Effects of Cancer Presence and Therapy on the Platelet Proteome

Platelets are involved in tumor angiogenesis and cancer progression. Previous studies indicated that cancer could affect platelet content. In the current study, we investigated whether cancer-associated proteins can be discerned in the platelets of cancer patients, and whether antitumor treatment may affect the platelet proteome. Platelets were isolated from nine patients with different cancer types and ten healthy volunteers. From three patients, platelets were isolated before and after the start of antitumor treatment. Mass spectrometry-based proteomics of gel-fractionated platelet proteins were used to compare patients versus controls and before and after treatment initiation. A total of 4059 proteins were detected, of which 50 were significantly more abundant in patients, and 36 more in healthy volunteers. Eight of these proteins overlapped with our previous cancer platelet proteomics study. From these data, we selected potential biomarkers of cancer including six upregulated proteins (RNF213, CTSG, PGLYRP1, RPL8, S100A8, S100A9) and two downregulated proteins (GPX1, TNS1). Antitumor treatment resulted in increased levels of 432 proteins and decreased levels of 189 proteins. In conclusion, the platelet proteome may be affected in cancer patients and platelets are a potential source of cancer biomarkers. In addition, we found in a small group of patients that anticancer treatment significantly changes the platelet proteome.

Fibrinolysis in Platelet Thrombi

The extent and duration of occlusive thrombus formation following an arterial atherothrombotic plaque disruption may be determined by the effectiveness of endogenous fibrinolysis. The determinants of endogenous fibrinolysis are the subject of much research, and it is now broadly accepted that clot composition as well as the environment in which the thrombus was formed play a significant role. Thrombi with a high platelet content demonstrate significant resistance to fibrinolysis, and this may be attributable to an augmented ability for thrombin generation and the release of fibrinolysis inhibitors, resulting in a fibrin-dense, stable thrombus. Additional platelet activators may augment thrombin generation further, and in the case of coronary stenosis, high shear has been shown to strengthen the attachment of the thrombus to the vessel wall. Neutrophil extracellular traps contribute to fibrinolysis resistance. Additionally, platelet-mediated clot retraction, release of Factor XIII and resultant crosslinking with fibrinolysis inhibitors impart structural stability to the thrombus against dislodgment by flow. Further work is needed in this rapidly evolving field, and efforts to mimic the pathophysiological environment in vitro are essential to further elucidate the mechanism of fibrinolysis resistance and in providing models to assess the effects of pharmacotherapy.

Hematological parameters and protein metabolism in the blood of pregnant rats under the effect of vanadium citrate

Dose-dependent changes in protein metabolism in the blood and hematological parameters of pregnant rats under the effect of vanadium citrate are presented in the article. The animals were divided into five groups: group I – non-pregnant females, II – pregnant females consuming pure water without additives, III, IV, V – females which during the mating and pregnancy period received the solution of vanadium citrate at concentrations of 0.03, 0.125 and 0.50 μg V/mL water. The research findings show that in pregnant animals of group II, the level of urea and alkaline phosphatase activity increased, meanwhile aspartate aminotransferase activity decreased, as compared to the non-pregnant females of group І. The levels of total protein and albumin decreased; however, the content of β-globulins increased in the pregnant animals of group II, as compared with that in group I. Also, in the rats of group II, there was a decrease in hemolysis time, total content of erythrocytes and hemoglobin, the content of old and mature erythrocytes, while the content of young erythrocytes increased, as compared to group I. The platelet content and thrombocrit in rats of group II increased in comparison with group I. The content of leukocytes and lymphocytes in pregnant animals of group II decreased, while the content of granulocytes increased, in contrast to non-pregnant rats. Under the effect of vanadium citrate at concentrations of 0.03–0.50 μg V/mL, there was a significant increase in the maximum number of prohemolized erythrocytes, the time of maximum hemolysis was delayed by 0.4–0.6 min, as compared with the pregnant rats of group II. This did not affect the time of total hemolysis in rats of groups III and V, as compared with the pregnant animals in group II. Under the effect of vanadium citrate, an increase in the content of young erythrocytes was observed, as compared with group II. The hemoglobin content decreased at the concentration of 0.125 μg V/mL, while at the concentration of 0.50 μg V/mL it increased, as compared to the pregnant animals of group II. Also, under the effect of vanadium citrate there was a decrease in the mean hemoglobin concentration in the erythrocyte. In pregnant animals fed with vanadium citrate solutions, the platelet content and thrombocrit, the relative width of platelet distribution by volume decreased, as compared with the pregnant rats of group II. The content of leukocytes, lymphocytes and granulocytes under the effect of vanadium citrate increased, as compared with the pregnant animals in group II. Under the effect of vanadium citrate at the concentration of 0.03 μg V/mL, the level of albumin, creatinine and aspartate aminotransferase activity increased in blood plasma in comparison with group II. Meanwhile, at the concentration of 0.125 μg V/mL, the relative content of γ-globulins and aspartate aminotransferase activity increased, alkaline phosphatase activity and urea level decreased in comparison with group II. However at the concentration of 0.50 μg V/mL, the relative α- and γ-globulins content and aspartate aminotransferase activity increased, at the same time, the relative β-globulins content and urea level decreased in comparison with group II. Therefore, vanadium citrate normalizes the indicators of protein metabolism during pregnancy, thus it can be considered as a potential dietary drug for the pregnant.

Quantitative Morphology of Cerebral Thrombi Related to Intravital Contraction and Clinical Features of Ischemic Stroke

Background and Purpose: The purpose was to assess quantitatively and qualitatively the composition and structure of cerebral thrombi and correlate them with the signs of intravital clot contraction (retraction), as well as with etiology, severity, duration, and outcomes of acute ischemic stroke. Methods: We quantified high-resolution scanning electron micrographs of 41 cerebral thrombi for their detailed cellular and noncellular composition and analyzed histological images for the overall structure with the emphasis on red blood cell compression, fibrin age, and the signs of inflammation. Results: Cerebral thrombi were quite compact and had extremely low porosity. The prevailing cell type was polyhedral compressed erythrocytes (polyhedrocytes) in the core, and fibrin-platelet aggregates were concentrated at the periphery; both findings are indicative of intravital contraction of the thrombi. The content of polyhedrocytes directly correlated with the stroke severity. The prevalence of fibrin bundles was typical for more severe cases, while the content of fibrin sponge prevailed in cases with a more favorable course. The overall platelet content in cerebral thrombi was surprisingly small, while the higher content of platelet aggregates was a marker of stroke severity. Fibrillar types of fibrin prevailed in atherothrombogenic thrombi. Older fibrin prevailed in thrombi from the patients who received thrombolytics, and younger fibrin dominated in cardioembolic thrombi. Alternating layers of erythrocytes and fibrin mixed with platelets were common for thrombi from the patients with more favorable outcomes. Thrombi with a higher number of leukocytes were associated with fatal cases. Conclusions: Most cerebral thrombi undergo intravital clot contraction (retraction) that may be of underestimated clinical importance. Despite the high variability of the composition and structure of cerebral thrombi, the content of certain types of blood cells and fibrin structures combined with the morphological signs of intravital contraction correlate with the clinical course and outcomes of acute ischemic stroke.

Effects of short-time cold impact on hematologic chrarcteristics of tundra vole (Microtus oeconomus PALLAS)

Проведено изучение воздействия низкой температуры (-50С) в течение двух часов на гематологические показатели самцов полевок-экономок (Microtus oeconomus Pallas) разного возраста (2, 4 и 10 месяцев). Обнаружена зависимость проявления эффекта от возраста животных. В большей степени на общее переохлаждение реагировали полевки старшей возрастной группы. Установлено, что действие холода вызывает умеренную лейкопению и лимфопению у половозрелых животных (4 и 10 месяцев), а также понижение содержания тромбоцитов в периферической крови у 10 месячных самцов. Полученные данные свидетельствуют о чувствительности показателей периферической крови самцов полевок-экономок к кратковременному действию низкой температуры и являются отражением неспецифичных клеточных реакций. We studied the effect of the 2-hours exposure to the low temperature (-50°C) on the hematological parameters of male tundra vole (Microtus oeconomus Pallas) of different ages (2, 4, and 10 months). We found the relation of the effect on the age of the animals. Voles of the older age group responded to a higher degree to general hypothermia. The action of the cold causes moderate leukopenia and lymphopenia in mature animals (4 and 10 months), as well as a decrease in platelet content in peripheral blood in 10-month-old males. The data obtained indicate the sensitivity of the peripheral blood indices of male tundra voles to the short-term effect of low temperature. They are a reflection of non-specific cellular reactions.

DYNAMICS OF SOME INDICATORS OF ENDOMETRITIS IN INFERTILE COWS

The article presents the results of the research obtained studying the dynamics of some indicators of endometritis in symptomatic infertility. The research was carried out at the Department of Obstetrics, Surgery and Physiology of Domestic Animals of the Don State Agrarian University in Persianovsky settlement, Oktyabrsky district, Rostov region. It was found that with latent endometritis, the most significant changes in the number of erythrocytes in 1 ml of blood were fixed with a minimum coefficient of variation (5.67–6.07); the platelet content in latent endometritis in cows compared to physiological norms for clinically healthy animals decreased by 26, 6%, and with chronic endometritis by 39.6%. The number of erythrocytes in cows with latent endometritis decreased by 1.1 times and in infertile cows with chronic endometritis – by 1.3 times compared to physiological norms typical for clinically healthy animals. Also, increased glucose content at this stage indicated a high level of bioenergetic processes in the body of cows, which proves its good protective forces. But a significant deviation was noted in the indicators of gluconeogenesis: with latent endometritis – by 1.15 times, with chronic endometritis – by 1.29 times (p < 0.01). The content of cholesterol in the blood was higher in cows with chronic endometritis, regardless of the form of its occurrence, in comparison with clinically healthy cows (p < 0.05), and the concentration of cholesterol in chronic endometritis was slightly higher than in latent endometritis (p < 0.05).

Platelets as a ‘natural factory’ for growth factor production that sustains normal (and pathological) cell biology

Platelets have attracted substantial attention in the current decade owing to their unexpected pleiotropic properties and conflicted functions. In fact, platelets participate in both health (hemostasis) and disease (thrombotic diseases). Much of the plasticity of platelets comes from the fact that platelets are the reservoir and the ‘natural factory’ of growth factors (GFs), with pivotal functions in wound repair and tissue regeneration. By combining the platelets’ plasticity and biotechnological processes, PlateInnove Biotechnology optimized the production of GFs in nanoparticle biointerfacing by platelet content, which opens an avenue of possibilities.

Abstract 147: Outcomes of the Stroke Thromboembolism Registry of Imaging and Pathology: A Multicenter International Study

Background: We performed a multicenter prospective clinical registry across 11 centers to study the association between histopathological characteristics of retrieved clots and imaging, stroke etiology and clinical outcomes. Materials and Methods: Following IRB approval at the 11 centers, patients were enrolled in the STRIP registry. All retrieved emboli were sent for histopathological analysis with H&E and MSB staining. Demographic variables, comorbidities, stroke etiology, imaging findings and procedural details were collected for each case. We studied the association between clot histopathology and imaging findings, stroke etiology and and revascularization outcomes. Student’s t-test was used for continuous variables and chi-squared testing for categorical variables. Results: To date, 1022 patients have been included. There was a significant correlation between platelet rich clots and the absence of hyperdensity on non-contrast CT [p=0.321, p=0.003) and a significant inverse correlation between the percentage of platelets and mean HU on NCCT (p=-0.243, p=0.025). The proportion of platelet-rich clots (55.0% versus 21.2%, p=0.005) and the percentage of platelet content (22.1% versus 13.9%, p=0.03) was significantly higher in patient with large artery atherosclerosis compared to those with a cardioembolic etiology. There was no correlation between RBC density, WBC density, fibrin density or platelet density and revascularization outcomes with stent-retrievers. However, we have found that with aspiration alone, patients with platelet rich clots are less likely to be fully revascularized (i.e. TICI 2c/3) than non-platelet rich clots (OR=0.36, 95%CI=0.12-0.81, P<.0001). Meanwhile, patients with RBC rich clots are more likely to be completely revascularized with aspiration alone than those with RBC poor clots (OR=2.71, 95%CI=1.25-3.24, P=0.02). Conclusions: Interim analysis of the STRIP registry suggests that the platelet content of a clot may be the most revealing factor in determining a clot’s etiology, imaging features and revascularization outcome. Platelet rich clots are less dense on NCCT, are associated with a large artery atherosclerosis source and are less likely to be completely revascularized with aspiration alone.