Expression pattern of the human ABC transporters in pluripotent embryonic stem cells and in their derivatives

Background: ATP-binding Cassette (ABC) transporters have key roles in various physiological functions as well as providing chemical defense and stress tolerance in human tissues. In this study we have examined the expression pattern of all ABC proteins in pluripotent human embryonic stem cells (hESCs) and in their differentiated progenies. We paid special attention to the cellular expression and localization of multidrug transporter ABC proteins. Methods: Stem cell differentiation was carried out without chemical induction or cell sorting, and specialized cell types were separated mechanically. Cellular features regarding pluripotency and tissue identity, as well as ABC transporter expression were studied by flow cytomtery, immuno-microscopy and qPCR-based low-density arrays. Results: Pluripotent hESCs and differentiated cell types (cardiomyocytes, neuronal cells and mesenchymal stem cells) were distinguished by morphology, immunostaining markers and selected mRNA expression patterns. We found that the mRNA expression levels of the 48 human ABC proteins also clearly distinguished the pluripotent and the respective differentiated cell types. When multidrug and lipid transporter ABC protein expression was examined by using well characterized specific antibodies by flow cytometry and confocal microscopy, the protein expression data corresponded well to the mRNA expression results. Moreover, the cellular localization of these important human ABC transporter proteins could be established in the pluripotent and differentiated hESC derived samples. Conclusions: These studies provide valuable information regarding ABC protein expression in human stem cells and their differentiated offspring. The results may also help to obtain further information concerning the specialized cellular functions of selected ABC transporter

DLQI-R scoring improves the discriminatory power of theDermatology Life Quality Index in patients with psoriasis,pemphigus and morphea

Background The Dermatology Life Quality Index (DLQI) rates ‘not relevant’responses (NRRs) as the item on the questionnaire having no impact on thepatients’ lives at all. The DLQI-Relevant (DLQI-R) is a recently developed scoringthat adjusts the total score of the questionnaire for the number of NRRs indicatedby a patient.Objectives To compare the discriminatory power of the original and DLQI-R scor-ing approaches in terms of absolute and relative informativity.Methods Cross-sectional data from 637 patients with morphea, pemphigus andpsoriasis were used for the analyses. To assess absolute and relative informativity,Shannon’s index and Shannon’s evenness index were calculated for the 10 itemson the questionnaire and for DLQI and DLQI-R total scores.Results Mean DLQI and DLQI-R scores of patients were 613 vs. 691. In the su b-set of patients wit h NRRs (n = 261, 41%), absolute informativity was higherwith the DLQI-R scoring for all eight items with NRR options in all three condi-tions. The DLQI-R exhibited a better relative informativity in 8, 8 and 6 items inpemphigus, morphea and psoriasis, respectively. The DLQI-R led to an improve-ment in average item-level informativity in all DLQI score bands up to 20 points.Regarding total scores, the DLQI-R produced both a higher absolute and relativeinformativity in all three conditions.Conclusions In patients with morphea, pemphigus and psoriasis, DLQI-R scoringimproves the discriminatory power of the questionnaire by benefiting from theadditional informa tion in NRRs. DLQI-R scoring may be us eful both in clinicalpractice and research. A scoring chart has been developed to aid physicians withscoring

Az életminőség mérése a bőrgyógyászatban: hazai tapasztalatok = Measuring quality of life in dermatological care : experiences from Hungary

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Analysis of Intracellular Calcium Signaling in Human Embryonic Stem Cells

Measurement of changes in intracellular calcium concentration is one of the most common and useful tools for studying signal transduction pathways or cellular responses in basic research and drug screening purposes as well. Increasing number of such applications using human pluripotent stem cells and their derivatives requires development of calcium signal measurements for this special cell type. Here we describe a modified protocol for analysis of calcium signaling events in human embryonic stem cells, which can be used for other pluripotent cell types (such as iPSC) or their differentiated offspring as well

Efficient generation of human embryonic stem cell-derived cardiac progenitors based on tissue-specific EGFP expression

Cardiac progenitor cells (CPCs) are committed to the cardiac lineage but retain their proliferative capacity before becoming quiescent mature cardiomyocytes. In medical therapy and research the use of human pluripotent stem cell-derived CPCs would have several advantages over mature cardiomyocytes, as the progenitors show better engraftment into existing heart tissues, and provide unique potential for cardiovascular developmental, as well as for pharmacological studies. Here we demonstrate that the CAG promoter-driven EGFP reporter system allows the identification and isolation of embryonic stem cell-derived CPCs. Tracing of CPCs during differentiation confirmed upregulation of surface markers, previously described to identify cardiac precursors and early cardiomyocytes. Isolated CPCs express cardiac lineage specific transcripts, still have proliferating capacity and can be re-aggregated into embryoid body-like structures (CAG-EGFPhigh rEBs). Expression of TROPONIN T and NKX2.5 mRNA are upregulated in long-term cultured CAG-EGFPhigh rEBs, in which more than 90% of the cells become Troponin I positive mature cardiomyocytes. Moreover, about one third of the CAG-EGFPhigh rEBs show spontaneous contractions. The method described here provides a powerful tool to generate expandable cultures of pure human CPCs which can be used to elucidate toxicological mechanisms during cardiac development, as well as to explore early markers of the cardiac lineage

Psoriasisban szenvedő betegek életminőségének vizsgálata Magyarországon [Assessment of health-related quality of life in psoriasis patients in Hungary]

Absztrakt: Bevezetés: Magyarországon a psoriasisos betegpopuláció egészségi állapota és életminősége kevéssé ismert. Célkitűzés: Kutatásunk célja a hazai psoriasisos betegek életminőségének vizsgálata és összehasonlítása az általános populációval, illetve az életminőséget befolyásoló tényezők azonosítása. Módszer: 2012 és 2016 között két keresztmetszeti kérdőíves felmérést végeztünk két hazai egyetemi klinika psoriasisos betegeinek bevonásával. Az életminőséget az EQ-5D-3L és a hozzá tartozó Vizuális Analóg Skála (EQ VAS), valamint a Bőrgyógyászati Életminőség Index (DLQI) segítségével vizsgáltuk. Az életminőséget befolyásoló tényezőket többváltozós lineáris regresszióval elemeztük. Eredmények: A kérdőívet 434 psoriasisos beteg töltötte ki (átlagéletkor 49 év, 65% férfi, 81% középsúlyos vagy súlyos psoriasis). A felmérés időpontjában a betegek 43%-a biológiai terápiában részesült. A mintában az EQ-5D-3L, az EQ VAS és a DLQI átlaga 0,74 ± 0,28, 69,06 ± 20,98, illetve 6,78 ± 7,38 volt. A fájdalom/rossz közérzet, szorongás/lehangoltság, mozgékonyság, szokásos tevékenységek és önellátás dimenziókban rendre a betegek 54, 43, 40, 32 és 15 százaléka jelzett problémát. Az EQ-5D-3L-index alapján a betegek életminősége rosszabb volt, mint a nemben és korban illesztett általános populációé, a különbség férfiaknál a 25–34 és 45–64 éves korcsoportokban, illetve nőknél a 18–64 éves korcsoportban volt statisztikailag szignifikáns (p<0,05). Szignifikánsan rosszabb volt a nők (p = 0,042), az arthritis psoriaticás (p<0,001) és a tenyéri/talpi psoriasisos (p = 0,031) betegek életminősége. Jobb életminőséget találtunk a felsőfokú végzettségű és a teljes vagy részmunkaidőben foglalkoztatott betegeknél (p<0,001). Következtetések: Kutatásunk az első kiterjedt életminőség-felmérés az EQ-5D-kérdőívvel psoriasisos betegek körében Magyarországon és tágabban egész Közép-Kelet-Európában. Az eredmények hasznosak lehetnek a psoriasis terápiáinak költséghatékonysági modellezésében, illetve egészségpolitikai döntések előkészítésében. Orv Hetil. 2018; 159(21): 837–846. | Abstract: Introduction: The health status and health-related quality of life (HRQoL) of patients with psoriasis in Hungary are understudied. Aim: To assess HRQoL in psoriasis patients, to compare HRQoL of psoriasis patients to that of the general public in Hungary and to identify predictors of HRQoL. Method: Between 2012 and 2016, two cross-sectional surveys were carried out among psoriasis patients at two academic dermatology departments. HRQoL was assessed by EQ-5D-3L, EQ Visual Analogue Scale (EQ VAS) and Dermatology Life Quality Index (DLQI). Predictors of HRQoL were analysed by multivariate linear regression. Results: 434 patients were enrolled (mean age 49 years, 65% male, 81% moderate-to-severe psoriasis, 43% treated with biologics). Mean EQ-5D-3L, EQ VAS and DLQI scores were 0.74 ± 0.28, 69.06 ± 20.98 and 6.78 ± 7.38, respectively. Overall, 54%, 43%, 40%, 32% and 15% of patients indicated at least some problems in pain/discomfort, anxiety/depression, mobility, usual activities and self-care. EQ-5D-3L index scores in patients were lower compared to the age- and gender-matched general population in Hungary. The difference was statistically significant for the age groups 25–34 and 45–64 in males, and 18–64 in females (p<0.05). Female gender (p = 0.042), psoriatic arthritis (p<0.001) and palmoplantar psoriasis (p = 0.031) were associated with lower EQ-5D-3L index scores. On the contrary, employed and highly educated patients reported higher EQ-5D-3L index scores (p<0.001). Conclusions: We were the first to assess HRQoL in psoriasis patients by using EQ-5D questionnaire in Hungary, and more broadly in Central and Eastern Europe. Our findings are useful for cost-effectiveness modelling of psoriasis treatments and decision-making in healthcare. Orv Hetil. 2018; 159(21): 837–846

Characterization of calcium signals in human embryonic stem cells and in their differentiated offspring by a stably integrated calcium indicator protein

Intracellular calcium signaling pathways play a major role in cellular responses such as proliferation, differentiation and apoptosis. Human embryonic stem cells (hESC) provide new possibilities to explore the development and differentiation of various cell types of the human body. Intracellular calcium responses to various ligands and the calcium signaling pathways, however, have not been thoroughly studied in embryonic stem cells and in their differentiated progenies. In our previous work we demonstrated that the use of the fluorescent calcium indicator Fluo-4 with confocal microscopy allows sensitive and reliable measurements of calcium modulation in human embryonic stem cells and stem-cell derived cardiomyocytes. Here we developed a human embryonic stem cell line stably expressing a genetically encoded Ca2+ indicator (GCaMP2) using a transposon-based gene delivery system. We found that the differentiation properties were fully preserved in the GCaMP2-expressing hESC lines and Ca imaging could be performed without the need of toxic dye-loading of the cells. In undifferentiated hES cells the calcium signals induced by various ligands, ATP, LPA, trypsin or angiotensin II were comparable to those in Fluo-4 loaded cells. In accordance with previous findings, no calcium signal was evoked by thrombin, histamine or GABA. Cardiomyocyte colonies differentiated from hES-GCaMP2 cells could be recognized by spontaneous contractions and Ca2+ oscillations. GCaMP2-expressing neural cells were identified based on their morphological and immuno-staining properties and Ca signals were characterized on those cells. Characteristics of both the spontaneous and ligand-induced Ca2+ signals, as well as their pharmacological modification could be successfully examined in these model cells by fluorescence imaging

Cystás fi brosissal élő betegek és gondozóik életminősége és betegségterhei: a BURQOL-RD európai felmérés magyarországi eredményei | Health related quality of life and disease burden of patients with cystic fibrosis and their caregivers: Results of the European BURQOL-RD survey in Hungary

Bevezetés: Magyarországon kevéssé ismertek a cystás fibrosis betegség egyéni és társadalmi terhei. Célkitűzés: A cystás fibrosissal élő betegek és gondozóik egészséggel összefüggő életminőségének és terheinek felmérése. Módszer: Az európai BURQOL-RD-projekt keretében keresztmetszeti kérdőíves felmérést végeztek EQ-5D-5L-kérdőívvel és retrospektíven vizsgálták az egészségügyi ellátások igénybevételét. Eredmények: 110 beteg (korcsoportok, év: 0–13, N = 48; 14–17, N = 12; ≥18, N = 50) vett részt a vizsgálatban, a diagnózis felállításakor az életkor medián 1 év volt. Az EQ-5D-5L-indexérték szignifikánsan alacsonyabb volt a 18–24 és 25–34 éves korosztályban a hazai populációs átlagnál (p<0,05). Tüdőgyógyászati szakvizsgálaton 75 beteg (68%) járt, kórházi felvételre 55 beteg (50%) került az elmúlt 6, illetve 12 hónapban, dornase alfa-kezelést 57 beteg (52%) kapott. Öt felnőtt beteget (10%) segített közeli családtag nem hivatásos gondozóként. Következtetések: A cystás fibrosis jelentős életminőség-csökkenéssel jár. A kutatás elsőként nyújt alapadatokat a kelet-közép-európai régióból a cystásfibrosis-ellátás egészség-gazdaságtani elemzéseihez. Orv. Hetil., 2014, 155(42), 1673–1684. | Introduction: Data on disease burden of cystic fibrosis in Hungary are scarce. Aim: To assess quality of life and resource utilisations of patients with cystic fibrosis. Method: In a cross-sectional survey (BURQOL-RD project), the EQ-5D-5L questionnaire was applied and healthcare utilisations were retrospectively surveyed. Results: 110 patients participated in the study (age-groups, year: 0–13, N = 48; 14–17, N = 12; ≥18, N = 50), median age at the diagnosis was 1 year. EQ-5D-5L score in age-groups 18–24 and 25–34 was significantly lower than in the general population (p<0.05). 75 patients (68%) attended pulmonology care, 55 patients (50%) were hospitalised in the past 6 and 12 months, respectively, and 57 patients (52%) were taking dornase alpha. Five adult patients (10%) received help from non-professional caregiver. Conclusions: Cystic fibrosis leads to significant deterioration of quality of life. This study is the first from the Central Eastern European region that provides basic inputs for further health economic evaluations of cystic fibrosis care. Orv. Hetil., 2014, 155(42), 1673–1684