The association between self-image and defence mechanisms in a group of adolescent patients receiving psychiatric treatment

Aim. The aim of the study was to explore the relationship between various areas of selfimage and defence mechanisms in adolescents. The study included a division into groups according to whether or not they were receiving psychiatric treatment. Methods. Data were obtained from two groups: a clinical group (30 persons), consisting of adolescent patients of the Adolescent Inpatient Ward of the Child and Adolescent Psychiatry Clinic and a control group (40 persons), adolescents attending upper secondary school. The Defence Style Questionnaire DSQ-40 and the Offer Self Image Questionnaire were used in the study. Results. Results showed no differences, in the maturity levels of the defence mechanisms, between the two groups. Subjects from the clinical group had a significantly lower self-image of themselves than subjects from the control group.. In both groups, the use of mature defence mechanisms was accompanied by a positive self-image, while the use of less mature defence mechanisms was associated with a lower self-image. Comparison of the groups revealed different relationships between the aspects of self-image and used defence mechanisms, in particular the mechanism of projection. Number of significant correlations was greater in the clinical group. Conclusions. In the context of lower self-image, the study revealed the importance of such defence mechanisms as projection, acting out, somatization or schizoid fantasies. The obtained results seem to confirm a hypothesis that the assessment of the maturity of defence mechanisms in the period of adolescence is less clear and clinically useful

Myoblast-conditioned media improve regeneration and revascularization of ischemic muscles in diabetic mice

INTRODUCTION: Diabetes is associated with reduced expression of heme oxygenase-1 (HO-1), a heme-degrading enzyme with cytoprotective and proangiogenic properties. In myoblasts and muscle satellite cells HO-1 improves survival, proliferation and production of proangiogenic growth factors. Induction of HO-1 in injured tissues facilitates neovascularization, the process impaired in diabetes. We aimed to examine whether conditioned media from the HO-1 overexpressing myoblast cell line can improve a blood-flow recovery in ischemic muscles of diabetic mice. METHODS: Analysis of myogenic markers was performed at the mRNA level in primary muscle satellite cells, isolated by a pre-plate technique from diabetic db/db and normoglycemic wild-type mice, and then cultured under growth or differentiation conditions. Hind limb ischemia was performed by femoral artery ligation in db/db mice and blood recovery was monitored by laser Doppler measurements. Mice were treated with a single intramuscular injection of conditioned media harvested from wild-type C2C12 myoblast cell line, C2C12 cells stably transduced with HO-1 cDNA, or with unconditioned media. RESULTS: Expression of HO-1 was lower in muscle satellite cells isolated from muscles of diabetic db/db mice when compared to their wild-type counterparts, what was accompanied by increased levels of Myf5 or CXCR4, and decreased Mef2 or Pax7. Such cells also displayed diminished differentiation potential when cultured in vitro, as shown by less effective formation of myotubes and reduced expression of myogenic markers (myogenic differentiation antigen - myoD, myogenin and myosin). Blood flow recovery after induction of severe hind limb ischemia was delayed in db/db mice compared to that in normoglycemic individuals. To improve muscle regeneration after ischemia, conditioned media collected from differentiating C2C12 cells (control and HO-1 overexpressing) were injected into hind limbs of diabetic mice. Analysis of blood flow revealed that media from HO-1 overexpressing cells accelerated blood-flow recovery, while immunohistochemical staining assessment of vessel density in injected muscle confirmed increased angiogenesis. The effect might be mediated by stromal-cell derived factor-1α proangiogenic factor, as its secretion is elevated in HO-1 overexpressing cells. CONCLUSIONS: In conclusion, paracrine stimulation of angiogenesis in ischemic skeletal muscle using conditioned media may be a safe approach exploiting protective and proangiogenic properties of HO-1 in diabetes

Vitamin D Depletion in Pregnancy Decreases Survival Time, Oxygen Saturation, Lung Weight and Body Weight in Preterm Rat Offspring

Animal studies suggest a role of vitamin D in fetal lung development although not studied in preterm animals. We tested the hypothesis that vitamin D depletion aggravates respiratory insufficiency in preterm rat offspring. Furthermore, the effects of vitamin D depletion on growth and lung surfactant were investigated. Female Sprague-Dawley rats were randomly assigned low vitamin D (VDL) or control diet before mating and followed with serum 25-hydroxyvitamin D (s-25(OH)D) determinations. After cesarean section at gestational day 19 (E19) or day 22 (E22), placental weight, birth weight, crown-rump-length (CRL), oxygenation (SaO2) at 30 min and survival time were recorded. The pup lungs were analyzed for phospholipid levels, surfactant protein A-D mRNA and the expression of the vitamin D receptor (VDR). S-25(OH)D was significantly lower in the VDL group at cesarean section (12 vs. 30nmol/L, p<0.0001). Compared to the controls, E19 VDL pups had lower birth weight (2.13 vs. 2.29g, p<0.001), lung weight (0.09 vs. 0.10g, p = 0.002), SaO2 (54% vs. 69%, p = 0.002) as well as reduced survival time (0.50 vs. 1.25h, p<0.0001). At E22, the VDL-induced pulmonary differences were leveled out, but VDL pups had lower CRL (4.0 vs. 4.5cm, p<0.0001). The phospholipid levels and the surfactant protein mRNA expression did not differ between the dietary groups. In conclusion, Vitamin D depletion led to lower oxygenation and reduced survival time in the preterm offspring, associated with reduced lung weight and birth weight. Further studies of vitamin D depletion in respiratory insufficiency in preterm neonates are warranted

Characterization of Microfibrillar-associated Protein 4 (MFAP4) as a Tropoelastin- and Fibrillin-binding Protein Involved in Elastic Fiber Formation

MFAP4 (microfibrillar-associated protein 4) is an extracellular glycoprotein found in elastic fibers without a clearly defined role in elastic fiber assembly. In the present study, we characterized molecular interactions between MFAP4 and elastic fiber components. We established that MFAP4 primarily assembles into trimeric and hexameric structures of homodimers. Binding analysis revealed that MFAP4 specifically binds tropoelastin and fibrillin-1 and -2, as well as the elastin cross-linking amino acid desmosine, and that it co-localizes with fibrillin-1-positive fibers in vivo. Site-directed mutagenesis disclosed residues Phe(241) and Ser(203) in MFAP4 as being crucial for type I collagen, elastin, and tropoelastin binding. Furthermore, we found that MFAP4 actively promotes tropoelastin self-assembly. In conclusion, our data identify MFAP4 as a new ligand of microfibrils and tropoelastin involved in proper elastic fiber organization

Surfactant Protein D Deficiency Aggravates Cigarette Smoke-Induced Lung Inflammation by Upregulation of Ceramide Synthesis

Cigarette smoke (CS) is the main cause of chronic obstructive pulmonary disease. Surfactant protein D (SP-D) is an important anti-inflammatory protein that regulates host immune defense in the lungs. Here, we investigated the role of SP-D in a murine model of CS-induced inflammation. Pulmonary SP-D localization and abundance was compared between smoker and non-smoker individuals. For in vivo studies, wildtype, and SP-D-deficient mice were exposed to CS for either 12 weeks or 3 days. Moreover, the effect of therapeutic administration of recombinant fragment of human SP-D on the acute CS-induced changes was evaluated. Pulmonary SP-D appeared with heterogenous expression in human smokers, while mouse lung SP-D was uniformly upregulated after CS exposure. We found that SP-D-deficient mice were more susceptible to CS-induced macrophage-rich airway inflammation. SP-D deficiency influenced local pro-inflammatory cytokine levels, with increased CCL3 and interleukin-6 but decreased CXCL1. Furthermore, CS exposure caused significant upregulation of pro-inflammatory ceramides and related ceramide synthase gene transcripts in SP-D-deficient mice compared to wildtype littermates. Administration of recombinant fragment of human SP-D (rfhSP-D) alleviated CS-induced macrophage infiltration and prevented induction of ceramide synthase gene expression. Finally, rfhSP-D treatment attenuated CS-induced human epithelial cell apoptosis in vitro. Our results indicate that SP-D deficiency aggravates CS-induced lung inflammation partly through regulation of ceramide synthesis and that local SP-D enrichment rescues CS-induced inflammation

Cross-species high-resolution transcriptome profiling suggests biomarkers and therapeutic targets for ulcerative colitis

Background: Ulcerative colitis (UC) is a disorder with unknown etiology, and animal models play an essential role in studying its molecular pathophysiology. Here, we aim to identify common conserved pathological UC-related gene expression signatures between humans and mice that can be used as treatment targets and/or biomarker candidates.Methods: To identify differentially regulated protein-coding genes and non-coding RNAs, we sequenced total RNA from the colon and blood of the most widely used dextran sodium sulfate Ulcerative colitis mouse. By combining this with public human Ulcerative colitis data, we investigated conserved gene expression signatures and pathways/biological processes through which these genes may contribute to disease development/progression.Results: Cross-species integration of human and mouse Ulcerative colitis data resulted in the identification of 1442 genes that were significantly differentially regulated in the same direction in the colon and 157 in blood. Of these, 51 genes showed consistent differential regulation in the colon and blood. Less known genes with importance in disease pathogenesis, including SPI1, FPR2, TYROBP, CKAP4, MCEMP1, ADGRG3, SLC11A1, and SELPLG, were identified through network centrality ranking and validated in independent human and mouse cohorts.Conclusion: The identified Ulcerative colitis conserved transcriptional signatures aid in the disease phenotyping and future treatment decisions, drug discovery, and clinical trial design

Impact of apolipoprotein E polymorphism on macrophage properties in vitro - implications in wound healing with focus on laser microdissection

Apolipoproteina E (ApoE) jest jednym z najważniejszych białek kontrolujących metabolizm lipidów w organizmie. Szeroko opisane zostały jej właściwości przeciwmiażdżycowe oraz przeciwzapalne. W populacji ludzkiej białko to występuje w postaci jednej z trzech izoform: ApoE2, ApoE3 lub ApoE4. Uważa się, że korzystne immunomodulacyjne działanie ApoE osłabione jest u osobników z izoformą ApoE4, prowadząc często do rozwoju chorób neurodegeneracyjnych czy chorób układu krążenia. Niektóre badania sugerują jednak, że allel ApoE4 może wywierać korzystny wpływ na ewolucyjnie pierwotne, wrodzone procesy takich jak np. gojenie ran. Ponieważ makrofagi wydają się być głównymi komórkowymi mediatorami zaangażowanymi we wszystkie fazy regeneracji uszkodzonej tkanki, celem badań było określenie wpływu polimorfizmu ApoE in vitro na własności proliferacyjne i migracyjne makrofagów, które mogą okazać się ważne dla procesu gojenia ran. Wyniki badań przeprowadzonych na linii makrofagów mysich RAW264.7 kontrolnych bądź ze stabilną nadekspresją ludzkiej ApoE3 albo ApoE4 wskazują, że różnice w genotypie ApoE nie mają wpływu na proliferację makrofagów, wywierają jednak silny wpływ na ich migrację. Dowiedziono, że potencjał migracyjny niestymulowanych makrofagów z nadekspresją ApoE4 jest znacząco zwiększony w stosunku do pozostałych dwóch linii. Nie zaobserwowano jednak różnic we wpływie izoform ApoE na tempo migracji po stymulacji komórek kontrolnych egzogennymi ludzkimi białkami ApoE3 bądź ApoE4.Aby zbadać wpływ polimorfizmu ApoE na status makrofagów in vivo, konieczna jest ich izolacja z preparatów tkankowych z użyciem mikrodysekcji laserowej (LMD). W celu optymalizacji tej metody przeprowadzono eksperyment gojenia ran na myszach typu dzikiego, a zabezpieczone fragmenty zranionej skóry poddano procesowi LMD. Optymalizacja procedury wymagała opracowania metod wstępnego przygotowania tkanki, właściwego etapu mikrodysekcji, izolacji RNA z pobranych wycinków oraz późniejszej analizy ekspresji genów. Ponadto przetestowano procedury barwień fragmentów skóry, w tym fluorescencyjne barwienie na F4/80 w celu wizualizacji makrofagów.Podsumowując, wyniki uzyskane w niniejszej pracy wskazują, że obecność genotypu ApoE4 odpowiada za zwiększone własności migracyjne makrofagów. Możliwe, iż makrofagi z ekspresją ApoE4 są odpowiedzialne in vivo za nasilenie fazy zapalnej procesu gojenia ran, co może się przyczyniać do szybszej regeneracji tkanki. LMD może stanowić idealną technikę umożliwiającą zbadanie roli takich makrofagów w gojeniu ran. Jej procedura jednak wciąż wymaga uważnej i precyzyjnej optymalizacji.Apolipoprotein E (ApoE) is a major protein controlling lipid homeostasis with known atheroprotective and anti-inflammatory activities. It exists in humans in three isoforms: ApoE2, ApoE3 and ApoE4. The beneficial immunomodulatory properties of ApoE are weakened in ApoE4 carriers, often leading to development of cardiovascular or neurodegenerative diseases. However, some studies suggest that ApoE4 genotype may provide protection in evolutionarily old, innate processes like wound healing. As macrophages are cells deeply involved in all stages of tissue regeneration, the aim of this study was to investigate the influence of ApoE polymorphism in vitro on their proliferation and migration, properties which may contribute to the wound healing response. Experiments performed on RAW264.7 murine macrophage cell line with or without stable overexpression of human ApoE3 or ApoE4 show that although differences in ApoE genotype has no effect on macrophage proliferation, cell migration is deeply affected. It was found that macrophages overexpressing ApoE4 has basal migratory potential considerably increased. However, differential effects between ApoE3 and ApoE4 were not visible after stimulation of cells lacking either isoform with exogenous human ApoE3 or ApoE4. In our future experiments we plan to explore the effects of ApoE polymorphism in vivo. Because of that, laser microdissection (LMD), a novel sophisticated technique allowing for isolation of distinct structures from tissue sections, was used. Wound healing model was performed on wild type mice, and collected wound tissues were subjected to LMD. The procedure was optimized in context of tissue preparation, dissection process itself, subsequent RNA isolation and analysis of gene expression. Furthermore, different stainings, including the visualization of macrophages, have been checked. Although the first trials were promising, the RNA isolation and staining methods were not fully successful.To conclude, obtained results indicate that ApoE4 genotype is responsible for higher migratory capacity of macrophages. It may suggest that ApoE4 macrophages in vivo are responsible for more pronounced inflammatory phase that may contribute to faster healing of the wound. LMD would be a perfect technique to assess the role of such macrophages in different phases of wound healing response. However, its procedure still requires attentive and accurate optimization before it will be fully suitable for experiments

Commensal Intestinal Protozoa&mdash;Underestimated Members of the Gut Microbial Community

The human gastrointestinal microbiota contains a diverse consortium of microbes, including bacteria, protozoa, viruses, and fungi. Through millennia of co-evolution, the host&ndash;microbiota interactions have shaped the immune system to both tolerate and maintain the symbiotic relationship with commensal microbiota, while exerting protective responses against invading pathogens. Microbiome research is dominated by studies describing the impact of prokaryotic bacteria on gut immunity with a limited understanding of their relationship with other integral microbiota constituents. However, converging evidence shows that eukaryotic organisms, such as commensal protozoa, can play an important role in modulating intestinal immune responses as well as influencing the overall health of the host. The presence of several protozoa species has recently been shown to be a common occurrence in healthy populations worldwide, suggesting that many of these are commensals rather than invading pathogens. This review aims to discuss the most recent, conflicting findings regarding the role of intestinal protozoa in gut homeostasis, interactions between intestinal protozoa and the bacterial microbiota, as well as potential immunological consequences of protozoa colonization

MFAP4-Mediated Effects in Elastic Fiber Homeostasis, Integrin Signaling and Cancer, and Its Role in Teleost Fish

Microfibrillar-associated protein 4 (MFAP4) is an extracellular matrix (ECM) protein belonging to the fibrinogen-related domain superfamily. MFAP4 is highly expressed in elastin-rich tissues such as lung, blood vessels and skin. MFAP4 is involved in organization of the ECM, regulating proper elastic fiber assembly. On the other hand, during pathology MFAP4 actively contributes to disease development and progression due to its interactions with RGD-dependent integrin receptors. Both tissue expression and circulating MFAP4 levels are associated with various disorders, including liver fibrosis and cancer. In other experimental models, such as teleost fish, MFAP4 appears to participate in host defense as a macrophage-specific innate immune molecule. The aim of this review is to summarize the accumulating evidence that indicates the importance of MFAP4 in homeostasis as well as pathological conditions, discuss its known biological functions with special focus on elastic fiber assembly, integrin signaling and cancer, as well as describe the reported functions of non-mammalian MFAP4 in fish. Overall, our work provides a comprehensive overview on the role of MFAP4 in health and disease