Exploratory Circular RNA Profiling in Adrenocortical Tumors

The histological differential diagnosis of adrenocortical adenoma and carcinoma is difficult and requires great expertise. Measures taken towards the distinction of adrenal tumors are of paramount importance. The non-coding circular RNAs (circRNAs) were shown to be expressed in a tissue and tumor specific manner. CircRNAs are investigated as a useful adjunct to the differential diagnosis of benign and malignant tumors of several organs, but they have not been investigated in adrenocortical tumors yet. Here, we have performed circRNA profiling in adrenocortical tumors by next-generation sequencing to detect already known and de novo circRNAs. Out of the five most differentially expressed circRNAs, circPHC3 could be confirmed by TaqMan RT-qPCR to be overexpressed in carcinoma and adenoma vs. healthy tissues in an independent validation cohort

Tissue miRNA Combinations for the Differential Diagnosis of Adrenocortical Carcinoma and Adenoma Established by Artificial Intelligence

SIMPLE SUMMARY: The histological differential diagnosis of adrenocortical adenoma and carcinoma is difficult and requires great expertise. MiRNAs were shown to be useful for the differential diagnosis of benign and malignant tumors of several organs, and several findings have suggested their utility in adrenocortical tumors as well. Here, we have selected tissue miRNAs based on the literature search, and used machine learning to identify novel clinically applicable miRNA combinations. Combinations with high sensitivity and specificity (both over 90%) have been identified that could be promising for clinical use. Besides being a useful adjunct to histological examination, these miRNA combinations could enable preoperative adrenal biopsy in patients with adrenal tumors suspicious for malignancy. ABSTRACT: The histological analysis of adrenal tumors is difficult and requires great expertise. Tissue microRNA (miRNA) expression is distinct between benign and malignant tumors of several organs and can be useful for diagnostic purposes. MiRNAs are stable and their expression can be reliably reproduced from archived formalin-fixed, paraffin-embedded (FFPE) tissue blocks. Our purpose was to assess the potential applicability of combinations of literature-based miRNAs as markers of adrenocortical malignancy. Archived FFPE tissue samples from 10 adrenocortical carcinoma (ACC), 10 adrenocortical adenoma (ACA) and 10 normal adrenal cortex samples were analyzed in a discovery cohort, while 21 ACC and 22 ACA patients were studied in a blind manner in the validation cohort. The expression of miRNA was determined by RT-qPCR. Machine learning and neural network-based methods were used to find the best performing miRNA combination models. To evaluate diagnostic applicability, ROC-analysis was performed. We have identified three miRNA combinations (hsa-miR-195 + hsa-miR-210 + hsa-miR-503; hsa-miR-210 + hsa-miR-375 + hsa-miR-503 and hsa-miR-210 + hsa-miR-483-5p + hsa-miR-503) as unexpectedly good predictors to determine adrenocortical malignancy with sensitivity and specificity both of over 90%. These miRNA panels can supplement the histological examination of removed tumors and could even be performed from small volume adrenal biopsy samples preoperatively

Az epesavak és a tripszin hatása a pankreász vezeték sejtek bikarbonát szekréciójára: klinikai és kísérletes vizsgálatok = The effects of bile acids and trypsin on pancreatic ductal bicarbonate secretion.

Az akut pankreatitisz egyik legyakoribb oka az epekövesség. A betegség kialakulásának pontos mechanizmusa azonban nem kellőképpen tisztázott. A projekt során célul tűztük ki az epesavak hatásának vizsgálatát pankreasz duktális epitél sejtekre (PDEC), illetve azok fő funkciójára, a bikarbonát szekrécióra. Azt tapasztaltuk, hogy a luminálisan adott kis dózisú nem konjugált kenodeoxikólsav (CDC) IP3-mediált kalcium szignalizáción keresztül stimulálja a bikarbonát szekréciót, míg a nagy dózis mind apikálisan, mind a bazalisan gátolta azt. A gátló hatás hátterében a CDC jelentős mitokondrium károsító hatása áll, mely következményesen ATP deplécióval jár. Kísérleteink során azt találtuk, hogy az ATP depléció önmagában is képes gátolni a bikarbonát szekréciót. A projekt másik célja az aktiválódott tripszin hatásának vizsgálata volt. Kísérleteinkben igazoltuk, hogy a luminálisan adott tripszin, vagy PAR2 aktiváló peptid (AP) gátolja a bikarbonát szekréciót. Kísérleteink alapján vaószínű, hogy , hogy a tripszin bikarbonát szekréciót gátló hatásának hátterében a luminális CFTR-ra kifejtett gátló hatás állhat. Jelen projekt eredményei nagy mértékben hozzájárulnak az akut pankreatitisz kialakulásának pontosabb megértéséhez. | Cholelithiasis is one of the most common reasons of acute pancreatitis. However the patomechanism of this disease is not well understood. Our aim was to investigate the effects of bile acids on pancreatic ductal epithelial cells (PDEC). We clearly showed that luminal admnistration of small doses chenodeoxycholate (CDC) stimulate bicarbonate secretion via IP3 mediated calcium signalisation. In addition. high doses of CDC administered either from the basolateral or the luminal membrane, inhibit bicarbonate secretion . CDC induced mitochondrial damage, wich evoked intracellular ATP depletion. We fuond, that ATP depletion by itself can inhibit pancreatic bicarbonate secretion. The other aim of the project was to investigate the effects of activated trypsin. We found, that luminal administration of trypsin, or PAR-2 activating peptide (AP) inhibit pancreatic bicarbonate secretion. Furthermore, patch clamp experiments revealed that the inhibitory effect of trypsin on bicarbonate secretion is caused via the inhibition of CFTR Cl- channel. Our results considerably contibute to the pathogenesis of acute pancreatitis

A D-vitamin metabolizmusa humán hepatocellularis carcinomában és az azt körülvevő tumormentes májszövetben

Absztrakt Bevezetés: Az 1,25-dihidroxi-D3-vitamin tumorellenes hatása hepatocellularis carcinomában már részben ismert. Célkitűzés: Az 1,25-dihidroxi-D3-vitamint inaktiváló CYP24A1-mRNS- és fehérjeexpresszió, az aktiváló CYP27B1- és a VDR-mRNS-expresszió mértékének összehasonlítása humán hepatocellularis carcinomában és az azt körülvevő tumormentes májszövetben. Módszer: 13 beteg friss fagyasztott májszövetmintáját a CYP24A1-mRNS- és fehérje-, 36 beteg paraffinba ágyazott májszövetmintáját használtuk a VDR- és a CYP27B1-mRNS-expresszió kimutatására. Az mRNS-expressziót RT-PCR-rel, a fehérjét immunhisztokémiai vizsgálatokkal mértük. Eredmények: A hepatocellularis carcinomaminták többségében kimutatható volt a CYP24A1-mRNS-expresszió, míg a nem tumoros májszövetminták egyikében sem. A CYP27B1- és VDR-expresszió szignifikánsan alacsonyabb hepatocellularis carcinomában a tumormentes májszövethez képest (p<0,05). A CYP24A1-mRNS-expressziót fehérjeszintézis követi. Következtetések: A CYP24A1 inaktiváló enzim jelenléte, az aktiváló CYP27B1 és a VDR csökkent expressziója humán hepatocellularis carcinomában a D-vitamin csökkent helyi aktivitására enged következtetni, mint egy menekülő mechanizmus a tumor részéről a D-vitamin antitumorhatása ellen. Orv. Hetil., 2016, 157(48), 1910–1918. | Abstract Introduction: 1,25-Dihydroxy vitamin D3 mediates antitumor effects in hepatocellular carcinoma. Aim: We examined mRNA and protein expression differences in 1,25-Dihydroxy vitamin D3-inactivating CYP24A1, mRNA of activating CYP27B1 enzymes, and that of VDR between human hepatocellular carcinoma and surrounding non-tumorous liver. Methods: Snap-frozen tissues from 13 patients were studied for mRNA and protein expression of CYP24A1. Paraffin-embedded tissues from 36 patients were used to study mRNA of VDR and CYP27B1. mRNA expression was measured by RT-PCR, CYP24A1 protein was detected by immunohistochemistry. Results: Expression of VDR and CYP27B1 was significantly lower in hepatocellular carcinoma compared with non-tumorous liver (p<0.05). The majority of the HCC samples expressed CYP24A1 mRNA, but neither of the non-tumorous liver. The gene activation was followed by CYP24A1 protein synthesis. Conclusions: The presence of CYP24A1 mRNA and the reduced expression of VDR and CYP27B1 mRNA in human hepatocellular carcinoma samples indicate decreased bioavailability of 1,25-Dihydroxy vitamin D3, providing an escape mechanism from the anti-tumor effect. Orv. Hetil., 2016, 157(48), 1910–1918

The Importance of Aquaporin 1 in Pancreatitis and Its Relation to the CFTR Cl- Channel

Aquaporins (AQPs) facilitate the transepithelial water flow involved in epithelial fluid secretion in numerous tissues;however, their function in the pancreas is less characterized. Acute pancreatitis (AP) is a serious disorder in which specific treatment is still not possible. Accumulating evidence indicate that decreased pancreatic ductal fluid secretion plays an essential role in AP;therefore, the aim of this study was to investigate the physiological and pathophysiological role of AQPs in the pancreas. Expression and localization of AQPs were investigated by real-time PCR and immunocytochemistry, whereas osmotic transmembrane water permeability was estimated by the dye dilution technique, in Capan-1 cells. The presence of AQP1 and CFTR in the mice and human pancreas were investigated by immunohistochemistry. Pancreatic ductal HCO3- and fluid secretion were studied on pancreatic ducts isolated from wild-type (WT) and AQP1 knock out (KO) mice using microfluorometry and videomicroscopy, respectively. In vivo pancreatic fluid secretion was estimated by magnetic resonance imaging. AP was induced by intraperitoneal injection of cerulein and disease severity was assessed by measuring biochemical and histological parameters. In the mice, the presence of AQP1 was detected throughout the whole plasma membrane of the ductal cells and its expression highly depends on the presence of CFTR Cl- channel. In contrast, the expression of AQP1 is mainly localized to the apical membrane of ductal cells in the human pancreas. Bile acid treatment dose- and time-dependently decreased mRNA and protein expression of AQP1 and reduced expression of this channel was also demonstrated in patients suffering from acute and chronic pancreatitis. HCO3- and fluid secretion significantly decreased in AQP1 KO versus WT mice and the absence of AQP1 also worsened the severity of pancreatitis. Our results suggest that AQP1 plays an essential role in pancreatic ductal fluid and HCO3- secretion and decreased expression of the channel alters fluid secretion which probably contribute to increased susceptibility of the pancreas to inflammation

Asszisztált autogén drenázs

Az expectoratio a cystás fibrosis (CF) fizioterápiás kezelés egyik sarokköve. Számos aktív és passzív technika és eszköz áll rendelkezésünkre. Az egyik legnehezebb az újszülöttek kezelése, ahol csak passzív technikákat alkalmazhatunk. Az idősebbeknél használt autogén drenázs passzív változatát, az asszisztált autogén drenázst alkalmazhatjuk újszülötteknél és nagyon fiatal CF betegeknél. Ennek során a kéz segítségével állítjuk be a megfelelő kilégzési áramlást és hosszt, kerülve a túlzott szorítás alkalmazását