Chronically Elevated Interleukin-6 Disturbs the Coagulation Cascade in Mice

Introduction: Pro-inflammatory cytokines play an essential role as activators of the hemostatic system and in the regulation of physiological antithrombotic mechanisms. Interleukin-6 (IL-6) influences platelet production and platelet activation. It was associated with accelerated clotting and intravascular coagulation in tissue factor (TF)-driven murine thrombosis models. However, the precise role of myeloid cell-derived IL-6 on thrombosis formation and the hemostatic system is still unknown. Methods and Results: To better understand the role of IL-6 in thrombosis and the hemostatic system, we developed a new mouse strain with Cre-recombinase driven constitutive IL-6 expression specifically in myeloid cells (LysM-IL-6 OE, Control mice: IL-6 OE). LysM-IL-6 OE mice had a prolonged tail bleeding time and lacked venous thrombus formation induced by inferior vena cava (IVC) stenosis. There were no differences in D-Dimer levels in LysM-IL-6 OE mice neither on baseline level nor after IVC ligation. However, we found unstoppable post-operative bleedings in LysM-IL-6 OE. They showed a prolonged aPTT, a significantly increased INR and a prolonged thrombin converting time. The Factor V and IX expression were reduced, but von Willebrand factor, antithrombin and fibrinogen expression were up-regulated and could not explain the missing thrombus formation. We found significantly elevated erythrocyte sedimentation in line with erythrocytes aggregates, which seemed to be mediated by IL-6 and α2M. Most importantly, hepatic levels of thrombin inhibitor α2 macroglobulin (α2M) mRNA and protein were increased in LysM-IL-6 OE/+ mice compared to control mice. In parallel, Platelet erythrocyte interaction seemed to be essential in the development of the bleeding phenotype. Conclusions: These findings show the role of chronically elevated IL-6 in driving the accumulation of A2m on the surface of erythrocytes, thereby mediating a phenotype of increased bleeding complications. This work was supported by the DFG KA4035/1-1 and by the German Ministry for Education and Research (BMBF 01EO1503) Disclosures Lämmle: Takeda: Membership on an entity's Board of Directors or advisory committees; Ablynx: Membership on an entity's Board of Directors or advisory committees, Other: Travel Support, Speakers Bureau; Baxter: Other: Travel Support, Speakers Bureau; Alexion: Other: Travel Support, Speakers Bureau; Siemens: Other: Travel Support, Speakers Bureau; Bayer: Other: Travel Support, Speakers Bureau; Roche: Other: Travel Support, Speakers Bureau; Sanofi: Other: Travel Support, Speakers Bureau. Ruf: ARCA bioscience: Consultancy, Patents & Royalties; ICONIC Therapeutics: Consultancy; MeruVasimmune: Current holder of individual stocks in a privately-held company.

GENDER-DEPENDENT HEMOSTASIS PROFILE IN APPARENTLY HEALTHY YOUNG PEOPLE

The objective of the study is to reveal the parameters of red blood cells, rheology, endothelin level, Willebrand factor, tPA-PAI-1 complex, antithrombin III, procoagulantive mechanisms of the hemostatic system and fibrinogen in medical students in the process of learning, taking into account gender differences. Materials and Methods. We examined 100 relatively healthy people (50 men and 50 women who did not have any complaints, were not under regular medical check-up, and did not have any proven diagnoses). The absence of diseases was confirmed by medical documentation, since all study participants underwent an annual preventive examination for admission to study. The volunteers were 18–30 years old. Results. Women had a higher endothelin level if compared with healthy men, which led to less powerful vasoconstriction and increased procoagulant blood activity. At the same time, higher levels of both procoagulant, anticoagulant and fibrinolytic blood activity were recorded. A significantly lower concentration of the tPA-PAI-1 complex and a change in the vWF index were also noted. In addition, relatively healthy women showed greater activity of VIIIa factor, higher resistance of Va factor to active protein C, and significantly higher fibrin content if compared with healthy men. Key words: sex differences, rheology, endothelin level, Willebrand factor, tPA-PAI-1 complex, AT III, procoagulantive mechanisms of the hemostatic system and fibrinogen. Цель. Выявить особенности показателей красной крови, реологии, уровня эндотелина, фактора Виллебранда, комплекса tPA-PAI-1, антитромбина III, прокоагулянтных механизмов системы гемостаза и фибриногена у студентов медицинских вузов с учетом половых различий в процессе обучения. Материалы и методы. Было обследовано 100 относительно здоровых (не предъявлявших жалоб, не состоявших на диспансерном учете и не имевших установленных диагнозов) человек (50 мужчин и 50 женщин). Отсутствие заболеваний подтверждалось медицинской документацией, так как все участники исследования проходили ежегодный профилактический осмотр для допуска к учебе. Возраст добровольцев составлял от 18 лет до 30 лет. Результаты. У женщин отмечался более высокий уровень эндотелина по сравнению со здоровыми мужчинами, что приводило к менее мощной вазоконстрикции и усилению прокоагулянтной активности крови. При этом были зарегистрированы более высокие значения как прокоагулянтной, так и противосвертывающей и фибринолитической активности крови. Одновременно с этим отмечена достоверно более низкая концентрация комплекса tPA-PAI-1, изменение показателя vWF. Кроме того, у относительно здоровых женщин была зафиксирована большая активность фактора VIIIа, более высокая резистентность фактора Vа к активному протеину С, а также достоверно более высокое содержание фибрина по сравнению со здоровыми мужчинами. Ключевые слова: половые различия, реология, уровень эндотелина, фактор Виллебранда, комплекс tPA-PAI-1, АТ III, прокоагулянтные механизмы системы гемостаза и фибриногена.

Anticoagulants and the Hemostatic System: A Primer for Occupational Stress Researchers

Anticoagulation, the body’s mechanism to prevent blood clotting, is an internal biomarker of an individual’s response to stress. Research has indicated that understanding the causes, processes, and consequences of anticoagulation can provide important insight into the experience of individuals facing emotional and occupational strain. Unfortunately, despite their importance, the mechanisms and implications of anticoagulation are unfamiliar to many researchers and practitioners working with trauma-exposed professionals. This paper provides an accessible primer on the topic of anticoagulation, including an overview of the biological process, the research connecting these processes with emotional and occupational functioning, as well as some potential methods for assessment.

Immunological and hemostasiological aspects of pathogenesis of obliterating atherosclerosis of arteries of the lower extremities

Objectives to study the changes in the immunological status and hemostatic system in patients with obliterating atherosclerosis of the lower limb arteries with the damage to the femoral-popliteal-tibial segment. Material and methods. A comprehensive study of the cytokine status and hemostatic system in patients with obliterating atherosclerosis of lower limb arteries with lesions of the femoral-popliteal tibia segment was carried out. Results. The progressive course of obliterating atherosclerosis is often associated with an increased content of cytokines (IL-1, IL-6, IL-8, TNF), as well as with the phenomena of hypercoagulation (increased platelet aggregation activity, shortened APTT, CT, TT, increased fibrinogen content), SFMCs, D-dimer). Conclusion. The study of the immunological status and hemostatic system should be performed in all patients with obliterating atherosclerosis of the lower limb arteries in order to adequately medically correct and prevent postoperative complications.

Hemostatic Biomarkers and Cancer Prognosis: Where Do We Stand?

Cancer patients are characterized by hypercoagulable state and an increased rate of thrombotic events, the most common being venous thromboembolism. Several hemostatic pathways that are significantly implicated in mechanisms of thromboembolic disease are also involved in growth, invasion, and metastatic spread of malignant cells as well in tumor-induced neo-angiogenesis. This close connection between cancer and the hemostatic system has prompted numerous studies on the role of alterations in the level plasma biomarkers of the different compartments of hemostasis in predicting cancer prognosis. In this review, we collect the results of several exemplificative studies that have evaluated clotting activation biomarkers in relation to different cancer outcomes with a final emphasis on current research and forthcoming directions in this field.

Blood Conservation and Hemostasis Management in Pediatric Cardiac Surgery

Pediatric cardiac surgery is associated with significant perioperative blood loss needing blood product transfusion. Transfusion carries serious risks and implications on clinical outcomes in this vulnerable population. The need for transfusion is higher in children and is attributed to several factors including immaturity of the hemostatic system, hemodilution from the CPB circuit, excessive activation of the hemostatic system, and preoperative anticoagulant drugs. Other patient characteristics such as smaller relative size of the patient, higher metabolic and oxygen requirements make successful blood transfusion management extremely challenging in this population and require meticulous planning and multidisciplinary teamwork. In this narrative review we aim to summarize risks and complications associated with blood transfusion in pediatric cardiac surgery and also to summarize perioperative coagulation management and blood conservation strategies.

CORRECTION OF DETOXIFICATION AND HEMOSTASIS-REGULATING LUNG ABILITY IN ACUTE PANCREATITIS

In recent years, the study of the functional state of the lungs has become a topical issue for urgent surgery. This is due to the fact that the lungs play an important role in the regulation of the inflammatory response, and in the body balance control. The aim of the study is to establish the detoxification and hemostasis-correcting lung ability in acute pancreatitis experimentally and clinically; to determine the effectiveness of quantum metabolic therapy in correcting lung functional status. Materials and Methods. The experiments were performed on adult dogs (n=30). The animals were divided into two groups: the first group (n=15) – a pancreatitis model, the second group (n=15) – a pancreatitis model with complex treatment, including quantum irradiation and remaxol. Forty patients with acute severe pancreatitis were examined clinically. They were also divided into two groups. Patients of the first group (n=20) were treated according to clinical guidelines, whereas patients of the second group (n=20) underwent complex treatment, including quantum irradiation and remaxol. The level of toxic products in arterial and venous blood and the state of the hemostatic system were determined. Results. Having analyzed the results of the study, the authors can claim that pancreas tissue injury in acute pancreatitis results in endogenous systemic inflammation, that is manifested in dysfunctional lung disorders, namely, a decrease in lung detoxification activity and impaired hemostasis-regulating ability. Complex treatment with quantum radiation and remaxol increases lung reserves, thus, resisting trigger damage agents. It is manifested in an increase in lung detoxification ability and less hemostasis defects. Conclusion. The inclusion of quantum radiation and remaxol in the complex therapy of acute pancreatitis is pathogenetically substantiated. Such a therapy decreases dysfunctional lung events, such as an increase in lung detoxification ability and a decrease in hemostasis defects. Keywords: acute pancreatitis, lungs, endotoxicosis, hemostasis. В последние годы изучение функционального состояния легких стало актуальным вопросом для ургентной хирургии. Это обусловлено тем, что легкие играют важную роль в регуляции воспалительного ответа, в поддержании гомеостатических констант. Цель исследования – в эксперименте и клинике при остром панкреатите установить детоксикационную и гемостазкорригирующую способность легких; определить эффективность квантово-метаболической терапии в коррекции их функционального статуса. Материалы и методы. Эксперименты поставлены на взрослых собаках (n=30). Две группы: первая (n=15) – модель панкреатита, вторая (n=15) – модель панкреатита, лечение комплексное, включающее квантовое облучение и ремаксол. Клинический материал – 40 пациентов с острым тяжелым панкреатитом. Первая группа (n=20) – лечение согласно клиническим рекомендациям, вторая группа (n=20) – лечение комплексное, включающее квантовое облучение и ремаксол. В артериальной и венозной крови определяли уровень токсических продуктов и состояние системы гемостаза. Результаты. Анализ результатов экспериментальной и клинической частей исследования дает основание утверждать, что поражение тканей поджелудочной железы при остром панкреатите приводит к явлениям эндогенного системного воспаления, проявлением которого являются дисфункциональные расстройства легких в виде снижения их детоксикационной активности и нарушения гемостазрегулирующей способности. Проведение комплексного лечения с включением квантового облучения и ремаксола увеличивает резервы легких для противостояния триггерным агентам повреждения, что проявляется повышением их детоксикационной способности и меньшими расстройствами в системе гемостаза. Выводы. Включение в комплексную терапию острого панкреатита квантового облучения и ремаксола патогенетически обосновано. При такого рода терапии дисфункциональные явления со стороны легких уменьшаются, частным проявлением является повышение их детоксикационной способности и уменьшение расстройств в системе гемостаза. Ключевые слова: острый панкреатит, легкие, эндотоксикоз, гемостаз.

Non-invasive imaging of gene expression and protein secretion dynamics in living mice

The liver is the largest organ and main source for secretory proteins with functions critical to health and disease. Tools to non-invasively study the fate of secretory proteins in vivo are scarce. Here we present a multimodal reporter mouse to query the expression and secretion dynamics of prothrombin, a prototypical liver-derived secretory protein. Using optical in vivo imaging, we confirm known modifiers of prothrombin expression and secretion. We discover extrahepatic prothrombin expression in multiple sites (including testes, placenta, brain, kidney, heart and lymphatic system) and in emerging tumors, resulting in significant amounts of tumor-derived prothrombin in the blood with procoagulant properties. Syngeneic cell lines from this mouse model enable unravelling regulatory mechanisms in high resolution, and in a scalable format ex vivo. Beyond discovering new functions in the hemostatic system, this model allows identifying rheostats in the cross-talk between gene expression and availability of a secretory protein. It is also a valuable resource for uncovering novel (tissue-specific) therapeutic vulnerabilities.

Effect of moderate physical exercises on the relationship of variability of the heart rhythm with the level of blood pressure and hemodynamic functions in women with essential hypertension

Цель исследования - изучение взаимосвязи между вариабельностью сердечного ритма (ВСР) системой гемостаза и гемодинамическими функциями у женщин больных эссенциальной гипертензией (ГБ) в зависимости от методов применяемой терапии. Методика. Под наблюдением находились 72 женщины, страдающие гипертонической болезнью. Обследовано 2 группы пациенток: 1-я группа (ГБ-1) находилась на медикаментозной терапии, 2-я (ГБ-2) - наряду с медикаментозной терапией регулярно на протяжении 2-3 лет проходила не менее 3 полуторамесячных курсов кинезитерапии. Для изучения гемодинамики был использован датчик динамического рассеяния света (miniature Dynamic Light Scattering - mDLS) от Elfi-Tech (Rehovot, Israel), измеряющий сигналы, инициированные кожным кровотоком и использующий методику разложения сигнала на частотные компоненты, связанные с разными гемодинамическими источниками. Из пульсовой компоненты mDLS сигнала извлекалась информация о вариабельности RR интервалов, и рассчитывались индикаторы вариабельности сердечного ритма. Изучали показатели свёртывающей системы крови: активированное парциальное тромбопластиновое время - (АПТВ), протромбиновое время (ПТВ) с вычислением МНО, тромбиновое время (ТВ), концентрация фибриногена, факторов II (протромбин), IIa (тромбин), IX и Х [10]. Кроме перечисленных методов исследования определялся пространственный рост фибринового сгустка, осуществляемый с помощью прибора «Регистратор Тромбодинамики Т-2». Достоинством способа является его объективность и то, что программой предусмотрена фоторегистрация роста сгустка через 5, 15 и 30 мин. Результаты. Как в группе ГБ-1, так и ГБ-2 выявлены многочисленные корреляционные связи между различными показателями ВСР, системы гемостаза и гемодинамических Данные представленные в виде матрицы свидетельствуют о том, что сдвиги в системе гемостаза и гемодинамики у больных ГБ-1, обусловленные деятельностью сердца и осуществляемые при участии как симпатического, так и парасимпатического отделов АНС способствуют возникновению тромботических осложнений. В то же время у больных ГБ-2 ВСР практически не коррелирует с показателями системы гемостаза и в меньшей степени связана с гемодинамические функции, благодаря чему состояние гемостаза и гемодинамики приближается к показателям здоровых женщин. Между тем, выявленные взаимосвязи между системой гемостаза и гемодинамическими функциями как у больных ГБ-1, так и ГБ-2 направлены на предотвращение внутрисосудистого свёртывания крови. Заключение. Применение систематической умеренной физической нагрузки на протяжении 2 - 3 лет способствует нормализации взаимоотношений между ВСР, системой гемостаза и гемодинамическими функциями у больных ГБ и способствует более устойчивой нормализации кровяного давления. Aim. To study the relationship between heart rate variability (HRV), hemostasis, and hemodynamic functions in women with essential hypertension (EH), depending on the method of therapy. Methods. 72 women with hypertension were monitored. Two patient groups were evaluated: the first group (EH-1) had recceived only drug therapy and the second group (EH-2), in addition to drug therapy, had regularly participated in at least three 1.5 mos long courses of kinesitherapy over a 2-3 years period. To study hemodynamics, we used a miniature Dynamic Light Scattering (mDLS) sensor from Elfi-Tech (Rehovot, Israel), which measures signals initiated by skin blood flow and uses a technique for decomposing the signal into frequency components associated with different hemodynamic sources. Information on the variability of RR intervals was extracted from the pulse component of the mDLS signal, and parameters of heart rate variability were calculated. Indexes of the coagulation system were studied, including activated partial thromboplastin time (APTT), prothrombin time (PTT) with calculation of INR, thrombin time (TT), concentrations of fibrinogen and factors II (prothrombin), IIa (thrombin), IX, and X [10]. In addition, spatial fibrin clot growth was determined with a Thrombodynamics Registrator T-2 apparatus. The method benefits are its objectivity and a possibility of photorecording of the clot growth at 5, 15 and 30 min. Results. Both in the EH-1 and EH-2 groups, numerous correlations were detected between various parameters of HRV, the hemostatic system, and hemodynamic functions, as well as between the parameters of the hemostatic system and hemodynamic functions. The data are presented in the form of a matrix. The data indicate that shifts in the hemostatic and hemodynamic systems of EH-1 patients induced by cardiac activity and resulting from activities of both the sympathetic and parasympathetic sections of the autonomic nervous system (ANS) are aimed at enhancing the hemostatic properties of blood, and this contributes to thrombotic complications. At the same time, in EH-2 patients, HRV had practically no effect on the hemostatic system and to a lesser extent was related with the hemodynamic function. Due to this, the state of hemostasis and hemodynamics in EH-2 patients approaches the state found in healthy women. In addition, the relationship between the hemostatic system and hemodynamic function in both EH-1 and EH-2 patients are aimed at preventing intravascular coagulation. Conclusion. The use of systematic, moderate physical activity for 2 to 3 years helps hypertensive patients to normalize the relationship between HRV, the hemostatic system, and hemodynamic functions. This contributes to a more normal and stable blood pressure.