[The interdisciplinary approach as tool of pharmaceutical education of children: from theory to practice].

The pharmaceutical literacy is a necessary element of ensuring quality of human life that is to be formed at early age. The article demonstrates that key direction of development of health literacy is pharmaceutical education involving pharmaceutical workers. The necessity of development of pharmaceutical literacy in children through involvement into process of pharmaceutical education pedagogues and parents/legal representatives of child. The article presents analysis of normative legal documents regulating strategic directions of state and international policy in the field of protection of health and rights of minor citizen/children that regulate organization of pharmaceutical and educational activities and requirements to pharmaceutical and pedagogical workers within the framework of their professional role. The problematic zones in organization of pharmaceutical counseling of minors citizen were discovered. The necessity to improve professional competence of pharmaceutical and pedagogical workers in organization of pharmaceutical education of children of preschool and school age is established. The results of sociological survey of minor citizen and their parents demonstrated inadequate level of pharmaceutical literacy of respondents. On the basis of research results structural model of interaction of participants of pharmaceutical education of children (pharmaceutical workers - parents - pedagogues). The communication relations at the stage of transferring pharmaceutical knowledge to minor personality were revealed. The main result of the study is original structural functional model of organization of pharmaceutical education of children implementing interdisciplinary approach in forming pharmaceutical knowledge in children of preschool and school age. The stages of interaction of participants and professional tasks of pharmaceutical and pedagogical specialists in process of teaching children skills of pharmaceutical safety are determined.

The Role of Cytokines in Maintaining the Dynamics of Cell-Cell Interaction between Natural Killer Cells and Trophoblast Cells.

We analyzed the effect of individual cytokines that are secretory products of placenta typical of the uteroplacental bed. The proinflammatory cytokines IL-6, IFNγ, and IL-1ß increased the expression of TGFßR2 molecule by trophoblast cells, while VEGF and PLGF increased the expression of CD45, CD29, and CD54 adhesion molecule by trophoblast cells. The antiinflammatory cytokine IL-4 increased LeptinR expression by trophoblast cells. PMA and TNFα also enhanced the adhesion of NK cells to trophoblast cells. Our findings suggest that NK cells involved CD11a, CD11b, and CD18 molecules during their transmigration through trophoblast, as well as during their transendothelial migration.

Methodological Approaches to Assessing the Size and Morphology of Microvesicles of Cell Lines.

Morphological properties and the size of microvesicles were assessed using atomic force microscopy, electron microscopy, and granulometric analysis. As these methods require significant numbers of microvesicles, we chose microvesicles derived from cell lines for our research.

Role of Caspases in the Cytotoxicity of NK-92 Cells in Various Models of Coculturing with Trophoblasts.

Studies of interactions between natural killer (NK) cells and trophoblasts and identification of conditions for the NK cells to perform their cytotoxic function are of fundamental and practical importance for understanding their role in the development of pathological processes and complications during pregnancy. In this study, we examined changes in the content of caspases and studied activation of these enzymes in Jeg-3 trophoblasts in various models of their coculturing with NK-92 cells and demonstrated the necessity of direct contact between these cell populations for the activation of caspase-8 and caspase-3 in the trophoblasts. Contact coculturing of the two cell lines resulted in the appearance of the cytotoxic protein granzyme B in Jeg-3 cells that was accompanied by a decrease in the content of this enzyme in NK-92 cells. Distant coculturing of NK-92 and Jeg-3 cells did not trigger initiator and effector caspases characteristic for the apoptosis development in Jeg-3 cells. The observed decrease in the content of procaspases in the trophoblasts may be associated with alternative non-apoptotic functions of these enzymes.

Interaction of NK Cells, Trophoblast, and Endothelial Cells during Angiogenesis.

We studied changes in angiogenesis during contact interaction of natural killer cells and endothelial cells in the presence of secretory products of trophoblast cells activated by various cytokines. Activated trophoblast regulates angiogenesis by producing soluble factors that affect endothelial cells either directly or indirectly through activation of proangiogenic activity of natural killer cells. A stimulating effect of the trophoblast supernatants activated by IL-1ß and an inhibitory effect of trophoblast supernatants activated by IL-6 and TGFß for the formation of tube-like structures by endothelial cells were revealed. During contact culturing, natural killer cells increased the length of tube-like structures formed by endothelial cells. The trophoblast activated by IL-1ß affects angiogenesis both directly through the production of proangiogenic factors and indirectly through activation of the proangiogenic potential of natural killer cells. Trophoblast activated by IFNγ affects angiogenesis only by stimulating the proangiogenic potential of natural killer cells. Under conditions of contact interaction of natural killer cells and endothelial cells, soluble factors of trophoblast activated by IL-6 or TGFß attenuated the angiogenesis-stimulating effect of natural killer cells.

Cytotoxic Activity of Peripheral Blood NK Cells towards Trophoblast Cells during Pregnancy.

We evaluated cytotoxic activity of peripheral blood NK cells towards trophoblast cells. NK cells either isolated or in the composition of mononuclear cell fraction, caused death of trophoblast cells. In women with physiological pregnancy, the cytotoxic effect of NK cells present in mononuclear cell fraction preincubated with IL-2 was lower than in nonpregnant women, who had never been pregnant previously, and in fertile women. Cytotoxic activity of isolated NK cells preincubated with IL-2 in fertile women was lower than in nonpregnant women, who had never been pregnant previously.

Interactions of NK Cells and Trophoblast Cells. Methodological Aspects.

NK cells present in different organs differ by their functional characteristics, in particular, proliferative activity. For studying tissue-resident NK cells, tissue-specific microenvironment should be reproduced. In case of decidual NK cells, this microenvironment is created by trophoblast cells. We developed a method for evaluation of proliferative activity of peripheral blood NK cells in the presence of trophoblast cells. Proliferative activity of peripheral blood NK cells was evaluated by the expression of protein Ki-67 after culturing with JEG-3 trophoblast cells. This method allows evaluating the functional state of NK cells in microenvironment specific for the decidua.

Effect of Cytokines on the Formation Tube-Like Structures by Endothelial Cells in the Presence of Trophoblast Cells.

Despite ample data on cytokine secretion in the uteroplacental interface, the influence of microenvironment cells, in particular, trophoblast cells on angiogenesis and the role of cytokines in this process remain poorly studied. We studied the influence of cytokines on the formation of tube-like structures by endothelial cells in the presence of trophoblast cells and showed that trophoblast cells suppressed the angiogenic potential of endothelial cells. Antiangiogenic cytokines IFN-γ, IL-10, TNF-α, and TGFß via modulation of trophoblast cells stimulated the formation of tube-like structures by endothelial cells. In the co-culture of endothelial and trophoblast cells, the effects of cytokines changed and they gained additional regulatory functions.

[Comparative research into sensitivity and specificity of immune-enzyme analysis with chemiluminescence and colorimetric detection for detecting antigens and antibodies to avian influenza viruses and newcastle disease].

The goal of this work was to demonstrate the results of the development of the enzyme-linked immunosorbent tests with chemiluminescence detection and colorimetric detection of specific viral antigens and antibodies for identifying the avian influenza and the Newcastle disease viruses: high sensitivity and specificity of the immuno- chemiluminescence assay, which are 10-50 times higher than those of the ELISA colorimetric method. The high effectiveness of the results and the automation of the process of laboratory testing (using a luminometer) allow these methods to be recommended for including in primary screening tests for these infectious diseases.

Detection of microparticles of leukocytic origin in the peripheral blood in normal pregnancy and preeclampsia.

Microparticles are microvesicles forming during cell activation and as a result of apoptotic cell death. Normal pregnancy is associated with apoptosis induction in active immune system cells, present in the decidual tissue. Preeclampsia is associated with activation of the peripheral blood leukocytes and more intense apoptosis of the trophoblast cells. As a result, the number of microparticles in the peripheral blood is changing in normal gestation and in preeclampsia. The content of the leukocytic microparticles in the peripheral blood is evaluated in normal pregnancy and preeclampsia. The content of neutrophilic and monocytic microparticles is higher than normally in preeclampsia, this indicating activation of these cells. The number of microparticles formed by NK cells is low in preeclampsia, which can reflect the incompetence of immunological tolerance mechanisms under these conditions.

[Effects of oxytocin pathway gene polymorphisms and adverse childhood experiences on emotion recognition in schizophrenia spectrum disorders]. / Effekty polimorfizma genov oksitotsinergicheskogo puti i neblagopriyatnogo detskogo opyta na raspoznavanie emotsii pri rasstroistvakh shizofrenicheskogo spektra.

OBJECTIVE: To study a role of the interaction of oxytocin pathway gene polymorphisms and adverse childhood experiences (ACE) in facial emotion recognition (FER) deficits in schizophrenia. MATERIAL AND METHODS: Patients with schizophrenia spectrum disorders (n=699) completed cognitive testing, which included a FER task. We determined patients' genotypes for common polymorphisms in three of the oxytocin pathway genes which were previously associated with face perception: OXTR (rs53576, rs7632287), CD38 (rs3796863) and ARNT2 (rs4778599). The presence of ACE in the patient's history was assessed via an analysis of medical records. RESULTS: In our sample, 49% of participants experienced ACE. ANCOVA adjusted for age and gender revealed a significant interaction effect of OXTR rs53576 with ACE on FER scores (F=11.51; p<0.001; η2p=0.02). The effect remained significant when accounting for cognitive functioning and negative symptoms. Carriers of the A allele without ACE recognized emotions worse than GG homozygotes without ACE (p=0.039) and carriers of the A allele with ACE (p=0.009). CONCLUSION: The results are consistent with the notion of the A (rs53576) allele's role in sensitivity to childhood experiences that influence the psychosocial development and can be used in further studies of the oxytocin treatment of social cognition and social adaptation of patients with schizophrenia.

[To the problem of somatic-hypochondriac paranoia]. / K probleme somaticheskoi-ipokhondricheskoi paranoii.

OBJECTIVE: To unite within the framework of a single clinical entity (based on the model of hypochondriacal paranoia) phenomena of the somatopsychotic and hypochondriacal range, which, in accordance with modern systematics, are classified as various categories of psychosomatic, affective disorders and personality disorders. MATERIAL AND METHODS: The sample for analysis consisted of 29 patients (with the diagnosis of delusional disorder (ICD-10; F22.0 in ICD-10), 10 men (34.5%) and 19 women (64.5%), the average age was 42.9±19.9 years; men - 10 nab. (34.5%), women - 19 nab. (64.5%). The average duration of the disease iswas 9.4±8.5 years. The psychopathological method was used as the main one. RESULTS: The article forms an alternative concept of somatic paranoia based on the model of hypochondriacal paranoia. The fundamental difference between the construct of somatic paranoia is an obligate connection between somatopsychic and ideational disorders. Somatopsychic (coenesthesiopathic) symptoms do not exist as an independent (equivalent to the structure of somatic clinical syndromes) dimensions and are formed exclusively with the participation of ideational phenomena. CONCLUSION: In accordance with the presented concept, coenesthesiopathic symptoms within the framework of somatic paranoia act as a somatic equivalent of delusional disorders.

[Associations between the oxytocinergic system genes, perinatal complications and interpersonal relationships in patients with schizophrenia]. / Issledovanie svyazi mezhdu genami oksitotsinergicheskoi sistemy, perinatal'nymi oslozhneniyami i formirovaniem mezhlichnostnykh otnoshenii u bol'nykh shizofreniei.

OBJECTIVE: To search for the associations between genes of the oxytocinergic pathway and psychosocial functioning in schizophrenia, namely, the ability of schizophrenic patients to form interpersonal relationships, taking into account the influence of such an environmental factor as perinatal complications. MATERIAL AND METHODS: The study included 383 people (140 women and 243 men, mean age 32.6±11.4 years), of whom 107 had a history of perinatal complications, and 276 did not. Psychosocial functioning was assessed using the Personal and social relationships domain of The Personal and Social Performance scale (PSP). Polymorphisms rs53576, rs4686302, rs1042778 in the oxytocin receptor gene (OXTR) and polymorphism rs3796863 in the transmembrane glycoprotein (CD38) gene were genotyped. RESULTS: There is the association between the OXTR rs53576 polymorphism and scores on the interpersonal relations domain (p=0.005). Significant differences are found between carriers of the GG genotype and carriers of the A allele (p=0.003). In the group without perinatal complications, the genotype does not have a significant effect on PSP score. There are no associations between other polymorphisms and the level of interpersonal relationships in any of the studied groups. CONCLUSION: The results are in accordance with the notions accepted on the basis of numerous evidences that link the genes of the oxytocinergic system with social behavior. We obtained new data on the influence of the known polymorphism OXTR rs53576 on the phenotype, which has not been studied previously in this aspect - the ability to form interpersonal relationships in patients with schizophrenia, while it was shown that the effect of the genotype depends on the environmental risk factor (perinatal complications).

Trophoblast cell influence on peripheral blood natural killer cell proliferation and phenotype in non-pregnant women and women in early pregnancy.

Natural killer (NK) cells are the main population of leukocytes in decidua during the first trimester of pregnancy. NK cells can have contact with trophoblast cells during pregnancy, which raises the possibility of mutual influence. This research aimed to evaluate the proliferation and phenotype of peripheral blood NK cells in the presence of trophoblast cells of the JEG-3 cell line. We showed that trophoblast cells of the JEG-3 cell line (American Type Culture Collection (ATCC), USA) produced TGFß. However, co-culturing of NK and trophoblast cells did not change the SMAD2/3 to pSMAD2/3 ratio within NK cells. These data indicate that the canonical signaling pathway from TGFß is not activated, but do not preclude activation of SMAD-independent signaling pathways through the effect of TGFß and/or other cytokines. We established that trophoblast cells inhibited both constitutive and IL-2-induced expression of Ki-67 proliferation marker by NK cells in vitro in both pregnant and non-pregnant women. Constitutive and induced Ki-67 expression by peripheral blood NK cells was increased in pregnant women compared with non-pregnant women. The influence of trophoblast cells on Ki-67 expression by NK cells was more pronounced in the presence of other mononuclear cells than in their absence. In the presence of trophoblast cells and IL-2, the number of NK cells with the CD16+CD57- phenotype in peripheral blood mononuclear cells (PBMCs) was increased in pregnant and non-pregnant women, compared with culturing with IL-2 only. This might reflect a decrease in the number of NK cells at the terminal stage of differentiation. We also revealed the increased content of NK cells with the CD16-CD56bright phenotype in PBMCs of pregnant women when incubated with trophoblast cells and IL-2, compared with culturing with trophoblast cells only. Our results suggest that NK cells need contact interactions with trophoblast cells and additional cytokine stimulation (IL-2, cytokines of other mononuclear cells) to acquire the CD56bright phenotype.

Changes in expression of Ki-67, CD16 and CD56 by natural killer cells from peripheral blood mononuclear cells in the setting of recurrent miscarriage after in vitro culturing in the presence of trophoblast cells and IL-2.

The aim of this research was to assess the proliferative activity of Natural Killer Cells (NK cells) from Peripheral Blood Mononuclear Cells (PBMCs) in the presence of trophoblast cells in women with a history of recurrent miscarriages. We examined the peripheral blood of women with recurrent miscarriage in the proliferative (n = 12) or secretory (n = 13) phase of their menstrual cycle, and pregnant women with a history of recurrent miscarriage at 6-7 weeks of their current pregnancy (n = 14). Controls were fertile non-pregnant women in the proliferative (n = 11) or secretory (n = 13) phase of their menstrual cycle, and pregnant women at 6-7 weeks of a physiologically normal pregnancy (n = 20). We used IL-2 as a factor maintaining PBMCs viability during long-term culturing. We established that culturing in the presence of IL-2 contributed to an increase in the number of CD56+CD16- NK cells and to a decrease in the number of CD56+CD16+ NK cells from PBMCs compared with these numbers before culturing in both healthy women and in women with recurrent miscarriage. After culturing of PBMCs in the presence of trophoblast cells and IL-2 (compared with culturing without trophoblast cells), the intensity of Ki-67 expression by NK cells was reduced in the whole NK cell population (CD3-CD56+), and in the CD56+CD16- and CD56+CD16+ populations of NK cells in women with recurrent miscarriage and in healthy controls. The intensity of CD56 expression was reduced in the presence of trophoblast cells and IL-2 in non-pregnant women with recurrent miscarriage in the secretory versus the proliferative phase of the menstrual cycle.

[Molecular genetic characteristics of the strain A/chicken/Moscow/2/2007 (H5N1) strain from a epizootic focus of highly pathogenic influenza A among agricultural birds in the neear-Moscow region (February 2007)].

Among agricultural birds in the near-Moscow Region (February 2007), local epizootics caused by the highly pathogenic avian influenza A/H5N1 virus seem to be of unintended manual origin. Such a situation may be considered to be model when the source of inoculation is elucidated in cases of potentially possible acts of bioterrorism. Molecular genetic analysis of isolated A/chicken/Moscow/2/2007 strain established its genetic similarity with the highly pathogenic strains detected in the Black-and-Caspian Sea region in 2006. At the same time, comparison of nucleotide sequences of the strain A/chicken/Moscow/2/2007 with the strains of Qinghai-Siberian genotype (CSG) for which the sequences of full-sized genomes are known in the international databases revealed a significant distinction of the near-Moscow strain from the earlier known analogues. The uniqueness of the primary structure of the PB1 gene is shown. The paper discusses the functional value of amino acid substitutions in the proteins of the strain A/chicken/Moscow/2/2007 and in other variants of CSG of the subtype H5N1.

[Identification and characterization of strains of Bacillus thuringiensis by genomic fingerprinting using biotinylated phage M13 DNA]. / Identifikatsiia i pasportizatsiia shtammov Bacillus thuringiensis metodom genomnoi daktiloskopii s ispol'zovaniem biotinilirovannoi DNK faga M13]

Genomic DNA of the entomopathogenic bacterium Bacillus thuringiensis was analyzed by the genomic fingerprinting technique. The biotin-labeled single-stranded DNA of the phage M13 was used as a marker of hypervariable sequences. A procedure for analyzing the differentiation among various Bacillus thuringiensis strains was developed. Characteristic patterns of fingerprints were obtained for several strains, the main representatives of subspecies that are most frequently used in the manufacture of bacterial insecticides, such as subsp. thuringiensis, subsp. kurstaki, and subsp. galleriae. Because no essential differences were revealed in band patterns upon comparing fingerprints of crystal-producing bacterial strains with those of acrystallic mutants, it was assumed that the loss of crystal-producing ability in the insect pathogen Bacillus thuringiensis is not connected with significant rearrangement of its genome.

[Long remissions in schizophrenia following single and serial episodes in adolescence (clinico-catamnestic study)]. / Dlitel'nye remissii pri shizofrenni posle odnokratnykh i seriinykh pristupov v iunosheskom vozraste (kliniko-katammesticheskoe issledovanie).

The report is related to a study of 56 cases of schizophrenia with protracted remissions, following acute psychotic attacks in adolescence. A relatively high frequency of such attacks was established in adolescent attack-like schizophrenia (26.9%). It was possible to determine the clinical traits of such cases and the differences in the premorbid personality. The conclusion is made on the necessity of differentiated evaluation of the criteria of social and working prognosis in such cases.

[Features of the clinical picture and course of ambulatory schizophrenia with onset in young adulthood (clinico-catamnestic study)]. / Osobennosti kliniki i techeniia ambulatornoi shizofrenii nachavsheisia v iunosheskom vozraste (kliniko-katamnesticheskoe issledovanie).

From a population of epidemiologically examined schizophrenics of three Moscow regions a group of 96 patients was selected. These patients have fallen ill in adolescence and youth and have never been hospitalized after their primary dispensary examination. A catamnestic examination of them carried out in 10 to 15 years has shown that the "ambulatory" schizophrenia of the juvenile age is characterized by prevalence of neurosis-like and effaced forms, and takes a course mainly in the form of lingering atypical pubertal "schube" (the so-called "state of juvenile asthenic inadequacy"). This state ends in regression of the symptoms and in the patients' social and working re-adaptation, as the juvenile age is past, however, shallow schizophrenic changes still remain. The question on the expedience of extramural treatment of such patients and expansion of indications for their social and working rehabilitation is discussed.