Emocionalna kompetencija i simptomi posttraumatskoga stresnog poremećaja kod veterana Domovinskog rata

This study examined the relationship between four factors of posttraumatic stress disorder symptoms – re-experiencing, avoidance, dysphoria and hyperarousal – and particular domains of emotional competence and emotional regulation and control among 215 Croatian Homeland War veterans. Cross-sectionally, emotional competence subscales – perceiving and understanding emotions, expressing and labelling emotions, managing and regulating emotions – were associated with dysphoria symptom cluster only, while emotional regulation and control subscales – influence of emotion and mood on memory and emotional reaction control – were associated with dysphoria and re-experiencing symptom clusters. The results of this study are consistent with the view that successful recovery from trauma requires adaptive emotion competence skills and that therefore difficulties in dealing with emotions (understanding, expressing or regulating) are a risk factor for the development and/or maintenance of posttraumatic stress disorder symptoms. It appears that interventions organized toward improving emotional competence and regulation may be useful as complementary or independent treatments for combat-related posttraumatic stress disorder.U ovom istraživanju ispitivali smo povezanost četiriju faktora posttraumatskoga stresnog poremećaja – ponovno proživljavanje traume, izbjegavanje, disforija i pojačana pobuđenost – i određenih aspekata emocionalne kompetencije i emocionalne regulacije i kontrole na uzorku od 215 branitelja Domovinskog rata. Podskale emocionalne kompetencije – percipiranje i razumijevanje emocija, izražavanje i imenovanje emocija, upravljanje i reguliranje emocijama – povezane su sa simptomima faktora disforije, koji uključuju ograničeni afekt, poteškoće u spavanju, osjećaj uskraćene budućnosti, otežano koncentriranje i iritabilnost, dok su podskale emocionalne regulacije i kontrole – utjecaj emocija i raspoloženja na pamćenje i kontrola emocionalnih reakcija – povezane sa simptomima disforije i simptomima ponovnoga proživljavanja traume, npr. aktualna i intruzivna uznemirujuća prisjećanja, aktualni uznemirujući snovi, ponašanja ili osjećaji kao da se događaji ponovno zbivaju te intenzivni psihološki distres. Rezultati istraživanja sugeriraju da su poteškoće u procesiranju i reguliranju emocija mogući rizični faktor za razvoj i/ili održavanje simptoma posttraumatskoga stresnog poremećaja

COPING STRATEGIES IN WAR VETERANS 20 YEARS AFTER THE EXPOSURE TO EXTREME STRESS

Many soldiers encounter difficulties while transitioning from military to civilian life. Such severe traumatic events may also have long-term effects. Previous studies have shown a strong relationship between coping strategies and posttraumatic stress disorder (PTSD) symptoms. The aim of this study was to investigate how veterans who were exposed to war trauma 20 years ago now deal with everyday life stressors, and how their current coping strategies relate to the four-factor model of PTSD. A total of 220 male Croatian Homeland War Veterans between the ages of 38 and 75 participated. Questionnaires included Combat Exposure Scale, Posttraumatic Stress Disorder Checklist – Military Version and Ways of Coping Questionnaire. Results showed positive association between dysphoria and escape-avoidance coping strategy and negative association between dysphoria and positive reappraisal coping strategy. Given that dysphoria symptoms are associated with the chronicity of PTSD and poorer response to PTSD therapy treatment in war veterans, our results underscore the importance of treating dysphoria symptoms and promoting engagement coping strategies for this population

The Role of Extracellular Vesicles and PIBF in Embryo-Maternal Immune-Interactions

Pregnancy represents a unique immunological situation. Though paternal antigens expressed by the conceptus are recognized by the immune system of the mother, the immune response does not harm the fetus. Progesterone and a progesterone induced protein; PIBF are important players in re-adjusting the functioning of the maternal immune system during pregnancy. PIBF expressed by peripheral pregnancy lymphocytes, and other cell types, participates in the feto-maternal communication, partly, by mediating the immunological actions of progesterone. Several splice variants of PIBF were identified with different physiological activity. The full length 90 kD PIBF protein plays a role in cell cycle regulation, while shorter splice variants are secreted and act as cytokines. Aberrant production of PIBF isoforms lead to the loss of immune-regulatory functions, resulting in and pregnancy failure. By up regulating Th2 type cytokine production and by down-regulating NK activity, PIBF contributes to the altered attitude of the maternal immune system. Normal pregnancy is characterized by a Th2-dominant cytokine balance, which is partly due to the action of the smaller PIBF isoforms. These bind to a novel form of the IL-4 receptor, and induce increased production of IL-3, IL-4, and IL-10. The communication between the conceptus and the mother is established via extracellular vesicles (EVs). Pre-implantation embryos produce EVs both in vitro, and in vivo. PIBF transported by the EVs from the embryo to maternal lymphocytes induces increased IL-10 production by the latter, this way contributing to the Th2 dominant immune responses described during pregnancy

Osteopontin–metallothionein I/II interactions in experimental autoimmunune encephalomyelitis

Osteopontin (OPN), an extracellular matrix (ECM) glyco-phosphoprotein, plays an important role in autoimmune-mediated demyelinating diseases, including multiple sclerosis and experimental autoimmune encephalomyelitis (EAE). As an integrin and CD44 binding protein it participates in bidirectional communication between the ECM and target cells and affects transduction pathways that maintain neuronal and immune cell homeostasis. Its biological activity is also heavily influenced by microenvironment, which stimulates the cleavage of OPN and changes its functions. In this study we estimated the expression profile of OPN in neural tissues of DA rats during the first relapse of chronic relapsing EAE and investigated the relationship of OPN to metallothionein I+II (MTs), which play pivotal role in zinc-related cell homeostasis and in protection of CNS against cytokine-induced injury. The data showed that in EAE rats OPN mRNA and protein levels increased concurrently with the transcription of MTs and that within the spinal cord (SC) lysates EAE-afflicted rats had a higher content of OPN fragments of low molecular weight than untreated and CFA-treated rats. The expression of OPN and MTs was upregulated on ependymal, lymphoid and astroglial cells and on multiple αvβ3+ neurons in SC and in the brain (cortex, white matter, hippocampus, and cerebellum). Besides, multiple cells co-expressed OPN and MTs. Granular OPN signals were detected in secretory vesicles of Golgy (αvβ3 neurons) and in patches adjacent to the plasma membrane (subventricular zone). The findings imply that in demyelinating lesions are generated proteolytic OPN fragments and that OPN/MT interactions contribute to tissue remodeling during an autoimmune attac

The Role of Extracellular Vesicles and PIBF in Embryo-Maternal Immune-Interactions

Pregnancy represents a unique immunological situation. Though paternal antigens expressed by the conceptus are recognized by the immune system of the mother, the immune response does not harm the fetus. Progesterone and a progesterone induced protein; PIBF are important players in re-adjusting the functioning of the maternal immune system during pregnancy. PIBF expressed by peripheral pregnancy lymphocytes, and other cell types, participates in the feto-maternal communication, partly, by mediating the immunological actions of progesterone. Several splice variants of PIBF were identified with different physiological activity. The full length 90 kD PIBF protein plays a role in cell cycle regulation, while shorter splice variants are secreted and act as cytokines. Aberrant production of PIBF isoforms lead to the loss of immune-regulatory functions, resulting in and pregnancy failure. By up regulating Th2 type cytokine production and by down-regulating NK activity, PIBF contributes to the altered attitude of the maternal immune system. Normal pregnancy is characterized by a Th2-dominant cytokine balance, which is partly due to the action of the smaller PIBF isoforms. These bind to a novel form of the IL-4 receptor, and induce increased production of IL-3, IL-4, and IL-10. The communication between the conceptus and the mother is established via extracellular vesicles (EVs). Pre-implantation embryos produce EVs both in vitro, and in vivo. PIBF transported by the EVs from the embryo to maternal lymphocytes induces increased IL-10 production by the latter, this way contributing to the Th2 dominant immune responses described during pregnancy

Mechanism of progesterone action in embryo implantation during early pregnancy in mice

Cilj istraživanja: Implantacija razvojno kompetentnog embrija u receptivan uterus je ključan proces tijekom uspostavljanja trudnoće u sisavaca. Prema današnjim saznanjima smatra se da je nepravilna implantacija uzrok 75 % neuspjelih trudnoća. Aktivnost steroidnih hormona, estrogena (E) i progesterona (P), preko njihovih jezgrinih receptora je ključna za proces implantacije. Mehanizam djelovanja E-a, a pogotovo P-a tijekom rane trudnoće još nije dovoljno razjašnjen. Glavni cilj ovog istraživanja bio je utvrditi ulogu progesteronskog receptora (PR) u mehanizmu implantacije blastociste in vivo. Ispitala sam: 1. vremenski i prostorni odnos između PR-a i njime reguliranih gena: Hand2 i Cox2 te ER i FGF9 u uterusu miša tijekom peri-implantacijskog razdoblja, 2. prostorne promjene izražaja PR-om reguliranih gena tijekom implantacije, 3. promjene izražaja Hand2, Cox2, FGF9 i PR gena u implantacijskom mjestu u odnosu na ne-implantacijsko mjesto i 4. regulaciju izražaja Hand2 i FGF9 s P-om i E-om u ovarijektomiziranim mišicama. Materijali i metode: Kao model za istraživanje mehanizama uključenih u trudnoću koristila sam BALB/c miševe. Metodom imunofluorescencije na rezovima tkiva odredila sam prostorni i vremenski raspored PR-a, ERα, COX2, HAND2 i FGF9, kao i njihov međusobni odnos tijekom rane trudnoće. Metodu lančane reakcije polimeraze u stvarnom vremenu (RT-PCR) koristila sam za određivanje izražaja Hand2 i FGF9 gena tijekom rane trudnoće. Razliku izražaja P-om reguliranih gena na implantacijskom u odnosu na ne-implantacijsko mjesto sam odredila RT-PCR metodom. Promjene izražaja bjelančevina Hand2 i FGF9 tijekom rane trudnoće odredila sam western blot metodom. Da bi odredila regulaciju izražaja Hand2 i FGF9 u uterusu koristila sam ovarijektomizirane WT mišice tretirane steroidnim hormonima. Ulogu PR izoformi u izražaju Hand2 u uterusu odredila sam koristeći ovarijektomizirane PRAKO i PRKO mišice tretirane egzogenim E-om i P-om. Rezultati: Tijekom peri-implantacijskog razdoblja dinamika izražaja i odnos PR-a i njime reguliranih gena, Hand2, COX2, ERα i FGF9 mijenja se tijekom procesa rane trudnoće u miševa i strogo je stanično specifičan. Transkripcijska aktivnost PR-a je neophodna za izražaj Hand2 i Cox2 u specifičnoj populaciji stromalnih stanica na implantacijskom mjestu. Na implantacijskom mjestu izražaj Hand2 i COX2 je značajno povećan u usporedbi s ne-implantacijskim mjestom. Izražaj Hand2 u mjestu je utišana. Ovim istraživanjem je po prvi puta pokazano da je FGF9 važan čimbenik u uspostavljanju komunikacije između kompetentne blastociste i receptivnog uterusa. Zaključak: Precizno usklađeni mehanizmi koje reguliraju P i E, preko PR-a i ERα su ključni za stvaranje pogodnog okoliša za implantaciju embrija. Transkripcijski čimbenici, PR-a i Hand2 zajedno potiču diferencijaciju stromalnih stanica na mjestu implantacije. Ta populacija stanica je mjesto biosinteze prostaglandina. Prostaglandini su medijatori angiogeneze i decidualizacije u uterusu. Čimbenik rasta, FGF9 je važan za proces implantacije i uspostavljanje trudnoće u miša.Aim of the study: Implantation of a developmentally competent embryo in a receptive uterus is an essential process during the establishment of pregnancy in mammals. Based on the current knowledge, it is assumed that improper implantation is the cause of 75% of failed pregnancies. Synchronized activity of steroid hormones estrogen (E) and progesterone (P) through their nuclear receptors is the key for successful pregnancy. Mechanisms of E and particularly P-regulated pathways during implantation still need to be clarified. The aim of this research was to determine progesterone receptor’s (PR) role in the establishment of pregnancy in vivo using a mouse model organism. I investigated: 1. spatiotemporal relationship between PR and its targets: Hand2, COX2, ER and FGF9 during the periimplantation period; 2. spatial changes in the expression of PR-regulated genes during implantation; 3. changes in expression of Hand2, COX2, FGF9 and PR genes at the implantation site in relation to the non-implantation site and 4. steroid hormone regulation of Hand2 and FGF9 expression. Material and methods: For these studies BALB/c mice were used. Immunofluorescence analyses of uterus tissue obtained from pregnant mice were used to study the spatiotemporal relationship between PR and PR-regulated genes during peri-implantation. Changes in the expression of PR regulated genes at the implantation site in relation to the non-implantation site were analyzed by using immunofluorescence on uterus tissue, as well as using quantitative real time-PCR (RT-PCR). Differences in Hand2 and FGF9 gene and protein expression during periimplantation were determined by using RT-PCR as well as by western blot. To study the regulation of Hand2 and FGF9 gene expression by steroid hormones in the uterus, ovariectomized WT mice were hormonally treated. The role of PR isoforms in Hand2 expression was studied by using E and P treated ovariectomized PRAKO and PRKO mice. Results: During peri-implantation period the expression of PR and PR regulated genes Hand2, COX2, ERα and FGF9 dynamically changes in the uterus and is highly cell specific. Transcriptional activity of PR is necessary for the expression of Hand2 and COX2 in the specific population of the stromal cells at the implantation site. At the implantation site the expression of Hand2 and Cox2 was significantly higher in comparison to the non-implantation site. The expression of Hand2 is regulated by the PR-A isoform in endometrium. Transcriptional activity of ERα is downregulated at the implantation site. For the first time, this study showed that FGF9 is an important factor for the establishment of communication between a competent blastocyst and a receptive uterus. Conclusion: Balance in P and E regulated activity, throughout their cognate receptors is crucial to establish a suitable environment for embryo implantation in mice. The interaction of transcription factors PR and Hand2 is necessary for differentiation of specific stromal cells at the site of implantation. In these cells PR and Hand2 stimulate production of prostaglandins, which are the key mediators of angiogenesis and decidualization in the uterus. Growth factor FGF9 is an important factor for implantation and the establishment of pregnancy in the mice

Spatial and Temporal Analyses of FGF9 Expression During Early Pregnancy

Background: Fibroblast growth factors (FGFs), in complex with their receptors (FGFRs), regulate a broad spectrum of biological functions including cellular proliferation, survival, migration, and differentiation. In human endometrial stromal cells, FGF9 is regulated with estrogen (E). Methods/Results: First, we report that in uterus tissue of ovariectomized wild type mice, FGF9 is present in three isoforms and is regulated with E. Second, we found that during peri-implantation, Fgf9 expression reached its peak at day 4.5 of pregnancy. Immunofluorescence analyses demonstrated overlapping FGF9 and COX2 expression surrounding the blastocyst attachment site. Next, we identified FGF9- and CD31-positive cells as a part of the microvessels; however, expression was localized to a distinct population of cells. Finally, our data showed synchronized, spatial expression of FGF9 on the luminal epithelium with FGFR2 present on the trophectoderm. Conclusion: Our data suggest that FGF9 is a crucial factor required to establish the appropriate microenvironment for successful implantation and the maintenance of pregnancy

Analysis of heart and neural crest derivatives- expressed protein 2 (HAND2)-progesterone interactions in peri-implantation endometrium†

Implantation is restricted to a narrow window when the local endometrial microenvironment is supportive of the invading embryo. The ovarian steroid hormones estrogen (E) and progesterone (P) are principal regulators of uterine receptivity. Suppression of E-dependent proliferation of luminal epithelium (LE) by P is mandatory for embryo implantation. Here, we report that the balance of E receptor α (ERα) and P receptors (PR) activity controls HAND2 expression, a key transcription factor that determines the fate of the implanting embryo and thereby pregnancy outcome. As a model, we used wild-type mice as well as mice in which either both PR isoforms or the A-isoform was genetically ablated (PRKO and PRAKO, respectively). Detailed spatiotemporal analyses of PR, HAND2, and ERα expression at implantation site demonstrated co-expression of HAND2 and PR but not ERα. Furthermore, in hormonally treated ovariectomized WT, PRAKO and PRKO mice, E suppresses endometrial HAND2 expression. Adding P together with E partially rescues HAND2 expression in WT, but not PRAKO and PRKO animals. Therefore, infertility in PRAKO mice is at least in part associated with the loss of PR-A-regulated HAND2 expression

The involvement of the progesterone receptor in PIBF and Gal‐1 expression in the mouse endometrium

Problem: The progesterone-regulated genes, PIBF and Gal‐1, are key players in the feto-maternal immunological interaction. This study aims to investigate the expression of PIBF and Gal‐1 in WT and progesterone receptor KO models as well as subsequent effects of PIBF on decidualization of stromal cells. Method of the study: PRAKO, PRBKO and PRKO BALB/c mice were used for assessing the role of PR isoforms in PIBF induction. PIBF‐ and Gal‐1 mRNA expression in the uterus was tested by real‐time PCR. The effect of PIBF on decidualization of endometrial stromal cells was verified by anti-desmin immunofluorescence. Immunohistochemistry was used for testing PIBF expression in the uterus. Gal‐1, ERα and PR positive decidual NK cells were detected by immunofluorescence. Results: PIBF mRNA was significantly increased in progesterone‐treated WT mice, but not in PRKO and PRAKO mice. PIBF protein expression was reduced in the endometria of PRKO and PRAKO, but not in PRBKO mice. During a 6‐day culture, PIBF induced decidual transformation of endometrial stromal cells. PIBF expression in the mouse uterus was highest during the implantation window, while Gal‐1 mRNA expression continuously increased between day 2.5 and day 11.5 of gestation. Decidual NK cells express Gal‐1 and ERα, but not PR at day 7.5 murine pregnancy. Conclusion: PIBF produced via engagement of PRA, is highly expressed in the endometrium during the implantation window, and plays a role in decidualization. The concerted action of PIBF and Gal‐1 might contribute to the low cytotoxic activity of decidual NK cells