TREATMENT OF FACIAL PAIN IN RHINOSINUSITIS

Purpose: to evaluate the eff ectiveness of treatment of facial pain based on the study of the mechanism of neurogenic pain symptoms in rhinosinusitis.Materials and methods: the study includes 380 patients with rhinosinusitis with varying severity of pain symptoms . Age subjects ranged from 15 to 81 years. Th e investigation of the level of substance P in plasma.Results: in excess of the high end - tration of substance P in the serum of patients with rhinosinusitis more than 2000 pg / ml refl ect participation in the infl ammatory process of neurogenic mechanisms and require pathogenetic therapy.Summary: treatment of neurogenic infl ammation was performed drugs prednisolone and Milgamma. Relief of neurogenic component of the infl ammatory process in rhinosinusitis improves the eff ectiveness of treatment and reduce the time to 3-5 days

CHANGES IN THE CAPILLARY AND VENOUS BLOOD CYTOKINE PROFILE OF PATIENTS WITH PSORIASIS DEPENDING ON THE TREATMENT

ABSTRACT. Psoriasis is a chronic autoimmune skin disease involving T-cell immunity. The interleukin (IL)-23/IL-17/IL-22 cytokine axis is key in the immunopathogenesis of psoriasis. The role of the IL-36 subfamily regulating inflammation in the skin is shown. Topical preparations are used to treat psoriasis. Objective: to study changes in the cytokine profile of venous and capillary blood taken near the focus of psoriatic inflammation, depending on the treatment with topical preparations. 40 patients with psoriasis, mean age 43.7 years, were examined. Group 1a (20 people) received local treatment with mometasone, Group 1b (20 people) received topical gel containing IL-36 receptor antagonist. 20 healthy people, mean age 46.6 years, consisted the control group 2. Capillary blood was collected from a finger, in patients near the lesion 200 μl in a microvette with EDTA. Venous blood was taken from the cubital vein 3 ml into a vacuum tube with EDTA. The concentration of 15 cytokines in blood plasma was tested by the multiplex method (MagPix, BioRad, USA). The effectiveness of therapy was assessed using the PASI and DLQI indices. At the end of treatment (day 14), the PASI and DLQI indices significantly decreased in both groups. On the 28th day, the PASI index in Group 1a returned to its original level, in group 1b it remained steadily reduced. Before treatment, the levels of all cytokines except IL-10 in the capillary blood of patients with psoriasis were significantly increased compared to Group 2, and the levels of 5 cytokines were increased in the venous blood. After 14 days in Group 1a, the levels of IL-1, IL-4, IL-6, IL-21, IL-22, IL-23, IL-25, IL-33 significantly decreased in capillary blood, and only IL-17F, IL-21, IL-33 and TNF in the venous blood. On the 28th day, the concentrations of almost all cytokines returned to their original level. In Group 1b, on the 14th day, the levels of IFN-γ, IL-1, IL-4, IL-17F, IL-21, IL-22, IL-23, IL-25, IL-33 significantly decreased in capillary blood, and in venous blood - IFN-γ, IL-21, IL-22, IL-23, IL-33. On the 28th day, the concentration continued to decrease, or the level of these cytokines remained reduced, and IL-6 significantly decreased in the vein. Thus, the method for determining the profile of capillary blood cytokines from the area of ​​psoriatic lesions can be used to monitor the effect of treatment in patients with psoriasis

Severity evaluation of bhronic polypous rhinosinusitis based on changes of IL-1β cytokine and IL-1ra concentrations in nasal secretions

We provide the data on distribution of IL-1β and IL-1ra cytokine concentrations in nasal secretions from the patients with different forms of chronic polypous rhinosinusitis. The control group (group 1) included healthy persons. The second group consisted of patients exhibiting chronic polypous rhinosinusitis (CRS-NP), free of clinical signs of purulent inflammation in paranasal sinuses and/or bronchial asthma. The patients of group 3 were diagnosed with chronic purulent polypous rhinosinusitis (CPRS-NP). Group 4 consisted of the patients with chronic polypoid rhinosinusitis complicated by bronchial asthma (CRS-NP + BA). Cytokine concentration was determined by means of ELISA tests for all the patient groups. We have found that the local concentrations of IL-1ra and IL-1β cytokines were significantly different (p < 0.05) from the group of healthy individuals, dependent on the clinical forms of chronic polyposis. IL-1β concentrations in the group of patients with CRS-NP (No. 2) were increased 5-fold, for group of CPRS-NP patients (No. 3), these levels showed a 15.5-fold increase. In the group of patients with CRS-NP + BA (No. 4), the Il-1 levels were 13fold higher in comparison with healthy controls. The IL-1ra concentration in the group of healthy people (No. 1) was 1825.64+463.70 pg/ml. Upon development of CRS-NP (No. 2), an increase in its values up to 3646.21±263.39 pg/ml was registered. Upon development of CPRS-NP (No. 3), an increase in appropriate values up to 2305.33±282.86 pg/ml was noted, and, in cases of CRS-NP + BA, a decrease in its values to 956.15±213.02 pg/ml was observed.Meanwhile, a conflicting character was revealed for the changes in these cytokines, thus preventing their usage as an indicator of polyposis severity. At the same time, estimation of the CIL-1ra/CIL- 1β ratio in nasal secretions of the patients in groups 1, 2, 3 and 4 based on two-dimensional projection of the spatial CIL-1ra/ CIL-1β distribution provides a basis for discrimination of the patient groups. The CIL-1ra/ CIL-1β ratio as assessed for nasal secretions of patients belonging to groups 1, 2, 3, and 4 shows a general, statistically significant decrease with increasing severity of chronic polyposis (p < 0.001), thus making this parameter indiformative for intensity of the pathological process. The resulting 95% confidence intervals confirm that the cytokine ratios are promising parameter in order to estimate the efficiency of immune therapy

Ноотропні властивості тетрапептиду acetyl-(D-lys)-lys-arg-arg-amide (KK-1) на моделі хвороби альцгеймера у щурів, зумовленої хронічним введенням скополаміну

Improvement of Alzheimer’s disease (AD) therapy is one of the most important tasks of the modern medicine and pharmacy. Creation and the pharmacological study of the original medicines for treating AD are promising. The most promising class of these drugs is neuroprotectors. In the Research Institute of Highly Pure Biopreparations the tetrapeptide acetyl-(D-Lys)-Lys-Arg-Arg-amide (the laboratory code – KK-1) has been synthesized. Under the conditions of the pre-clinical study it has demonstrated the pronounced neuroprotective and nootropic properties on the model of cerebral ischemia. The aim of the current work is to study the nootropic properties of KK-1 on the model of AD in rats. The scopolamine-induced AD in rats was reproduced. For 10 days the rats with AD model were treated with KK-1 (0.1 mg/kg intranasally once a day) or with the reference medicine donepezil (1 mg/kg orally once a day). After the therapy the functional state of the rats’ CNS was evaluated using the open-field test (OFT), extrapolation escape task (EET) and the test of the conditioned reflex of passive avoidance (CRPA). The level of acetylcholine (Ach) and the activity of acetylcholinesterase (ACE) were measured using spectrophotometry in the whole brain homogenate and brain synaptosoms. The peptide KK-1 studied reduced the stress-related anxiety in OFT in rats, increased their exploratory activity (by 1.4 times, р<0.05 compared to the pathology group) and decreased the emotional response on the stress (by 44%, р<0.05). In EET the tetrapeptide KK-1 stimulated the rats’ cognitive functions, decreased the time of escape by 4.7 times (р<0.05 compared to the pathology group). Under the conditions of the CRPA test the high anti-amnestic activity of KK-1 was found (56.5% on the 1st day after the therapy and 81.5% on the 10th day). According to all indices the activity of KK-1 exceeded the reference medicine donepezil. The mechanism of the KK-1 nootropic action is in the increase of the Ach level in the rats’ brain synaptosoms probably due to stimulation of its synthesis. The peptide KK-1 moderately inhibits the ACE leading also to the Ach level increase in the cholinergic synapse synaptic cleft. It is likely that the peptide KK-1 has an effect on the Gq/G11-peptide-coupled M1-, M3-, and M5-cholinergic receptors as a positive allosteric modulator.Усовершенствование терапии болезни Альцгеймера (БА) – одно из наиболее актуальных заданий современной медицины и фармации. Перспективно создание и фармакологическое исследование оригинальных лекарственных средств для ее терапии. Перспективным классом таких средств являются нейропротекторы. В НИИ Особо чистых биопрепаратов создан пептид acetyl-(D-Lys)-Lys-Arg-Arg-amide (лабораторный шифр КК-1). В эксперименте на модели церебральной ишемии КК-1 демонстрирует выраженные нейропротекторные и ноотропные свойства. Цель исследования – изучить ноотропные свойства КК-1 на модели БА. У крыс моделировали скополамин-обусловленную БА. На протяжении 10 дней осуществляли терапию фармакологическим препаратом КК-1 (0,1 мг/кг интраназально 1 раз в день) или препаратом сравнения (донепезил, 1 мг/кг внутрижелудочно 1 раз в день). После терапии оценивали функциональное состояние животных по тестам открытого поля (ОП), экстраполяционного избавления (ЭИ) и условного рефлекса пассивного избегания (УРПИ). В гомогенате целого головного мозга (ГМ) и синаптосомах ГМ спектрофотометрически определяли уровень ацетилхолина (АХ) и активность ацетилхолинэстеразы (АХЭ). Исследуемый препарат КК-1 уменьшает стресс-обусловленную тревожность крыс с моделью БА в тесте ОП, повышая их ориентировочно-исследовательскую активность (в 1,4 раза, р<0,05) и уменьшая уровень эмоциональных реакций (на 44%, р<0,05). В тесте ЭИ тетрапептид КК-1 повышает когнитивные функции крыс, уменьшая время подныривания под край цилиндра в 4,7 раза (р<0,05 с группой патологии). По тесту УРПИ установлена высокая антиамнестическая активность КК-1 (56,5% и 81,5% на первые и десятые сутки соответственно). По всем показателям КК-1 превышает активность препарата сравнения донепезила.  Механизм ноотропного действия КК-1 на модели БА заключается в повышении уровня АХ в синаптосомах ГМ, вероятно, за счет стимуляции его синтеза. КК-1 умеренно угнетает АХЭ, чем дополнительно способствует накоплению АХ в синаптической щели холинергического синапса. Вероятно также взаимодействие пептида КК-1 с Gq/G11-связанным М1-, М3-, и М5-холинергическими рецепторами по принципу позитивной аллостерической модуляции.Удосконалення терапії хвороби Альцгеймера (ХА) – одне з найактуальніших завдань сучасної медицини та фармації. Перспективним є створення та фармакологічне вивчення оригінальних лікарських засобів для її терапії. Перспективним класом таких засобів є нейропротектори. У НДІ Особливо чистих біопрепаратів створено тетрапептид acetyl-(D-Lys)-Lys-Arg-Arg-amide (лабораторний шифр КК-1). В експерименті на моделі церебральної ішемії КК-1 виявляє потужні нейропротекторні та ноотропні властивості. Мета дослідження – дослідити ноотропні властивості КК-1 на моделі ХА. У щурів моделювали скополамін-зумовлену ХА. Протягом 10 днів їх лікували пептидом КК-1 (0,1 мг/кг інтраназально, 1 раз на день) або препаратом порівняння (донепезил, 1 мг/кг внутрішньошлунково, 1 раз на день). Після терапії оцінювали функціональний стан ЦНС щурів за тестами відкритого поля (ВП), екстраполяційного вивільнення (ЕВ) та умовного рефлексу пасивного уникнення (УРПУ). У гомогенаті цілого головного мозку (ГМ) та синаптосомах ГМ спектрофотометрично визначали рівень ацетилхоліну (АХ) та активність ацетилхолінестерази (АХЕ). Досліджуваний фармакологічний препарат КК-1 зменшує стрес-зумовлену тривожну поведінку щурів із модельною ХА за тестом ВП, підвищуючи їх орієнтовно-дослідницьку активність (у 1,4 рази, р<0,05) та зменшуючи емоційні реакції (на 44%, р<0,05). В тесті ЕВ пептид КК-1 покращує когнітивні функції щурів, зменшуючи час пірнання під край циліндра у 4,7 рази (р<0,05). За тестом УРПУ встановлено високу антиамнестичну активність досліджуваного препарату (56,5% та 81,5% на 1 та 10 день після терапії відповідно). За усіма показниками КК-1 перевершує активність донепезилу. Механізм ноотропної дії КК-1 на моделі ХА полягає у підвищенні рівня АХ в синаптосомах М-холінергічних рецепторів, імовірно, шляхом стимуляції його синтезу. КК-1 помірно пригнічує АХЕ, чим додатково сприяє накопиченню АХ в синаптичній щілині холінергічного синапсу. Можливою є також взаємодія тетрапептиду КК-1 із Gq/G11-зв’язаними М1-, М3-, та М5- холінергічними рецепторами за принципом позитивної алостеричної модуляції

Influence of VEGF deprivation upon vascular formation by endothelium in the presence of macrophages

Development of angiogenesis depends on the functional state of endothelial cells, as well as on the balanced secretion of cytokines, growth factors and chemokines by endothelial cells and cells of microenvironment. Macrophages represent an essential component of the microenvironment and take part in the formation of blood vessels both due to the production of cytokines and due to contact interactions with endothelial cells. VEGF is among the most important cytokines that control angiogenesis at all its stages. Currently, the role of VEGF in the intercellular interactions of endothelial cells and macrophages is not well described. The aim of our study was to investigate the effect of VEGF deprivation using monoclonal antibodies on angiogenesis under conditions of co-cultivation of endothelium and macrophages. Materials and methods: monoclonal antibodies to VEGF-A were used for VEGF deprivation in monoculture of endothelial cells and in co-culture of endothelial cells with macrophages. The IL-1β, IL-6 and TNFα cytokines were used as inducers. When VEGF-A was removed from the medium, endothelial cells show plasticity and form longer vessels, they modify the expression of VEGF receptors. Macrophages regulate endothelial cell activity through the secretion of cytokines, including VEGF, and through contact interactions with endothelial cells. THP-1 cells increase the sensitivity of endothelial cells to VEGF by stimulating the VEGFR1 and VEGFR3 expression, this effect is VEGF-A-independent. The IL-1β, IL-6, TNFa cytokines independently stimulate non-branching angiogenesis, increasing the length of the vessels. At the same time, IL-ip increases the VEGFR1 expression on the surface of endothelial cells. In contrast, IL-6 and TNFα decrease it, thereby regulating the sensitivity of endothelial cells to VEGF. The effects of these cytokines are not dependent on VEGF-A. The IL-1β, IL-6, TNFα cytokines promote acquisition of anti-angiogenic properties by THP-1 cells that is independent on VEGF-A, as well as on expression of its receptors by endothelial cells. Thus, VEGF is an important, but not the sole factor controlling angiogenesis. Under conditions of VEGF-A deficiency, either endothelial cells or microenvironment cells are able to compensate for its functional load due to the production of other growth factors

Effect of immune drugs to treat acute viral nasopharyngitis

The task in treating acute nasopharyngitis (ANP) deals with reducing the disease symptoms and the risk of complications. The lack of reliable antiviral drugs makes it important to search for appropriate medicines among other pharmacotherapeutic groups.The study involves a comparative analysis of the efficiency and estimates potential: the recombinant interferon α2b and the compound containing fungal β-D-glucans used in treat ANPThe studies involved patients with ANP from 18 to 55 years old. As many as 152 people were examined including the following: 38 were practically healthy people (group 1); and 114 patients wuth ANP: 38 people (group 2) was subject to a standard therapy (vasoconstrictor nasal drops, nasal cavity irrigation using 0.1% Miramistine solution, gargling using the Furacilin solution); forty people (group 3) were administered application of intranasal interferon α2b of 105 IU, it was delivered with a spray into each nasal passage twice a day; 36 people (group 4) were administered an immunotropic drug containing β-D-glucans orally twice a day. The duration of drug administration lasted 7 days. Polymerase chain reaction (PCR) was used to identify the ANP etiological factor. Concentrations of cytokines IL-1β, IL-1ra were estimated using enzyme immunoassay (ELISA) technique. Clinical efficiency was assessed through score approach. The following symptoms were taken into account: general malaise, sore throat, character of nasal discharge, and the difficulty of nasal breathing. The results of the study were analyzed using parametric and nonparametric statistical methods. In 60.0% the nasal secretions of patients revealed RV. The distribution of cytokine concentrations in nasal secretions in group 1 indicated that the concentration of IL-1β was in the range of 20.0-25.0 pg/ml, and the concentration of IL-1ra was about 1250.0-2500.0 pg/ml. Developing ANP stimulated an increase in IL-1β concentration up to 30.0-70.0 pg/ml in nasal secretions of patients without affecting IL-1ra concentrations. On day 7 of treatment, the cytokine concentrations among the patients treated using the immunotropic drugs were the same as in the group of healthy individuals. There were no significant changes in cytokine production on day 7 in the group of patients undergoing the standard treatment. Application of proposed immunobiological medicines to ANP does not result in overproduction of proinflammatory cytokine IL-1β in nasal secretion. This confirms that these drugs are promising in the treating strategy including reduction of the risk of developing complications

Study of local inflammatory response in different clinical forms of chronic polypous rhinosinusitis

Chronic rhinosinusitis (CRS) affects 5-12% of the world’s adult population. Chronic rhinosinusitis with nasal polyps (CRSwNP) accounts for 25-30% of all cases of CRS. CRSwNP-associated inflammatory process in nasal mucosa and paranasal sinuses depends on the characteristics of local immunity, including expression of a number of cytokines. The aim of this work was to investigate the parameters of local immunity in various clinical forms of CRSwNP. In this work, the concentrations of pro-inflammatory cytokines, i.e., interleukin-1β (IL-1β) and IL-8, antimicrobial function of neutrophils from the nasal cavity was evaluated, along with histological and immunohistochemical studies of polyposis tissue. The study included 4 groups of patients: a control group of practically healthy individuals, patients with CRSwNP, clinical cases with chronic purulent rhinosinusitis and nasal polyps (CPRSwNP), and patients with CRSwNP complicated by bronchial asthma (CRSwNP + BA), including the cases with asthmatic triad (CRSwNP + intolerance to NSAIDs + BA). The patients were classified on the basis of their clinical characteristics and severity of the course of the disease. Interleukin-1β (IL-1β) and IL-8 concentrations in nasal secretions were determined by enzyme-linked immunosorbent assay (ELISA). To assess functional activity of neutrophils, a lysosomal cationic test was used on the smears from mucous surface of inferior turbinate. Histological examination of the polypous tissue biopsies was performed in slices stained with Carazzi’s hematoxylin and eosin. IL-1β and IL-8 location in the polypous tissue were detected by indirect immunohistochemistry. In all groups of the patients, IL-1β and IL-8 concentrations exceeded those in the control group. The levels of IL-1β in the groups with CPRSwNP, CRSwNP + BA were significantly increased as compared with the CRSwNP group. IL-8 concentrations in the CRSwNP and CPRSwNP groups were significantly higher than in the CRSwNP + BA group. When analyzing antimicrobial function of neutrophils, the decreased average values of cytochemical coefficient were shown in CPRSwNP and CRSwNP + BA groups, compared with the control group and CRSwNP. In all clinical forms of CPMS, complex histopathological changes were observed, including leukocyte infiltration, fibrosis, edema, and collagen depositions. In addition, the integrity of epithelial layer was found to be damaged in polyposis, epithelial metaplasia is detected as well as increased mucus production. These disorders lead to a decrease in muco-ciliary clearance in nasal cavity. The most significant pathomorphological changes occur in CRSwNP + BA, especially in cases of asthmatic triad. According to immunohistochemical data, in various forms of CRSwNP, IL-1β- and IL-8-positive leukocytes, predominantly macrophages, are detected in the polypous tissue both subepithelially and in the connective tissue stroma of the polyps. Changed concentrations of pro-inflammatory cytokines in nasal secretion of the patients, altered antimicrobial activity of mucosal neutrophils, and characteristic pathomorphological disorders in polypous tissue of patients with CRSwNP are associated with severity of inflammatory process and clinical course of the disease. The results obtained are essential to understanding the mechanisms of pathogenesis in various subgroups of CRSwNP, assessing severity of the disease and efficiency of the treatment applied

Cytokine Gene Polymorphisms in Chronic Adenoiditis

The aim of our research was to study the multiphase response in a system of pro-inflammatory and anti-inflammatory cytokines due to the additive contribution of homozygous and heterozygous genotypes for the polymorphic allelic variants of the interleukin-1β (IL-1β) and interleukin-4 (IL-4) genes in patients with chronic adenoiditis (CA). Materials and Methods: The study included 388 children with CA. Associations between the IL1B gene (rs1143634) (C+3954T) SNP and the IL-4 gene (rs2243250) (C-589T) SNP and the clinical manifestations and clinical outcome of CA were investigated. Genotyping for the studied SNPs was performed using real-time PCR. The study of genotype-associated cytokine production in accordance with the level of concentration of IL-1β, IL-4 in blood serum with the method of solidphase EIA using horseradish peroxidase as an indicating enzyme was carried out. Results: The presence of homozygous or heterozygous genotypes of the studied SNPs of the IL-1β and IL-4 genes was characterized with genetically determined cytokine-production forming the phenotypical polymorphism. The conducted research into congenital immunity factors with an assessment of genetically determined cytokine production has revealed 5 options of the cytokine response and their corresponding frequencies. We extrapolated the results on clinical and functional outcomes of chronic adenoiditis, which allowed us to identify non-randomness in the nature of chronic adenoiditis as a multifactorial disease. Conclusion: The obtained data are evidence of the phenotypic-genetic heterogeneity of CA

PREPARATION AND PROPERTIES OF MURINE ANTI-IDIOTYPIC MONOCLONAL ANTIBODIES RECOGNIZING PRIMARY RABBIT POLYCLONAL ANTIBODIES AGAINST MORPHINE DERIVATIVES

Anti-idiotypic antibodies (Ab2), according to the network theory of Jerne, are second-generation immunoglobulins that are produced against the idiotype of an antibody to a specific antigen. Despite the large number of works devoted to the study of the properties of these proteins, their role in the regulation of the immune system is not fully known. It may consist in maintaining or blocking a minimal immune response to the antigen. The study of Ab2 is of great practical and scientific importance. The special properties of Ab2, namely, the ability to partially reproduce the structure of the primary antigen and, upon immunization, induce the appearance of tertiary antibodies, which, like first-generation antibodies, can bind to the antigen, have found application in the development of Ab2-based vaccines, in particular, for the treatment of tumors. In view of the presence of a number of limitations on research related to psychoactive substances, the development of Ab2- based vaccines against drug addiction also seems promising. To example, anti-idiotypic antibodies obtained for this purpose possessing a cocaine-like structure are described in the literature. In this work, murine monoclonal anti-idiotypic antibodies (mAb2) mimicking the structure of various morphine derivatives were obtained. Rabbit polyclonal antibodies to the 6-hemisuccinyl derivative of morphine conjugated with bovine serum albumin isolated by affinity chromatography were used as primary antibodies for immunization. Four hybridoma clones were obtained as a result of the fusion of immunized mice lymphocytes with mouse Sp2/0 mouse myeloma cells by the Milstein-Köhler method. After growth in animals, mAb2 produced by hybridoma cells were affinity purified. We investigated the physicochemical and antigenic properties of the isolated antibodies. It was shown that the obtained mAb2 differ in immunological specificity, competing in different degree with morphine derivatives for binding to first-generation antibodies. We tested the possibility of using the obtained mAb2 as antigen analogues in the solid-phase enzyme-linked immunosorbent assay to determine the titer of primary antibodies against morphine in the blood serum of laboratory animals immunized with morphine derivatives. Based on the obtained anti-idiotypic antibodies, it is proposed to develop test systems to determine the serum opiate-specific antibodies in people after specific vaccination for therapeutic or prophylactic purposes to avoid the use of drugs as antigens immobilized on the solid phase in the analysis