Congenital hyperinsulinism caused by a de novo mutation in the ABCC8 gene: a case report

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Thrombocyta aktiváció során bekövetkező változások biokémiai és sejtbiológiai jellemzés = Biochemical and cell biological changes during platelet activation

Ex vivo mintákon végzett analízisek közül elvégeztük 30 db stent beültett beteg esetében perifáriás vérminták analízisét az alábbi thrombocyta aktivációs markerekre: thrombocyta P-selectin, solubilis P-selectin, CD63 expresszió, heterotipikus aggregátumok aránya, mikropartikula mennyiség. Fenti eredményeket korreláltattuk a betegek előzetes kezelési protokolljával (csak ASA, vagy ASA+clopidogrel). A XIII-as faktorral (F XIII) kapcsolatos vizsgálatok során megállapítást nyert, hogy thrombocyták TRAP aktiválása, az FXIII expresszió fokozódásával jár. Azonban kimutattuk, hogy ez a plazmában lévő FXIII kötődése és nem az intracellluláris FXIII felszíni megjelenése miatt jön létre. Specifikus fibrinogének felhasználásával tisztáztuk az FXIII kötődés biokémiai jellemzőit. A thrombocyta foszfatáz inhibitorokkal végzett kísérletek során PRP-ben bizonyítottuk, hogy az 1 és 2A típusú foszfatázoknak csak együttes gátlása hozza létre a thrombocyta aktiválás során észlelhető hatást. TRAP aktivált mintákon bizonyítottuk, hogy mind az aggregáció, mind a P-selectin expresszió dózisfüggő módon gátlódik calyculin (CLA) hatására. Ugyanakkor nem aktivált mintákon azt találtuk, hogy a CLA dózisfüggő módon alakváltozást hoz létre a thrombocytákban, mely aggregometriával, valamint áramlási citometriai fényszórási és antigén expressziós vizsgálatokkal bizonyítható volt. | On ex vivo samples we examined 30 cases of stent implanted patients where in peripheral blood samples the following platelet activation markers were investigated: paletelt P-selectin, soluble P-selectin, CD63, heterotypic aggregates, microparticles. The above data were correlated with previous treatment protocol of the patients (ASA only or ASA and clopidogrel). Regarding studies with factor XIII we identified that in TRAP-activated platelets FXIII is elevated on the platelet surface. However, we proved, that FXIII is not derived from intracellular source but is attached from the plasma. By using specific fibrinogens we identified the biochemical requirements for FXIII binding to platelets. The manuscript was submitted to Thrombosis and Haemostasis where a minor revision was required. Regarding phosphatase inhibitors we found that phosphatase 1 and 2A types are both involved in platelet activation, since their simultaneous inhibition by calyculin is required to prevent TRAP-elicited P-selectin axpression and platelet aggregation. At the same time on non-activated platelet samples we found that calyculin dose-dependently inhibited platelet shape-change as measured by aggregometry and flow cytometry

Sejt- és molekuláris biológiai tulajdonságok vizsgálata gyermekkori akut lymphoblastos leukaemiában. A minimális reziduális betegség nyomonkövetése, új kezelés stratégia kialakítása = Cellular and molecular biological properties of childhood acute lymphoblastic leukemia. Assessment of minimal residual disease and development of novel therepautic strategies

A gyermekkor leggyakoribb neoplasztikus elváltozásai a rosszindulatú vérképzőszervi betegségek. Fő célunk a gyermekkori akut lymphoblastos leukaemia (ALL), az akut myeliod leukaemia (AML), valamint a myelodysplasiás szindróma (MDS) kezelési eredményeinek és a tartós életminőség javítása volt új sejt- és molekuláris biológiai diagnosztikai és terápiás eljárások bevezetésével. Elemeztük a kombinált citosztatikus ALL-BFM-95 protokollal elért kezelési eredményeket gyermekkori ALL-ben. Tanulmányoztunk új leukaemia-asszociált marker (FXIII-A) kifejeződését AML és ALL blasztokban és négyszínű immunfluoreszcens jelölés alkalmazási lehetőségeit. Ezen módszerek alkalmasak a leukaemiás sejtpopuláció megbízható azonosítására és a minimális maradék betegség (MRD) meghatározására, ezért szerepet játszhatnak a rizikóbecslésben. Elemeztük serkentő és gátló hatású citokinek szabályozó hatásait gyermekkori ALL-ben, MDS-ben, essentiális thrombocythemiában és összehasonlításként lobos folyamatokban. Vizsgáltuk a hagyományos citosztatikus kezelés kiegészítésére lehetőséget teremtő innovatív terápiás eljárásokat, rituximab, interferon-alfa, cisz-retinsav és bcl-2 antisense oligonukleotid preparatum in vitro és in vivo terápiás effektusait. A sejtterápiás eljárások fejlesztése céljából modellként tanulmányoztuk Glanzmann thrombasthaeniás beteg rendellenességét komplex molekuláris módszerekkel, valamint a kóros és a vad típusú gének transzfektálásának hatékonyságát emlős sejtekben. | Hematopietic malignancies represent the most frequent form of cancer in children. The major aim of the present project was to improve treatment results and quality of life in children with acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS) by introducing novel cellular and molecular biological diagnostic and therapeutic methods. Nationwide results in children with ALL treated with the conventional combined cytostatic ALL-BFM-95 protocol were analyzed retrospectively. Expression of a novel leukemia-associated marker (FXIII-A) was studied in AML és ALL blasts and four color flow cytometric immunophenotyping was introduced. These methods allow the exact identification of the leukemic cell population rendering them powerful tools for assessing minimal residual disease and risk estimation. Regulatory effects of stimulatory and inhibitory cytokines were analyzed in childhood ALL, MDS and essential thrombocythemia as compared to inflammatory lesions. Innovative adjuvant treatment approaches, such as in vitro and in vivo therapeutic effects of rituximab, interferon-alpha, cis-retinoic acid and bcl-2 antisense oligonuleotide preparation were studied. As a molecular model for developing cell therapeutical methods, aberration of a patient with Glanzmann thrombasthenia was assessed by applying complex molecular methods and transfecting the mutant and wild-type genes into mammalian cells

Comparison of a New P2Y12 Receptor Specific Platelet Aggregation Test with Other Laboratory Methods in Stroke Patients on Clopidogrel Monotherapy

BACKGROUND: Clinical studies suggest that 10-50% of patients are resistant to clopidogrel therapy. ADP induced platelet aggregation, a widely used test to monitor clopidogrel therapy, is affected by aspirin and is not specific for the P2Y12 receptor inhibited by clopidogrel. OBJECTIVES: To develop a P2Y12-specific platelet aggregation test and to compare it with other methods used for monitoring clopidogrel therapy. PATIENTS/METHODS: Study population included 111 patients with the history of ischemic stroke being on clopidogrel monotherapy and 140 controls. The effect of clopidogrel was tested by a newly developed ADP(PGE1) aggregation test in which prostaglandin E1 treated platelets are used. Results of conventional ADP induced platelet aggregation, VerifyNow P2Y12 assay and ADP(PGE1) aggregation were compared to those obtained by flow cytometric analysis of vasodilator stimulated phosphoprotein (VASP) phosphorylation. Reference intervals for all assays were determined according to the guidelines of Clinical Laboratory Standards Institute. RESULTS: The P2Y12-specificity of ADP(PGE1) test was proven by comparing it with ADP aggregation in the presence of P2Y1 antagonist, adenosine 3', 5'-diphosphate. The method was not influenced by aspirin treatment. Approximately 50% of patients were clopidogrel resistant by conventional ADP aggregation and VerifyNow tests. The ADP(PGE1) method and the VASP phosphorylation assay identified 25.9% and 11.7% of patients as non-responders, respectively. ADP(PGE1) aggregation showed good correlation with VASP phosphorylation and had high diagnostic efficiency. CONCLUSION: The new ADP(PGE1) method is a reliable test for monitoring P2Y12 receptor inhibition by platelet aggregation. As a subset of patients are non-responders, monitoring clopidogrel therapy by adequate methods is essential

Low thrombin generation predicts poor prognosis in ischemic stroke patients after thrombolysis.

Thrombolysis by intravenous recombinant tissue plasminogen activator (rt-PA) is an effective therapy in acute ischemic stroke (AIS). Thrombin generation test (TGT) is a global hemostasis test providing information about the speed and amount of generated thrombin in plasma. Here we aimed to find out whether results of this test before the initiation of thrombolysis might predict outcomes. Study population included 120 consecutive AIS patients, all within 4.5 hours of their symptom onset, who underwent thrombolysis by rt-PA. Blood samples were collected from all patients upon admission and TGT was performed using platelet poor plasma. Clinical data of patients including the NIHSS were registered at admission, day 1 and 7 after therapy. The ASPECT score was assessed using CT images taken before and 24 hours after thrombolysis. Long-term functional outcome was defined 3 months after the event by the modified Rankin Scale. Endogenous Thrombin Potential (ETP) and Peak Thrombin were significantly lower in patients with cardioembolic IS. Symptomatic intracranial hemorrhage (SICH) was found in 6 patients and was significantly associated with low ETP and Peak Thrombin levels. A multiple logistic regression model revealed that an ETP result in the lower quartile is an independent predictor of mortality within the first two weeks (OR: 6.03; 95%CI: 1.2-30.16, p<0.05) and three months after the event (OR: 5.28; 95%CI: 1.27-21.86, p<0.05). Low levels of ETP and Peak Thrombin parameters increase the risk of therapy associated SICH. A low ETP result is an independent predictor of short- and long-term mortality following thrombolysis

Analytical parameters and validation of homopolymer detection in a pyrosequencing-based next generation sequencing system

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Impaired Immunosuppressive Effect of Bronchoalveolar Mesenchymal Stem Cells in Hypersensitivity Pneumonitis: preliminary findings

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Use of Hemadsorption in a Case of Pediatric Toxic Shock Syndrome

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