (URLLT) and to establish the mechanisms for maintaining inflammation after the withdrawal of NSAIDs. MATERIAL AND METHODS: The study included 21 patients who underwent contact ureterolithotripsy (URLT). The severity of leukocyturia was assessed 1 day after URLT, 2 days (the last appointment of NSAIDs, the total duration of the drug was 9 days) and 3 days (24 hours after NSAID discontinuation). The number of circulating platelet-leukocyte aggregates (PLA) was calculated by microscopy of stained blood smears. Analysis of the functional activity of platelet receptors involved in the modulation of the acute inflammatory response was performed by the turbidimetric method on a ChronoLog analyzer (USA).Statistical analysis was performed using the MedCalc package. RESULTS: After URSL, when NSAIDs were prescribed to patients, the level of leukocyturia decreased (p<0.05) compared to that at the time of hospitalization. A similar dynamics was found by analyzing the amount of TLA in the blood. Similar dynamics was found in the analysis of the amount of TLA in the blood. After 24 hours of NSAIDs cancellation, an increase in the severity of leukocyturia was detected (p<0.001). At the same time, normoreactivity of the 2-adrenergic receptor, GPVI receptor, AT1 receptor, PAT receptor, P2X1 receptor and A2A receptor, as well as hyporeactivity of the 2-adrenergic receptor and P2Y receptors, were revealed. An analysis of correlations made it possible to establish that the 2-adrenoreceptor, AT1 receptor, and GPVI receptor play a key role in the formation of TPA. Incubation of blood cells in vitro with agonists made it possible to establish that the maximum effect of TLA formation was reproduced during the interaction of the 2-adrenergic receptor and the AT1 receptor. CONCLUSION: With the abolition of NSAIDs, activation of the sympathetic-adrenal and renin-angiotensin systems, as well as remodeling of the basement membrane of the vascular wall are risk factors for the development of acute pyelonephritis after URLS.

[Extracellular matrix remodeling as a risk factor for the progression of cerebrovascular pathology]. / Remodelirovanie vnekletochnogo matriksa kak faktor riska progressirovaniya tserebrovaskulyarnoi patologii.

In recent years, attention of experts has been attracted to remodeling of the brains' extracellular matrix in connection with analysis of alterations' causes of the blood-brain barrier (BBB), development of neuroinflammation and neovasculogenesis, as well as the possibility of regeneration of nervous tissue in cerebrovascular pathology. The review discusses the molecular mechanisms of increasing BBB permeability in stroke associated with the interaction of structural elements of the capillary wall (endothelium and pericytes), glial cells and perivascular leukocytes. The role of brain extracellular matrix remodeling in the pathogenesis of neuroinflammation and participation of pericytes in this process are discussed. Management of remodeling process of structural elements in extracellular matrix may be a promising new technology in the treatment of patients with cerebrovascular pathology.

[Risks of progression of cerebrovascular pathology associated with the activity of the brain purinergic system]. / Riski progressirovaniya tserebrovaskulyarnoi patologii, svyazannye s aktivnost'yu purinergicheskoi sistemy mozga.

Until now, there is no understanding of the relationship between risk factors and the progression of cerebrovascular pathology. The review presents facts that confirm the involvement of various subtypes of purine P2 receptors in neuron activation, growth and myelination of axons, migration and microglia phagocytosis, astrogliosis, regulation of vascular tone, thrombosis and angiogenesis, neuroinflammation and immune responses. The data suggest the possibility of the activation of purinergic system of the brain during the development of main risk factors for cerebrovascular pathology (age, arterial hypertension, diabetes), as a stereotypical mechanism that can affect the homeostasis of the ensemble "neuron-glia-capillary". Purinergic P2 receptors may be a potential target for the development of pharmacological methods to limit the progression of cerebrovascular pathology.

[Activity of α2-adrenergic and PAF receptors of platelets as risk factor of acute pyelonephritis during urolithiasis in eldery women.]

The aim of the study is to identify possible mechanisms that could provoke a recurrence of chronic obstructive pyelonephritis (COP) in eldery women, thus to find out - why age and sex can be risk factors for acute pyelonephritis in urolithiasis. The results of 43 women over 65 years of age (mean age 71±1,5 years) are presented in study. The 26 patients were examined in remission phase of COP; 17 patients were examined in recurrence phase. For platelet stimulation epinephrine and platelet activating factor (PAF) were used at an effective concentration (EC50). The formation of platelet-leukocyte aggregates (PLA) was modeled in vitro by incubation of stimulated platelets (epinephrine and PAF) and intact leukocytes. The number of intact PLA was examined after blood smears staining according to Romanovsky-Giemsa method. In women older than 65 years realization of acute inflammation in urinary tract with urolithiasis occurs on the background of normal sensitivity of α2-adrenergic receptors and hyperreactivity of the PAF-receptors of platelets. In remission phase of COP the hyporeactivity of α2-adrenergic and PAF receptors of platelets was revealed but under interaction of epinephrine and PAF activation of platelets are possible which accompanied by formation of PLA. In women over 65 years hypoactivity of platelets in remission phase of COP can be a risk factor for the acute pyelonephritis development, since the simultaneous action of epinephrine and PAF increases the number of PLAs that recruit leukocytes for the realization of acute inflammatory reaction in urolithiasis.

[Interaction of epinephrine and ADP in regulation of platelet function in chronic cerebral ischemia ].

The purpose is devoted to test of hypothesis that patients with chronic cerebral ischemia (CCI) have decreased secretion of platelet ADP as the reason of platelet aggregation restriction in response to stimulation of adrenaline. Methods. We used platelet-rich plasma which was separated by centrifugation from peripheral blood of 55 patients with a diagnosis of CCI of stage 1-2. Platelets aggregation was studied on aggregometer Chrono - Log (USA). ADP and Epinephrine were used for platelet stimulation at effective concentration (EC50). Modulatory role of ADP subthreshold doses (0.5 mM) in platelet activation was analyzed with its addition to a suspension of platelets stimulated by agonists (EC50). Results. In 35 patients (group 1) with platelet hyperreactivity to ADP (EC50) response of platelets to Epinephrine was heterogeneous: in 17 cases (48.6%) there was high response (50%) and in 18 cases (51.4%) there was low platelet response to Epinephrine. 20 patients (group 2) had hyporesponsiveness of platelets upon stimulation by both agonists. It was established that the low initial response of platelets to Epinephrinе in vitro might be due to reduced secretion of ADP, i.e. limited adaptive response since administration of ADP subthreshold doses enhances adrenoreactivity of platelets. If pathochemical violations underlying the formation of platelet disadaptation are reversible, it is possible to recover the reaction of platelets to Epinephrine. Conclusion. In reducing the functional response of platelets to Epinephrinе key issue is establishing the reversibility of violations of platelets adaptive response of platelets.

[Possibilities to Control Early Stages of Inflammation in and of Thrombus Formation in Cardiovascular Pathology].

Circulating platelet-leukocyte aggregates (PLA) are present in patients with various diseases of the cardiovascular system and are considered as sensitive markers of platelet activation. At the same time characteristics of PLA in the circulating blood continue to be discussed in the context of the search for early markers of inflammation, which would bring us close to ability to limit the development of inflammatory response at the earliest stage. This review is devoted to analysis of studies of LA in the context of assessment of functional activity of platelets and possibilities of cooperation of blood cells during activation of leukocytes at early stage of realization of inflammation process. Platelets are considered as important amplifiers of inflammation. Moreover, the extent and specificity of platelet-leukocyte interactions are determined by the spectrum of receptors on the blood cell surface triggering a variety of intracellular signaling pathways. The facts that the use of known pharmacological agents can result in inhibition of stimulated platelets, blocking the binding of activated platelets with leukocytes are provided. Accumulated knowledge creates the preconditions for simultaneous control of inflammatory and thrombotic processes in various diseases of cardiovascular system.

[ROLE α2-ADRENERGIC RECEPTORS IN REGULATION PLATELET REACTIVITY IN THE ELDERLY AT CHRONIC OBSTRUCTIVE PYELONEPHRITIS].

Objective of the research was to determine involvement of platelets and the role of adrenaline in chronic inflammation maintaining and the initiation of acute inflammatory response in elderly patients with chronic obstructive pyelonephritis against this background. The study includes 60 patients with chronic obstructive pyelonephritis (COPN), which are distributed into two groups: basic - 22 elderly patients (age 73±1,5 years) and the comparison group - 38 middle-aged patients (52,5±2,4 years). The study excluded patients who took antiplatelet drugs and non-selective blockers of α adrenergic receptors at least 1 week before the study. Analysis of platelets adrenoreactivity in vitro was carried out at the time of hospitalization before the start of conservative therapy. Platelet-rich plasma was isolated from peripheral blood by centrifuging. ADP and epinephrine were used in the effective (EC50) and sub-threshold (EC10) concentrations to stimulate platelets. The formation of platelet-leukocyte aggregates was reproduced in vitro upon incubation of stimulated platelets (at a concentration of adrenaline EC50) and intact leukocytes isolated from patient peripheral blood. The study of platelet reactivity revealed that in elderly patients acute inflammatory response realization (relapse of COPN) is against optimal functioning of platelets α2 adrenergic receptors. Significant increase in the number of platelet-leukocyte aggregates is possible. Remission of COPN (the presence of chronic inflammation) in the examined patients of various ages was associated with platelet hypoadrenoreactivity. Increased platelet adrenoreactivity during transition from remission to relapse of COPN in the elderly patients is possible if adequate synthesis of ADP in platelets and its secretion from dense granules are preserved. The observed interaction of adrenaline and ADP with stimulated platelet hyporesponsiveness probably ensures adaptive response aimed at acute inflammatory response in COPN.

[Molecular mechanisms of individual platelet reactivity in hematuria secondary to lithotripsy].

AIM: To investigate the mechanisms of individual platelet reactivity to ADP and adrenaline associated with the variability of hematuria after lithotripsy in patients with chronic obstructive pyelonephritis (COPN). MATERIALS AND METHODS: The study included 41 COPN patients admitted to the Department of Urology for lithotripsy (LT). The contact ultrasonic LT was performed using the Karl Storz Calcuson Ultrasonic Lithotripsy System. Postoperative hematuria was assessed by microscopic red blood cell count. Platelets were separated from the citrated peripheral blood by centrifugation. Platelet aggregation was measured by Chrono-log aggregometer using agonists (ADP, adrenaline) at a concentration of EC50 and EU10. RESULTS: There were three types of platelet functional response to ADP and adrenaline after LT (increased, unchanged and decreased aggregation), but the predominant type of individual response was increased platelet aggregation. Testing 24 hours after LT revealed 7 platelet phenotypes differing in functional activity of 2-adrenoceptor agonist and purine receptors (R2Y1 and R2Y12). Normal purine receptor activity was associated with the ability of platelets to respond to adrenaline by increasing the functional activity aimed at limiting hematuria. Reduced platelet response to ADP after LT reaching the level of hyporesponsiveness may be viewed as a predictor of severe hematuria after surgery. CONCLUSION: Individual platelet reactivity, manifested by the interaction of ADP and adrenaline agonist, determines the effectiveness of the increase in pro-aggregation capacity of platelets in developing postoperative hematuria.

[Thromboxane A2: Mechanisms of Synthesis and Intracellular Signaling System of Realization].

One of major complications of ischemic heart disease is myocardial infarction, which develops as a result of thrombosis at the site of ruptured atherosclerotic plaque. Platelets activation and aggregation are the key events of this process. The efficiency of aspirin and/or clopidogrel use is limited by residual platelet reactivity what indicates the need to explore its mechanisms. This review covers intracellular signaling systems involved in realization of effects of the main platelet agonists in order to specify new molecules for the target therapy in case of aspirin resistance.

[The recovery of the function of hyporesponsive platelets during the antiplatelet therapy in patients with chronic cerebral ischemia]. / Vosstanovlenie funktsional'noĭ aktivnosti giporeaktivnykh trombotsitov pri antiagregantnoĭ terapii u patsientov s khronicheskoĭ ishemieĭ golovnogo mozga.

AIM: To establish the ability of recovery of platelet proagregation status in patients with chronic cerebral ischemia receiving standard medical therapy. MATERIAL AND METHODS: Thirty patients were studied. Platelets were isolated from peripheral blood by centrifugation. For in vitro platelet stimulation we used ADP, epinephrine and PAF. Platelet aggregation was recorded on aggregometer. RESULTS AND CONCLUSION: 24 hours after the beginning of standard therapy, the patients had 13 platelet phenotypes related to the functional state of three main receptors (purine receptors P2Y1 and P2Y12, α2-adrenergic receptors, and PAF- receptors. After 9-21 days of treatment (before discharge from the hospital), 15 patients (50%) had platelet phenotypes that did not change, i.e. standard medical therapy did not affect the performance of at least one of the receptor clusters, so the risk for initiation of thrombogenesis still remained. Remaining 15 patients had a phenotype with low platelet reactivity to all three agonists studied - cluster [ADP (↓) epinephrine (↓) PAF (↓)] indicating the achievement of antiplatelet effect. The given phenotype is characterized by summation or potentiation of the effects of agonists - ADP and epinephrine in 6 cases (40%), epinephrine and PAF in 6 (40%), ADP and PAF in 3 (20%), which means that conditions for recovery of functional activity of hyporeactive platelets could be further created.

[Interaction of the Humoral Agonist During the Platelets Activation in Patients with Chronic Cerebral Ischemia].

OBJECTIVE: The aim of the study was to determine the effect of the standard drug therapy of 95 patients with chronic cerebral ischemia (CCI) on the functional status of platelets as possible participants of microcirculation of the brain. METHODS: Platelets were isolated from peripheral blood by centrifugation and examined at 24 hours after initiation of standard medical therapy including aspirin (100 mg). The cells were stimulated in vitro using adenosine diphosphate (ADP), epinephrine, platelet activating factor (PAF) and serotonin in an effective concentration (EC50) inducing in healthy individuals platelet aggregation in the range of 50 ± 5%. A study of platelet aggregation was carried out on aggregometer Chrono-Log (USA). RESULTS: Research included 95 patients with CCI 1-2 Stage--82 patients taking antiplatelet and antihypertensive drugs before hospitalization (main group), 13--did not receive these drugs during 7 days prior to hospitalization (comparison group). After start conservative treatment, only ADP induced platelet aggregation, which was similar (p > 0.05) with values in healthy individuals. The pharmacological inhibition of the functional activity of platelets other investigated agonists reproduced hyporesponsiveness of platelets. Against this background, in 34 (41.5%) patients at 24 hours after initiation of therapy occurred potentiation effects of PAF and epinephrine in the test in vitro; whereas the summation of the effects of serotonin and epinephrine on platelets was detected in 12 (14.6%) patients. The basis of this phenomenon may be strengthening effect of ADP secreted from dense-granules, additional stimulation by Gi-protein signaling system by epinephrine and Gq-protein by the action of PAF and serotonin. CONCLUSION: Response to the combined action of platelet agonists may be predictor of the risk of thrombogenesis at CCI.

[The age characteristics of reaction of leukocytes in males under chronic obstructive pyelonephritis].

The leucocytosis and increase of numbers of neutrophils are two indicators commonly applied to evaluate acute inflammatory reaction. At that, the normality reference range is a standard of comparison. Considerably more often occurs the need to evaluate severity of patient condition or expression of inflammation determined by individual reactivity of organism. In this context the reaction of leukocytes was analyzed in 80 males of three age categories: younger than 55 years, 55-65 years and older than 65 years. All participants of study had verified diagnosis of chronic obstructive pyelonephritis. The comparison of leukopoiesis and erythrocyte sedimentation rate at the phase of remission and recurrence of chronic obstructive pyelonephritis of each age category made it possible to single out groups of patients with different reactivity of organism. In perspective, a possibility appears to individualize tactics of conservation therapy under chronic obstructive pyelonephritis and to evaluate its effectiveness.

[Purine receptors and associated signaling systems in regulation of platelet function].

Purine receptors of platelets play key role in hemostasis and thrombogenesis and they are used as targets for antiplatelet therapy. Platelets express three subtypes of purine receptors--P2X1, P2Y1 and P2Y12. The expression, ligands and sensitivity of purine receptors of platelets and their role in thrombogenesis are discussed in this article. Authors debate information about basic molecular mechanisms, links with intracellular signaling and biological effects of purine receptors activation. In conclusion, understanding of signaling mechanisms of purine receptors is a basis for further pharmacological strategy and effective antiplatelet therapy development.

[The impact of antisecretory therapy on gastroduodenal ulcers healing after acute bleeding].

In this paper we compared the efficacy of proton pump inhibitors (PPIs) and H2 receptor antagonists on the morphogenesis of the marginal zone of gastric and duodenal ulcers in 56 patients withacute gastroduodenal bleeding. It is shown that the antisecretory drugs in the treatment of patients with acute ulcerative bleedingnot only affect on the secretory activity of the glands in gastroduodenal zone, but it also modulates inflammatory reparative process and the status of mucous and bicarbonate barrier. A greater anti-inflammatory effect of PPI in comparison with H2-receptor antagonists has been proved. Appointment of PPIs had more pronounced stimulation of angiogenesis and cell proliferation of the surface epithelium.

[Age-related features of peripheral blood cell adaptive reaction in women with chronic obstructive pyelonephritis].

The aim of the study was to evaluate the adaptive changes of peripheral blood cells of recurrence inflammation in aged women with chronic obstructive pyelonephritis (COPN). An analysis of clinical and laboratory data was carried out on 50 women who were distributed into two age periods: under 55 years (the comparison group) and over 65 years. In women of 65+, during recurrence of COPN, the degree of leukocyte increase, neutrophils, monocytes and erythrocyte sedimentation rate (ESR) reached 90,1, 20,8, 71,4 and 31,6%, whereas lymphocyte count was less by 2,8 times in comparison with remission. In women under 55 years (in recurrence of COPN), degree of leukocyte increase was 87% (p<0,001), neutrophils--4, 8% (p<0,05), monocytes--in 2,04 times (p<0,001), ESR was less in 3,1 times (p<0,001) and lymphocyte count was less than 19,5% (p>0,05). The manifestation of neutrophilocytosis and lymphocytopenia might be due to age-related features of adaptive reaction in peripheral blood cells during the COPN recurrence.

[Molecular mechanisms of thrombogenesis].

Platelets are the key elements of hemostasis and optimal model for analysis of normal and altered mechanisms of thrombogenesis. Adhesion of platelets is initiated by von Willebrand factor, collagen, fibronectin, thrombospondin and laminin through GP VI, GPIb-V-IX complex, integrine receptors α(2)ß(1), α(IIb)ß, and Fc(γ)-receptors IIA. Their activation is accompanied with signalosome formation, increase of intracellular Ca2+ level, proteins phosphorylation, adhesion and degranulation of platelets. Major stimulators of progression phase are humoral factors and factors accumulated in dense granules of platelets, which include ADP, TA2, epinephrine, serotonin and thrombin. Their action is mediated by stimulation of G-protein coupled receptors: Gq, Gi, Gs, Gz, G12, G13, which can modulate activity of adenilate cyclase, phospholipase C, phosphoinositid-3-kinase, p115-RhoGEF and promote recruitment of platelets in the zone of injury. The phase of stabilization is due to contact-dependent signaling by integrins - predominantly α(IIb)ß(3), which ligand is fibrinogen, inducing reorganization of cytoskeleton in platelets, formation of strong intercellular junctions and retraction of thrombus.

[The role of purine receptors of thrombocytes in regulation of hemostasis].

The article discusses the role of ADP and ATP and their receptors in regulation of functional state of thrombocytes, maintenance of vessel wall homeostasis and coordination of inflammatory reparative process. The thrombocytes carry three modes of purine receptors: P2X1 (cationic channel activated by ATP) P2Y1 and P2Y12 (associated with G-proteins and activated by ADP). The article considers intracellular mechanisms of implementation of effects of ADP and ATP in thrombocytes6 mechanisms of their desensitization, relationship with metabolism of arachidonic acid and tyrosine kinase and role in regulation of intracellular concentration of Ca2+. The data concerning polymorphism of receptors P2Y is presented. The role of this phenomenon in thrombogenesis disorders, development of pathology of cardio-vascular system and resistance to anti-thrombocyte therapy is discussed.

[Interdisciplinary integration as an instrument for development a standard of medical education].

The formation of a competent doctor must be based on teaching standards for each discipline, the development of which is possible only as a result of provision of a real interdepartmental integration. The fundamental subjects must be taught not on the assumption of narrow professional interests, but in accordance with the requirements and demands of the clinical disciplines. The fulfillment of this task requires the development of a single computer base "Standards of Medical Education" and elaboration of the interdisciplinary modules. These must be based on the analysis of the "demand" of clinical Departments for new information blocks, the distribution of these blocks into different parts of the syllabus, their filling with the appropriate content, development of the adequate forms and methods of teaching.

[Functional activity of monocytes and mechanisms of iNOS intracellular regulation during wound process].

To investigate the dynamics and mechanisms of iNOS activity during wound process, the peripheral blood monocytes of 22 patients with acute foot wounds were analyzed. The basal and LPS-stimulated nitrites production was estimated at 1, 3-4, 7-10 and 14-18 days of wound process. iNOS activity and its molecular regulation was estimated by the inhibitory analysis. It was shown that since 1 to 3-4 day of wound process the basal and LPS-stimulated activity of iNOS, PDE and 5-LOX were elevated initially and than decreased. The COX and PkC activities increased after 3 days and reached the maximum level at days 7-10. The activity of PkA, which inhibits iNOS, intensively increased form 7-10th to 14-18th days of healing, and was accompanied by arginase-1 stimulation. Thus monitoring of intracellular signaling system can be used for diagnostics and correction of wound healing.

[Blood monocytic L-arginine metabolic changes in diabetic foot syndrome].

An inhibition test was used to study mechanisms responsible for L-arginine metabolic disturbances in the blood monocytes of patients with diabetic foot syndrome (DFS). It showed enhanced baseline iNOS activity and inhibition of the arginase pathway with lower nitrite production in response to the administration of lipopolysaccharide in the monocytes of patients with DFS. Impaired L-arginine metabolism was related to the higher activities of protein kinase C (PKC), phosphodiesterase (PDE), and 5-lipoxygenase (5-LO) along with decreased cyclooxygenase activity and drastic protein kinase A (PKA) inhibition. Within the first week, no changes in the wound process were associated with persistent metabolic disturbances of arachidonic acid and serine-threonine kinases with the higher sensitivity of AT1 receptors. In patients with DFS, the condition for wound process termination was decreased baseline iNOS activity and enhanced arginase-1 activity during PKA stimulation with the lower activity of 5-LO, PDE, and PKS. However, impaired mechanisms in the regulation of monocytic L-arginine metabolism persisted even a month later, which predetermines skin remodeling disturbance and the likelihood of recurrent DFS