Introduction. The state of the blood flow within the capillaries and close blood vessels is highly important in practice for the revealing of pathogenetic mechanisms of both systemic and local circulatory disorders. Aim of the study was to define the parameters of microcirculation and the level of blood flow fluctuations (flux) in the distal segments of upper and lower limbs (in fingers of hands and toes of feet) in children of 6–7 years old; and to describe the possible differences in the mechanisms of blood flow modulation in boys and girls. Materials and methods. Skin microcirculation was assessed in middle fingers of hands and great toes of feet in children of 6-7 years old (14 girls and 7 boys in prone position) by means of laser doppler flowmetry. Results. The ranges for parameters of microcirculation (PM) for distal segments of upper and lower limbs in children of mentioned age group were defined, also it was shown that the PM are significantly lower in the lower limbs comparing to those of the upper limbs (both in groups of girls and boys). Asymmetry of PM in the feet was not found; the features of right hand-left hand asymmetry for PM in girls and boys are described. The analysis of modulation of blood flow fluctuations (fluxmotions) of different frequencies showed the profound role of vasomotor (myogenic) rhythm for regulation of microcirculation. Conclusion. Increased neurogenic influences on the modulation of fluxmotions in girls of 6-7 years old may be an evidence of the ongoing development of the mechanisms of blood flow regulation, particularly the association with the growth rate of girls is possible.

BackgroundAutoinflammatory component rather than autoimmune one is to be leading mechanism in the pathogenesis of the systemic juvenile idipathic arthritis (sJIA). Macrophage activation sydrome (MAS) ifatal complication of sJIA also is a result of a hyperactivation of monocytes/macrophages [1, 2]. Given those data, macrophage (M1 and M2) reprogramming becomes a very promising strategy to achieve remission of the disease. Existing animal models of arthritis (particularly collagen-induced arthritis) are not adequate for experimental implementation of this approach, because a key element of their pathogenesis is the formation of autoaggressive antibodies. For a long time, adjuvant arthritis in rats was meant to be the most close (but still not optimal) animal model to investigate the pathogenesis of rheumatoid arthritis and juvenile idiopathic arthritis [3].ObjectivesTo create the model of arthritis with the typical systemic manifestations caused by the use of a macrophage M1 phenotype differentiation stimulus in Wistar rats.MethodsDay 0: Rats (8 male Wistar rats, 8 month old) were injected intracutaneously (right footpads) with 0,1 ml of complete Freund's adjuvant (CFA, 5 mg/ml, DIFCO LABORATORIES, Michigan, USA) and injected intraperitoneally (i.p.) with 5 μg/kg body weight of lipopolysaccharide (LPS, 10 μg/ml, Medgamal, Russia). In control groups equivalent volumes of physiological 0.9% sodium chloride solution were used. Day 18: rats were reinjected i.p. with LPS (10 μg/kg body weight). Day 39: rats were reinjected intracutaneously (left footpads) with 0,1 ml of CFA and reinjected i.p. with LPS (10 μg per animal).ResultsThe temperature over the hind paw joints was found to significantly increase (p<0,05) for at least 5 days in the group of LPS+CFA (main group) in comparison with the group of CFA only. This effect was confirmed to depend on the degree of inflammation in the joints as significant differences (p=0,019) were observed between left and right hind paws in the main group on the Day 42.Metatarsal joint diameters were increased significantly (p<0,05) for at least 5 days (after Day 39) in the main group and remained increased (non-significantly) until the end of the study.Pain assessment was performed by the method [4, 5], the severity of local changes in the region of inflamed joints [4]. More pronounced pain and arthritis symptoms after Day 39ConclusionsOur novel animal model of arthritis (combined stimulation by M1 stimulus LPS and CFA) is cheap and easy to reproduce. The model allows to investigate systemic mechanisms of arthritis that pathogenetically are close to the systemic manifestations of arthritis in humans.ReferencesSikora K.A., Grom A.A. Curr Opin Pediatr. 2011; 23 (6): 640–646Paediatric rheumatology ATLAS. Editors: A. A. Baranov, E. I. Alekseeva, Gedike. - Reed Elsevier. Moscow.-2010.-P.22–42.Vercoulen Y. et al. Arthritis Research & Therapy. 2009; 11: 231Darvish S.F. et al. PLoS One. 2013; 8 (11): e79284Bush K.A. et al. Rheumatology (Oxford). 2001; 40 (9): 1013–1021Disclosure of InterestNone ...

Roles of Different Macrophage Phenotypes in the Pathogenesis of Some Human Diseases Macrophages have recently been shown to play a key role in promoting of recovery after some diseases as well as in aggravation of inflammatory responses, all the functions being resulted from microenvironmental conditions and therefore phenotypes acquired by macrophages in these conditions. In this article some protective functions of macrophages during infectious and oncologic diseases as well as pathogenic roles in a number of inflammatory diseases are reviewed. Much attention is devoted to opportunities of macrophage reprogramming.

BackgroundThe systemic juvenile idiopathic arthritis (sJIA) is a problem with high social significance all over the world. Targeted cell reprogramming becomes one of the most important lines for modern medicine and can be regarded as an origin for new highly improved treatment strategy of sJIA. As rat monocytes exhibited CD163 expression in a similar level with that shown for peritoneal macrophages [1], we decided to use CD163 as a key factor indicating cell reprogramming in the study.ObjectivesTo investigate the dynamical changes in subpopulations of peripheral blood mononuclear cells (PBMC) and to assess doxycycline and dexamethasone effects in a model of arthritis with the systemic manifestations.MethodsAnimal model [2] was adapted in 24 Wistar rats (males, 6 month old). On the day of the last stimulation all the rats were divided into 3 equal groups and additional subcutaneous (s.c.) injections were performed as follows: DOXY-group – doxycycline (50 mg/kg, Saratov, Russia), DEXA-group – dexamethasone (4 mg/kg, KRKA, Slovenia), control group – 0,9% sodium chloride solution (Belarus). The s.c. injections were repeated on Day 54. Time points were 0, 21, 41, and 55 Days. PBMC were assessed by flow cytometry (BD FACSCanto II, USA) according to manufacturer's instructions. Staining was performed with FITC Anti-Rat CD11b (BD Pharmingen), anti-rat CD68 RPE (Serotec, UK), anti-rat CD163 ALEXA FLUOR 647 (Serotec, UK). CD11b+CD68+ and CD11b-CD68+ cells were regarded as monocytes and circulating dendritic cells consequently. At the termination animal organ masses were measured.ResultsUp to Day 55 proportions of CD163+ in CD11b+CD68+ population changed synchronically in all groups. On Day 55 the proportions (in comparison with the data of Day 41) were significantly higher in DEXA-group (p<0,05) but didn't change in DOXY- and control groups (p>0,05). Cell reprogramming was also observed in population of CD11b-CD68+. So, on Day 41 the proportions of CD163+ cells in CD11b-CD68+ population were significantly increased (in comparison with the data of Days 0 and 21) in DOXY- and DEXA-groups (p<0,05) but not in control group (p>0,05). Mentioned changes were subsequent with other parameters of inflammation. We observed significantly lower heart masses in DOXY- and DEXA-groups (median=0,63 and 0,635g consequently) in comparison with control group (median=0,74g) (p<0,05), but no difference between DOXY- and DEXA- groups (p>0,05).ConclusionsIn a model of arthritis with the systemic manifestations in Wistar rats we demonstrated that subpopulations of PBMC (CD11b-CD68+ and CD11b+CD68+) underwent reprogramming. Doxycycline and dexamethasone modyfied the dynamics of the reprogramming. In DOXY- and DEXA-groups there were lower heart masses than in the control group, the last fact is subsequent with the data by De P. et al. [3]. We can speculate that monocytes and dendritic cells undergo reprogramming (CD163neg and CD163+) in a similar way with M1 and M2 macrophages.References Moghaddami M, Cleland LG, Mayrhofer G. Int Immunol. 2005; 17(8)...

Цель исследования - изучение перепрограммирования мононуклеарных лейкоцитов на модели системного ювенильного идиопатического артрита (сЮИА), воспроизводимой у крыс Wistar с использованием полного адъюванта Фрейнда и липополисахарида. Методика. сЮИА воспроизведен у 6-месячных крыс-самцов Wistar. На 40-е сут. эксперимента животные были разделены на 3 группы: 1-я группа - контроль; 2-я - группа доксициклина; 3-я - группа дексаметазона. Взятие проб крови у животных проводили на нулевые, 41-е и 55-е сут. Мононуклеарные клетки периферической крови выделяли гравиметрически, после чего окрашивали их на маркеры и внутриклеточные цитокины. Дифференцировали моноциты (CD3-CD4+) и Т-хелперы (CD3+CD4+). Анализировали динамику внутриклеточной экспрессии интерлейкина IL-4 (рассматривали как маркер про-М2 фенотипа, так как в случае выделения из клетки ИЛ-4 служит стимулятором М2 поляризации макрофагов) и IFN-g (как маркер про-М1 фенотипа) по данным проточной цитофлуориметрии. Применяли непараметрический статистический тест Mann-Whitney-Wilcoxon в программе R для статистической обработки данных. Результаты и заключение. При моделировании сЮИА выявлено закономерное изменение фенотипа моноцитов. Применение же доксициклина и дексаметазона приводило к более ранней поляризации их по про-М2-пути в отношении моноцитов (на 41-е сут.) в сравнении с контролем. Про-М1 эффект (на 55-е сут., в сравнении с контролем) выявлен также в группах доксициклина и дексаметазона. У животных разных групп обнаружены характерные динамические изменения внутриклеточной экспрессии цитокинов. Важно, что различная направленность поляризации фенотипа при сЮИА и применении препаратов наблюдается не только у моноцитов, но и у Т-хелперов. The study objective was to evaluate targeted reprogramming of peripheral blood mononuclear cells in systemic juvenile idiopathic arthritis (sJIA) modeled in 6-month-old male Wistar rats by co-administration of complete Freund’s adjuvant and lipopolysaccharide. Methods. On day 40 of the experiment, rats were divided into three groups: control, doxycycline, and dexamethasone groups. Blood samples were collected on days 0, 41, and 55. Peripheral blood mononuclear cells were isolated gravimetrically and stained for markers and cytokines. Monocytes (CD3-CD4+) and T-helpers (CD3+CD4+) were differentiated as target cells. IL-4 was considered a marker for the pro-M2 phenotype since IL-4 can activate M2 macrophage polarization; IFN-g was considered a marker for the pro-M1 phenotype. Time-related changes in the intracellular expression of IL-4 and IFN-g were studied using flow cytometry. Data were analyzed using nonparametric statistical tests (Mann-Whitney-Wilcoxon) in the R environment for statistical computing. Results and conclusions. Monocytes (like macrophages) underwent reprogramming during the development of modeled sJIA disease. In monocytes of doxycycline and dexamethasone treatment groups, pro-M2 effects were observed earlier (day 41) than in the control group. Pro-M1 effects were observed in monocytes of doxycycline and dexamethasone groups on day 55, as compared with the control group. Characteristic time-related changes of intracellular cytokine expression were described for different groups. Importantly, the differently directed phenotype polarization was observed in sJIA and treatment groups for both monocytes and T-helpers.