3-Amidinophenylalanine-Derived Matriptase Inhibitors can Modulate Hepcidin Production in vitro

Matriptase-2 (MT-2) is a type II transmembrane serine protease and predominantly attached to the surface of hepatocytes. MT-2 decreases the production of hepcidin, a key regulator of iron homeostasis. In this study, the effects of four 3-amidinophenylalanine-derived combined matriptase-1/matriptase-2 (MT-1/2) inhibitors (MI-432, MI-441, MI-460, and MI-461) on hepcidin production were investigated in hepatocyte mono- and hepatocyte-Kupffer cell co-cultures. In MI-461-treated cell cultures, the extracellular hydrogen peroxide contents and the interleukin-6 and -8 (IL-6 and IL-8) levels were determined and compared to controls. Hepcidin overproduction was observed in hepatocytes upon treatment with MI-432, MI-441 and MI-461 at 50 mu M. In contrast, extracellular hydrogen peroxide levels were not elevated significantly after matriptase inhibition with MI-461. Furthermore, MI-461 did not induce increases in IL-6 and IL-8 levels in these hepatic models. A model of the binding mode of inhibitor MI-461 in complex with MT-2 revealed numerous polar contacts contributing to the nanomolar potency of this compound. Based on the in vitro data on hepcidin regulation, treatment with MI-461 might be valuable in pathological states of iron metabolism without causing excessive oxidative stress

The Impact of Acute Matriptase Inhibition in Hepatic Inflammatory Models

Purpose. Dysfunction of matriptase-2 can be involved in iron regulatory disorder via downregulation of hepcidin expression. In the present study, we investigated the effects of 3-amidinophenylalanine-derived matriptase inhibitors on porcine hepatic inflammatory cell models. Methods. Hepatocyte-Kupffer cell cocultures (ratio of 2 : 1 and 6 : 1) were treated with four structurally related matriptase inhibitors at 50 μM. Cell cytotoxicity and relative expressions of IL-6 and IL-8 and the levels of hepcidin were determined by MTS and porcine-specific ELISA. The extracellular H2O2 contents were analyzed by Amplex Red method. Results. Matriptase inhibitors at 50 µM for 24 h did not increase cell death rate. The elevated ROS production observed after short-term application of inhibitor MI-441 could be correlated with lowered hepcidin expression. MI-460 could significantly enhance hepcidin levels in the supernatants of cocultures (by 62.21±26.8% in hepatocyte-Kupffer cell, 2 : 1, and by 42.6±14.3% in hepatocyte-Kupffer cell, 6 : 1, cocultures, resp.). No significant changes were found in IL-6 and IL-8 levels in cocultures exposed to matriptase inhibitors. Conclusions. Based on in vitro findings, administration of MI-460 via modulation of hepcidin expression without cytotoxic and oxidative stress inducing properties might be a reliable alternative to treat iron overload in human and veterinary clinical practice

The Impact of Acute Matriptase Inhibition in Hepatic Inflammatory Models

Purpose. Dysfunction of matriptase-2 can be involved in iron regulatory disorder via downregulation of hepcidin expression. In the present study, we investigated the effects of 3-amidinophenylalanine-derived matriptase inhibitors on porcine hepatic inflammatory cell models. Methods. Hepatocyte-Kupffer cell cocultures (ratio of 2 : 1 and 6 : 1) were treated with four structurally related matriptase inhibitors at 50 μM. Cell cytotoxicity and relative expressions of IL-6 and IL-8 and the levels of hepcidin were determined by MTS and porcine-specific ELISA. The extracellular H2O2 contents were analyzed by Amplex Red method. Results. Matriptase inhibitors at 50 µM for 24 h did not increase cell death rate. The elevated ROS production observed after short-term application of inhibitor MI-441 could be correlated with lowered hepcidin expression. MI-460 could significantly enhance hepcidin levels in the supernatants of cocultures (by 62.21±26.8% in hepatocyte-Kupffer cell, 2 : 1, and by 42.6±14.3% in hepatocyte-Kupffer cell, 6 : 1, cocultures, resp.). No significant changes were found in IL-6 and IL-8 levels in cocultures exposed to matriptase inhibitors. Conclusions. Based on in vitro findings, administration of MI-460 via modulation of hepcidin expression without cytotoxic and oxidative stress inducing properties might be a reliable alternative to treat iron overload in human and veterinary clinical practice

Atípusos haemorrhagiás bőrtünetek hátterében igazolt hajas sejtes leukaemia poliszenzibilizált betegen

A nemszteroid gyulladáscsökkentők (NSAID-ok) lokális és szisztémás formában történő alkalmazása egyre elterjed- tebbé vált, az ezzel kapcsolatos adverz reakciók gyakoriak. A szerzők egy 49 éves férfi beteg esetét ismertetik, aki lábszárak fájdalma miatt etofenamáttartalmú lokális készítményt alkalmazott, majd az alkalmazás helyén, UV-expozíciót követően, haemorrhagiás, atípusos, kontaktdermatitisnek megfelelő bőrtünetek jelentek meg. Tüneteinek rapid terjedése miatt sürgősségi fekvőbeteg-ellátásra szorult. Fizikális vizsgálata során az etofenamátexpozíció helyén, mindkét lábszáron, valamint azon túlmenően a karokon, valamint a törzsön és az arcon bevérzett, konfluáló, erythemás seropapulákat és maculákat láttunk, melyek mellett testszerte és a buccalis nyálkahártyán petechiák voltak megfigyelhetők. A nagy kiterjedésű és aspecifikus bőrtünetek, a fizikális vizsgálattal észlelt splenomegalia, valamint a perifériás vérképeltérés miatt indított onkohematológiai kivizsgálás során hajas sejtes leukaemia diagnózisa igazolódott. Epicutan tesztelés (ET) során az etofenamáttartalmú gélre adott, erősen pozitív reakció mellett fakátrány-, propilén- glikol-, propolisz-, ’fragrance mix I’, metilizotiazolinon-, benzoesav- és perubalzsam-poliszenzibilizáció volt verifi- kálható. Az etofenamát hatóanyaggal elvégzett lymphocytatranszformációs teszt (LTT) és CD69-expresszió-vizsgálat negatív eredményt adott

Progresszív kórlefolyást mutató szisztémás mastocytosis

Authors report on a case of a male patient of systemic mastocytosis that was associated with extensive cutaneous lesions. Chronic diarrhoea worsening his quality of life was well managed by the administration of antihistamines. The pleural fluid recurrence soon after drainage has been controlled by the administration of alpha interferon. 40 years after the onset of the first skin signs progression has been manifested in the development of "B" (bone marrow infiltration rate >30%, dysmyelopoiesis, serum tryptase >20 mug/L, hepato- and splenomegaly) and "C" symptoms (liver function test abnormalities, cytopenia, malabsorption, osteoporosis). The patient died at age of 87. The authors' aim was to attract attention on this rare disease and emphasize that symptomatic therapy with antihistamines and drugs available based on customised rights by the National Health Insurance Fund might provide good quality of life. Orv Hetil. 2018; 159(5): 192-196

Identification of miRSNPs associated with the risk of multiple myeloma

Accepted articleMultiple myeloma (MM) is a malignancy of plasma cells usually infiltrating the bone marrow, associated with the production of a monoclonal immunoglobulin (M protein) which can be detected in the blood and/or urine. Multiple lines of evidence suggest that genetic factors are involved in MM pathogenesis, and several studies have identified single nucleotide polymorphisms (SNPs) associated with the susceptibility to the disease. SNPs within miRNA-binding sites in target genes (miRSNPs) may alter the strength of miRNA-mRNA interactions, thus deregulating protein expression. MiRSNPs are known to be associated with risk of various types of cancer, but they have never been investigated in MM. We performed an in silico genome-wide search for miRSNPs predicted to alter binding of miRNAs to their target sequences. We selected 12 miRSNPs and tested their association with MM risk. Our study population consisted of 1,832 controls and 2,894 MM cases recruited from seven European countries and Israel in the context of the IMMEnSE (International Multiple Myeloma rESEarch) consortium. In this population two SNPs showed an association with p<0.05: rs286595 (located in gene MRLP22) and rs14191881 (located in gene TCF19). Results from IMMEnSE were meta-analyzed with data from a previously published genome-wide association study (GWAS). The SNPs rs13409 (located in the 3UTR of the POU5F1 gene), rs1419881 (TCF19), rs1049633, rs1049623 (both in DDR1) showed significant associations with MM risk. In conclusion, we sought to identify genetic polymorphisms associated with MM risk starting from genome-wide prediction of miRSNPs. For some mirSNPs, we have shown promising associations with MM risk. What's new? Even though deregulation of miRNA expression has been associated with human cancers little information is available regarding their relation with MM susceptibility. We performed an in silico genome-wide search for miRSNPs and selected the most promising ones for an association study. The SNPs with the strongest associations with MM risk are localized in genes which have never been related with MM.This work was partially funded by: intramural funds of German Cancer Research Center (DKFZ), Grant ref. HUS412A1271 from the “Gerencia Regional de Salud de la Junta de Castilla y Léon”. This work was supported by grants from the Instituto de Salud Carlos III (Madrid, Spain; PI12/02688). Catalan Government DURSI grant 2014SGR647 and Instituto de Salud Carlos III, co7funded by FEDER funds –a way to build Europe– grants PI11701439 and PIE13/00022info:eu-repo/semantics/publishedVersio

Cardiopoietic cell therapy for advanced ischemic heart failure: results at 39 weeks of the prospective, randomized, double blind, sham-controlled CHART-1 clinical trial

Cardiopoietic cells, produced through cardiogenic conditioning of patients' mesenchymal stem cells, have shown preliminary efficacy. The Congestive Heart Failure Cardiopoietic Regenerative Therapy (CHART-1) trial aimed to validate cardiopoiesis-based biotherapy in a larger heart failure cohort

3D-s fiziológiás és gyulladásos májsejt modell alkalmazása hatóanyagjelöltek tesztelésére

A matriptáz-2 (MT-2) a májsejtek felületén lévő transzmembrán szerin proteáz, amely a hepcidin termelését, a szervezet vasháztartását szabályozza. A matriptáz gátlás in vitro tanulmányozására adhat lehetőséget a hidrogél segítségével előállított 3D-s szerkezetbe ágyazott májsejt és májsejt- Kupffer sejt ko-kultúra. A 3D-s sejttenyésztés előnye a 2D-s technikákhoz képest, hogy a sejtek in vivo mikrokörnyezetének tulajdonságai jobban érvényesülnek, ezáltal fenotípusok, funkciójuk megtartottabb. Kísérleteink során két, 3-amidinofenilalanin szerkezetű matriptáz inhibitor (MI-441 és MI-461) hatásait vizsgáltuk májsejt mono- és Kupffer sejtekkel kiegészített ko- kultúrán (6:1 sejtaránnyal). Immunfluoreszcens festéssel nyomon követtük egy tight junction fehérje, az occludin elhelyezkedési mintázatát a két sejttenyészetben. Az extracelluláris hidrogén-peroxid szintet Amplex Red reagenssel történő mérésével határoztuk meg. Az esetleges gyulladásos válaszok kialakulásának vizsgálatára az interleukin-6 és az interleukin-8 (IL-6, -8) képződését és a gátlószerek hepcidin szintekre gyakorolt hatását sertés specifikus ELISA kittel határoztuk meg

Treatment options and new perspectives in iron homeostasis disturbances Literature review

SUMMARY The authors summarize in their study the causes and the symptoms of iron toxicosis and iron deficiency anaemia and the diagnostic and treatment options of disturbances in iron homeostasis. Plasma iron level can be a reliable indica tor of iron deficiency anaemia, however, the low iron level can also be caused by chronic inflammation, infections and cancers. The iron supplementation of neonatal suckling piglets is of high veterinary importance, which is the most appropriate treatment for prevention of iron deficiency anaemia. The authors point out that iron toxicosis developed after administration of excessive iron can be compensated through parenteral deferoxamine, however other chelators applied mainly in human medicine such as deferiprone or deferasirox might be also effective in veterinary field. In this study the role of hepcidin in iron homeo stasis of the body is also discussed. The maintenance of physiological iron lev els in spite of rapid turnover of the iron suggests the regulation of hepcidin by plasma iron. In case of iron deficiency, the hepcidin amount is lowered, which then facilitates the rate of iron transport into the blood. In contrast, if the iron storage is saturated, the level of hepcidin produced by liver becomes elevated. Hepcidin oversecretion can also be induced by infection and inflammation with excessive interleukin-6 production. The authors also describe the compounds acting via hepcidin regulation which can be therapeutically beneficial in the treatment of iron homeostatic disturbances. The pharmacological intervention of the interplay between hepcidin and matriptase-2 has been one of the most recently discovered research fields which might involve the introduction of mat riptase modulators into the drug therapy of iron disorders