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Rearrangement of the immune system during pregnancy is a strictly controlled, dynamic process in which the first and third trimesters are, respectively, pro-inflammatory, and anti-inflammatory periods. However, monocyte involvement in regulating the pro/anti-inflammatory balance remains poorly understood. The functional phenotype of monocytes is known to depend on their subsets assessed by CD14 and CD16 expression, and is associated with expression of M1(CCR2)- and M2(CD206) molecules, associated with pro- and anti-inflammatory activity, respectively. Here we have investigated the expression of CCR2 and CD206 in classical (CD14++CD16- , cMo), intermediate (CD14++CD16+, iMo), and non-classical monocytes (CD14+CD16++, nMo) in pregnant women at different gestational ages in comparison with nonpregnant women. The study included 14 pregnant women in the first trimester, 20 in the second trimester, 26 in the third trimester, and 29 fertile non-pregnant women. One-way analysis of variance in these groups revealed significant differences CCR2 and CD206 expression (more pronounced in classical and intermediate monocytes and stronger in relation to CD206 expression). Overall, monocytes from pregnant women had decreased CCR2- and increased CD206 expression, suggesting a shift towards an anti-inflammatory profile. These changes appeared in the first trimester (increased CD206 mean fluorescence intensity [MFI] in cMo and iMo, p < 0.05) and reached their maximum in the second trimester, manifested by significant increase in CD206 and decrease in CCR2 expression (% of cells, MFI) in all monocyte subsets. In the third trimester, CD206+ cMo decreased, as compared to the second trimester (p < 0.05), and the percentage of CCR2+ cMo and iMo increased. Of note, these changes in the first and third trimesters were combined with increased pro-inflammatory expression profile of non-classical monocytes which was restricted by the non-classical monocyte subpopulation in the first trimester, then being mediated by intermediate and non-classical monocytes in the third trimester. The data obtained suggest involvement of monocytes in regulation of the pro- and anti-inflammatory balance during pregnancy, with predominant development of the M2 profile in classical monocytes during the first and third trimesters, and in all monocyte subsets over second trimester, along with increase in the M1 proinflammatory profile of intermediate and non-classical monocytes in the first and third trimesters.

Objective: to summarize the available data on the features of vascularization of endometrioid adenocarcinoma (EAC). Material and Methods. The search for relevant sources was performed in the “PubMed” database using the keywords “endometrium + cancer + angiogenesis”, “endometrium + cancer + angiogenesis + lymph”. Of the selected sources, 78 were included in this review. Results. Angiogenesis is an important and necessary stage in the pathogenesis of the appearance, progression and metastasis of EAC and, thus, the study of tumor vascularization provides an opportunity to improve diagnosis and personalized approach to treatment. Vascular density correlates with advanced stage of EAC, high grade of malignancy, myometrial invasion, cervical and adnexal lesions, vascular invasion, metastases to lymph nodes (LN), the presence of cancer cells in the peritoneal fluid, low overall survival and survival without tumor progression. There are publications that deny the connection of vascularization with the histological type of tumor, its grade, lymphovascular invasion, lymph node metastases, the depth of myometrial invasion, and these publications even prove that microvessel density is not an independent prognostic factor. So, there is still no consensus and final opinion, as evidenced by low or high vascularization of EAC. Recently, there are many drugs that affect both the processes of angiogenesis directly and the inducers and factors that control vascular growth. Unfortunately, all such drugs have a fairly high toxicity, and resistance to them very quickly develops. Conclusion. Despite numerous results of studies devoted to the study of the formation of blood vessels and isolated data on lymphangiogenesis in EAC, there is no data in the literature on studying changes in the vascularization of LN in gynecological cancer. However, proangiogenic and antiangiogenic factors are disseminated throughout the body and must exert their effects in distant organs and tissues. Based on changes in the vascularization of LN, it will apparently become possible to predict the activity of angiogenesis in the primary tumor, assess the prognosis of the disease, and the effectiveness of the treatment. In addition, significant expression of the vascular network in an enlarged lymph node biopsied for diagnosis may be a symptom of the development of a malignant tumor in the lymph collection region, even in the absence of metastases.

Photosynthetic water oxidation by Photosystem II (PSII) is a fascinating process because it sustains life on Earth and serves as a blue print for scalable synthetic catalysts required for renewable energy applications. The biophysical, computational, and structural description of this process, which started more than 50 years ago, has made tremendous progress over the past two decades, with its high-resolution crystal structures being available not only of the dark-stable state of PSII, but of all the semi-stable reaction intermediates and even some transient states. Here, we summarize the current knowledge on PSII with emphasis on the basic principles that govern the conversion of light energy to chemical energy in PSII, as well as on the illustration of the molecular structures that enable these reactions. The important remaining questions regarding the mechanism of biological water oxidation are highlighted, and one possible pathway for this fundamental reaction is described at a molecular level.

Background. New implantation methods are of great importance due to the development of endoprostheses in traumatology and orthopedics, restorative medicine and dentistry. Equally important is the early detection and description of the implant-associated complications. The aim of the study is to find and describe thrombi and emboli in the heart and lungs formed after experimental implantation of metallic devices in the peripheral part of limb using extracellular vesicles of mesenchymal stromal cells. Methods. Outbred rabbits of both genders at the age from 4 to 6 months and of weight from 3 to 4 kg underwent experimental implantation. The study enrolled 57 species in total. They were divided into two groups: 30 animals underwent implantation of metallic devices using extracellular vesicles of mesenchymal stromal cells (EV MSCs), 27 — without their use. The rabbits’ hearts and lungs were studied by light microscopy methods at different stages after integration of screw titanium implants into the proximal condyle of the tibia using EV MSCs. Results. After implantation of metallic devices into the proximal condyle of the tibia, we detected fibrin, detritus and even the red bone marrow structures (various blast forms of hematopoietic cells: megakaryocytes, cells of the erythroid and myeloid lineages) in the right cavities of the heart. In the pulmonary arteries, we also found thrombi and emboli, which either led to the obliteration of the involved vessel or to gradual lysis, not disappearing completely within 10 days of follow-up. Conclusions. After intraosseous implantation of the metallic devices, there is an embolism risk in the right atria and ventricle of the heart and the pulmonary arteries and veins due to the debris migration with the bloodstream from the surgery site. At the same time, one cannot exclude a thrombotic risk in the heart and pulmonary arteries as a reaction to the presence of detritus. It is advisable to take measures aimed at preventing both debris releasing into the bloodstream and pulmonary embolism during any implantations into the bone tissues, even of relatively small devices. Using EV MSCs to affect the implant engraftment processes has no significant effect on the severity and frequency of thromboembolic complications.

Macrophages are key regulatory cells of fibrogenesis. They can have pro- or antifibrotic activity due to their plasticity and heterogeneity. Some studies have shown the antifibrotic effect of macrophages on dermal fibroblasts, but the effect of macrophages on the lung fibroblast functions remains unexplored. Therefore, the purpose of this study was to examine the influence of conditioned media of human macrophage differentiated by M-CSF/GM-CSF and further dexamethasone polarized/unpolarized on the TGF-β-induced lung fibroblast differentiation. Macrophages were derived from peripheral blood monocytes of healthy donors. Monocytes were differentiated by M-CSF or GM-CSF for 7 days. On day 5, dexamethasone was added to generation of polarized macrophages M-M(Dex) and GM-M(Dex). Polarized macrophages were compared with non-polarized M-M0 and GM-M0, to which dexamethasone was not added. Next, the conditioned medium of these macrophage subtypes was collected and tested for inhibition the lung fibroblast differentiation (HLF210 cell line). To do this, TGF-β (inducing differentiation factor) and conditioned macrophage medium were added to fibroblast cultures. Effectiveness of differentiation was estimated by the expression of the myofibroblast marker, α-smooth muscle actin (α-SMA), and the production of extracellular matrix protein, collagen I. The expression of α-SMA was determined using flow cytometry. The concentration of collagen I was measured by ELISA. Since our data indicates that spontaneous activation of fibroblasts occurs during standard cultivation, the α-SMA expression was investigated in 3D culture of fibroblasts. Notably, the content of α-SMA-positive cells in 3D cultures was significantly reduced, indicating more physiological growth cells. Regardless of the differentiation stimulus, the conditioned media of dexamethasone-polarized macrophages do not affect the level of collagen I production or the α-SMA expression. On the contrary, M-M0 showed a strong inhibitory effect that reduced the amount of collagen I in the fibroblast cultures and the expression of marker myofibroblasts by fibroblasts. Interesting, GM-M0 had no such effect and did not prevent lung fibroblast differentiation like polarized cells. Taken together, the findings suggest that M-M0 macrophages may have antifibrotic properties. Furthermore, the lack of this effect in GM-M0 macrophages indicates that the differentiation factor plays a significant role in the development of the antifibrotic macrophage phenotype.

Varicose veins of the lower extremities is one of the most common diseases on Earth and its combination with other diseases is inevitable. The combination of diseases aggravates the course of each other. Rheumatoid arthritis worsens the course of varicose veins with trophic disorders – both independently – an increase in pro-inflammatory factors, a change in microcirculation, deformation of the limb axis with a violation of musculoskeletal function, and treatment of rheumatoid arthritis reduces the immune response of the body reducing the reparative capabilities of the body. Clinical case: a patient with a long course of rheumatoid arthritis and varicose veins. For a long time he used cytostatic drugs with a subsequent transition to hormonal drugs. For about 5 years, he noted the appearance of ulcers on both shins, which took a circular shape, which significantly reduced the quality of life and did not respond to conservative treatment. He repeatedly underwent inpatient treatment with intravenous administration of general restorative and vascular drugs. He was also constantly observed in the polyclinic at the place of residence with the implementation of local treatment of ulcerative surfaces. The patient was successively treated for varicose veins by endovenous laser coagulation of the trunks of large subcutaneous veins. The second stage was performed debridement of the wound surface and skin grafting with a free perforated flap. Between the stages, the appointment of drug therapy according to clinical recommendations. Rheumatoid arthritis and varicose veins aggravate each other, especially trophic manifestations. Treatment of such patients should be carried out by a team ofspecialists of various profiles together. Step-by-step treatment ofsuch patients with careful preparation for each of them shows excellent results. A special place is occupied by the patients’ commitment to treatment.

Introduction SARS-CoV virus showed transneuronal penetration through the olfactory bulb resulting in the rapid intracranial spread. So, olfactory dysfunction is an early marker of COVID-19 infection. However, individuals may develop chronic olfactory impairment for more than six months in 1–10% of cases. Objectives The study’s objective was to evaluate the efficacy and safety of intranasal immunotherapy using bioactive substances produced by M2 macrophages for the treatment of people with long-term post-COVID-19 hyposmia. Methods Seven individuals with long-term persistent hyposmia (7 to 24 months), associated with PCR-confirmed coronavirus infection were evaluated for olfactory function at baseline, one, and six to twelve months after therapy. Results The intranasal inhalation of M2 macrophage conditioned medium (one time per day for 28-30 days) was well tolerated. Furthermore, olfactometry demonstrated that the patients restored their capacity to perceive (Kruskal-Wallis H test 14.123, p = 0.0009) and recognize odors (H = 11.674, p = 0.0029). In addition, the subjective evaluation of smell significantly improved (H = 11.935, p = 0.0026). At the 6- to 12-month follow-up, the majority of patients (5/7) reported extremely high levels of satisfaction with the outcomes, and the remaining two patients also felt generally positive about the therapy’s success. Conclusions Overall, our study showed that the use of intranasal inhalations as a method of delivering bioactive factors and the conditioned medium of M2 macrophages as a therapeutic agent are both safe, well tolerated and, according to preliminary data, clinically effective in the treatment of patients with long-term post-COVID-19 hyposmia. Disclosure of Interest None Declared

Introduction Chronic inflammation in depression is associated with decreased levels of neurotrpohic factors and suppressed neurogenesis. We have previously shown that intranasal therapy with soluble factors from M2 macrophages polarized by interaction with apoptotic cells in serum deprivation conditions (M2(LS); LS - low serum) and characterized by anti-inflammatory and pro-regenerative activity leads to the correction of the behavioral pattern in mice with a depression-like state. Objectives The present study focuses on the effect of M2(LS) macrophages on neuronal density in the frontal cortex and hippocampus of depression-like mice. Methods Depressive-like state was formed in passive male mice (CBAxC57Bl/6)F1) as a result of repeated experience of defeat in agonistic interactions with aggressive partner during 20 days (the sensory contact model). Depression-like mice were then treated intranasally with M2(LS) macrophages conditioned medium for 7 days. After that, the number of mature neurons in the frontal cortex and hippocampus was assessed using Nissl staining. Results The neuronal density in the pyramidal layer of the frontal cortex was significantly lower in depression-like mice than that in the intact control group of mice (p=0,047). At the same time, the number of neurons in the experimental group of mice that received soluble M2(LS) factors, was higher than that in depressive-like untreated control mice (p=0,003) and was comparable to that in the intact group of mice. At the same time the neuronal density in the CA1 and CA3 hippocampal areas did not change in depression-like mice following intranasal treatment with conditioned medium of M2(LS) macrophages. Conclusions The data obtained may indicate the neuroprotective effect of M2(LS) macrophages in the stress-induced depression model, which is realized through soluble factors and manifests itself in an increase of the pyramidal neurons density in the frontal cortex. Disclosure of Interest None Declared

Abstract Green organisms evolve oxygen (O2) via photosynthesis and consume it by respiration. Generally, net O2 consumption only becomes dominant when photosynthesis is suppressed at night. Here, we show that green thylakoid membranes of Scots pine (Pinus sylvestris L) and Norway spruce (Picea abies) needles display strong O2 consumption even in the presence of light when extremely low temperatures coincide with high solar irradiation during early spring (ES). By employing different electron transport chain inhibitors, we show that this unusual light-induced O2 consumption occurs around photosystem (PS) I and correlates with higher abundance of flavodiiron (Flv) A protein in ES thylakoids. With P700 absorption changes, we demonstrate that electron scavenging from the acceptor-side of PSI via O2 photoreduction is a major alternative pathway in ES. This photoprotection mechanism in vascular plants indicates that conifers have developed an adaptative evolution trajectory for growing in harsh environments.

During pregnancy, the maternal immune system must maintain tolerance to paternal antigens, at the same time being able to eliminate pathogens, which is achieved by the weakening of adoptive immunity and the activation of innate immunity, in particular, monocytes. However, the question about the functional phenotype of monocytes, having not only pro-inflammatory, but also anti-inflammatory activity, remains open. In the given work, we have investigated the expression of M2-associated suppressive markers Arg1 and MerTK in monocyte subpopulations during uncomplicated pregnancy. Fifty-three pregnant women with uncomplicated gestation were recruited, including 14 pregnant in the 1st trimester, 20 – in the 2nd and 19 – in the third pregnancy trimester. The comparison group consisted of 15 fertile unpregnant women without aggravated somatic anamnesis, with a history of at least one childbirth. The findings showed that in the unpregnant group circulating Mo express Arg1 and MerTK, and the most relative number of Arg1+ and MerTK+ cells is concentrated in intermediate and nonclassic monocytes. During pregnancy the expression of researched molecules in monocytes reliably increases. An increase in MerTK expression is manifested by a simultaneous increase in the number of MerTK+ cells and the mean fluorescence intensity of this marker; it is observed in the 1st and 2nd trimesters and registered in all three monocyte subpopulations. At the same time, an increase in Arg1 expression is manifested either by an enhancement of Arg1+ cells, or an increase in receptor density; it is registered throughout pregnancy, including the 3rd trimester, and is maximally expressed in classic monocytes. There is a direct correlation between the number of Arg1+ and MerTK+ cells in intermediate Mo, which increases with the progression of pregnancy, and in the 3rd trimester is also detected in classical and non-classical Mo. In general, the revealed increase in the expression of Arg1 and MerTK by monocytes indicates an increase in the anti-inflammatory potential of monocytes during pregnancy, and the involvement of monocytes in the regulation of the inflammatory process at the system level. Moreover, the features of Arg1 and MerTK expression in various monocyte subpopulations during pregnancy suggest that monocytes expressing Arg1 and MerTK can mediate different mechanisms of immune adaptation during pregnancy.

Recently, there has been a steady increase in depressive disorders, which occupy an important place in the structure of the causes of disability. In the pathogenesis of depression, an important role is played by neuroinflammation, which is associated with impaired adult neurogenesis. Notably, neuroinflammation is partially reversible, and the leading role in the initiation and regulation of neuroregeneration is given to macrophages. Opposite states of macrophage activation are classically activated M1 and alternatively activated M2 macrophages, characterized, respectively, by pro- and anti-inflammatory activity. A balance shift towards M2 macrophages has been considered as a new therapeutic strategy of psycho-neurological disorders. One of the inducers of the M2 phenotype is the efferocytosis. We have previously developed an original protocol for the generation of human macrophages under conditions of deficiency of growth / serum factors, in which M2 phenotype is formed through efferocytosis. Macrophages (M2(LS), LS – Low Serum) obtained according to this protocol express M2-associated markers, and are characterized by high production of growth and pro- angiogenic factors (IGF-1, VEGF, BDNF, EGF, FGF-basic, etc.), which can suppress inflammation and stimulate neuroregeneration / neuroplasticity. In the model of stress-induced depression, the antidepressant effect of soluble factors of M2(LS) macrophages was shown, accompanied by a decrease in the level of pro- inflammatory cytokines in certain brain structures. However, the effect of M2(LS) factors on neurogenesis remained unexplored. In the present work, which is a continuation of the aforementioned study, we analyzed the effect of intranasal administration of M2(LS) soluble factors on neuronal density in different brain areas – the frontal cortex and hippocampus – of depression-like mice. The results obtained showed that neuronal density in the frontal cortex, CA1 and CA3 zones of the hippocampus, was significantly higher in mice with intranasal administration of M2(LS) conditioned medium than in depression-like mice, and reached the level of neuronal density in intact animals. These results may indicate the neuroregenerative activity of M2(LS) macrophages in the model of stress-induced depression, which is mediated through soluble factors and manifests itself in an increase in the density of neurons in the brain.

T cells expressing checkpoint receptors PD-1, TIM-3 etc., are potential targets for monoclonal antibody immunotherapy in multiple myeloma (MM). However, checkpoint expressing T cell compartment includes different subsets, and their dysregulation following anti-checkpoint therapy can lead to the development of adverse events.The aim of this study was to evaluate activation markers – homeostatic cytokine receptors and transcription factors expressed by PD-1and TIM-3-positive T cells.Material and Methods. Relative counts of circulating PD-1and/or TIM-3-positive and negative T cells expressing common ɣ-chain cytokine receptors CD25, CD122, CD127, phosphorylated STAT5, and transcription factor EOMES associated with T cell exhaustion were studied using flow cytometry in 17 healthy donors, 22 MM patients with remission and 7 MM patients with progressive disease.Results. T cells expressing PD-1 and/or TIM-3 inhibitory checkpoint receptors in MM patients consisted of CD25+EOMESactivated cells, CD4+CD25+CD127-FOXP3+ regulatory T cells (Treg), CD4+CD25-EOMES+ dysfunctional cells. CD25+ T cells from healthy donors and MM patients, regardless of the expression of the studied checkpoint receptors, were EOMES-negative. No such association was found for CD122 and CD127 cytokine receptors. EOMES is a marker of T cell exhaustion for CD4+ T cells, but not for CD8+ T cells, in which it is more associated with activation. The proportion of CD4+ Tregs among circulating PD-1+ and TIM-3+ T cells was relatively low. A higher content of cytokine receptors in the population of TIM-3+ T cells may indicate the predominant involvement of TIM-3 in the control of homeostatic proliferation of mature T cells under lymphopenic conditions, while the expression of PD-1 may be more associated with the regulation of activation through T cell receptor. PD-1+ and/or TIM-3+ levels of activated, dysfunctional, and regulatory T cells can potentially be used to predict the safety and efficacy of targeted immunotherapy.

In humans circulating monocytes include classical (CD14++CD16- ), intermediate (CD14++CD16+) and non-classical/alternative (CD14+CD16++) monocytes, which in turn can be activated via the classical or alternative pathway. Pregnancy is accompanied by significant changes in the monocyte compartment, which is manifested by an increase in the number of circulating monocytes, including the proportion of intermediate monocytes, and a change in their function. However, the functional properties of monocyte subsets during gestation remain largely unexplored. We hypothesized that circulating monocytes may be activated in an alternative pattern and acquire features of M2 polarization (anti-inflammatory / immunosuppressive properties). The aim of the investigation was to study M2-associated markers that characterize the anti-inflammatory and immunosuppressive potential of myeloid cells in subpopulations of circulating monocytes in fertile nonpregnant women and women with uncomplicated pregnancy in the 2nd trimester. It was shown that in fertile non-pregnant women intermediate and non-classical monocytes are characterized by a higher expression of M2-associated markers (CD206, Arginase 1, MerTK) compared to classical monocytes. In the 2nd trimester of pregnancy, the expression of these molecules on monocytes increases significantly, which is manifested by 1) an increase in the proportion of CD206+ cells in subpopulations of classical and intermediate monocytes, 2) an increase in the mean fluorescence intensity of Arginase 1 in all monocyte subsets, 3) an increase in the proportion of MerTK+ cells in subpopulations of classical and intermediate monocytes and mean fluorescence intensity across all monocyte subsets. The highest content of CD206+ and MerTK+ cells in pregnant women is detected in the subpopulation of intermediate monocytes, and the highest values of the mean fluorescence intensity of Arginase 1 and MerTK – in the subpopulations of intermediate and non-classical monocytes. The data obtained demonstrate that monocytes of pregnant women in the 2nd trimester of pregnancy are characterized by signs of M2 polarization. This is confirmed not only by an increase in the expression of the M2-associated mannose receptor CD206, but also by an increase in the expression of Arginase 1 and MerTK, which mediate the immunosuppressive activity of myeloid cells and, in particular, macrophages of the M2 phenotype. Further studies of M2-associated markers in monocyte subpopulations during gestation will allow a more detailed characterization of the regulatory role of circulating myeloid cells during pregnancy.

GM-CSF and M-CSF, the hematopoietic colony-stimulating factors, induce various phenotypic changes in macrophage lineage populations and promote cell differentiation, respectively, into M1- and M2-like macrophages. The pro- and anti-inflammatory properties of macrophages generated by these colony-stimulating factors are well described, but the contribution of differentiation and polarization signals to the fibromodulatory activity of macrophages remains unexplored. To clarify the differences in the fibrogenesis regulation mechanisms inherent in differently activated macrophages, we studied the effects of macrophage-conditioned media on proliferation and differentiation of dermal fibroblasts. In this study, the human macrophages generated from peripheral blood monocytes were investigated. They were induced for differentiation by M-CSF or GM-CSF, being further polarized in the M1 direction with lipopolysaccharide and, in the M2 direction, with IL-4 or dexamethasone. Proliferative response of the fibroblasts was determined radiometrically by [3H]-thymidine incorporation. Differentiation into myofibroblasts was determined with flow cytometry technique, as expression of a specific marker α-smooth muscle actin (α-SMA). The level of macrophage TGF-β1 production was assessed using an appropriate ELISA kit. The data obtained indicate that the macrophages differentiated under the influence of “homeostatic” M-CSF are characterized by a moderate stimulating effect upon fibroblast proliferation, and the effects of M2 (IL-4) and M2 (Dex) macrophages exceed that of M1 (LPS), but do not differ significantly from each other. The M-CSF-induced M1 (LPS) and M2 (IL-4) macrophages, but not M2 (Dex), enhance the fibroblast differentiation and show similar level of stimulation. In contrast to M-CSF, the macrophages induced by “pro-inflammatory” GM-CSF exhibit a pronounced stimulatory effect on fibroblast proliferation, and the effects of M2 macrophages exceed those of M1 cells, being most pronounced for M2 (Dex). At the same time, only GM-CSF-induced M2 (IL-4) macrophages enhance fibroblast differentiation. Dexamethasone-polarized macrophages do not significantly affect fibroblast differentiation regardless of the CSF used (M-CSF or GM-CSF). The content of TGF-β1 in the supernatants of differently activated macrophages does not correlate with the level of stimulating effect of macrophage-conditioned media upon fibroblast differentiation. In general, the data obtained suggest the involvement of differentiation and polarization signals into modulation of pro- and anti-fibrogenic properties of macrophages.

Introduction. Neuromodulation has proven itself in the treatment of patients suffering from idiopathic overactive bladder and non-obstructive urinary retention, who are resistant to conservative therapy. The possible use of the method in the population of patients with neurogenic lower urinary tract dysfunction (NLUTD) is of undoubted clinical interest.Objective. To analyze the current possibilities and features of neuromodulation in a cohort of patients with NLUTD.Materials and methods. Original research materials published in the PubMed, eLibrary, SciVerse (ScienceDirect), Scopus, Medline, EMBASE databases, websites of professional associations without restrictions on the date of publication were used. Sixty sources were selected for citation, with preference given to systematic reviews, meta-analyses and RCTs .Results. In relation to NLUTD, transcranial and peripheral magnetic stimulation, intravesical electrical stimulation, tibial, pudendal electrical stimulation, and stimulation of the dorsal pudendal nerve, as well as sacral and epidural methods of neurostimulation are considered.Conclusion. The current literature optimistically presents the experience of using neuromodulation in the NLUTD patient population with the largest evidence base for invasive sacral and tibial stimulation. The studies are based on heterogeneous populations, limited by small sample sizes with insufficient descriptive part of the degree and severity of neurological diseases, and it should be considered when forming guidelines. However, the lack of other suitable therapies and promising initial results indicate the importance of further efforts to improve the applied methods of neuromodulation. Further studies are needed with larger sample sizes, better classification of diseases, and controlled study design

Viral hepatitis remains the most common cause of liver cirrhosis (LC). Monocytes, capable of migrating to the liver and participating in inflammation and fibrogenesis, play an important role in the LC pathogenesis as confirmed by the association of certain monocyte subsets with the disease severity and mortality in alcoholic and biliary LC. However, the clinical and prognostic relevance of monocytes in viral LC remains poorly investigated. This study was aimed to investigate the disturbances in circulating monocytes including classical (CD14++CD16, cMo), intermediate (CD14++CD16+, iMo) and non-classical monocytes (CD14+CD16++, nMo) in patients with viral LC, as well as their correlation with viral characteristics, LC severity and progression of the disease 12 months after combination therapy. A significant increase in iMo and nMo, cMo level tended to decrease, and a two-fold decline in cMo/iMo ratio was revealed in patients with viral LC vs. healthy donors. These changes in monocyte pattern did not depend on the type of virus (HCV vs HBV/HDV) or its replication (replication vs the integrative phase), but were associated with the LC severity. The iMo level was positively correlated with laboratory indicators of liver damage, ChildPugh (rS = 0.57; P = 0.001) and MELD score (rS = 0.41; P = 0.033). ROC analysis showed that the cMo/iMo ratio at 9.5 allowed to predict the risk of LC progression with a sensitivity of 83.3% and a specificity of 76.2%. Of note, in comparison groups patients with alcoholic or biliary/autoimmune LC also demonstrated increased frequencies in iMo and nMo and decreased cMo/iMo ratio. However, in this case, the LC severity was negatively correlated with CD16+monocytes, particularly with the iMo and nMo subset, respectively, in alcoholic and biliary LC, evidencing the protective role of such cell subsets. Thus in viral LC the changes in circulating monocyte profile to increased iMO and nMo as well as decreased cMo are not associated with the virus type and replication; in contrast to the alcoholic and biliary/autoimmune LC, the level of iMo directly correlates with the indicators of liver damage and LC severity; cMo/iMo ratio is a biomarker of therapy response/disease progression.

Background: Immune checkpoint ligand-receptor interactions appear to be associated with multiple myeloma (MM) progression. Simultaneously, previous studies showed the possibility of PD-1 and TIM-3 expression on T cells upon stimulation with common γ-chain family cytokines in vitro and during homeostatic proliferation. The aim of the present work was to study the impact of homeostatic proliferation on the expansion of certain T cell subsets up-regulating PD-1 and TIM-3 checkpoint molecules. Methods: The expression of CD25, CD122, CD127 common γ-chain cytokine receptors, phosphorylated signal transducer and activator of transcription-5 (pSTAT5) and eomesodermin (EOMES) was comparatively assessed with flow cytometry in PD-1- and TIM-3-negative and positive T cells before the conditioning and during the first post-transplant month in peripheral blood samples of MM patients. Results: Substantial proportions of PD-1- and TIM-3-positive T lymphocytes expressed common γ-chain cytokine receptors and pSTAT5. Frequencies of cytokine receptor expressing cells were significantly higher within TIM-3+ T cells compared to PD-1+TIM-3− subsets. Considerable proportions of both PD-1-/TIM-3-negative and positive CD8+ T cells express EOMES, while only moderate frequencies of CD4+ PD-1+/TIM-3+ T cells up-regulate this transcription factor. Besides, the surface presence of CD25 and intranuclear expression of EOMES in CD4+ T cells were mutually exclusive regardless of PD-1 and TIM-3 expression. The stimulation with common γ-chain cytokines up-regulates PD-1 and TIM-3 during the proliferation of initially PD-1/TIM-3-negative T cells but fails to expand initially PD-1+ and TIM-3+ T cell subsets in vitro. Conclusions: Both PD-1 and TIM-3 expressing T cells appear to be able to respond to homeostatic cytokine stimulation. Differences in common γ-chain cytokine receptor expression between PD-1+ and TIM-3+ T cells may reflect functional dissimilarity of these cell subsets. Checkpoint blockade appears to alleviate lymphopenia-induced proliferation of PD-1+ T cells but may raise the possibility of immune-mediated adverse events. [ABSTRACT FROM AUTHOR]

Inspired by the period-four oscillation in flash-induced oxygen evolution of photosystem II discovered by Joliot in 1969, Kok performed additional experiments and proposed a five-state kinetic model for photosynthetic oxygen evolution, known as Kok's S-state clock or cycle1,2. The model comprises four (meta)stable intermediates (S0, S1, S2 and S3) and one transient S4 state, which precedes dioxygen formation occurring in a concerted reaction from two water-derived oxygens bound at an oxo-bridged tetra manganese calcium (Mn4CaO5) cluster in the oxygen-evolving complex3-7. This reaction is coupled to the two-step reduction and protonation of the mobile plastoquinone QB at the acceptor side of PSII. Here, using serial femtosecond X-ray crystallography and simultaneous X-ray emission spectroscopy with multi-flash visible laser excitation at room temperature, we visualize all (meta)stable states of Kok's cycle as high-resolution structures (2.04-2.08 Å). In addition, we report structures of two transient states at 150 and 400 µs, revealing notable structural changes including the binding of one additional 'water', Ox, during the S2→S3 state transition. Our results suggest that one water ligand to calcium (W3) is directly involved in substrate delivery. The binding of the additional oxygen Ox in the S3 state between Ca and Mn1 supports O-O bond formation mechanisms involving O5 as one substrate, where Ox is either the other substrate oxygen or is perfectly positioned to refill the O5 position during O2 release. Thus, our results exclude peroxo-bond formation in the S3 state, and the nucleophilic attack of W3 onto W2 is unlikely.

Macrophages are the major cells of innate immunity with a wide range of biological effects due to their great plasticity and heterogeneity. Macrophages play a key role in neuroregeneration following nervous tissue injury. However, the neuroregenerative potential of various macrophage phenotypes, including those polarized by efferocytosis, remains unexplored. The aim of this study was to compare the neuroregenerative and neuroprotective activity of soluble factors secreted by variously activated human macrophages on the functions of neural progenitors in an in vitro model of ischemia or ischemia/hypoxia. Macrophages were polarized by interferon-γ (M1), IL-4 (M2a), or interaction with apoptotic cells (M2(LS)). The effect of macrophages conditioned media on the proliferation, differentiation, and survival of SH-SY5Y cells damaged by serum deprivation alone (ischemic conditions) or in combination with CoCl2 (ischemic/hypoxic conditions) was assessed. All studied macrophages stimulated the proliferation and differentiation of SH-SY5Y cells. On day 3, the pro-proliferating effect of M1 and M2 was similar and did not depend on the severity of the damaging effect (ischemia or ischemia/hypoxia), while on day 7 and under ischemic/hypoxic conditions, the effects of M2(LS) exceeded those of M1 and M2a cells. The prodifferentiation effects of macrophages were manifested in both short- and long-term cultures, mainly under ischemic/hypoxic conditions, and were most characteristic of M2(LS) cells. Importantly, the ischemia/hypoxia model was accompanied by the pronounced death of SH-SY5Y cells. Only macrophages with the M2 phenotype demonstrated antiapoptotic activity, and the effect of M2(LS) was higher than that of M2a. The results obtained indicate that human macrophages have neuroprotective and neuroregenerative activity, which is mediated by soluble factors, is most characteristic for macrophages activated by efferocytosis (M2(LS)), and is most prominent under in vitro conditions simulating the combined effect of ischemia/hypoxia.

On the occasion of the 90th birthday of Govindjee, Professor Emeritus of Plant Biology, Biochemistry, and Biophysics, the University of Illinois at Urbana-Champaign (UIUC), over 100 celebrants have sent felicitations and messages to thank him for mentoring, nurturing, and building the community of photosynthesis researchers belonging to four generations; and in making the scientific knowledge accessible to students and young researchers via his monumental writings and editorial contributions. Govindjee joined UIUC in September of 1956 to study as a graduate student in the laboratory of Robert Emerson. In 1961, he joined UIUC as an Assistant Professor and retired as a full professor in 1999. He is well-known for pioneering work in oxygenic photosynthesis, leading to the current Z-scheme, and for his breakthrough advances concerning light harvesting, primary charge separation, the role of bicarbonate on the two-electron gate of photosystem II, water oxidation, nonphotochemical quenching, and the use of biophysical techniques, such as prompt fluorescence, delayed fluorescence, thermoluminescence, and nuclear magnetic resonance. Today, despite his retirement, Govindjee continues to explore several important questions in the field of photosynthesis and documents the history of science. This tribute, in turn, attempts to capture scientific collaborations, as well as scholarly and personal contributions made by Govindjee to the lives of hundreds of scholars and students worldwide.

The aim of the present study was to assess safety and clinical efficacy of inhalation immunotherapy based on intranasal administration of bioactive factors produced by the M2 phenotype macrophages in children with language impairments, as well as to study the effect of inhalation immunotherapy on the cytokine profile in the patients' blood serum. The study was carried out according to the NCT04689282 protocol (www.ClinicalTrials.gov) and included 14 children (9 boys / 5 girls), aged 3 to 8 years, with language impairments associated with perinatal or postnatal CNS lesions of various origin. The children recruited into the study were assessed by a neurologist and speech therapist before the therapy, at the end of the course (1 month), and 6 months later. Serum samples for cytokine analysis were obtained before and 1 month after therapy. The course of intranasal inhalations by the conditioned M2 media (2 ml one time per day for 28-30 days) was safe and well tolerated. None of the 14 treated children had significant adverse reactions and severe undesirable events. Intranasal immunotherapy led to a decrease in the severity of language problems, which manifested by improved speech understanding by 45%; the sensorimotor level of speech, by 51%; word formation skills, by 72%, as well as a twofold increase in general and fine motor skills. In children with signs of autism spectrum disorders, along with a language improvement, a decrease in the severity of autistic symptoms was registered, as evidenced by statistically significant decrease in the CARS score from 42.5 to 38.5 after 1 month, and to 33 points after 6 months (p < 0.05). The clinical effect was revealed rather soon, i.e., within a month after the first procedure, being maintained or intensified during a follow-up for 6 months. At the same time, two-thirds of the children showed a clear clinical improvement, with insignificant effect in the rest of patients. Comparative analysis of the serum cytokine levels in these subgroups showed that children with a pronounced positive response to inhaled immunotherapy differed in the following parameters: (1) initially higher level of VEGF and IGF-1, and (2) decrease the level of TNFα in response to intranasal immunotherapy. In summary, we first tested a fundamentally new approach based on the use of soluble factors from M2-type macrophages and intranasal route of their administration in order to treat the children with severe language impairments, demonstrating safety and obtained preliminary data on effectiveness of such approach.

Macrophages play a key role in triggering and regulation of neuroregeneration. The characteristic feature of macrophages is pronounced plasticity, which manifests itself in the ability of macrophages to change their functional phenotype depending on the micromilieu. Apoptotic cell clearance (efferocytosis) is an important inducer of a macrophage polarization to M2 phenotype under pathological settings. Previously, we have developed an original protocol for the generation of M2-like macrophages, polarized by efferocytosis under serum-deprived conditions (M2 (LS), Low Serum). The present study was aimed to assess a neuroregenerative potential of M2 (LS) macrophages. We studied their effect on the differentiation of SH-SY5Y cells in comparison with retinoic acid (RA). As the morphological criteria of differentiation we have assessed the relative content of differentiated cells, i.e., cells with a neurite length exceeding the cell body length, and the average neurite length on days 3, 7, and 13. The ratio of neuron-like (N-type) and epithelial-like (S-type) cells in cultures was also assessed. SH-SY5Y cells were characterized by a low level of spontaneous differentiation, both under standard conditions (10% FBS) and serum deprivation (1% FBS). Upon RA treatment, SH-SY5Y cells stopped proliferating and underwent neuronal differentiation. Cultivation of SH-SY5Y cells in the presence of M2 (LS) conditioned medium also led to a significant increase in the relative content of differentiated cells, the average length of neurite-like processes, as well as a change in the balance of S- and N-type cells towards a pronounced predominance of the latter. The morphological features of differentiation were significantly less pronounced at early stage (day 3) of differentiation as compared with the RA-induced changes and reached the level of positive control only at later stages (day 13) (p < 0.05). In contrast to retinoic acid, M2 (LS) conditioned medium induced neuronal differentiation of SH-SY5Y cells without suppressing their proliferative activity. The data obtained may indicate a high neuroregenerative potential of M2 macrophages in vitro, which is realized through soluble factors and manifests itself in promoting SH-SY5Y differentiation.

Apoptosis and subsequent removal of dead cells are an essential part of wound healing. Macrophages phagocytize apoptotic cells (efferocytosis) and contribute to the resolution of inflammation. However, their participation in fibrogenesis and the mechanisms of influence on this process remain unclear. In the present study, we focused on the fibrogenic properties of human monocyte-derived macrophages polarized in the M2 direction by interaction with apoptotic cells. We studied their influence on the proliferation ([3H]-thymidine incorporation), differentiation (by the expression of α-SMA, a myofibroblast marker) and collagen-producing activity (ELISA) of dermal fibroblasts compared to classically (LPS) and alternatively (IL-4) activated macrophages. Macrophages polarized by the interaction with apoptotic cells had a unique phenotype and profile of produced factors and differed from the compared macrophage subtypes. Their conditioned media promoted the proliferation of dermal fibroblasts and the expression of α-SMA in them at the level of macrophages stimulated by IL-4, while the stimulating effect on the collagen-producing activity was more pronounced compared to that of the other macrophage subtypes. Moreover, they are characterized by the high level of production of pro-fibrotic factors such as TIMP-1, TGF-β1 and angiogenin. Taken together, M2-like macrophages polarized by efferocytosis demonstrate in vitro pro-fibrotic activity by promoting the functional activity of dermal fibroblasts and producing pro-fibrotic and pro-angiogenic factors.

This article is the second communication in a series of articles devoted to the effects of a domestic preparation of macrophage-activating factor (GcMAF-RF) and assessment of its biological properties. The aim of this work was to study the effect of the GcMAF-RF upon M0 → M1 polarization of macrophages (Mph), and activation of the professional properties of ex vivo generated antigen-presenting dendritic cells (DC), as well as on ex vivo production of pro-inflammatory (TNFα, IL-1β, IL-6, IFNγ, IL-17, IL-18) and anti-inflammatory (TGF-β, IL-4, IL-10) cytokines, growth factors (IL-2, GM-CSF, G-CSF, VEGF) and chemokines (MCP, IL-8) by the whole blood cells from healthy donors. Mph and DC were generated from the monocytes (3 to 5×106 /ml) derived from adherent fraction of peripheral blood mononuclear cells (MNC) of healthy donors. Granulocyte/macrophage colony-stimulating factor (rhGM-CSF) was used to obtain Mph, whereas DC production was induced by GM-CSF and interferon-α. To provide M1 polarizing signals, bacterial lipopolysaccharide (LPS from E. coli 0114:B4) was used in controls. In experimental series, GcMAF-RF was added 48 h before the end of culture. The stimulating effect of the obtained Mph and DC upon cell proliferation was assessed in allogeneic mixed culture of leukocytes (alloMLC) using radiometric technique, by 3 H-thymidine incorporation. The influence index (IR) of Mph or DC upon allo-SCL was calculated as the ratio of the proliferative response of MNCs in the presence of Mph, or DC to the level of spontaneous MNC proliferation. To determine the cytokine production by human whole blood cells ex vivo, peripheral blood samples from 3 donors with two replicate GcMAF-RF preparations were used, at a total of 6 points. All variants of the study were carried out with mitogen-activated and non-activated blood cells. The cytokine content was determined by the ELISA assays. The effects of GcMAF-RF were quantified as a fold increase (FI), i.e., the ratio of cytokine production in the presence of GcMAF-RF to the level of their spontaneous production. It was shown that the GcMAF-RF preparation was as effective, as lipopolysaccharide (LPS), the standard Mph and DC activator which induces polarization of differentiated M0-macrophages into M1 cells and final maturation of DCs, manifesting by a significant increase in their allo-stimulatory activity in a mixed leukocyte culture (allo-MLC). Moreover, GcMAF-RF stimulates production of numerous cytokines and chemokines (TNFα, IL-1β, IL-6, IL-18, IL-4, IL-10, GM-CSF, G-CSF, VEGF, IL-8), by blood cells (granulocytes, lymphocytes, monocytes), thus indicating direct participation of the macrophage activator GcMAF-RF in various immune processes. The domestic GcMAF-RF drug induces polarization of macrophages M0 → M1, final maturation of DCs and allostimulating activity of Mf and DCs, and is also able to effectively stimulate circulating blood cells to synthesize cytokines/chemokines with pro-inflammatory and immunoregulatory activities.

Light-induced oxidation of water by photosystem II (PS II) in plants, algae and cyanobacteria has generated most of the dioxygen in the atmosphere. PS II, a membrane-bound multi-subunit pigment protein complex, couples the one-electron photochemistry at the reaction centre with the four-electron redox chemistry of water oxidation at the Mn4CaO5 cluster in the oxygen-evolving complex (OEC). Under illumination, the OEC cycles through five intermediate S-states (S0 to S4), in which S1 is the dark-stable state and S3 is the last semi-stable state before O-O bond formation and O2 evolution. A detailed understanding of the O-O bond formation mechanism remains a challenge, and will require elucidation of both the structures of the OEC in the different S-states and the binding of the two substrate waters to the catalytic site. Here we report the use of femtosecond pulses from an X-ray free electron laser (XFEL) to obtain damage-free, room temperature structures of dark-adapted (S1), two-flash illuminated (2F; S3-enriched), and ammonia-bound two-flash illuminated (2F-NH3; S3-enriched) PS II. Although the recent 1.95 Å resolution structure of PS II at cryogenic temperature using an XFEL provided a damage-free view of the S1 state, measurements at room temperature are required to study the structural landscape of proteins under functional conditions, and also for in situ advancement of the S-states. To investigate the water-binding site(s), ammonia, a water analogue, has been used as a marker, as it binds to the Mn4CaO5 cluster in the S2 and S3 states. Since the ammonia-bound OEC is active, the ammonia-binding Mn site is not a substrate water site. This approach, together with a comparison of the native dark and 2F states, is used to discriminate between proposed O-O bond formation mechanisms.

Modulation of glycoregulation in agriculture is reviewed here with emphasis on the elucidation of previously unknown pathways involving vacuolar lectins as well as a bypass of lectins that direct free sugars toward productivity. The reversible binding sequences of the endogenous lectin cycle are compared to an induced lectin bypass, as follows. (1) In the cycle, carbohydrate ligands, with similar binding specificities that compete for binding sites on lectins, are involved in the natural cycle of sugar exchanges. (2) For the bypass, tightly bound ligands that occupy lectins prevent free sugars from binding, making them available for productivity. This bypass is induced by methyl-α-D-mannopyranoside, a biochemical plant growth regulator for photosynthesis. Integration of this novel technology, with structural elements crucial for ligand binding by the lectins and with nitrogen assimilation, provides the basis for successful modulation of glycoregulation in crops for enhancement of quality and quantity.

Macrophages (Mφ) play a key role in regulation of fibrogenesis, including proliferation of fibroblasts and myofibroblasts, differentiation of progenitor cells into myofibroblasts, as well as synthesis and secretion of the extracellular matrix, mainly collagen. The direction of the Mφ effects (stimulation or suppression) is determined by a number of factors, including the stage of the fibrotic process and the Mφ functional phenotype dependent on the signals of microenvironment. One of the feasible ways of the fibrogenesis regulating is the secretion of pro- or antifibrotic factors such as matrix metalloproteinases, inhibitors of metalloproteinases and some cytokines. However, existing data on ability to secrete these factors by various subpopulations of human Mφ are rare and controversial. The aim of this study was to characterize the ability of human M1, M2a, and M2c Mφ differentiating in the presence of GM-CSF to produce matrix metalloproteinases (MMP-9) and their tissue inhibitors (TIMP-1), as well as some cytokines and growth factors. As compared to M2 macrophages, the M1 macrophages polarized by lipopolysaccharide produced significantly more TNFα, IL-6 and IL-2 that have pro-inflammatory activity and are able to initiate a fibrotic process. In turn, M2a Mφ stimulated by IL-4 were characterized by a high level of VEGF production and, at the same time, low levels of TNFα and IL-6, which may determine the important role of these cells at the proliferative stage of fibrosis and stimulation of extracellular matrix deposition. Finally, M2c Mφ polarized by dexamethasone, exhibited the М2а-like cytokine profile, i.e., VEGF was actively produced against the background of low TNFα and IL-6 synthesis. Moreover, all three Mφ subpopulations did actively secrete MMP-9 and TIMP-1, without significant difference in production of these factors. However, M2c Mφ differed by a significantly higher MMP-9/TIMP-1 ratio index compared to M1 and M2a Mφ, and it is crucial at the rearrangement stage of the fibrotic process. Thus, the production of MMP-9 and TIMP-1, together with other pleiotropic cytokines and growth factors by various Mφ subtypes may reflect their role in regulation of fibrotic process at various stages.

Early prediction for ischemic stroke (IS) outcome is a major challenge since it may help to optimize treatment program and to make it more personalized. Since T cells with regulatory activity are involved in different pathophysiological processes in brain stroke, including inflammation, immune suppression, brain damage and repair, the study of T cells as potential biomarkers has essential importance. The present work aimed to study the circulating T cell subsets with phenotype of type 1 T helper cells (Th1) and regulatory T cells (Treg), and their ratio during the acute phase of IS, depending on stroke severity, inflammatory response and 3-month outcome (according to modified Rankin scale, mRs). Patients and methods. The study included 61 patients with a newly diagnosed IS (severity according to NIHSS ≥ 5), in the first 24-48 h after stroke onset, and 20 age/sex-related healthy donors. Laboratory examination included assessment of leukocytosis, neutrophillymphocyte ratio (NLR) and CRP concentration. Mononuclear cells were isolated from peripheral blood to study T cell subsets. Th1 and Tregs were measured by FACS analysis as CD4+IFNγ+ and CD4+CD25hiT cells, respectively. During the first 24-48 h after stroke, the patients had elevated values of leukocyte counts, NLR and CRP. Higher levels of these parameters in severe stroke compared with mild stroke, as well as direct correlation of NIHSS with NLR and CRP evidenced that the stroke severity was associated with more pronounced inflammatory response. Patients were also characterized by a significant decrease in CD4+IFNγ+Th1 cells, an increase in CD4+CD25hiTreg, and a marked decrease in Th1/Treg ratio. Furthermore, in patients with NIHSS ≥ 8 (moderate and severe stroke), the percentage of CD4+IFNγ+T cells was in direct correlation, and the number of CD4+CD25hiT cells was inversely related to CRP and NLR values. The changes of T cell subsets were more pronounced in patients with a favorable 3-month outcome (mRs > 3). As a result, the patients with poor outcome (mRs ≤ 3) had higher CD4+IFNγ+T cell proportion, lower CD4+CD25hiT cell percentage and 4-fold higher CD4+IFNγ+/CD4+CD25hi ratio compared with opposing group. ROC analysis revealed a “good” quality of prognosis based on evaluation of the CD4+IFNγ+/CD4+CD25hi ratio as a monopredictor of adverse outcome (AUC = 0.75) and “very good” quality of prognosis when the indicated ratio was combined with NIHSS scale (AUC = 0.82). The data obtained suggest that a decrease of Th1/Тreg ratio, due to a decrease in CD4+IFNγ+ and increased CD4+CD25hiT cell counts during the acute phase of ischemic stroke is a compensatory reaction directed at inhibition of inflammatory response, and has a prognostic significance as early predictor of the outcome at 3 months.