Condensation of 4-hydroxy-2-thiazolines with 1,2-phenylenediamine as a novel effective route to thiazolo[3,4-a]quinoxalines

Thiazolo[3,4-a]quinoxalin-4-ones were prepared in two steps starting from methyl phenylchloropyruvate using a new strategy for the construction of the ring system. A key step in this new method involves the reaction of 4-hydroxytetrahydrothiazoles with 1,2-phenylendiamines

Purinergic Profiling of Regulatory T-cells in Patients With Episodic Migraine

Objectives: Immune responses in migraine are poorly characterized, yet implicated in the disease pathogenesis. This study was carried out to characterize purinergic profiles of T-cells in patients with episodic migraine without aura (MWoA) to provide mechanistic evidence for ATP and adenosine involvement in modulation of immune regulation in migraine.Methods: Peripheral blood samples were obtained from patients with migraine (n = 16) and age-matched control subjects (n = 21). Subsets of T-cells were identified by flow cytometry based on specific membrane markers.Results: Migraine patients showed reduced total T-cell counts in the peripheral blood. Whereas the total number of CD3+CD4+, CD3+CD8+, or regulatory T lymphocytes (Treg) was not changed, the proportion of Treg CD45R0+CD62L– and CD45R0–CD62L– cells was increased. Interestingly, in migraine, less Treg cells expressed CD39 and CD73 suggesting disrupted ATP breakdown to adenosine. The negative correlations were observed between the duration of migraine and the relative number of CD73+CD39– Tregs and total number of CD73-positive CD45R0+CD62L+ Tregs.Conclusion: Obtained data indicate that T-cell populations are altered in episodic migraine and suggest the involvement of Tregs in the pathophysiology of this disorder. Reduced expression of CD39 and CD73 suggests promotion of ATP-dependent pro-inflammatory and reduction of adenosine-mediated anti-inflammatory mechanisms in migraine

Activation of P2X7 Receptors in Peritoneal and Meningeal Mast Cells Detected by Uptake of Organic Dyes: Possible Purinergic Triggers of Neuroinflammation in Meninges

Extracellular ATP activates inflammasome and triggers the release of multiple cytokines in various immune cells, a process primarily mediated by P2X7 receptors. However, the expression and functional properties of P2X7 receptors in native mast cells in tissues such as meninges where migraine pain originates from have not been explored. Here we report a novel model of murine cultured meningeal mast cells and using these, as well as easily accessible peritoneal mast cells, studied the mechanisms of ATP-mediated mast cell activation. We show that ATP induced a time and dose-dependent activation of peritoneal mast cells as analyzed by the uptake of organic dye YO-PRO1 as well as 4,6-diamidino-2-phenylindole (DAPI). Both YO-PRO1 and DAPI uptake in mast cells was mediated by the P2X7 subtype of ATP receptors as demonstrated by the inhibitory effect of P2X7 antagonist A839977. Consistent with this, significant YO-PRO1 uptake was promoted by the P2X7 agonist 2',3'-O-(benzoyl-4-benzoyl)-ATP (BzATP). Extracellular ATP-induced degranulation of native and cultured meningeal mast cells was shown with Toluidine Blue staining. Taken together, these data demonstrate the important contribution of P2X7 receptors to ATP-driven activation of mast cells, suggesting these purinergic mechanisms as potential triggers of neuroinflammation and pain sensitization in migraine.Peer reviewe

Activation of P2X7 Receptors in Peritoneal and Meningeal Mast Cells Detected by Uptake of Organic Dyes: Possible Purinergic Triggers of Neuroinflammation in Meninges

Extracellular ATP activates inflammasome and triggers the release of multiple cytokines in various immune cells, a process primarily mediated by P2X7 receptors. However, the expression and functional properties of P2X7 receptors in native mast cells in tissues such as meninges where migraine pain originates from have not been explored. Here we report a novel model of murine cultured meningeal mast cells and using these, as well as easily accessible peritoneal mast cells, studied the mechanisms of ATP-mediated mast cell activation. We show that ATP induced a time and dose-dependent activation of peritoneal mast cells as analyzed by the uptake of organic dye YO-PRO1 as well as 4,6-diamidino-2-phenylindole (DAPI). Both YO-PRO1 and DAPI uptake in mast cells was mediated by the P2X7 subtype of ATP receptors as demonstrated by the inhibitory effect of P2X7 antagonist A839977. Consistent with this, significant YO-PRO1 uptake was promoted by the P2X7 agonist 2′,3′-O-(benzoyl-4-benzoyl)-ATP (BzATP). Extracellular ATP-induced degranulation of native and cultured meningeal mast cells was shown with Toluidine Blue staining. Taken together, these data demonstrate the important contribution of P2X7 receptors to ATP-driven activation of mast cells, suggesting these purinergic mechanisms as potential triggers of neuroinflammation and pain sensitization in migraine

Purinergic profiling of regulatory T-cells in patients with episodic migraine

© 2018 Nurkhametova, Kudryavtsev, Khayrutdinova, Serebryakova, Altunbaev, Malm and Giniatullin. Objectives: Immune responses in migraine are poorly characterized, yet implicated in the disease pathogenesis. This study was carried out to characterize purinergic profiles of T-cells in patients with episodic migraine without aura (MWoA) to provide mechanistic evidence for ATP and adenosine involvement in modulation of immune regulation in migraine. Methods: Peripheral blood samples were obtained from patients with migraine (n = 16) and age-matched control subjects (n = 21). Subsets of T-cells were identified by flow cytometry based on specific membrane markers. Results: Migraine patients showed reduced total T-cell counts in the peripheral blood. Whereas the total number of CD3+CD4+, CD3+CD8+, or regulatory T lymphocytes (Treg) was not changed, the proportion of Treg CD45R0+CD62L– and CD45R0–CD62L– cells was increased. Interestingly, in migraine, less Treg cells expressed CD39 and CD73 suggesting disrupted ATP breakdown to adenosine. The negative correlations were observed between the duration of migraine and the relative number of CD73+CD39– Tregs and total number of CD73-positive CD45R0+CD62L+ Tregs. Conclusion: Obtained data indicate that T-cell populations are altered in episodic migraine and suggest the involvement of Tregs in the pathophysiology of this disorder. Reduced expression of CD39 and CD73 suggests promotion of ATP-dependent pro-inflammatory and reduction of adenosine-mediated anti-inflammatory mechanisms in migraine

Novel capsaicin-induced parameters of microcirculation in migraine patients revealed by imaging photoplethysmography

© 2018, The Author(s). Background: The non-invasive biomarkers of migraine can help to develop the personalized medication of this disorder. In testing of the antimigraine drugs the capsaicin-induced skin redness with activated TRPV1 receptors in sensory neurons associated with the release of the migraine mediator CGRP has already been widely used. Methods: Fourteen migraine patients (mean age 34.6 ± 10.2 years) and 14 healthy volunteers (mean age 29.9 ± 9.7 years) participated in the experiment. A new arrangement of imaging photoplethysmography recently developed by us was used here to discover novel sensitive parameters of dermal blood flow during capsaicin applications in migraine patients. Results: Blood pulsation amplitude (BPA) observed as optical-intensity waveform varying synchronously with heartbeat was used for detailed exploration of microcirculatory perfusion induced by capsicum patch application. The BPA signals, once having appeared after certain latent period, were progressively rising until being saturated. Capsaicin-induced high BPA areas were distributed unevenly under the patch, forming “hot spots.” Interestingly the hot spots were much more variable in migraine patients than in the control group. In contrast to BPA, a slow component of waveforms related to the skin redness changed significantly less than BPA highlighting the latter parameter as the potential sensitive biomarker of capsaicin-induced activation of the blood flow. Thus, in migraine patients, there is a non-uniform (both in space and in time) reaction to capsaicin, resulting in highly variable openings of skin capillaries. Conclusion: BPA dynamics measured by imaging photoplethysmography could serve as a novel sensitive non-invasive biomarker of migraine-associated changes in microcirculation

Риск развития кровотечений, требующих гемотрансфузии, после транскатетерной имплантации аортального клапана у пациентов с сопутствующей ишемической болезнью сердца

Introduction. Perioperative bleeding is a potentially life-threatening complication of surgical intervention. The incidence of perioperative bleeding during transcatheter aortic valve implantation (TAVI) reaches 6%. An increased risk of perioperative bleeding is noted in patients requiring constant antiplatelet therapy, which includes patients with concomitant coronary artery disease (CAD).Aim. The study of risk factors of bleeding, which require blood transfusion in candidates for TAVI with concomitant CAD.Materials and methods. A retrospective study. The patients with concomitant CAD who underwent TAVI from 2016 to 2021 with hemodynamically significant coronary artery stenosis (n = 270) were included in this study. The incidence of early postoperative bleeding requiring blood transfusions was analyzed. Risk factors of bleeding development were evaluated.Results. The average age of the patients was 77.7 ± 7.2 years, the number of male and female patients was comparable (45.9 and 54.1%).The majority of patients were diagnosed with chronic heart failure (90.4%), more than half of the patients suffered from pulmonary hypertension (51.9%). The incidence of bleeding that required blood transfusion was 9.3%. The risk factors of the bleeding were dual antiplatelet therapy, baseline anemia (hemoglobin less than 120 g/l), history of stroke, chronic renal failure and critical aortic stenosis. The risk factors were included in the predictive model.Conclusion. Bleeding requiring blood transfusion in patients with severe aortic stenosis and CAD after TAVR occurs in 9.3%. It can be partially predicted using the predictive model. The using of the predictive model may be useful in determining the predominant risk of thrombotic or bleeding events after surgery.Введение. Периоперационное кровотечение является потенциально жизнеугрожающим осложнением хирургического вмешательства.Встречаемость периоперационных кровотечений при транскатетерной имплантации аортального клапана (ТИАК) достигает 6%. Повышенный риск развития периоперационных кровотечений отмечается у пациентов, требующих постоянной антиагрегантной терапии, к которой относятся пациенты с сопутствующей ишемической болезнью сердца (ИБС).Цель. Изучить факторы риска развития кровотечений, требующих гемотрансфузии, у пациентов, госпитализированных для проведения ТИАК, при сопутствующей ИБС.Материалы и методы. Проведено ретроспективное исследование. Рассмотрены пациенты, перенесшие ТИАК в период с 2016 по 2021 г., при сопутствующей ИБС с гемодинамически значимым поражением коронарных артерий (n = 270). Изучена частота развития ранних послеоперационных кровотечений, требующих проведения гемотрансфузий. Выполнена оценка факторов риска развития кровотечений.Результаты. Средний возраст исследуемых составил 77,7 ± 7,2 года, количество пациентов мужского и женского пола было сопоставимо – 45,9 и 54,1%. У большинства пациентов была диагностирована хроническая сердечная недостаточность (90,4%), у половины – легочная гипертензия (51,9%). Частота развития кровотечений, потребовавших проведения гемотрансфузии, составила 9,3%. Повышение риска развития кровотечений, требующих гемотрансфузии, ассоциировано с наличием следующих факторов риска: двойная антиагрегантная терапия, исходная анемия (гемоглобин менее 120 г/л), инсульт в анамнезе, хроническая почечная недостаточность и критический аортальный стеноз. Полученные факторы риска включены в прогностическую модель.Заключение. У 9,3% пациентов с тяжелым аортальным стенозом и сопутствующей ИБС при проведении ТИАК развиваются кровотечения, требующие гемотрансфузии. Развитие кровотечения можно частично спрогнозировать с помощью предложенной в исследовании модели. Ее использование в клинической практике может быть полезным для определения превалирующего риска тромботических или геморрагических событий в послеоперационном периоде

Mast Cell Mediators as Pain Triggers in Migraine: Comparison of Histamine and Serotonin in the Activation of Primary Afferents in the Meninges in Rats

© 2020, Springer Science+Business Media, LLC, part of Springer Nature. The meninges around the brain are characterized by an abundant blood supply, a high density of sensory nerves, and large numbers of mast cells. In migraine, the commonest neurological disorder, activation of trigeminal nerve fibers in the meninges is the initial trigger mechanism for generating pain signals. The recently suggested concept of the neuroimmune synapse suggests that mast cell transmitters can activate receptor proteins in close-lying nerve endings, leading to the generation of nociceptive spike activity. Serotonin and histamine, presumptive triggers for migraine, are classical transmitters released on activation of mast cells. Our recent research has identifi ed powerful activation of primary afferents by serotonin, mediated mainly via 5-HT3 receptors. However, the role of histamine in meningeal neuroimmune synapses has received little study. The present study therefore used recording of spike activity from primary afferents in the meninges in rats to study the role of histamine as a possible trigger for pain in migraine. Results from testing a wide range of histamine concentrations identified only a minimal (about 12%) effect with 10 μM histamine on the nociceptive activity of the trigeminal nerve. More detailed cluster analysis showed that the proportion of fibers reacting to histamine was no more than 29%, increases in spike activity in these fibers being significantly lower than on exposure to serotonin. Longer (4 h) exposure to histamine also produces no significant change in trigeminal nerve activity. The results do not exclude a stimulatory role for histamine in migraine but suggest that this mast cell transmitter has an action other than activation of the trigeminal nerve

Purinergic profiling of regulatory T-cells in patients with episodic migraine

© 2018 Nurkhametova, Kudryavtsev, Khayrutdinova, Serebryakova, Altunbaev, Malm and Giniatullin. Objectives: Immune responses in migraine are poorly characterized, yet implicated in the disease pathogenesis. This study was carried out to characterize purinergic profiles of T-cells in patients with episodic migraine without aura (MWoA) to provide mechanistic evidence for ATP and adenosine involvement in modulation of immune regulation in migraine. Methods: Peripheral blood samples were obtained from patients with migraine (n = 16) and age-matched control subjects (n = 21). Subsets of T-cells were identified by flow cytometry based on specific membrane markers. Results: Migraine patients showed reduced total T-cell counts in the peripheral blood. Whereas the total number of CD3+CD4+, CD3+CD8+, or regulatory T lymphocytes (Treg) was not changed, the proportion of Treg CD45R0+CD62L– and CD45R0–CD62L– cells was increased. Interestingly, in migraine, less Treg cells expressed CD39 and CD73 suggesting disrupted ATP breakdown to adenosine. The negative correlations were observed between the duration of migraine and the relative number of CD73+CD39– Tregs and total number of CD73-positive CD45R0+CD62L+ Tregs. Conclusion: Obtained data indicate that T-cell populations are altered in episodic migraine and suggest the involvement of Tregs in the pathophysiology of this disorder. Reduced expression of CD39 and CD73 suggests promotion of ATP-dependent pro-inflammatory and reduction of adenosine-mediated anti-inflammatory mechanisms in migraine