Double-Stranded RNA Attenuates the Barrier Function of Human Pulmonary Artery Endothelial Cells

Circulating RNA may result from excessive cell damage or acute viral infection and can interact with vascular endothelial cells. Despite the obvious clinical implications associated with the presence of circulating RNA, its pathological effects on endothelial cells and the governing molecular mechanisms are still not fully elucidated. We analyzed the effects of double stranded RNA on primary human pulmonary artery endothelial cells (hPAECs). The effect of natural and synthetic double-stranded RNA (dsRNA) on hPAECs was investigated using trans-endothelial electric resistance, molecule trafficking, calcium (Ca2+) homeostasis, gene expression and proliferation studies. Furthermore, the morphology and mechanical changes of the cells caused by synthetic dsRNA was followed by in-situ atomic force microscopy, by vascular-endothelial cadherin and F-actin staining. Our results indicated that exposure of hPAECs to synthetic dsRNA led to functional deficits. This was reflected by morphological and mechanical changes and an increase in the permeability of the endothelial monolayer. hPAECs treated with synthetic dsRNA accumulated in the G1 phase of the cell cycle. Additionally, the proliferation rate of the cells in the presence of synthetic dsRNA was significantly decreased. Furthermore, we found that natural and synthetic dsRNA modulated Ca2+ signaling in hPAECs by inhibiting the sarco-endoplasmic Ca2+-ATPase (SERCA) which is involved in the regulation of the intracellular Ca2+ homeostasis and thus cell growth. Even upon synthetic dsRNA stimulation silencing of SERCA3 preserved the endothelial monolayer integrity. Our data identify novel mechanisms by which dsRNA can disrupt endothelial barrier function and these may be relevant in inflammatory processes

Endothelial-Mesenchymal Transition of Brain Endothelial Cells: Possible Role during Metastatic Extravasation

Cancer progression towards metastasis follows a defined sequence of events described as the metastatic cascade. For extravasation and transendothelial migration metastatic cells interact first with endothelial cells. Yet the role of endothelial cells during the process of metastasis formation and extravasation is still unclear, and the interaction between metastatic and endothelial cells during transendothelial migration is poorly understood. Since tumor cells are well known to express TGF-beta, and the compact endothelial layer undergoes a series of changes during metastatic extravasation (cell contact disruption, cytoskeletal reorganization, enhanced contractility), we hypothesized that an EndMT may be necessary for metastatic extravasation. We demonstrate that primary cultured rat brain endothelial cells (BEC) undergo EndMT upon TGF-beta 1 treatment, characterized by the loss of tight and adherens junction proteins, expression of fibronectin, beta 1-integrin, calponin and a-smooth muscle actin (SMA). B16/F10 cell line conditioned and activated medium (ACM) had similar effects: claudin-5 down-regulation, fibronectin and SMA expression. Inhibition of TGF-beta signaling during B16/F10 ACM stimulation using SB-431542 maintained claudin-5 levels and mitigated fibronectin and SMA expression. B16/F10 ACM stimulation of BECs led to phosphorylation of Smad2 and Smad3. SB-431542 prevented SMA up-regulation upon stimulation of BECs with A2058, MCF-7 and MDA-MB231 ACM as well. Moreover, B16/F10 ACM caused a reduction in trans-endothelial electrical resistance, enhanced the number of melanoma cells adhering to and transmigrating through the endothelial layer, in a TGF-beta-dependent manner. These effects were not confined to BECs: HUVECs showed TGF-beta-dependent SMA expression when stimulated with breast cancer cell line ACM. Our results indicate that an EndMT may be necessary for metastatic transendothelial migration, and this transition may be one of the potential mechanisms occurring during the complex phenomenon known as metastatic extravasation

Loci Memoriae Hungaricae

Vorwort (Miklós Takács) - 7 ; Teil I: Galeerensklaverei László Zsigmond Bujtás: Die ungarischen Galeerensklaven-Prediger als Erinnerungsort in der Niederlande im 17–18. Jahrhundert - 11 ; Gábor Pusztai: Michiel de Ruyter in der ungarischen Erinnerung - 44 ; Márta Zsuzsanna Pintér: Theatrum Eperjesiensis. Das Schultheater in Preschau und die Darstellung der Galeerensklaven auf der Bühne - 58 ; Eva Kowalská: Exulanten und Galeerensklaven in den Dokumenten und als Erinnerungsort der slowakischen Lutheraner - 76 ; Dávid Csorba: Die Rezeption des Martyriums der ungarischen calvinistischen Galeerensklaven - 91 ; Gergely Tamás Fazakas: Die Erinnerung an die protestantischen Galeerensträflinge des 17. Jahrhunderts nach dem Freiheitskampf 1848/1849 in Debrecen - 104 ; Teil II: 1848 Pál S. Varga: „Die Menschensaaten wurden zu nichte“ – ZumVerhältnis von individuellen und kollektiven Erfahrungen in der ungarischen Lyrik nach dem Freiheitskampf 1848/49 - 131 ; Péter Bényei: Kollektives Trauma, kommunikatives Gedächtnis: 1848/49 als Erinnerungsort im Novellenzyklus von Mór Jókai Schlachtenbilder und Scenen aus Ungarns Revolution 1848 und 1849) - 139 ; Orsolya Manhercz: Das Gedächtnis von 1848–1849 im Spiegel der Kaiserreise von 1852 - 161 ; Kálmán Kovács: Ban Joseph Jellačić von Bužim als „treuer Diener“ der Stephanskrone. Das reichspatriotische Festspiel Croatiens Jubel von Jellačić - 174 ; Wolfgang Häusler: Wien und Ungarn 1848/49 – Denkmäler und Stätten revolutionärer Erinnerung - 193 ; AutorInnen - 240 ; Namensregister - 24

Ser80Ile mutation and a concurrent Pro25Leu variant of the VHL gene in an extended Hungarian von Hippel-Lindau family

Von Hippel-Lindau disease (VHL) is a rare autosomal dominant disease characterized by development of cystic and tumorous lesions at multiple sites, including the brain, spinal cord, kidneys, adrenals, pancreas, epididymis and eyes. The clinical phenotype results from molecular abnormalities of the VHL tumor suppressor gene, mapped to human chromosome 3p25-26. The VHL gene encodes two functionally active VHL proteins due to the presence of two translational initiation sites separated by 53 codons. The majority of disease-causing mutations have been detected downstream of the second translational initiation site, but there are conflicting data as to whether few mutations located in the first 53 codons, such as the Pro25Leu could have a pathogenic role. In this paper we report a large Hungarian VHL type 2 family consisting of 32 members in whom a disease-causing AGT80AAT (Ser80Ile) c.239G>A, p.Ser80Ile mutation, but not the concurrent CCT25CTT (Pro25Leu) c.74C>T, p.Pro25Leu variant co-segregated with the disease. To our knowledge, the Ser80Ile mutation has not been previously described in VHL type 2 patients with high risk of pheochromocytoma and renal cell cancer. Therefore, this finding represents a novel genotype-phenotype association and VHL kindreds with Ser80Ile mutation will require careful surveillance for pheochromocytoma. We concluded that the Pro25Leu variant is a rare, neutral variant, but the presence such a rare gene variant may make genetic counseling difficult

Abstracts from the 20th International Symposium on Signal Transduction at the Blood-Brain Barriers

https://deepblue.lib.umich.edu/bitstream/2027.42/138963/1/12987_2017_Article_71.pd

The comparative histomorphology and corticosteroid profile of adrenal glands in some African antelopes /

Adrenal glands from five species of South African antelope; cape eland (Taurotragus o.oryx), gemsbok (Oryx g.gazella), southern greater kudu (Tragelaphus s.strepsiceros), red hartebeest (Alcelaphus buselaphus caama), springbok (Antidorcas marsupialis hofmeyri), were collected from 43 trophy-hunted males for histology and corticosteroid analysis. The gross anatomy of the adrenal glands are species-specific, with the left gland being most variable. There were differences found in the number of cortex capsular layers and zona glomerulosa between species. Extensive capsular trabeculae penetrates deep into the cortex in only the largest antelope, i.e. eland and gemsbok, and are representative of these species. In all species the zona glomerulosa form variations in types of cellular cord structures, with the greater kudu having the most unique architecture of horizontally stratified, highly columnar cells that form winding cords which arches at the capsular end, and resemble those observed in equine species. Medullary capsules were observed in the eland, and incomplete capsules in the gemsbok and greater kudu. The medulla is characterized by an outer, adrenaline secreting zone that encapsulates a inner noradrenaline secreting zone in all species. The corticosteroid patterns are typical of bovids, with cortisol and corticosterone present, however significantly larger amounts of 18-hydroxy-corticosterone were found in all species of antelope. The total identified corticosteroid contents had interspecies differences, which are possibly based on species body size

Volumetric hemodynamic changes and postoperative complications in hypothermic liver transplanted patients

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