Funkcionális genomikai rendszer kidolgozása gyermekkori acut lymphoid leukémiában DNS és cDNS chip analízis valamint immunfenotipizálás alapján = Development of a novel functional genomic system in pediatric acute lymphoid leukemia based on DNA and cDNA chip analysis as well as immunophenotyping

Olyan molekuláris markerek genomikai identifikálását tűztük ki célul, melyek a gyermekkori akut limfoid leukémia (ALL) patomechanizmusának felderítésében, osztályzásának pontosításában és a kezelésre adott válasz predikciójában szerepet játszhatnak. A blaszt-minták gyűjtésének akadályozottsága miatt projektünkben a génpolimorfizmusok vizsgálata kapott fő hangsúlyt. Létrehoztunk egy nemzetközi viszonylatban is kimagaslóan nagynak számító biobankot ALL-es és osteosarcomás gyermekek továbbá egészséges véradók genomi DNS-mintáiból. Az ABCB1 1236C/T, 2677G/T,A és 3435T/C, ABCC1 2012G/T, ABCC2 1249G/A és 3563T/A, ABCC3 -211C/T, ABCG2 34G/A és 421C/A, SLC19A1 80G/A és GGH -401C/T génpolimorfizmusok meghatározását végeztük, és korreláltattuk a betegek adataival. Eredményeink szerint az ABCB1 és ABCG2 vizsgált polimorfizmusai nem hajlamosítanak ALL kialakulására. Megállapítottuk, hogy az ABCB1 3435T/C genotípus prediktív értékkel bír a vizsgált kemoterápiás protokoll kapcsán kialakuló immunszuppresszió vonatkozásában. Az ABCB1 3435TT genotípus és az ABCG2 421A allél hordozása szinergisztikus interakció révén hajlamosít a citosztatikus szerek központi idegrendszeri mellékhatásaira ALL esetében. A GGH 401TT genotípusú betegek metotrexát-eliminációja gyorsabbnak bizonyult mind az ALL-es, mind az osteosarcomás betegek adagolási sémája esetén, és súlyos szövődmények is ritkábban lépnek fel. Ezen eltérések alapját képezhetik a terápia-individualizációt előkészítő további tanulmányoknak | Our objective was to identify molecular markers in the genome that play important role in the pathomechanism, accurate classification and response to therapy in childhood acute lymphoblastic leukemia (ALL). Given the fact that the collection of malignant blast samples was highly hampered, we mainly concentrated on studying somatic gene polymorphisms. A large DNA-biobank was established consisting of samples from pediatric ALL, osteosarcoma and healthy blood donors. The ABCB1 1236C/T, 2677G/T,A and 3435T/C, ABCC1 2012G/T, ABCC2 1249G/A and 3563T/A, ABCC3 -211C/T, ABCG2 34G/A and 421C/A, SLC19A1 80G/A, GGH -401C/T polymorphisms were genotyped and the association of genotype and clinical data were examined. In our study, the polymorphisms of ABCB1 and ABCG2 were not related to susceptibility to ALL. We found that the ABCB1 3435T/C genotype is predictive for the severe immunosuppression developed during the given ALL chemotherapy protocol. The ABCB1 3435TT genotype and carrying ABCG2 421A allele exert synergistic interaction in predisposing patients to the central nervous system adverse effects of cytostatic drugs in ALL. The elimination of methotrexate proved to be faster in patients of GGH -401TT genotype in case of both chemotherapy regimens used in ALL and osteosarcoma, and severe side effects of methotrexate was also more rare in this group. These results may lay the foundation of further studies to develop individual drug dosing protocols

Spectral density of Barkhusen noise in FINEMET-type materials

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Altered agonist sensitivity of a mutant V2 receptor suggests a novel therapeutic strategy for nephrogenic diabetes insipidus.

Loss of function mutations of the type 2 vasopressin receptor (V2R) in kidney can lead to nephrogenic diabetes insipidus (NDI). We studied a previously described, but uncharacterized mutation of V2R (N321K missense mutation) of an NDI patient. The properties of the mutant receptor were evaluated. We constructed a highly sensitive Epac based BRET (bioluminescence resonance energy transfer) biosensor to perform real-time cAMP measurements after agonist stimulation of transiently transfected HEK293 cells with V2Rs. beta-arrestin binding of the activated receptors was examined with luciferase-tagged beta-arrestin and mVenus-tagged V2Rs using BRET technique. Cell surface expressions of HA-tagged receptors were determined with flow cytometry using anti-HA-Alexa488 antibodies. Cellular localization examinations were implemented with fluorescent tagged receptors visualized with confocal laser-scanning microscopy. The effect of various vasopressin analogues on V1R was tested on mouse arteries by wire myography. N321K mutant V2R showed normal cell surface expression but the potency of AVP for cAMP generation was low, while the clinically used desmopressin (dDAVP) was not efficient. The beta-arrestin binding and internalization properties of the mutant receptor were also different compared to the wild type. Function of the mutant receptor can be rescued with administration of V2R receptor agonist dVDAVP, which had no detectable side effects on V1R in the effective cAMP generating concentrations. Based on the findings we could propose a therapeutical strategy for NDI patients carrying the N321K mutation, since our in vivo experiments suggest that dVDAVP could rescue the function of the N321K-V2R without significant side effect on V1R

What have we learned from two-pore potassium channels? Their molecular configuration and function in the human heart

Two-pore domain potassium channels (K2P) control excitability, stabilize the resting membrane potential below firing threshold, and accelerate repolarisation in different cells. Until now, fifteen different genes for the six K2P channel subfamily were cloned. The pore-forming part is translated from two genes and they are built up from a dimer of two two-unit transmembrane domains functioning with a wide spectrum of physiological profiles. K2P ion channels were discovered in the last two decades and gave novel opportunity to recognize the complex molecular mechanism of the potassium ion flux, and may lead to the design of individual drug targeting in the future. In this review, we summarise the structure, function, channelopathies and pharmacological silhouette of the two-pore potassium channels in the human tissues. In addition, we present the computer model of the partially reconstructed wild type K2P1/TWIK1 lacking the intracellular C and N terminal loop

dUTPase expression correlates with cell division potential in Drosophila melanogaster

dUTPase is a dNTP sanitizing enzyme that prevents the appearance of the potentially harmful uracil bases in DNA by hydrolyzing cellular dUTP. This function of dUTPase is found to be essential in many organisms including Drosophila melanogaster. Previously we showed that the expression pattern of dUTPase determines the extent of uracil accumulation in the genome of different tissues. We wished to reveal the regulatory mechanism that eventually leaves a set of tissues to have uracil-free and intact genome. We found that the expression pattern established by the promoter of Drosophila dUTPase overlaps with mRNA and protein expression pattern, excluding the involvement of other posttranscriptional contribution. This promoter was found to be active in primordial tissues, such as in imaginal discs of the larvae, in the larval brain and in reproductive organs. In the case of brain and imaginal tissues, we observed that the promoter activity depends on DRE motifs, the docking site of DREF, which is known as a transcriptional activator of genes involved in replication and proliferation. These results suggest that dUTPase expression is fine-tuned to meet the requirements of DNA synthesis, in tissues where the maintenance of genome integrity is of high importance. This article is protected by copyright. All rights reserved

Expression of Tight Junction Components in Hepatocyte-Like Cells Differentiated from Human Embryonic Stem Cells

Human embryonic stem cells can be differentiated in vitro into a wide variety of progeny cells by addition of different morphogens and growth factors. Our aim was to monitor the expression pattern of tight junction (TJ) components and various cellular markers during differentiation of stem cell lines toward the hepatic lineage. Human embryonic stem cell lines (HUES1, HUES9) were differentiated into endoderm-like cells, and further differentiated to hepatocyte-like cells. Gene expressions of Oct3/4, Nanog, alpha-fetoprotein, albumin, cytokeratins (CK-7, CK-8, CK-18, CK-19), ATP-binding cassette (ABC) transporters (ABCC2, ABCC7, ABCG2), and various TJ components, including claudin-1, claudin-4, claudin-5, claudin-7, and tricellulin, as well as an extracellular matrix component, agrin were monitored during hepatic differentiation by real-time quantitative PCR. The differentiated cells exhibit epithelial morphology and functional assessments similar to that of hepatocytes. The expression level of stem cell marker genes (Oct3/4 and Nanog) significantly and gradually decreased, while liver-associated genes (alpha-fetoprotein, albumin) reached their highest expression at the end of the differentiation. The endoderm-like cells expressed claudin-1, which declined eventually. The expression levels of cholangiocyte markers including claudin-4, CK-7, CK-19, and agrin gradually increased and reached their highest level at the final stage of differentiation. In contrast, these cells did not express notable level of claudin-7, CK-8 and tricellulin. The marker set used for monitoring differentiation revealed both hepatocyte and cholangiocyte characteristics of the differentiated cells at the final stage. This is the first report describing the expression level changes of various TJ components, and underlining their importance in hepatic differentiation. © 2015 Arányi Lajos Foundatio

A növénytermesztési folyamatok modellezése = Modelling of crop production processes

Az elvégzett feladatokat négy nagyobb csoportba lehet sorolni: 1. Intelligens információs rendszerek építése és folyamatos karbantartása 2. Növényi növekedési modellek összegyűjtése, adaptálása, fejlesztése 3. Kockázatelemzés 4. Döntéstámogató rendszerek építése 1. Az információs rendszerek témakör a következő létrehozott fontos adatbázisokat tartalmazza: meteorológiai adatbázis historikus mért adatokra, meteorológiai adatbázis szcenáriókra, tartamkísérletek információi, talajtani információs rendszer, növényi termésátlagok megyei idősorai, zöldségtermesztési agroökológiai információs rendszer. 2. A növényi növekedési modellekkel kapcsolatban foglalkoztunk az agroökológiai táplálékhálózati rendszer populációdinamika biomassza modelljének a fenológiai általánosításával, nemzetközileg elfogadott modellek összegyűjtésével és összehasonlító tesztelésével, a növénytermesztés és időjárás sztochasztikus kapcsolatának elemzésével, a búza és kukorica modellek adaptálásával és hasznosítási lehetőségük feltárásával. 3. A kockázat mértékének növekedését az általánosított sztochasztikus dominancia-módszer egy közelmúltban egyszerűsített változatával, a kockázati averziótól is függő sztochasztikus hatásossági kritérium segítségével igazoltuk. | The completed tasks can be classified into four groups: 1. Constructing and continious maintainence of intelligent information systems: a. Database of historical meteorological data b. Database of climate scenario meteorological data c. Information of field experiments d. Soil information system e. Regional time series of plant yield averages f. Agroecological information system of vegetables 2. Collecting, adapting and development of plant growth models a. Phenological generalisation of the biomass model of the agroecological food web population dynamics. b. Collecting and comparative testing of internationally accepted models. c. Analysis of the stochastic relationship of the crop production and weather. d. Adaptation and revealing of the utilization possibilities of the wheat and maize models. 3. Risk analysis a. We justified the increasing of risk with a recently simplified version of the stochastic dominance method, namely with the stochastic effectiveness criteria depending on risk aversion. 4. Development of decision support systems: a. Planning of environmentally sparing agriculture, sustainable management, study of agrometeorological and pest dynamical conditions