Genomiális eltérések és génexpresszió közötti kapcsolat vizsgálata, melanomák metasztázisképzésre jellemző genetikai markerek kutatása = Association between genomic alterations and gene expression, search for genetic markers in association with melanoma metastasis formation

A projekt keretében melanomák genom és génexpressziós eltéréseit analizáltuk. Q-PCR-el gyakori mutációt találtunk a BRAF és NRAS onkogénekre. BRAF mutációval asszociálódott jellegzetes eltérések a 7p22, 7q21, 7q31 szakaszok többlete és a 10q21, 10q26 lókuszok vesztése voltak. Hierachikus klaszter analizissel két jellegzetes molekuláris alosztályt figyeltünk meg, amely a kevésbé aggresszív tumoroktól az agresszív daganatokat egyértelműen elválasztották. 1095 eltérő expressziójú gén az ulcerációval asszociálódott. Közülük 1021 gén alulregulált volt a rosszprognózisú csoportban, és érdekes, hogy ugyanezek a gének magas expressiójuak voltak a másik csoportban. Metasztázisok génexpressziója az ulcerációval mutatott azonos mintázatot. Az ulcerált felszínű melanomákban csökkent expressziójú gének funkcióját vizsgálva kimutattuk, hogy többségük a bőr- és szőrfejlődési funkció, dermatológiai betegségek, daganatfejlődés, sejtosztódás a sejtciklus, sejt-sejt interakció és sejtmotilitás szabályozó folyamatokhoz tartozik, a p53, ERK/MAP, IP3/AKT és WNT/? katenin, Nf-?B molekulási útvonalakon hatva. Interfázisos FISH analízissel kimutattuk, hogy az EGFR génkópiaszám többlete, primer melanomákban rossz prognózissal társul. A 9p21-es lokusz delécióját mind a korai, mind a késői melanomákban megfigyeltük. Nem találtunk szoros korrelációt a 9p21-es lokusz genetikai eltérései és a daganatok klinikopatológiai tulajdonságai között. | Within the framework of this project using Q-PCR we found frequent mutation of the BRAF and NRAS oncogenes in primary melanomas. Genom alterations exclusively associated with BRAF mutation were gains of 7p22, 7q21, 7q31, and losses of 10q21, 10q26. Unsupervised cluster analysis of geneexpression data revealed two characteristic molecular subclasses of melanoma, segregating aggressive tumors from less aggressive ones. Differentially expressed genes (1095) were associated with ulceration. Majority these (1021) were downregulated in subclass with bad prognosis and upregulated in the less aggressive group. Geneexpression signature of metastastases displayed similar signature, majority of the genes defined for the ulcerated lesions showed reduced expression in metastatic tumors. Using functional annotations five networks were identified belonging into the hair and skin development-, cancer-, cellular growth- and proliferation and affect the p53, Wnt/?-catenin and Nf-?B signaling pathways. By FISH we demonstrated that elevated copies of EGFR gene is associated with poor prognosis. Outcome of patients whose primary tumors had highly amplified EGFR gene was poor. Correlation between the gene amplification, mRNA level and protein expression was not linear. 9p21 deletion was present in early and late stages of the disease with similar frequency. We could not find strong correlation the 9p21 status of melanomas and the patients? clinical parameters

Prognosis in human glioblastoma based on expression of ligand growth hormone-releasing hormone, pituitary-type growth hormone-releasing hormone receptor, its splicing variant receptors, EGF receptor and PTEN genes

Purpose G lioblastoma (GB) is the most frequent brain tumor. Despite recent improvement in therapeutic strategies, the prognosis of GB remains poor. Growth hormone-releasing hormone (GHRH) may act as a growth factor; antagonists of GHRH have been successfully applied for experimental treatment of different types of tumors. The expression profile of GHRH receptor, its main splice variant SV1 and GHRH have not been investigated in human GB tissue samples. Methods We examined the expression of GHRH, fulllength pituitary-type GHRH receptor (pGHRHR), its functional splice variant SV1 and non-functional SV2 by RTPCR in 23 human GB specimens. Epidermal growth factor receptor (EGFR) and phosphatase and tensin homolog gene (PTEN) expression levels were also evaluated by quantitative RT-PCR. Correlations between clinico-pathological parameters and gene expressions were analyzed. Results E xpression of GHRH was found to be positive in 61.9 % of samples. pGHRH receptor was not expressed in our sample set, while SV1 could be detected in 17.4 % and SV2 in 8.6 % of the GB tissues. In 65.2 and 78.3 % of samples, significant EGFR over-expression or PTEN under-representation could be detected, respectively. In 47.8 % of cases, EGFR up-regulation and PTEN down-regulation occurred together. Survival was significantly poorer in tumors lacking GHRH expression. This worse prognosis in GHRH negative group remained significant even if SV1 was also expressed. Conclusion Our study shows that GHRH and SV1 genes expressed in human GB samples and their expression patterns are associated with poorer prognosis

Concurrence of chromosome 3 and 4 aberrations in human uveal melanoma

Uveal melanoma (UM) is the most common primary intraocular malignancy with a very poor prognosis. The most frequent chromosome aberration in UM is the monosomy of chromosome 3. Previously, we demonstrated that ~50% of UMs express type-I receptor for luteinizing hormone-releasing hormone (LH-RH-R). The gene encoding LH-RH-R is located in chromosome 4 (location: 4q21.2); however, the occurrence of numerical aberrations of chromosome 4 have never been studied in UM. In the present study, we investigated the abnormalities of chromosome 3 and 4, and the possible correlation between them, as well as with LH-RH-R expression. Forty-six specimens of UM were obtained after enucleation. Numerical aberrations of chromosome 3 and 4 were studied by fluorescence in situ hybridization (FISH). Chromosome 4 was detected in normal biparental disomy only in 14 (30%) samples; however, 32 cases (70%) showed more than 2 signals/nucleus. Monosomy of chromosome 3 could be found in 16 (35%) samples. In 6 specimens (13%), more than 2 copies of chromosome 3 were found, while normal biparental disomy was detected in 24 (52%) samples. Statistical analysis indicated a statistically significant (p<0.05) correlation between the copy number of chromosome 3 and 4. Moreover, moderate difference was revealed in the survival rate of the UM patients with various pathological profiles. No correlation was found between chromosome aberrations and LH-RH-R expression. Our results clearly demonstrate abnormalities in chromosome 3 and 4 and the incidence of the monosomy of chromosome 3 in human UM. In summary, our results provide new incite concerning the genetic background of this tumor. Our findings could contribute to a more precise determination of the prognosis of human UM and to the development of new therapeutic approaches to this malignancy

Use of vitamin K antagonists for secondary stroke prevention depends on the treating healthcare provider in Germany – results from the German AFNET registry

Background Anticoagulation using vitamin K antagonists (VKAs) significantly reduces the risk of recurrent stroke in stroke patients with atrial fibrillation (AF) and is recommended by guidelines. Methods The German Competence NETwork on Atrial Fibrillation established a nationwide prospective registry including 9,574 AF patients, providing the opportunity to analyse AF management according to German healthcare providers. Results On enrolment, 896 (9.4 %) patients reported a prior ischaemic stroke or transient ischaemic attack. Stroke patients were significantly older, more likely to be female, had a higher rate of cardiovascular risk factors, and more frequently received anticoagulation (almost exclusively VKA) than patients without prior stroke history. Following enrolment, 76.4 % of all stroke patients without VKA contraindications received anticoagulation, which inversely associated with age (OR 0.95 per year; 95 % CI 0.92–0.97). General practitioners/internists (OR 0.40; 95 % CI 0.21–0.77) and physicians working in regional hospitals (OR 0.47; 95 % CI 0.29–0.77) prescribed anticoagulation for secondary stroke prevention less frequently than physicians working at university hospitals (reference) and office-based cardiologists (OR 1.40; 95 % CI 0.76–2.60). The impact of the treating healthcare provider was less evident in registry patients without prior stroke. Conclusions In the AFNET registry, anticoagulation for secondary stroke prevention was prescribed in roughly three-quarters of AF patients, a significantly higher rate than in primary prevention. We identified two factors associated with withholding oral anticoagulation in stroke survivors, namely higher age and—most prominently—treatment by a general practitioner/internist or physicians working at regional hospitals

Psychosocial factors and major adverse cardiac and cerebrovascular events after cardiac surgery

Our aim was to prospectively examine the association of psychosocial factors with adverse outcome after cardiac surgery. One hundred and eighty cardiac surgery patients were enrolled and contacted annually by mail. Depression [Beck depression inventory (BDI)], anxiety [state anxiety subscale in Spielberger State-Trait Anxiety Inventory (STAI-S) and trait anxiety subscale in Spielberger State-Trait Anxiety Inventory (STAI-T)] were investigated annually, social support, negative affectivity, social inhibition (SI), illness intrusiveness, self-rated health and sleeping disorders were investigated by standardized tests at the second and fifth year. The end-point was the major adverse cardiac and cerebrovascular event (MACCE) including death. Twenty-eight (15.5%) patients died by the end of the fifth year. At the end of the second and fifth years, 146 (81.1%) and 118 (65.5%) patients fulfilled the tests, respectively. At the end of the second year after adjustment for medical and perioperative factors worse self-rated health [adjusted hazard ratio (AHR): 0.67, P=0.006], sleeping disorders (AHR: 1.14, P=0.001), higher illness intrusiveness (AHR: 1.03, P=0.018), higher BDI (AHR: 1.12, P=0.001), STAI-S (AHR: 1.09, P=0.001) and higher STAI-T scores (AHR: 1.08, P=0.002) showed higher risk for MACCE. Significant individual elevation in scores of sleeping disorders, illness intrusiveness and SI were observed over the three-year period in the MACCE group. Assessment of psychosocial factors could help in identifying patients at high-risk for MACCE after cardiac surgery

Crosstalk between reactive oxygen species and pro-inflammatory markers in developing various chronic diseases: a review

The inflammation process in the human body plays a central role in the pathogenesis of many chronic diseases. In addition, reactive oxygen species (ROS) exert potentially a decisive role in human body, particularly in physiological and pathological process. The chronic inflammation state could generate several types of diseases such as cancer, atherosclerosis, diabetes mellitus and arthritis, especially if it is concomitant with high levels of pro-inflammatory markers and ROS. The respiratory burst of inflammatory cells during inflammation increases the production and accumulation of ROS. However, ROS regulate various types of kinases and transcription factors such nuclear factor-kappa B which is related to the activation of pro-inflammatory genes. The exact crosstalk between pro-inflammatory markers and ROS in terms of pathogenesis and development of serious diseases is still ambitious. Many studies have been attempting to determine the mechanistic mutual relationship between ROS and pro-inflammatory markers. Therefore hereby, we review the hypothetical relationship between ROS and pro-inflammatory markers in which they have been proposed to initiate cancer, atherosclerosis, diabetes mellitus and arthritis