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Schizophrenia is a disorder of strange and disordered thoughts, delusions, hallucinations, inappropriate influences, and disorders of psychosocial functioning. One of the management of schizophrenia therapy is the administration of atypical antipsychotic drugs, namely clozapine. Clozapine is an antagonist of serotonin (5-HT2) and dopamine type 2 (D2) receptors. However, the risk of using clozapine provides a risk of side effects in the form of metabolic syndromes such as impaired blood glucose regulation and disorders of blood elements such as leukocytes and platelets. This study aims to determine the effect of using clozapine on blood glucose, leukocyte, and platelet levels in schizophrenia patients at Banyumas General Hospital. This research method was carried out by cross-sectional study in outpatient schizophrenia at Banyumas General Hospital who received clozapine therapy and had met the inclusion and exclusion criteria, taken by consecutive sampling, then examined blood glucose levels, leukocyte counts, and platelet counts before and after giving clozapine therapy. Samples were taken on day 1 and day 10 of drug use. Paired t-test statistical analysis was used to see the effect of using clozapine on glucose, leucocyte, and platelet levels. In the T-test, it was found that there was a relationship between the use of clozapine on the patient's blood glucose levels, the use of clozapine did not have a significant effect on the increase in leukocyte levels and there was also a relationship between the use of clozapine and the patient's platelet levels. The results showed that the patient had an increase in blood glucose levels by a percentage (100%), the patient had leukopenia by a percentage (63.5%), the patient had thrombocytopenia by a percentage (75%), while the other patients had thrombocytosis. Patients with hyperglycemia, leucopenia, and thrombocytopenia were those who used clozapine> 12 months

BackgroundAdult primary immune thrombocytopenia (ITP) is a rare bleeding disorder of unknown cause. Recent estimates of its incidence and trend over time were acquired for England.MethodThe primary ITP population (using ICD 10 code D693 and excluding secondary ITP cases; positive predictive value: 82.6%) was sourced from NHS Digital inpatient and outpatient. Incidence rate (IR) for England and by age groups, sex, and regions were calculated and trends were assessed using average annual percent change (AAPC).ResultsA total of 25 805 patients (mean age 59 years; females 57.8%) diagnosed between 2003 and 2014 was identified. IRs increased from 4.2/100 000 to 6.4/100 000 over this period (AAPC:4.3%). For all sex-specific age groups, the IRs significantly increased over time, except 18–29 years males. The greatest increase was among females aged 30–39 (AAPC:8.7%). In contrast, among ≥70 years, ITP was more common in males (highest IR among ≥80 years males: 23.9/100 000). England's average annual IR was 6.1/100 000 for 2010–14. An estimated 2.5/100 000 (based on UKITP Registry data) was estimated to require 1st line treatment whereas 2.4/100 000 would have 1st and 2nd line treatments within 6 months from diagnosis. IRs for London and East Midlands were the highest (6.5/100 000).ConclusionsThis study found a rising incidence of primary ITP, with sharp increases among young women and elderly men. These findings put in context the impact of ITP on patients' lives and the healthcare services in England, especially with 17%–50% who may develop chronic ITP and require long-term care.

Peritonitis and peritonitis-associated sepsis are characterized by an increased formation of platelet–neutrophil complexes (PNCs), which contribute to an excessive migration of polymorphonuclear neutrophils (PMN) into the inflamed tissue. An important neutrophilic mechanism to capture and kill invading pathogens is the formation of neutrophil extracellular traps (NETs). Formation of PNCs and NETs are essential to eliminate pathogens, but also lead to aggravated tissue damage. The chemokine receptors CXCR4 and CXCR7 on platelets and PMNs have been shown to play a pivotal role in inflammation. Thereby, CXCR4 and CXCR7 were linked with functional adenosine A2B receptor (Adora2b) signaling. We evaluated the effects of selective CXCR4 and CXCR7 inhibition on PNCs and NETs in zymosan- and fecal-induced sepsis. We determined the formation of PNCs in the blood and, in addition, their infiltration into various organs in wild-type and Adora2b−/− mice by flow cytometry and histological methods. Further, we evaluated NET formation in both mouse lines and the impact of Adora2b signaling on it. We hypothesized that the protective effects of CXCR4 and CXCR7 antagonism on PNC and NET formation are linked with Adora2b signaling. We observed an elevated CXCR4 and CXCR7 expression in circulating platelets and PMNs during acute inflammation. Specific CXCR4 and CXCR7 inhibition reduced PNC formation in the blood, respectively, in the peritoneal, lung, and liver tissue in wild-type mice, while no protective anti-inflammatory effects were observed in Adora2b−/− animals. In vitro, CXCR4 and CXCR7 antagonism dampened PNC and NET formation with human platelets and PMNs, confirming our in vivo data. In conclusion, our study reveals new protective aspects of the pharmacological modulation of CXCR4 and CXCR7 on PNC and NET formation during acute inflammation.

Background: Inflammation is important in the development of atherosclerosis. Research suggested sex-dependent differences for the value of inflammatory markers for risk stratification of stroke patients with internal carotid artery stenosis (ICAS). We investigated whether leukocytes and thrombocytes were associated with ≥50% ICAS in acute stroke and whether this was sex-dependent.Methods: Patients included in the PASS were evaluated for the predictive value of leukocytes and thrombocytes for ICAS. In PASS patients with ischemic stroke or TIA were randomized between four days of ceftriaxone intravenously or standard stroke care alone. Logistic regression analysis was performed adjusting for NIHSS and other covariates.Results: 2550 patients were included in PASS. 1413 of 2550 patients (55%) were evaluated in this sub study. Female patients showed a mean of 8.55 x 109/L for leukocytes and 259 x 109/L for thrombocytes. Men showed a mean of 8.29 x 109/L for leukocytes and 224 x 109/L for thrombocytes. Multivariate logistic regression analysis showed that leukocytes were independently associated with ICAS ≥50% in male patients (OR 1.094, p=0.008), but not in female patients (OR 1.041, p=0.360). Thrombocytes were not associated with ICAS.Conclusions: We conclude that blood leukocyte count independently predicts ICAS in men after acute stroke, but not in women. Clinical Trial Registration-URL: https://doi.org/10.1186/ISRCTN66140176 Unique identifier: ISRCTN66140176

Aim . To assess the P2Y 12 inhibitors switch in patients ST segment elevation myocardial infarction (STEMI) in real-life’ clinical practice, evaluate the functional activity of thrombocytes and thrombocytopoiesis and determine the clinical and prognostic value of P2Y 12 inhibitors switch in the framework of dual antiplatelet therapy in patients with STEMI. Material and methods . We conducted local, stratified, prospective study in which were involved 101 patients, hospitalized no later than 12 hours after the STEMI manifestation. The antiplatelet therapy (APT), prescribed by the physicians at the pre-hospital and inpatient phases of treatment, was analyzed. Functional activity of thrombocytes, levels of thrombopoietin (THPO), stromal cell-derived factor 1 (SDF1) and thrombopoietic receptor (MPL) were investigated. The minimum observation period was 2 years. Death and repeated hospitalizations due to cardiovascular causes were monitored. Results . P2Y 12 inhibitors were switched in 32,7% of patients with STEMI. In the hospital, clopidogrel, which was prescribed at the prehospital phase, was replaced with ticagrelor (early APT escalation) — 22,8%. Patients with early APT escalation by the seventh day had significantly greater inhibition of platelet aggregation activity parameters (slope of the aggregation curve, latent aggregation time and area under the aggregation curve). Activation of the collagen-induced platelet aggregation was detected. With the early escalation of APT, the THPO level was statistically significantly higher, both on the second and on the 7th day measurements: 256,2 (209,0; 396,8) pg/ml vs 137,5 (105,7; 179,1) pg/ml (p=0,000) and 283,4 (228,9; 334,3) pg/ml vs 226,5 (163,2; 287,3) pg/ml (p=0,045), respectively. The frequency of reaching the combined endpoint (death + re-hospitalization) was 7,9% in patients who had a P2Y12 switch, and 28,1% in patients who did not change the P2Y 12 blocker. Conclusion . In actual clinical practice, patients with STEMI had the most frequent early APT escalation, which was characterized by a more significant suppression of adenosine diphosphate-induced platelet aggregation and secretion than in patients without P2Y 12 inhibitors switch, but with activation of collagen-induced aggregation. An increase in thrombocytogenesis was revealed in early replacement of clopidogrel by ticagrelor. Intrahospital replacement of the P2Y 12 inhibitor in patients with STEMI was accompanied by a decrease in the two-year death risk and repeated hospitalizations.

Platelets play an important part in the progression and pathological angiogenesis of brain glioma because of the different granules content and release of microvesicles that are the source of numerous mediators and bioactive substances, which probably provides a "strategy" for the tumour survival. The objective of study was exploring the effect of platelet-released secretion products of patients with brain glioma on the experimental model of tumour growth in vitro. For this purpose, the cells of glioma C6 were cultured for 72 hours under the addition of modified media containing platelet-released secretion products or conditioned media of peripheral blood cells of patients with glioma as well as persons of the comparison group without rough somatic pathology. In control glioma C6 cultures in standard conditions cell clusters were formed by the type of "spheroids", from which radial cell migration occurred, a tense cellular or reticular growth zone was formed, and tumour cells preserved their ability to mitotic division. Under the influence of platelet-released secretion products of patients with glioma, differently directed effects on cell mitotic activity and the number of cell clusters in glioma C6 cultures were detected depending on the degree of tumour malignancy: stimulating effect under the influence of platelet factors of patients with high-malignancy glioma (G4) and inhibitory effect – due to the influence of platelet factors of patients with differentiated glioma (G2). In contrast to the thrombocyte-released factors, the conditioned media of a common pool of peripheral blood cells of patients with G4 glioma suppressed the mitotic activity of tumour cells and did not affect the number of cell clusters. No changes in glioma C6 cultures were revealed after the influence of platelet-released secretion products of persons of the comparison group. The obtained data confirm the important role of platelets in the pathogenesis of brain glioma, pointing to the fundamental difference in the spectrum of biologically active molecules that are released by platelets of patients depending on the degree of tumour malignancy and are able to regulate the cell cycle and proliferative activity of the glioma tumour cells, which may have application as a diagnostic marker as well as predictive marker of response to antitumour therapy.

The article contains current data on complications developing in the wake of employing thrombocytes concentrate in clinical settings. Complication patterns are submitted, causes there of are analyzed. Technical aspects are discussed of optimum measures to be implemented in transfusions of the thrombocytes concentrate.

Grass carp reovirus (GCRV) causes serious losses to the grass carp industry. At present, infectious tissues of GCRV have been studied, but target cells remain unclear. In this study, peripheral blood cells were isolated, cultured, and infected with GCRV. Using quantitative real-time polymerase chain reaction (qRT-PCR), Western Blot, indirect immunofluorescence, flow cytometry, and transmission electron microscopy observation, a model of GCRV infected blood cells in vitro was established. The experimental results showed GCRV could be detectable in leukocytes only, while erythrocytes and thrombocytes could not. The virus particles in leukocytes are wrapped by empty membrane vesicles that resemble phagocytic vesicles. The empty membrane vesicles of leukocytes are different from virus inclusion bodies in C. idella kidney (CIK) cells. Meanwhile, the expression levels of IFN1, IL-1β, Mx2, TNFα were significantly up-regulated in leukocytes, indicating that GCRV could cause the production of the related immune responses. Therefore, GCRV can infect leukocytes in vitro, but not infect erythrocytes and thrombocytes. Leukocytes are target cells in blood cells of GCRV infections. This study lays a theoretical foundation for the study of the GCRV infection mechanism and anti-GCRV immunity.

Zhehestovska D. V., Hrebenyk M. V. Diagnostic and prognostic utility of the platelet parameters in patients with acute myocardial infarction. Journal of Education, Health and Sport. 2021;11(03): 67-76. eISSN 2391-8306. DOI http://dx.doi.org/10.12775/JEHS.2021.11.03.008 https://apcz.umk.pl/czasopisma/index.php/JEHS/article/view/JEHS.2021.11.03.008 https://zenodo.org/record/4632272 The journal has had 5 points in Ministry of Science and Higher Education parametric evaluation. § 8. 2) and § 12. 1. 2) 22.02.2019. © The Authors 2021; This article is published with open access at Licensee Open Journal Systems of Nicolaus Copernicus University in Torun, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author (s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non commercial license Share alike. (http://creativecommons.org/licenses/by-nc-sa/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 15.02.2021. Revised: 26.02.2021. Accepted: 23.03.2021. DIAGNOSTIC AND PROGNOSTIC UTILITY OF THE PLATELET PARAMETERS IN PATIENTS WITH ACUTE MYOCARDIAL INFARCTION D. V. Zhehestovska, M. V. Hrebenyk I. Horbachevsky Ternopil National Medical University, Faculty of Postgraduate Education, Department of Surgery, Ternopil, Ukraine Diana V. Zhehestovska – PhD student, I. Horbachevsky Ternopil National Medical University, Ternopil, Ukraine ORCID 0000-0001-6305-4143, e-mail: zhehestovska_dv@tdmu.edu.ua Maryan V. Hrebenyk – Professor, Head of the Department of Therapeutics and Family Medicine, I. Horbachevsky Ternopil National Medical University, Ternopil, Ukraine ORCID 0000-0002-8118-5742, e-mail: hrebenyk@tdmu.edu.ua Abstract Background. Complete blood count (CBC) is a simple and affordable diagnostic tool that is routinely used in therapeutic practice. At the same time, its diagnostic and prognostic potentials in terms of management of acute myocardial infarction (AMI) are currently limited. Based on a number of previous studies, we suggest that the analysis of platelet count and indices among AMI patients may have significant diagnostic and prognostic value, as well as dynamic changes in these parameters during the treatment course. Objectives. To evaluate the value of platelet indices and ratios in terms of their diagnostic and prognostic potential among patients with AMI at the time of hospitalization and on the 7th day of treatment. Materials and methods. The study involved 152 patients diagnosed with AMI (Group 1), 30 patients diagnosed with the chronic coronary syndrome (Group 2) and 24 healthy volunteers (Group 3). CBC was performed for all participants to determine all platelet indices. Also, anamnestic and clinical data were evaluated, as well as results of the coronary angiography. GRACE score was calculated. Results. The highest diagnostic value was shown by the WBC/MPV ratio (AUC 0,798; sensitivity 83,1 %; specificity 65,1 %) and PLT/L ratio (AUC 0,644; sensitivity 62,7 %; specificity 53,5 %), as opposed to traditionally used MPV (AUC 0,426; sensitivity 47,5 %; specificity 48,8 %) and PLT (AUC 0,484; sensitivity 60,8 %; specificity 58,1 %). We were unable to establish a relationship between CBC platelet counts at the time of hospitalization and the risk of complications according to the GRACE score. However, those changes according to CBC were obtained on the 7th day of treatment. In particular, the number of platelets was associated with the risk of hospital (r = 0,287; p = 0,016) and 6-month mortality (r = 0,260; p = 0,30), the duration of ischemia (r = 0,238; p = 0,016) and the number of affected vessels according to the results of angiography (r = 0,395; p = 0,002). Similarly, P-LCC correlated with the risk of hospital (r = 0,483; p = Conclusion. We found that combined ratios such as WBC/MPV and PLT/L had a higher diagnostic value compared to MPV, PLT and P-LCR. The prognostic potential of platelet indices was significantly higher on the 7th day of hospital stay. In particular, elevated levels of platelets, platelets, and P-LCR were correlated with a higher risk of hospital and 6-month mortality. Key words: inflammation; acute myocardial infarction; complete blood count; platelet; thrombocytes

Multiple non-aggregatory functions of human platelets (PLT) are widely acknowledged, yet their functional examination is limited mainly due to a lack of standardized isolation and analytic methods. Platelet apheresis (PA) is an established clinical method for PLT isolation aiming at the treatment of bleeding diathesis in severe thrombocytopenia. On the other hand, density gradient centrifugation (DC) is an isolation method applied in research for the analysis of the mitochondrial metabolic profile of oxidative phosphorylation (OXPHOS) in PLT obtained from small samples of human blood. We studied PLT obtained from 29 healthy donors by high-resolution respirometry for comparison of PA and DC isolates. ROUTINE respiration and electron transfer capacity of living PLT isolated by PA were significantly higher than in the DC group, whereas plasma membrane permeabilization resulted in a 57 % decrease of succinate oxidation in PA compared to DC. These differences were eliminated after washing the PA cells with phosphate buffer containing 10 mmol·L-1 EGTA, suggesting that several components, particularly Ca2+ and fuel substrates, were carried over into the respiratory assay from the serum in PA. A simple washing step was sufficient to enable functional mitochondrial analysis in subsamples obtained from PA. The combination of the standard clinical PA isolation procedure with PLT quality control and routine mitochondrial OXPHOS diagnostics meets an acute clinical demand in biomedical research of patients suffering from thrombocytopenia and metabolic diseases.

The paper presents the influence of eimeriostatics on morphological parameters of blood at experimental eimeriocis in checkens. According to a number of resolutions and instructions of the President, the poultry industry has been developed in dehkan farms and personal subsidiary plots, and the population has been tasked to produce dietary chicken and vitamin-rich eggs

CD4+FoxP3+ regulatory T cells (CD4+ Tregs) are known to dampen inflammation following severe trauma. Platelets were shown to augment their posttraumatic activation in burn injury, but the exact mechanisms remain unclear. We hypothesized that platelet activation mechanisms via GPIIb/IIIa, fibrinogen, and PAR4 have an immunological effect and modulate CD4+ Treg activation early after trauma. Therefore, C57Bl/6 N mice were injected with tirofiban (GPIIb/IIIa inhibition), ancrod (fibrinogen splitting enzyme), or tcY-NH2 (selective PAR4 antagonist peptide) before inducing a third-degree burn injury of 25% of the total body surface area. Changes in coagulation, and local and systemic CD4+ Treg activity were assessed via rotational thromboelastometry (ROTEM®) and phospho-flow cytometry 1 h post intervention. The inhibition of GPIIb/IIIa and fibrinogen locally led to a higher basic activity of CD4+ Tregs compared to non-inhibited animals. In contrast, PAR4 disruption on platelets locally led to an increased posttraumatic activation of CD4+ Tregs. Fibrinogen led to complete elimination of coagulation, whereas GPIIb/IIIa or PAR4 inhibition did not. GPIIb/IIIa receptor and fibrinogen inhibition increase CD4+ Tregs activity independently of trauma. Both are crucial for thrombus formation. We suggest platelets trapped in thrombi are unable to interact with CD4+ Tregs but augment their activity when circulating freely. In contrast, PAR4 seems to reduce CD4+ Treg activation following trauma. In summary, GPIIb/IIIa-, PAR4-, and fibrinogen-dependent pathways in platelets modulate CD4+ Treg baseline activity, independently from their hemostatic functionality. PAR4-dependent pathways modulate the posttraumatic interplay of platelets and CD4+ Tregs.

Background : Infant mortality in Indonesia is at the highest level among the countries in South East Asia. Asphyxia in newborn has become the second risk factor for infant mortality. Objective : The objective of this study was to investigate the correlation between the level of hemoglobin, hematocrit, and thrombocytes of the mothers and asphyxia in the newborns at Panembahan Senopati Bantul Hospital, Indonesia. Methods : The study employed case-control method with 282 samples, which 141 assigned in the case and control group. This study used medical record of the patients as the secondary data. Chi-square, multiple logistic regression, and Odds Ratio Calculation (OR) were used for data analysis. Results : There was a meaningful correlation between the level of hemoglobin (OR=2.3, 95%CI=1.3-4), hematocrit (OR=4.3, 95%CI=1.4-13.4, and thrombocytes (OR=5.3, 95%CI=1.14-24.6) of the mothers and asphyxia in newborns. Thrombocytes was the most dominant factor related to asphyxia in newborns. Conclusion : Mothers who have hemoglobin, hematocrit and thrombocytes levels under the normal have more than two times risk of asphyxia in their newborn than mothers who have normal hemoglobin, hematocrit and thrombocytes levels.

In recent decades, significant progress has been made in understanding the mechanisms of platelet function and platelet hemostasis correction. Platelets are considered as the most important participants of both normal, and pathological thrombotic process characteristic of the most different diseases and states. In the present review pathophysiological mechanisms of platelet synthesis of various mediators with paracrine effects, which can influence the function of other cells, are consecrated. The physiology of platelets was considered in detail. The leading role of platelets in pathogenesis of the majority of diseases of cardiovascular system as modulators of inflammatory reactions of the immune response which are considered as the leading mechanism of development of atherosclerosis was shown. The ability of platelets to encode inflammatory proteins allowing them to influence adaptive immunity functions was discussed. The role of platelets as a key component of the innate immune system was presented, which is confirmed by the presence of Tolllike receptors (TLR) and glycoproteins, such as integrin αIIbβ3, glycoprotein Ib-IX and FcγRIIa, involved in interaction with bacterial cells. The pathogenesis of the formation of platelet-leukocyte aggregates due to the rapid reversible interaction of P-selectin (CD62P) on the platelet surface with ligand-1 glycoprotein P-selectin (PSGL-1) on the plasma of leukocytes and the mechanism of extracellular neutrophil traps (NETs), as well as the influence of platelets on the function of lymphocytes was presented. The role of platelets in cancer progression, metastasis and thrombosis is considered, and the interrelation of thrombosis and metastasis in malignant diseases was analyzed. The efficiency of the use of antithrombotic drugs in the prevention of thrombosis and, as a consequence, in the prevention of cardiovascular diseases and cancer was discussed.

The ficoll density gradient method, the most widely used method for isolating avian peripheral blood mononuclear cells (PBMCs), results in significant contamination with nucleated thrombocytes. This thrombocyte contamination can affect both immunophenotyping and functional assays of T cells. We used full blood samples from ten vaccinated male layer hybrid chickens for isolating PBMCs, and have developed a low cost, combined dextran and ficoll density gradient separation method which, in comparison to the standard ficoll density gradient separation, markedly reduces thrombocyte contamination of the isolated PBMCs. An ex vivo T cell proliferation assay showed an overall reduced background proliferation in unstimulated and stimulated samples after combined dextran and ficoll separation of PBMCs, resulting in a statistically significant difference upon in vitro NDV challenge, Metoda razdvajanja gradijentom gustoće pomoću fikola najčešće je korištena metoda za izdvajanje leukocita iz periferne krvi ptica, ali koja rezultira značajnom kontaminacijom trombocitima. Trombociti u populaciji izdvojenih stanica utječu na rezultate fenotipskih (imunofenotipizacija), ali i funkcionalnih testova T limfocita. Koristeći uzorke pune krvi od 10 cijepljenih muških pilića komercijalnog hibrida nesilica razvili smo metodu izdvajanja leukocita kombiniranom primjenom dekstrana i gradijenta gustoće pomoću fikola CD8αex vivo test proliferacije T limfocita pokazao je smanjenu pozadinsku proliferaciju nestimuliranih stanica nakon izdvajanja leukocita kombiniranom primjenom dekstrana i fikola.

In vitro pharmacodynamic models are used to optimize in vivo dosing regimens in antimicrobial drug development. One limiting factor of such models is the lack of host factors such as corpuscular blood components as erythrocytes which have already been shown to impact activity of antibiotics and/or growth of the pathogen. However, the impact of thrombocytes has not previously been investigated. We set out to investigate if the addition of thrombocytes (set to physiological concentrations in blood of healthy human, i.e., 5 × 105 thrombocytes/μL standard growth media Mueller Hinton Broth, MHB) has an influence on bacterial growth and on the efficacy of antibiotics against Gram+ and Gram− bacteria. Growth assays and time-killing-curves (TKC) were performed with ATCC-strains of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa in triplicate over 24 h. The same approach was followed for 5 clinical isolates of Escherichia coli. Meropenem, ciprofloxacin, and tigecycline were tested as representatives of broad-spectrum antibiotics, and concentrations several-fold above and below the minimal inhibitory concentration (MIC) were simulated. No significant impact of thrombocytes was found on bacterial growth or antimicrobial stability for the investigated agents. Bacteria reduced thrombocyte content to different degree, indicating direct interaction of pathogens and thrombocytes. Impact on bacterial killing was observed but was not fully reproducible when thrombocytes from different donors where used. While interaction of bacteria and thrombocytes was evident in the present study, interaction between antibiotic activity and thrombocytes seems unlikely. Whether variability was caused by different thrombocyte concentrates needs further investigation.

Meta-analyses and Mendelian randomization (MR) may clarify the associations of smoking, blood cells and myeloproliferative neoplasms (MPN). We investigated the association of smoking with blood cells in the Danish General Suburban Population Study (GESUS, n = 11 083), by meta-analyses (including GESUS) of 92 studies (n = 531 741) and MR of smoking variant CHRNA3 (rs1051730[A]) in UK Biobank, and with MPN in a meta-analysis of six studies (n (total/cases):1 425 529/2187), totalling 2 307 745 participants. In the meta-analysis the random-effects standardized mean difference (SMD) in current smokers versus non-smokers was 0·82 (0·75–0·89, P = 2·0 * 10−108) for leukocytes, 0·09 (−0·02 to 0·21, P = 0·12) for erythrocytes, 0·53 (0·42–0·64, P = 8·0 * 10−22) for haematocrit, 0·42 (0·34–0·51, P = 7·1 * 10−21) for haemoglobin, 0·19 (0·08–0·31, P = 1·2 * 10−3) for mean corpuscular haemoglobin (MCH), 0·29 (0·19–0·39, P = 1·6 * 10−8) for mean corpuscular volume (MCV), and 0·04 (−0·04 to 0·13, P = 0·34) for platelets with trends for ever/ex-/current smokers, light/heavy smokers and female/male smokers. Analyses presented high heterogeneity but low publication bias. Per allele in CHRNA3, cigarettes per day in current smokers was associated with increased blood cell counts (leukocytes, neutrophils), MCH, red cell distribution width (RDW) and MCV. The pooled fixed-effects odds ratio for MPN was 1·44 [95% confidence interval (CI): 1·33–1·56; P = 1·8 * 10−19; I2 = 0%] in current smokers, 1·29 (1·15–1·44; P = 8·0 * 10−6; I2 = 0%) in ex-smokers, 1·49 (1·26–1·77; P = 4·4 * 10−6; I2 = 0%) in light smokers and 2·04 (1·74–2·39, P = 2·3 * 10−18; I2 = 51%) in heavy smokers compared with non-smokers. Smoking is observationally and genetically associated with increased leukocyte counts and red blood cell indices (MCH, MCV, RDW) and observationally with risk of MPN in current and ex-smokers versus non/never-smokers.

The modern sights on etiology, pathogeneses, diagnosis, differential diagnosis and treatment of the thrombocytosis are described

Platelets play an important role in initiating atherothrombosis, i.e. the formation of blood clots inside a blood vessel at areas of atherosclerotic vascular injury. The functional (prothrombotic) activity of platelets significantly varies both in healthy individuals and in patients with cardiovascular diseases. The increased platelet production and turnover may be one of the reasons for promoting platelet activity. Stimulating thrombocytopoiesis results in large and reticular (with an increased amount of RNA) "young" platelets in the bloodstream. These platelets contain more adhesive receptors, more secretory granules and have an increased aggregation capacity. The review provides data indicating that large and reticular platelets are not only markers, but also predictors of atherothrombotic events, and primarily of acute coronary syndrome. An increase in such platelet count in patients receiving antiplatelet drugs is associated with a decrease in effectiveness of their antiplatelet action. It is assumed that the appearance of large and reticular platelets in the blood of patients with atherosclerosis and atherothrombosis may be a consequence of an increase in the thrombopoietic activity of megakaryocytes in these pathological conditions.

Thrombocytes can be concentrated in blood derivatives and used as autologous transplants e.g. for wound treatment due to the release of growth factors such as platelet derived growth factor (PDGF). Conditions for processing and storage of these platelet-rich blood derivatives influence the release of PDGF from the platelet-bound α-granules into the plasma. In this study Platelet rich plasma (PRP) and Platelet concentrate (PC) were produced with a fully automated centrifugation system. Storage of PRP and PC for 1 h up to 4 months at temperatures between −20°C and +37°C was applied with the aim of evaluating the influence on the amount of released PDGF. Storage at −20°C resulted in the highest release of PDGF in PRP and a time dependency was determined: prolonged storage up to 1 month in PRP and 10 days in PC increased the release of PDGF. Regardless of the storage conditions, the release of PDGF per platelet was higher in PC than in PRP.

Equilibria between the membranes of erythrocytes as well as thrombocytes and solution ions in fatal accidental hypothermia were analyzed using a theoretical four-equilibria model. The model was developed to determinate parameters characterizing cell membrane-surrounding ion interactions: the total surface concentrations of both acidic and basic groups C A, C B, and association constants K AH, K BOH. Knowledge of these parameters was necessary to calculate the theoretical values of surface charge density. The model was validated by curve-fitting the experimental data points to simulated data generated by the model. The experimental and theoretical surface charge density values agree at pH 2-8, at higher pH the deviation was observed.

Imaging living cells by atomic force microscopy (AFM) promises not only high-resolution topographical data, but additionally, mechanical contrast, both of which are not obtainable with other microscopy techniques. Such imaging is however challenging, as cells need to be measured with low interaction forces to prevent either deformation or detachment from the surface. Off-resonance modes which periodically probe the surface have been shown to be advantageous, as they provide excellent force control combined with large amplitudes, which help reduce lateral force interactions. However, the low actuation frequency in traditional off-resonance techniques limits the imaging speed significantly. Using photothermal actuation, we probe the surface by directly actuating the cantilever. Due to the much smaller mass that needs to be actuated, the achievable measurement frequency is increased by two orders of magnitude. Additionally, photothermal off-resonance tapping (PORT) retains the precise force control of conventional off-resonance modes and is therefore well suited to gentle imaging. Here, we show how photothermal off-resonance tapping can be used to study live cells by AFM. As an example of imaging mammalian cells, the initial attachment, as well as long-term detachment, of human thrombocytes is presented. The membrane disrupting effect of the antimicrobial peptide CM-15 is shown on the cell wall of Escherichia coli. Finally, the dissolution of the cell wall of Bacillus subtilis by lysozyme is shown. Taken together, these evolutionarily disparate forms of life exemplify the usefulness of PORT for live cell imaging in a multitude of biological disciplines.

WOS: 000456836500002, PubMed ID: 30040071, Objective: To examine granulocytic and non-granulocytic cells in children with severe congenital neutropenia (SCN) and their non-neutropenic parents. Materials and Methods: Fifteen patients with SCN and 21 non-neutropenic parents were evaluated for a) CD95, CD95 ligand, annexin V, propidium iodide, cell cycle, and lymphocyte subsets by flow cytometry; b) rapid cell senescence (of leukocytes) by senescence-associated beta-galactosidase stain; c) aggregation tests by aggregometer; d) in vitro bleeding time by PFA-100 instrument; e) mepacrine-labeled dense granule number of thrombocytes by fluorescence microscope; and f) hematomorphology by light and electron microscope. HAX1, ELANE, G6PC3, CSF3R, and JAGN1 mutations associated with SCN were studied in patients and several parents. Results: Significant increase in apoptosis and secondary necrosis in monocytes, lymphocytes, and granulocytes of the patients and parents was detected, irrespective of the mutation type. CD95 and CD95 ligand results implied that apoptosis was non-CD95-mediated. Leukocytes of 25%, 12.5%, and 0% of patients, parents, and controls showed rapid cell senescence. The cell cycle analysis testable in four cases showed G1 arrest and apoptosis in lymphocytes of three. The patients had HAX1 (n=6), ELANE (n=2), G6PC3 (n=2), and unidentified (n=5) mutations. The CD3, CD4, and NK lymphocytes were below normal levels in 16.6%, 8.3%, and 36.4% of the patients and in 0%, 0%, and 15.4% of the parents (controls: 0%, 0%, 5.6%). The thrombocytes aggregated at low rates, dense granule number/thrombocyte ratio was low, and in vitro bleeding time was prolonged in 37.5%-66.6% of patients and 33.3%-63.2% of parents (vs. 0% in controls). Under electron and/or light microscope, the neutrophils, monocytes, lymphocytes, and thrombocytes in the peripheral blood of both patients and parents were dysplastic and the bone marrow of patients revealed increased phagocytic activity, dysmegakaryopoiesis, and necrotic and apoptotic cells. Ultrastructurally, thrombocyte adhesion, aggregation, and release were inadequate. Conclusion: In cases of SCN, patients' pluripotent hematopoietic stem cells and their non-neutropenic parents are both affected irrespective of the genetic defect., Turkish Society of Hematology [06-B], This project was supported by the Turkish Society of Hematology (Project No: 06-B). We would like to thank all the patients and their families for kindly participating in our study. We are grateful to Prof. Nejat Akar, Aysel Pekel MD, Prof. Birkay Basturk, and Associate Prof. Yunus Kasim Terzi for their valuable help and comments. We also thank Didem Ozkazanc and Ahmet Gul from Istatistik Dunyasi for doing the statistical analysis; Eren Cimen MA, Selma Sar, Hulya Dalgali, Gul Ece Ulusan, and Esra Basyigit for technical assistance; Murat Teberoglu for his secretarial contribution; and our nurses for drawing blood from the patients.

Silymarin is a traditional drug and food supplement employed for numerous liver disorders. The available studies indicate that its activities may be broader, in particular due to claimed benefits in some cardiovascular diseases, but the contributions of individual silymarin components are unclear. Therefore, we tested silymarin flavonolignans as pure diastereomers as well as their sulfated metabolites for potential vasorelaxant and antiplatelet effects in isolated rat aorta and in human blood, respectively. Eleven compounds from a panel of 17 tested exhibited a vasorelaxant effect, with half maximal effective concentrations (EC50) ranging from 20 to 100 &micro, M, and some substances retained certain activity even in the range of hundreds of nM. Stereomers A were generally more potent as vasorelaxants than stereomers B. Interestingly, the most active compound was a metabolite&mdash, silychristin-19-O-sulfate. Although initial experiments showed that silybin, 2,3-dehydrosilybin, and 2,3-dehydrosilychristin were able to substantially block platelet aggregation, their effects were rapidly abolished with decreasing concentration, and were negligible at concentrations &le, 100 &micro, M. In conclusion, metabolites of silymarin flavonolignans seem to have biologically relevant vasodilatory properties, but the effect of silymarin components on platelets is low or negligible.

There are a lot of natural resources that potentially used as herbal medicines in Indonesia. Some of the herbs, like Dewa leaves (Gynura pseudochina (L) D.C) and temu ireng rhizome (Curcuma aeruginosa Roxb.), were ussually used to increase number of thrombocytes in dengue patient. This current study was to determine the potential of dewa leaves (Gynura pseudochina (L) D.C) and temu ireng rhizome (Curcuma aeruginosa Roxb.) against the number of trombocytes, erythrocytes, and hematocrits level on male white rats (Rattus novergicus) by using Heparin Induction. Dosages of the extracts were 250 mg/kg BW and 500 mg/kg BW used for both dewa leaves and temu ireng rhizome. Extracts were given orally to the tested animals for seven continuous days. The number of trombocytes, erythrocytes, and hematocrites level were examine at day 7 using sysmex XE-5000 hematology analyzer. The result showed that the rat groups which were treated with dewa leaves extract at 500 mg/kgBW; temu ireng Rhizomes extract at 250 mg/kgBW and 500 mg/kgBW dosages had significant differences (α = 0.05) on number of thrombocytes. The percentages of thrombocytes number enhancement for temu ireng rhizomes extract 500 mg/kgBW, temu ireng rhizomes extract 250 mg/kgBW, dewa leaves extract 500 mg/kgBW were 26.98%, 24.48%, 19.44% respectively. The rats group which were treated with temu ireng Rhizomes extract at 500 mg/kgBW, dewa leaves extract at 250 mg/kgBW and 500 mg/kgBW had significant differences (α = 0.05) on erythrocytes number and hematocrits level enhancement. The percentages of erythrocytes number enhancement for dewa leaves extract at dose of 500 mg/kgBW and 250 mg/kgBW, temu ireng rhizomes extract 500 mg/kgBW were 9.59%, 9.11%, 9.02% respectively. The highest percentages of hematocrits level enhancement was given by dewa leaves extract at dose of 500 mg/kgBW (10,97%), suggesting that the extracts treatment would not trigger plasma leakage. Thus, dewa leaves extract at 500 mg/kgBW; temu ireng rhizomes extract at 250 mg/kgBW and 500 mg/kgBW dosages are potential as candidates to be medicinal herbs to increase numbers of thrombocytes on dengue fever treatment

Бахарева Анастасия Сергеевна, кандидат биологических наук, доцент кафедры спортивного совершенствования, Южно-Уральский государственный университет. 454080, г. Челябинск, проспект Ленина, 76. E-mail: bakharevaas@susu.ru, ORCID: 0000-0003-0518-7751. Исаев Александр Петрович, доктор биологических наук, профессор, директор научно- исследовательского центра «Спортивная наука», Южно-Уральский государственный университет. 454080, г. Челябинск, проспект Ленина, 76. E-mail: isaevap@susu.ru, ORCID: 0000-0002-4488-0228. Малеев Дмитрий Олегович, кандидат педагогических наук, доцент, заведующий кафедрой лыжного спорта, Тюменский государственный университет. 625003, г. Тюмень, ул. Володарского, 6. E-mail: massport@mail.ru, ORCID: 0000-0003-4254-1705. Аминов Альбер Сибагатуллович, кандидат биологических наук, доцент, заведующий кафедрой спортивного совершенствования, Южно-Уральский государственный университет. 454080, г. Челябинск, проспект Ленина, 76. E-mail: aminovas@susu.ru, ORCID: 0000-0003-0440-6553. A.S. Bakhareva1, bakharevaas@susu.ru, ORCID: 0000-0003-0518-7751, A.P. Isaev1, isaevap@susu.ru, ORCID: 0000-0002-4488-0228, D.O. Maleev2, massport@mail.ru, ORCID: 0000-0003-4254-1705, A.S. Aminov1, aminovas@susu.ru, ORCID: 0000-0003-0440-6553 1South Ural State University, Chelyabinsk, Russian Federation, 2Tyumen State University, Tyumen, Russian Federation Цель работы. Цель настоящего исследования заключалась в выявлении физиологических маркеров, определяющих скорость сокращения и расслабления мышечного волокна I типа лыжников-гонщиков на этапе подготовки спортивного совершенствования. Материалы и методы. Контингент исследования составили лыжники-гонщики мужского состава в возрасте 21–23 года, спортивной квалификации КМС (14 человек) и МС (6 человек). Для достижения поставленной цели в работе использовались инструментальные методы и методы математической статистики. Результаты. Установлено, что замедление времени конца свертывания крови от подготовительного периода к соревновательному на 5,04 % (р = 0,02) на фоне повышения гематокрита на 19,12 % (р = 0,035) происходит рост скорости сокращения мышечного волокна I типа, а увеличение уровня фермента ацетилхолинэстеразы (АХЭ) на 4,07 % (р = 0,015) – одновременно указывает на рост скорости его раслабления. Заключение. По результатам исследования выявлена перестройка в работе скелетных мышц лыжников-гонщиков с высоким уровнем спортивной результативности. Увеличение активности АТФ-азы актомиозина мышечных волокон I типа у лыжников- гонщиков к периоду стартов в сочетании с ростом уровня фермента ацетилхолинэстеразы (АХЭ) определяет интенсификацию спортивной результативности. Aim. The aim of this study is to define physiological markers, determining the speed of muscle contraction and relaxation of type I muscle fibers in the racing skiers of high qualification. Material and methods. We studied male racing skiers aged 21–23 with the qualification of the Candidate for Master of Sport (14 persons) and the Master of Sport (6 persons). In order to reach the aim of the study we used instrumental methods and mathematical statistics techniques. Results. We registered that the slowdown of the end of blood coagulation from the training stage to a competition one by 5.04 % (р = 0.02) against the increase in hematocrit by 19.12 % (p = 0.035) results in the increase in the speed of type I muscle contraction. At the same time, the increase in the acetylcholinesterase level (ACH) by 4.07 % (p = 0.015) indicates the increase in the speed of muscle relaxation. Conclusion. As a result of the study, we found out changes in the muscle performance of racing skiers with a high level of sport efficiency. The increase in the activity of actomyosin ATPase of type I muscle fiber in racing skiers before competitions together with the increase in the acetylcholinesterase enzyme (ACH) determine the intensification of sport performance. Статья выполнена при поддержке Правительства РФ (Постановление № 211 от 16.03.2013 г.), соглашение № 02.A03.21.0011. The work was supported by Act 211 Government of the Russian Federation, contact №02.A03.21.0011.

The preparation of thrombocyte concentrates with filtration before storage (in-line) makes it possible to avoid the presence of mononuclear cells in the concentrate and proinflammatory cytokines. Therefore, this filtration may result with decreased activation of trombocyte receptors in vitro, which may improve therapeutic efficiancy. Methods. We compared two groups, each with 30 therapeutic doses of concentrated thrombocytes. We prepared the first group using the classic model from the buffy coat and the other with concentrated thrombocyte samples filtrated during sampling, so-called in-line, with the WBC filter Imuflex (Terumo). Mononuclear cells (MNC), thrombocyte, and erythrocyte counts in the units of concentrated thrombocytes were obtained on an automatic cell counter, and we used flow cytometry to measure the expression of surface thrombocyte receptors. The results demonstrated that the trombocytes prepared with pre-storage filtration contained a very low level of mononuclear cells and markedly reduced trombocyte receptors. Conclusion. The number of MNC and expression of surface thrombocyte receptors were markedly lower in the concentrated thrombocyte units prepared with in-line filtration. The thrombocytes prepared in this way contain fewer mononuclear cells, are of higher quality, are more functional, and may produce a better therapeutic effect in vivo.

Background Use of Carica papaya leaf extracts, reported to improve thrombocyte counts in dengue patients, demands further analysis on the underlying mechanism of its thrombopoietic cytokines induction Methods In vitro cultures of peripheral blood leukocytes (PBL) and stem cells from human exfoliated deciduous teeth (SHED) were treated with unripe papaya pulp juice (UPJ) to evaluate its potential to induce thrombopoietic cytokines (IL-6 and SCF) Results In vitro scratch gap closure was significantly faster (p

Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid that acts either as an intracellular messenger or as a ligand for its membrane receptors. S1P is a normal constituent of blood, where it is found both in plasma and blood cells. Compared with other cell types, sphingolipid metabolism in erythrocytes and platelets has unique features that allow the erythrocytes and platelets to accumulate S1P. In plasma, S1P is bound mainly to HDLs and albumin. Of note, metabolism and biological activity of S1P is to a large extent affected by the type of its carrier. Plasma S1P is characterized by a short half-life, indicating rapid clearance by degradative enzymes and the presence of high-capacity sources involved in maintaining its high concentration. These sources include blood cells, vascular endothelium, and hepatocytes. However, the extent to which each of these contributes to the plasma pool of S1P is a matter of debate. Circulating S1P plays a significant physiological role. It was found to be the key regulator of lymphocyte trafficking, endothelial barrier function, and vascular tone. The purpose of this review is to summarize the present state of knowledge on the metabolism, transport, and origin of plasma S1P, and to discuss the mechanisms regulating its homeostasis in blood.

The effect of fatal carbon monoxide poisoning on equilibria between cell membranes and surrounding ions was described using a theoretical four-equilibria model. The model was developed to obtain parameters characterizing the interactions between solution ions and erythrocyte or thrombocyte membrane surface. The parameters are the total surface concentrations of both acidic and basic groups C A, C B and their association constants with solution ions K AH, K BOH. These parameters were used to calculate the theoretical values of surface charge density. The model was validated by comparison of these values to experimental data, which were determined from the electrophoretic mobility measurements of the blood cells. The experimental and theoretical surface charge density values agree at pH 2-8, and at higher pH, the deviation was observed.

Platelets from human blood can be isolated in high yield by centrifugation of whole blood over an iodixanol density barrier of 1.063 g/ml. The separation from all of the blood cells (which form a pellet) is based on the slower sedimentation velocity of the smaller platelets.

The objective of the investigation was evaluation of postmortem changes of electric charge of human erythrocyte and thrombocyte membranes after sudden unexpected death. The surface charge density values were determined on the basis of the electrophoretic mobility measurements of the cells carried out at various pHs of electrolyte solution. The interactions between both erythrocyte and thrombocyte membranes and electrolyte ions were studied. Values of parameters characterizing the membrane--that is, the total surface concentrations of both acidic and basic groups and their association constants with solution ions--were calculated on the basis of a four-equilibria mathematical model. The model was validated by comparison of these values to experimental data. We established that examined electric properties of the cell membranes are affected by sudden unexpected death. Postmortem processes occurring in the cell membranes can lead to disorders of existing equilibria, which in turn result in changes in values of all the above-mentioned parameters.

В статье рассматриваются вопросы изменения морфологии тромбоцитов на фоне хронического описторхоза с разной степенью инвазированности. В результате анализа полученных данных выявлено, что хроническая описторхозная инвазия приводит к ускорению непрерывно протекающего свертывания крови, вызывая гиперкоагуляцию и активацию тромбоцитов в зависимости от степени заражения, и подвергается наибольшим изменениям при максимальной инвазии., The article describes some questions of changes in platelet morphology against the background of chronic opistorchosis with different invasion extent. The resultant data analysis reveals acceleration of continuous blood coagulation leading to hypercoagulation and platelet activation dependant on invasion extent and is subjected to the greatest changes at maximum invasion

В экспериментах на сирийских хомяках показано, что хроническая описторхозная инвазия уменьшает толерантность свертывающей системы крови к воздействию прокоагулянтов, усугубляя активацию и потребление тромбоцитов, а также плазменных ферментных систем. В экспериментах на сирийских хомяках показано, что хроническая описторхозная инвазия уменьшает толерантность свертывающей системы крови к воздействию прокоагулянтов, усугубляя активацию и потребление тромбоцитов, а также плазменных ферментных систем. In experiments on syrian hamsters it is shown that chronic opistorhtosis reduces to tolerance of hemostasis system to influence of procoagulants, aggravating activation and consumption of thrombocytes, as well as blood enzymatics systems. Соловьёв В.Г., заведующий кафедрой медицинской и биологической химии Ханты-Мансийской государственной медицинской академии, доктор медицинских наук (Ханты-Мансийск);тел. 8(34673)93435; e-mail: vg_solovev@mail.ru Solovyev V.G., manager of the chair of medical and biological chemistry of Khanti-Mansiisky state medical academy, doctor of medical sciences (Khanty-Mansisk);tel. 8(34673 )93435; e-mail: vg_solovev@mail.ru. Зиновьева А.В., ассистент кафедры медико-биологических дисциплин Ханты-Мансийской государственной медицинской академии (Ханты-Мансийск);тел. 8(34673)93436; e-mail: zinchl60290@mail.ru Zinovieva A.V., assistant of biomedical disciplines of Khanti-Mansiisky state medical academy (Khanty-Mansisk); tel. 8(34673)93436; e-mail:zinchl60290@mail.ru

Background: In addition to the regulation of calcium homeostasis, vitamin D affects the cellular immune system, targets the TNF-α pathway and increases vasoconstrictor response to angiotensin II. We therefore examined the effect of 1,25-dihydroxy-vitamin D3 on coagulation and organ failure in experimental sepsis in the rat. Methods: Three series of placebo-controlled studies were conducted. All rats were pre-treated with daily SC injections of 1,25-dihydroxy-vitamin D3 100 ng/kg or placebo vehicle for 3 days. In study 1, sepsis was accomplished by abdominal surgery comprising a coecal ligation and puncture with a 1,2 mm needle, or sham surgery. In study 2, the rats had a single IP injection of lipopolysaccharide from E. Coli 0111:B4 (LPS) 8 mg/kg, or placebo. In study 3, an hour-long IV infusion of LPS 7 mg/kg, or placebo was given. Results: All three models of sepsis showed significant effects on coagulation and liver function with reduced thrombocyte count and prothrombin time together with elevated ALT and bilirubin (p

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:21:01Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:45:21Z : No. of bitstreams: 1 2-s2.0-33645529163.pdf: 376234 bytes, checksum: 24d772915ba79189cd790de247ada82d (MD5) Made available in DSpace on 2014-05-27T11:21:01Z (GMT). No. of bitstreams: 0 Previous issue date: 2004-01-01 This research studied the condition (Kn) relative factor, the hepatosomatic (HSR), the splenosomatic (SSR) relation, the erythrocytes, thrombocytes and leukocytes number, the glicogen locality by PAS method in thrombocytes and leucocytes and the total protein and electrolytes serices levels in both Cyprinus carpio L. sex. The female Kn showed higher value than the males. The female HSR average values were lower than the ones observed in males, while the leukocytes, lymphocytes and neutrophils percentage number were the highest. However, the males magnesium and choride serices levels were higher than the females. The HRS average values, hematocrit, haemoglobin concentration, average corspucular volume (ACV), average corpuscular haemoglobin concentration (ACHC), erythocytes number, trombocytes, monocytes, oesinophils, positive PAS leukocyte granular (PAS-LG), and total protein, sodium, potassium and calcium serices levels did not show significant statistics difference (p> 0.05) between males and females. The PAS method showed glicogen granules into the thronbocytes, eosinophils and neutrophils cytoplasm. Nevertheless, in the PAS-LG cytoplasm, neutral glicoprotein granules were also observed. Monocytes and lynfocytes showed negative reaction to PAS. In the teleosts, the gonodas matureness presents higher relation with the factor of condition. During the reprodution period it is unlikely to find the sexual effects isolated in the hemogram, because most of the time that is not the only factor to cause dimorphism. Centro de Aqüicultura da Unesp (Caunesp), Via Prof. Paulo Donato Castellane, 14884-900, Jaboticabal, Sao Paulo Departamento de Patologia Animal FCAV-Unesp, Via Prof. Paulo Donato Castellane, 14884-900, Jaboticabal, Sao Paulo Piscicultura São Geraldo, Caixa Postal 518, Sertãozinho, São Paulo Laboratório de Análises Clínicas Hospital Veterinário FCAV-Unesp, Via Prof. Paulo Donato Castellane, 14884-900, Jaboticabal, Sao Paulo Centro de Aqüicultura da Unesp (Caunesp), Via Prof. Paulo Donato Castellane, 14884-900, Jaboticabal, Sao Paulo Departamento de Patologia Animal FCAV-Unesp, Via Prof. Paulo Donato Castellane, 14884-900, Jaboticabal, Sao Paulo Laboratório de Análises Clínicas Hospital Veterinário FCAV-Unesp, Via Prof. Paulo Donato Castellane, 14884-900, Jaboticabal, Sao Paulo

Growing evidence supports the substantial pathophysiological impact of platelets on the development of acute lung injury. Methods for studying these cellular mechanisms in vivo are not present yet. The aim of this study was to develop a model enabling the quantitative analysis of platelet kinetics and platelet-endothelium interaction within consecutive segments of the pulmonary microcirculation in vivo. New Zealand White rabbits were anesthetized and ventilated. Autologous platelets were separated from blood and labeled ex vivo with rhodamine 6G. After implantation of a thoracic window, microhemodynamics and kinetics of platelets were investigated by intravital microscopy. Velocities of red blood cells (RBCs) and platelets were measured in arterioles, capillaries and venules, and the number of platelets adhering to the microvascular endothelium was counted. Kinetics of unstimulated platelets was compared with kinetics of thrombin-activated platelets. Velocity of unstimulated platelets was comparable to RBC velocity in all vessel segments. Unstimulated platelets passed the pulmonary microcirculation without substantial platelet-endothelial interaction. In contrast, velocity of activated platelets was decreased in all vascular segments indicating platelet margination and temporal platelet-endothelium interaction. Thrombin-activated platelets adhered to arteriolar endothelium; in capillaries and venules adherence of platelets was increased 8-fold and 13-fold, respectively. In conclusion, using intravital microscopy platelet kinetics were directly analyzed in the pulmonary microcirculation in vivo for the first time. In contrast to leukocytes, no substantial platelet-endothelium interaction occurs in the pulmonary microcirculation without any further stimulus. In response to platelet activation, molecular mechanisms enable adhesion of platelets in arterioles and venules as well as retention of platelets within capillaries.

In this paper, we show that a mouse monoclonal antibody, 111-427, specific for the Vß5.3 chain of the human T-cell receptor (TCR) for antigen, also reacts with chicken hematopoietic cells. Our data indicate that the majority of 111-427 positive cells among peripheral blood leucocytes (PBL) are thrombocytes. This antibody also recognizes two in vitro cell lines, III-C5, an IL-2-dependent T-cell-line and HD11, a macrophage cell line. In addition, erythrocytes and a minor subpopulation of thymus and spleen cells are also stained by the monoclonal antibody (mAb). No specific immunoprecipitation could be detected from125I radiolabeled cell lysates. By Western blotting techniques, the 111- 427 mAb identifies a single band of apparent molecular weight 91 kD, unaffected by reduction, from III-C5 and HD11 cell lysates. This band is absent in negative cell control lysates. On thrombocytes, the apparent molecular weight of the band is shifted to 87 kD. These results indicate that the mAb does not recognize the chicken T-cell receptor for antigen, but a cell surface marker shared primarily between thrombocytes and erythrocytes. This new chicken cell marker is compared to other cell surface markers in avian or mammalian species that present some analogies in their tissue distribution.

En este trabajo se evaluó el efecto del cadmio y de la temperatura sobre la fragilidad osmótica y el número de células sanguíneas, en el pez Colossoma macropomum. Los organismos fueron expuestos a 0,5 mg/L de cloruro de cadmiodurante 24 días y a dos temperaturas (25 y 30ºC). A los 0; 10 y 24 días de exposición, se les tomó muestras de sangre para determinar la curva de fragilidad osmótica, el conteo diferencial de células blancas y el conteo de reticulocitos. Los eritrocitos presentaron principios de hemólisis a 90 mmol/L de NaCl y hemólisis total a 10 mmol/L. El porcentaje de linfocitos fue >62%, incrementándose en los peces expuestos a cadmio, no obstante, el porcentaje absoluto (controles y expuestos) de linfocitos, disminuyó a los 24 días de experimentación, siendo más significativo a la temperatura de 30°C. Después de los neutrófilos, los trombocitos fueron el tercer tipo celular encontrado, sobre los cuales, ni el cadmio ni la temperatura, tuvieron efecto significativo. Sin embargo, el número absoluto de estas células aumentó en los organismos mantenidos a 25°C/24 días; contrario a lo observado en los peces a 30°C/24 días, quienes registraron un descenso de 4% en su abundancia. Los peces expuestos a Cd/30°C revelaron un significativo aumento de trombocitos en comparación con los expuestos Cd/25ºC. Los reticulocitos disminuyeron por la exposición al metal. El cadmio y la temperatura ejercen un efecto sinérgico sobre el número de trombocitos y afectan individualmente la producción de reticulocitos en el pez. In this work, the cadmium (CD) and temperature effect on the red cell osmotic fragility and blood cells count in the tropical fish Colossoma macropomum were evaluated. The fishes were exposed to 0.5 mg/L of cadmium chloride during 24 days and two temperatures (25 and 30°C). At 0; 10 and 24 days of Cd exposure, blood samples were taken from fishes to determine the osmotic fragility curve, the differential white cell and reticulocyte count. The erythrocytes osmotic fragility began at 90 mmol/L of NaCl and total hemolysis was observed at 10 mmol/L of NaCl. Lymphocytes were the more abundant cells (>62%). These cells increased in Cd-exposed fish; moreover, the lymphocytes absolute percentage (control and exposed) decrease at 24 days of experimentation, being more significant at 30°C. After the granulocytes acidophils, trombocytes were the third cellular type, they were not affect by Cd neither temperature. However, trombocyte absolute number increased than fishes at 25°C/24 d as fishes at 30°C/24. On the other hand, the Cd/30°C exposed fishes showed a significant increase of thrombocytes. Both, cadmium and temperature affect thrombocyte number. The reticulocytes decrease due to cadmium exposed fishes. A synergic effect is produced by Cd and temperature on trombocytes; however, these factors exert an individual effect in fish reticulocytes.

Aim. Isolation and study ofbiological activity of antimicrobial peptides from chickens thrombocytes. Materials and methods. Peptides from chickens thrombocytes, obtained by reverse-phase high-performance liquid chromatography method with stepped and linear gradients of concentration increase of the organic solvent were used in the study. Their antimicrobial activity was determined by microtitration method in broth; mechanism of biological effect - by using fluorescent spectroscopy method with DNA-tropic dyes. Results. Individual fractions of peptides were isolated from chickens thrombocytes, that possess antimicrobial activity against Staphylococcus aureus P209 and Escherichia coli K12. A disruption of integrity of barrier structures of microorganisms under the effect of thrombocyte antimicrobial peptides and predominance of cells with damaged membrane in the population of E. coli was established. Conclusion. The data obtained on antimicrobial activity and mechanism of bactericidal effect of the peptide fractions from chickens thrombocytes isolated for the first time expand the understanding of functional properties of chickens thrombocytes and open a perspective for their further study with the aim of use as antimicrobial means.