Growth-Stimulating Effect of Platelet Preparations Obtained by Different Methods on Human Fibroblast Culture.

We studied the growth-stimulating effect of platelet-based preparations obtained by different methods on cultured human fibroblasts M-22. Platelet lysates prepared from platelet-rich plasma, platelet-poor plasma, concentrated suspension of platelets washed from plasma, and platelet-rich plasma activated with calcium chloride (ActPRP) were used. The volume of the platelet preparations was 10-500 µl per 104 cells. The most effective dose of platelet-rich and platelet-poor plasma in cell culture was 20 µl, whereas for ActPRP, the most effective dose was 500 µl. Lysates of platelet-rich and platelet-poor plasma in doses of 100-500 µl inhibited fibroblast growth and disturbed their structural integrity. At the same time, lysates of washed platelets in doses of 10-500 µl stimulated cell growth and preserved their viability. An inverse correlation was found between the number of cells in culture and the level of proinflammatory cytokines in the platelet preparations.

Experimental Study of the Effect of Biological Matrixes with Stabilized and Non-Stabilized Platelets on Reparative Process in the Wound Equivalent to Deep Burn.

We studied reparative effect of platelet-filled biological matrixes in the treatment of mice with wounds equivalent to deep burn. The wound coatings were based on decellularized dermal matrix without platelets (control), with native platelets, and with platelets stabilized with 2.5 µM nanosilver. In 3 days, the epithelial layer and derma were absent in all groups and extensive scab was formed. Dermal matrix with platelets simulated intensive migration of macrophages and fibroblasts to the wound bottom; in the control group, this migration was absent. In 14 days, granulation tissue appeared in the wound bottom in animals of all groups; in the experimental groups, the number of vessels was 2-4-fold higher than in the control, though the number of inflammatory cells in experimental groups remained high. On day 21, the scab on the most of the wound area was absent in all animals of the experimental groups and epithelialization and hair growth were pronounced, comparing to control. Nevertheless, in experiment dermal layer was not already completed, inflammation reaction remained.

[Blood pressure variability in hemodialysis patients: prognostic significance and treatment possibilities].

A present review is devoted to the current state of the problem of blood pressure variability (BPV) in hemodialysis patients. The BPV classification and clinical significance of BPV metrics are discussed. The results of cohort and randomized studies on the high BPV influence on outcomes in hemodialysis patients, as well as on the possibilities of antihypertensive drugs in the treatment of high BPV in dialysis patients, are presented.

Interaction of Magnesium-Based Materials with Human Blood Cells and Culture of Human Diploid Cells In Vitro.

We studied morphofunctional properties of human blood cells and diploid culture cells exposed to different types of magnesium materials: pure magnesium (Mg), magnesium-yttrium-neodymium-zirconium alloy (Mg-Y-Nd-Zr) and magnesium-zinc-calcium alloy (Mg-Zn-Ca). The materials were incubated with donor blood and mesenchymal multipotent stromal cells over 3 days. The studied materials did not induce massive lysis of human erythrocytes and leukocytes in vitro, but gradually impaired their structural integrity. In all cases, spontaneous platelet aggregation was observed in 6 h. In the presence of pure Mg and Mg-Zn-Ca alloy, this was accompanied by a decrease in the number of platelets with granules. In 24 h, substantial platelet degranulation occurred in all cases and in 72 h, the platelets did not contain granules. In parallel, the formation of large aggregates (60 µ) was observed. In the culture of stromal cells, all Mg-based materials reduced structural integrity of cells in 24 h, but did not significantly inhibit cell proliferation. Structural integrity of stromal cells partially recovered by day 3 in culture. The studied materials (Mg, Mg-Y-Nd-Zr, and Mg-Zn-Ca) seemed to be low-toxic for human cells during short-term contact, but could stimulate platelet aggregation and spontaneous degranulation and reduced the viability of diploid cells in vitro.

Growth-Stimulating Effect of Human Platelets Stabilized with Silver Nanoparticles.

We studied proliferative activity of multipotent mesenchymal stromal cells on collagen matrices containing platelets stabilized with silver nanoparticles. Dose-dependent stimulation of the growth of stromal cells without their structural damage was observed in the presence of stabilized platelets in proportion of 20-120 mln per 105 multipotent mesenchymal stromal cells. The same doses of non-stabilized platelets produced less pronounced stimulation effect. In higher doses (≥180 mln) stabilized platelets inhibited cell proliferation and induced their damage. Stabilized platelets enhanced migration of multipotent mesenchymal stromal cells; after formation of the monolayer, they actively colonized deep layer of the collagen matrix. The formed monolayer of multipotent mesenchymal stromal cells survived over 14 days without appreciable cell damage.

[Prevalence of Markers of Chronic Kidney Disease in Patients With Arterial Hypertension: Results of Epidemiological Study CHRONOGRAF].

In an noninterventional observational open multicenter program CHRONOGRAF the prevalence of markers of chronic kidney disease (CKD) was studied in 1600 patients with arterial hypertension with and without type 2 diabetes mellitus. Glomerular filtration rate (GFR) was calculated (CKD­EPI formula) and albuminuria (AU) was determined as albumin/creatinine (A/Cr) ratio in the morning portion of urine. In 49.4% of patients decreased GFR 30 mg/g was detected. Decrease of GFR 30 mg/g - in 32.6%, combination of reduced GFR and high/very high AU - in 18%. High or very high level of combined risk of progression to CKD and cardiovascular complications had 28.5% of patients.

Morphofunctional Properties of Human Platelets Treated with Silver Nanoparticles.

We studied structural and functional properties of human platelets in the presence of nanosilver particles. Incubation with 0.05-5 µM silver nanoparticles suppressed platelet adhesion in a dose-dependent manner without affecting internal platelet structure; during adhesion, some granules were not exocytized. Spontaneous platelet activation was observed at nanoparticle concentrations 15-100 µM. Addition of 1-5 µM nanosilver to cells undergoing adhesion blocked massive platelet degranulation, but did not prevent the formation of lamellopodia. The maximum number of preserved granules in platelets was revealed in the presence of 2.5-5 µM silver nanoparticles: 50% after platelet preincubation with silver nanoparticles and 75-77% after stabilization of adherent platelets with silver nanoparticles.

[ACUTE HEPATIC LESION ASSOCIATED WITH MYOCARDIAL INFARCTION: CLINICAL SIGNIFICANCE AND MODERN DIAGNOSTICS].

This review of the literature is devoted to the problem of acute kidney injury (AKI) in patients with acute myocardial infarction (MI). We discuss the occurrence of AKI in patients with MI, mechanisms of its development and modern diagnostic methods. The article examines biomarkers of kidney injury that may be useful for early diagnostics of AKI.

[Diurnal rhythm of arterial pressure in patients with acromegaly].

24 hr monitoring of arterial pressure (AP) in 46 patients with acromegaly in active phase revealed systolic-diastolic arterial hypertension. Analysis of diurnal profile showed the absence of the normal nocturnal fall in AP (non-dipper profile) and the significant rise in the number of patients with elevated nocturnal AP (night-peakers).

[Therapy of Arterial Hypertension in Patients With Chronic Heart Failure and Signs of Chronic Kidney Disease With Fixed Perindopril/Amlodipine Combination].

PURPOSE: to assess efficacy of a fixed combination of perindopril arginine and amlodipine besylate in the treatment of hypertensive patients with chronic heart failure (CHF) and signs of chronic kidney disease (CKD). MATERIAL AND METHODS: Persons with CKD (n = 53, age 64.5 ± 8.2 years) were selected from 118 patients with grade 2-3 essential hypertension with CHF. Presence of CKD was confirmed by elevated blood levels of cystatin C and lowered glomerular filtration rate (GFR). Used doses of fixed perindopril/amlodipine combination were 5/5, 5/10, 10/10 mg. Efficacy was assessed after 2 months of therapy on the basis of results of the examination which included clinical blood pressure (BP), BP monitoring, 6-min walk test. Blood levels of creatinine, urea, cystatin C and GFR were also measured. RESULTS: Target BP was achieved in 92.6% of patients. Clinical BP was significantly decreased. Daily average systolic BP decreased by 14.7%, diastolic BP--by 14.4%. Systolic and diastolic BP load also decreased. Clinical condition improved. Tolerance to physical work increased. Level of cystatin C significantly decreased while GFR increased. CONCLUSION: Fixed perindopril/amlodipine combination produced good therapeutic effect in hypertensive patients with CHF and CKD.

[EFFECTIVENESS OF FIXED COMBINATION OF PERINDOPRILARGININE AND AMLODIPINE FOR THE TREATMENT OF ARTERIAL HYPERTENSION WITH CONCOMITANT CHRONIC CARDIAC INSUFFICIENCY AND SIGNS OF CHRONIC KIDNEY DISEASE].

AIM: to study effectiveness of a fixed combination of perindopril arginine and amlodipine besylate for the treatment of arterial hypertension with concomitant chronic cardiac insufficiency and signs of chronic kidney disease Materials and methods: 53 (44.9%) patients aged 64.5±8.2 yr with signs of chronic kidney disease (CKD) were selected from 118 subjects with grade II-III essential AH and chronic cardiac insufficiency They had an increased blood cystatin C level (mean 1.4±0.3 mg/l) and/or decreased glomerular filtration rate (GFR) calculated by 3 formulas: 77.6±15.9 (MDRD), 79.3±17.1 (CKD-EPI) and 57.7±15.2 ml/min/1. 73 m3 (from cystatin C level). Fixed combinations of perindopril arginine and amlodipine besylate (5/5, 5/10, 10/10 mg) were used to treat the patients. Their effectiveness was evaluated 2 months after the onset of therapy based on results of AP measurement, 24 hr AP monitoring, assessment of clinical conditions and 6 min walk test. Blood creatinine, urea and cystatin C levels and GFR were measured. RESULTS: Target AP was achieved in 82.6% of the patients. Systolic AP (SAP) decreased significantly from 172±11.2 to 135.7±8.2 mmHg (p<0.01), diastolic AP (DAP)from 110±6.9 to 85.3±7.2 mm Hg (p<0.5). Mean daily SAP and DAP decreased by 14.7 and 14.4% respectively. SAP and DAP load also decreased Clinical conditions estimated in terms of the CCI functional class improved (a decrease from 6.2±1.1 to 4.2±1.3 scores (p<0.01)). Tolerance of physical activity increased from 157.2±12.6 to 320.4±32.2 m (p<0.05). Blood cystatin C level decreased and GFR increased. CONCLUSION: The fixed combination of perindopril arginine and amlodipine besylate ensures high therapeutic effect in patients with arterial hypertension and concomitant chronic cardiac insufficiency having signs of chronic kidney disease.

Optimization of the Method of Buccal Epithelial Cell Isolation and Culturing on Collagen Substrate for Ophthalmologic Application.

We performed morphofunctional analysis of cultured rabbit buccal epithelial cells in the primary culture, during passaging, and upon interaction with collagen substrates. Three different morphological types were identified. It was demonstrated that precursors of rabbit buccal epithelium were characterized by high proliferative activity, retain their biological properties throughout long-term culturing, and adhere to various collagen substrates.

[Therapy of Arterial Hypertension in Patients With Chronic Heart Failure and Signs of Chronic Kidney Disease With Fixed Perindopril/Amlodipine Combination].

PURPOSE: to assess efficacy of a fixed combination of perindopril arginine and amlodipine besylate in the treatment of hypertensive patients with chronic heart failure (CHF) and signs of chronic kidney disease (CKD). MATERIAL AND METHODS: Persons with CKD (n=53, age 64.5+/-8.2years) were selected from 118 patients with grade 2-3 essential hypertension with CHF. Presence of CKD was confirmed by elevated blood levels of cystatin C and lowered glomerular filtration rate (GFR). Used doses of fixed perindopril/amlodipine combination were 5/5, 5/10, 10/10mg. Efficacy was assessed after 2 months of therapy on the basis of results of the examination which included clinical blood pressure (BP).

Experimental substantiation and clinical use of a combination of dermal matrix with allogenic or autologous cells for the treatment of extensive traumatic wounds.

We carried out a comparative experimental and clinical study of the efficiency of the combination of dermal matrix with autologous and allogenic cells for creation of an optimal biological wound coating. Experiments on outbred mice have demonstrated that the use of dermal matrix in combination with allogeneic or autologous cells shortens the inflammation phase and accelerated maturation of the granulation tissue. Clinical application of biological wound dressing based on dermal matrix in combination with autologous or allogeneic mesenchymal multipotent stromal cells prevented septic complications and shortened the time of preparing extensive traumatic wounds with skin defect to autodermoplasty.

Morphofunctional analysis of human platelets by vital staining.

We developed a method for differential staining of human platelets preserving their functional activity based on vital fl uorochrome stains trypafl avin and acridine orange. Platelets stained with trypafl avin and acridine orange exhibited under a fl uorescent microscope green fl uorescence of the cytoplasm and red-orange fl uorescence of the granules. Morphofunctional analysis of platelets was carried out on the cells from donor blood, donor concentrated platelets, and cells from hematological patients and patients with thromboembolic complications. Populations with low (16 %) and high (2 %) morphofunctional activities of platelets were detected among donors. The morphofunctional parameters of platelets were sharply reduced in hematological patients with the hemorrhagic syndrome and elevated signifi cantly in patients with thromboembolic complications in comparison with donors. The method seemed to be effective for evaluating the platelet quality in donor and patients' blood components.

[Vegetative regulation of the cardiovascular system in patients with chronic heart failure with chronic renal insufficiency].

This review deals with vegetative regulation of cardiovascular system in patients with chronic heart failure (CHF) and signs of chronic renal insufficiency. CHF is currently regarded as a disturbance of neurohumoral mechanisms controlling blood circulation. At the same time, both prognosis and outcome of CHF depend on the presence of chronic renal insufficiency whose pathogenesis is poorly understood The authors emphasize the importance of elucidation of common pathogenetic mechanisms of these mutually complicating conditions.

[The morphologic evaluation of adhesive activity].

The morphologic evaluation of adhesive activity of human thrombocytes using both vital dye of cells and registration of adhesion process with fluorescent microscope is proposed. The regression dependence between adhesive activity of human thrombocytes and morpho-functional and aggregative activity of thrombocytes is established. The blood samples of 100 blood donors, 100 patients with hematologic pathology, 110 patients of traumatology, 50 patients with acute exogenous intoxications and 20 patients with chronic trophic ulcers were analyzed. The drastic decrease of adhesive activity of human thrombocytes and number of adhesively active thrombocytes was established in circulating blood of patients with bleedings of various etiology and hematologic patients. The significant increase of adhesive activity of human thrombocytes and number of adhesively active thrombocytes in patients with thrombosis is established. The morphologic evaluation of quality of thrombocytes is effective in case of analysis of biologic value of all population of circulating cells.

Effect of concentration of platelet-derived growth factor on proliferative activity of human fibroblasts.

Serum concentration of platelet-derived growth factor was measured in patients with trophic ulcers and proliferative activity of human fibroblasts was measured in wells with different content of added serum platelet-derived growth factor. In parallel, morphofunctional analysis of platelets from the whole blood of these patients was performed. A close correlation was found between morphofunctional parameters of blood platelets and serum concentration of platelet-derived growth factor. Platelet-derived growth factor in concentrations below 150 pg per well stimulated fibroblast proliferation and preserved their viability. Platelet-derived growth factor in concentrations >200 pg per well suppressed fibroblast proliferation and impaired their viability.

Sexual intercourse with leukocytospermic men may be a possible booster of oxidative stress in female partners of infertile couples.

Human semen has undoubtedly significant influence on the organism of the female counterpart. At the same time there are no studies in English literature investigating the influence of sexual intercourse on oxidative stress level in women's organism. Seventeen infertile couples where male partners were with (n = 5) or without (n = 12) leukocytospermia were enrolled in the study. Systemic oxidative stress levels were measured, whereby twice in female partners - before and 8-12 h after sexual intercourse. The men with leukocytospermia were characterized by oxidative stress that was substantially transferred to their partners during sexual intercourse as revealed by increase in 8-isoprostanes level (median 32.7 vs. 70.4 ng/mmol creatinine, p = 0.006). Sexual intercourse with male partner having leukocytospermia increases the oxidative stress level in the women's organism that may interfere with fertilization.

[Selective adsorption of endotoxin in the complex treatment of patients with severe sepsis].

Severe sepsis and septic shock remain the most serious problem of critical care medicine today with a mortality of 25-80%. Bacterial endotoxin is of considerable importance in the pathogenesis of sepsis. A selective hemosorbent in which endotoxin adsorption is accomplished via its binding to a synthetic peptide (LPS-A) has been recently designed in Sweden. The purpose of the study was to evaluate the efficiency of using the LPS-A in the complex treatment of patients with severe sepsis. Eight patients in whom the course of the underlying disease was complicated by the development of gram-negative sepsis in the postoperative period were examined. The patients had an APACHE II score of 20.4 +/- 4.5 LPS-A was performed using the hemosorbent Alteco LPS (Lund, Sweden) and an Aquarius hemoprocessor (Edwards Lifesciences) (U.S.A.). The use of LPS-A caused a significant reduction in endotoxin levels by 2.9 times. The level of procalcitonin was increased in all patients; after a LPS-A session, it was significantly decreased by 1.9 times. The use of LPS-A resulted in diminished tachycardia, elevated blood pressure and could reduce the doses of vasopressor amines. There was an improvement in lung gas exchange parameters. A study of the effect of LPS-A on the immune system showed a significant reduction (36.6%) in neutrophil oxygen-dependent metabolism according to the data of the spontaneous HCT test, a 2.4-fold decrease in large circulating immune complexes, and a rise in the count of mononuclear cells that died both by necrosis and apoptosis, the count of the cells significantly increased by 5.3 times in late apoptosis. A LPS-A session caused a significant drop in the blood level of endotoxin and procalcitonin and it was accompanied by a trend toward better hemodynamics and lung gas exchange and alleviated organ dysfunction. At the same time LPS-A showed the ambiguous effect on immune system parameters, which calls for further study.