A PIBF klónozása, immunológiai jellemzése és alkalmazási területei a terhességi és tumordiagnosztikában

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Role of Actinomyces spp. and related organisms in the development of medication-related osteonecrosis of the jaw (MRONJ) : Clinical evidence based on a case series

Medication-related osteonecrosis of the jaw (MRONJ) is an increasingly common consequence of antiresorptive treatment, which often leads to the development of necrotic exposed bone surfaces with inflammatory processes affecting the jawbone. Although the development of MRONJ is often associated with the inflammatory response or infections caused by the colonizing members of the oral microbiota, the exact pathogenesis of MRONJ is still not fully understood. In the present paper, we aimed to provide additional, microbiological culture-supported evidence, supporting the "infection hypothesis" that Actinomyces spp. and related organisms may play an important pathogenic role in the development of MRONJ and the resulting bone necrosis. In our case series, all patients presented with similar underlying conditions and anamnestic data, and have received antiresorptive medications (bisphosphonates or a RANK ligand (RANKL) inhibitor) to prevent the occurrence or progression of bone metastases, secondary to prostate cancer. Nevertheless, a few years into antiresorptive drug therapy, varying stages of MRONJ was identified in the mentioned patients. In all three cases, quantitative microbiological culture of the necrotic bone samples yielded a complex microbiota, dominated by Actinomyces and Schaalia spp. with high colony counts. Additionally, our followed-up case series document the treatment of these patients with a combination of surgical intervention and long-term antibiotic therapy, where favourable clinical responses were seen is all cases. If the "infection hypothesis" is valid, it may have significant consequences in the preventative and therapeutic strategies associated with this disease

Peripheral blood TIM-3 positive NK and CD8+ T cells throughout pregnancy : TIM-3/galectin-9 interaction and its possible role during pregnancy

Abstract Problem: The T-cell immunoglobulin and mucin domain (TIM) family is a relatively newly described group of molecules with a conserved structure and important immunological functions. Identification of Galectin-9 as a ligand for TIM-3 has established the Galectin-9/TIM-3 pathway as an important negative regulator of Th1 immunity and tolerance induction. Data about the TIM-3/Gal-9 pathway in the pathogenesis of human diseases is emerging, but their possible role during human pregnancy is not precisely known. The aim of our study was to investigate the number, phenotype and functional activity of TIM-3+ peripheral blood mononuclear cells during healthy human pregnancy. Methods of Study: 57 healthy pregnant women [first trimester (n = 16); second trimester (n = 19); third trimester (n = 22)] and 30 non-pregnant controls were enrolled in the study. We measured the surface expression of TIM-3 by cytotoxic T cells, NK cells and NK cell subsets as well as Galectin-9 expression by regulatory T cells by flow cytometry. We analyzed the cytokine production and cytotoxicity of TIM3+ and TIM3- CD8 T and NK cells obtained from non-pregnant and healthy pregnant women at different stages of pregnancy by flow cytometry. Serum Galectin-9 levels were measured by ELISA. Results: Our results show that the numbers of peripheral NK and cytotoxic T cells and their TIM-3 expression do not change between the first, second and third trimesters of pregnancy. Compared to non-pregnant individuals, regulatory T cells show higher level of Galectin-9 expression as pregnancy proceeds, which is in line with the level of Galectin-9 in the patients sera. Cytotoxic T cells, NK cells and NK cell subsets expressing TIM-3 molecule show altered cytokine production and cytotoxicity during pregnancy compared to non-pregnant individuals. Conclusion: Our results indicate that Galectin-9 expressing regulatory T cells, TIM-3+ cytotoxic T cells and NK cells could play an important role in the maintenance of healthy pregnancy

A PIBF receptor szerepe a terhesség fennmaradásában és a tumornövekedésben = The role of PIBF receptors in pregnancy and tumor development.

H2O2 hatására idő- és hőmérsékletfüggően csökken a membránfluiditás, nő a raftok száma. A PIBF receptorkötése után a membrán rigiddé válik, és megnő a raftok mérete, míg számuk csökken, H2O2 jelenlétében azonban csökken a PIBF receptorhoz való kötődése, nő membrán fluiditása és nem jön létre a lipid raftok clusteresedése. Oxidativ stresszben a membran lipid organizáció változásai gátolják a PIBF receptor aktiválódását és így a PIBF biológiai hatásának érvényesülését. A PIBFR izolálása A PIBF ligandot affinitás-cross linking technikával a PIBF receptorhoz kötöttük, a sejteket lizáltuk, majd raft frakciókat izoláltunk. Az immunizálás sikertelen volt. Újabb preparálás folyamatban. PIBF knock down hatására trophoblast (TR) sejtek MMP2 és MMP9 aktivitása, valamint inváziója fokozódott, míg a tumor (TU) sejteké csökkent. PIBF kezelt TR-ban AKT és ERK azonnal-, TU sejtekben ugyanezek, valamint a STAT3 csak hat óra múlva, de tartósan aktiválódtak. A PIBF kezelt TU sejtekben a kezeletlen sejtekhez képest több (pl. EGF és HB-EGF) molekula differenciáltan expresszált. PIBF kezet HB-EGF deficiens sejtekben STAT3 nem aktiválódik, tehát a PIBF-indukálta HB-EGF saját receptorán elindított jelátviteli úton aktiválja STAT3-t. A PIBF TR és TU sejtekben kapcsolódik a HB-EGF promoter regiójához, de a protein-DNS complex csak TU sejtekben tartalmazta a nuclearisan lokalizált teljes hosszúságú PIBF-t. PIBF az invazív gének repressziója-, ill. indukciója révén szabályozza a TR- és a TU inváziót. | Oxidative stress induces a time- and temperature-dependent decrease in membrane fluidity together with increased number of rafts. Ligand binding of the PIBFR results in rigidification of the membrane and raft clustering. In H2O2 treated cells receptor binding of PIBF is inhibited, membranes become more fluid and lipid raft clustering is absent. In oxidative stress conditions, altered membrane lipid organization prevents the activation of the PIBF receptor, thereby inhibiting the biological effects of PIBF. Isolation of the PIBFR. PIBF was irreversibly cross-linked with its receptor, the cells were lysed, and raft fractions were isolated. Immunization was not successful. Preparation of new antigens in progress. Invasiveness as well as MMP2 and MMP9 activity of PIBF knock down trophoblast (TR) cells increased, while those of tumor (TU) cells decreased. In TR PIBF induced an immediate and transient AKT and ERK activation, while in TU cells the same molecules plus STAT3 were activated after 6h. PIBF induced cells several molecules, e.g., EGF and HB-EGF in TU cells. In HB-EGF deficient cells, PIBF fails to activate STAT3, suggesting that PIBF-induced HB-EGF signals via its own receptor to activate STAT3. PIBF binds to the promoter region of HB-EGF both in TR and TU however, the full length PIBF (localized to the nucleus) is only present in the protein-DNA complex from TU cells. PIBF regulates TR and TU invasion by repressing or inducing invasive genes

A PIBF molekula tumornövekedésben, ill. a terhesség fenntartásában szerepet játszó régióinak azonosítása = Localization of the active centers accounting for the biological activity of the PIBF molecule

Progesteron-Indukált Blokkoló Factor (PIBF) in vivo a cytokintermelés befolyásolásán keresztül valósulnak meg. A molekula fenti hatásokért felelős regióinak azonosítása céljából a PIBF különböző részeit reprezentáló konstrukciókat állítottunk elő és vizsgáltunk ezek biológiai hatásait. A PIBF az IL-4 kiváltotta jelátvitelhez hasonlóan STAT6 valamint Jak1 foszforilációt indukál és gátolja a STAT4 foszforilálódását. STAT6 deficiens sejtekben a PIBF cytoknitermelésre kifejtett hatásai nem érvényesülnek. Megállapítottuk, hogy a PIBF-indukálta jelátvitel feltétele az IL-4R alpha lánca és a GPI-horgonyzott PIBF receptor által alkotott receptorkomplex megléte. A molekula N terminális részén található PN1 peptid a PIBF-hez hasonló jelátvitelt indukált és szignifikánsan fokozta a lymphocyták IL-10 termelését. Az NK aktivitást kizárólag a 48 kDa PIBF szakasz csökkentette szignifikáns mértékben. Egereket immunizáltunk PIBF-el, ill. A peptidekkel, majd a megfelelő ellenanyagszint kialakulása után az egereket a pároztattuk, vagy intraperitonealisan SP-2 myelomasejtekkel oltottuk őket. A terhesség kimenetelére gyakorolt hatást a beágyazódott, peték számával-, a tumornövekedésre kifejtett hatást pedig a hasüregből kinyerhető élő, ill. elpusztult tumorsejtek arányával jellemeztük. PIBF neutralizáló ellenanyagok jelenléte gátolja a pete beágyazódását. A PN1 és PN3 peptiddel történő immunizálás hatására szignifikánsan fokozódott a daganatsejtek pusztulása. | The Progesterone-Induced Blocking Factor (PIBF) exerts its in vivo effects by acting on cytokine production. In attempt to identify the biologically active regions of PIBF, five costructs (PN1, PN2, PN3, PN4 and PN5), representing different parts of of the molecule were designed for further functional testing. We have shown that PIBF signaling depends on the presence of a functional heterodimer receptor, consisting of the alpha chain of the IL-4 receptor and the GPI-anchored PIBF receptor. PIBF induces the phophorylation of STAT6 and of Jak1, while the phosphorylation of STAT4 is inhibited. The cytokine effects of PIBF are significantly reduced in STAT6 deficient cells. The N-terminal PN1 peptide induced STAT6 phosphorylation, and IL-10 production of peripheral lymphocytes was significantly increased in the presence of PN1. NK activity was not affected by either of the peptides. Mice were immunized with PIBF and the constructs. When neutralizing antibodies developed, the mice were mated, or injected i.p. with SP-2 myeloma cells. The effect of anti-PIBF or anti-peptide antibodies on pregnancy was determined by counting implantation sites, whereas that on tumor cell growth- by determining the ratio of dead and viable tumor cells recovered from the peritoneal cavity. We have shown that PIBF neutralizing antibodies interfered with implantation. Anti PN1 and PN3 neutralizing antibodies significantly increased the number of dead tumor cells

Influence of Galectin-9 Treatment on the Phenotype and Function of NK-92MI Cells in the Presence of Different Serum Supplements

Galectins are one of the critical players in the tumor microenvironment–tumor crosstalk and the regulation of local immunity. Galectin-9 has been in the limelight in tumor immunology. Galectin-9 possesses its multiplex biological functions both extracellularly and intracellularly, plays a pivotal role in the modulation of adaptive and innate immunity, and induces immune tolerance. NK-92MI cell lines against different malignancies were extensively studied, and recently published trials used genetically chimeric antigen receptor-transfected NK-92MI cells in tumor immunotherapy. Besides the intensive research in tumor immunotherapy, limited information is available on their immune-checkpoint expression and the impact of checkpoint ligands on their effector functions. To uncover the therapeutic potential of modulating Galectin-9-related immunological pathways in NK-cell-based therapy, we investigated the dose-dependent effect of soluble Galectin-9 on the TIM-3 checkpoint receptor and NKG2D, CD69, FasL, and perforin expression of NK-92MI cells. We also examined how their cytotoxicity and cytokine production was altered after Gal-9 treatment and in the presence of different serum supplements using flow cytometric analysis. Our study provides evidence that the Galectin-9/TIM-3 pathway plays an important role in the regulation of NK cell function, and about the modulatory role of Galectin-9 on the cytotoxicity and cytokine production of NK-92MI cells in the presence of different serum supplements. We hope that our results will aid the development of novel NK-cell-based strategies that target Galectin-9/TIM-3 checkpoint in tumors resistant to T-cell-based immunotherapy

Multiple divergent picobirnaviruses with functional prokaryotic Shine- Dalgarno ribosome binding sites present in cloacal sample of a diarrheic chicken

Picobirnaviruses (PBVs) of family Picobirnaviridae have bisegmented (S1 and S2 segments), doublestranded RNA genomes. In this study a total of N = 12 complete chicken PBVs (ChPBV) segments (N = 5 of S1 and N = 7 of S2, Acc. Nos.: MH425579-90) were determined using viral metagenomic and RTPCR techniques from a single cloacal sample of a diarrheic chicken. The identified ChPBV segments are unrelated to each other and distant from all of the currently known PBVs. In silico sequence analyses revealed the presence of conserved prokaryotic Shine-Dalgarno-like (SD-like) sequences upstream of the three presumed open reading frames (ORFs) of the S1 and a single presumed ORF of the S2 segments. According to the results of expression analyses in E. coli using 6xHis-tagged recombinant ChPBV segment 1 construct and Western blot these SD-like sequences are functional in vivo suggesting that S1 of study PBVs can contain three ORFs and supporting the bacteriophage-nature of PBVs

PACAP-38 and PAC1 Receptor Alterations in Plasma and Cardiac Tissue Samples of Heart Failure Patients

Pituitary adenylate cyclase activating polypeptide-38 (PACAP-38) is a multifunctional neuropeptide, which may play a role in cardioprotection. However, little is known about the presence of PACAP-38 in heart failure (HF) patients. The aim of our study was to measure the alterations of PACAP-38 like immunoreactivity (LI) in acute (n = 13) and chronic HF (n = 33) and to examine potential correlations between PACAP-38 and HF predictors (cytokines, NT-proBNP). Tissue PACAP-38 LI and PAC1 receptor levels were also investigated in heart tissue samples of patients with HF. Significantly higher plasma PACAP-38 LI was detected in patients with acute HF, while in chronic HF patients, a lower level of immunoreactivity was observed compared to healthy controls (n = 13). Strong negative correlation was identified between plasma PACAP-38 and NT-proBNP levels in chronic HF, as opposed to the positive connection seen in the acute HF group. Plasma IL-1 β, IL-2 and IL-4 levels were significantly lower in chronic HF, and IL-10 was significantly higher in patients with acute HF. PACAP-38 levels of myocardial tissues were lower in all end-stage HF patients and lower PAC1 receptor levels were detected in the primary dilated cardiomyopathy group compared to the controls. We conclude that PACAP-38 and PAC1 expression correlates with some biomarkers of acute and chronic HF; therefore, further studies are necessary to explore whether PACAP could be a suitable prognostic biomarker in HF patients

Szervezeti kultúra az AIESEC debreceni helyi közösségében

Diplomamunkám témájának a szervezeti kultúrát választottam az AIESEC Debreceni Helyi Bizottságának bemutatásán keresztül. A dolgozatomat a szervezet elméleti áttekintésével kezdtem: szervezetelméleti irányzatok, szervezet működésének formái. Ezt követően a kultúrát általános értelemben, majd a szervezeti kultúra jellemzőit mutattam be az AIESEC nemzetközi szervezeten keresztül.BSc/BAAndragógiaB