Release of annexin V-binding membrane microparticles from cultured human umbilical vein endothelial cells after treatment with camptothecin

BACKGROUND: Elevated plasma counts of endothelial microparticles (MP) have been demonstrated in various diseases with a vascular injury component. We used flow cytometry to study the MP-release from cultured human umbilical vein endothelial cells (HUVEC) stimulated by various agonists. MP-release by a topoisomerase I inhibitor camptothecin has been studied in detail. RESULTS: Overnight stimulation of HUVEC with either LPS or TNFα, or 30 min stimulation with thrombin, phorbol-myristate-acetate, tissue plasminogen activator, or angiotensin-II did not cause a significant release of annexin V-binding MP. In contrast, induction of apoptosis with 5 μM camptothecin, documented by 60–70% desquamation of HUVEC culture, annexin V-binding to the cells and DNA-fragmentation, led to a release of annexin V-binding microparticles (~80,000 MP/10(3) cells). This microparticle-release was prevented by Z-Val-Ala-Asp-fluoromethyl-ketone (ZVAD). Lower concentration of camptothecin (500 nM) induced comparable microparticle-release without loss of the culture confluence and without increase in annexin V-binding to the cells or DNA-fragmentation. Analyzed microparticles were free of nucleic acids and 95% of microparticles were 0.3–1 μm in size. Double-labeling flow cytometry assay showed that all annexin V-binding Microparticles expressed CD59 but only approximately 50% of these also expressed CD105. CONCLUSIONS: Camptothecin treated HUVEC released different populations of annexin V-binding membrane microparticles at early stage after proapoptotic stimulation before detection of phosphatidylserine exposure on the cells or DNA fragmentation. The microparticle-release was ZVAD sensitive but was not enhanced at the executive phase of apoptosis. These observations offer a new insight into microparticle-release as a marker of endothelial stimulation and injury

Antidepressant Use as an Example of the Medicalization and Pharmaceuticalisation of Life

Pojem medikalizace označuje proces, skrze nějž je určitý dříve nemedicínský problém nově transformován v problém medicínský. Cílem této práce je ukázat, jak lze pomocí tohoto konceptu přispět k lepšímu pochopení fenoménu současné masové spotřeby antidepresiv. Pokouším se zde demonstrovat, že nárůst v užívání antidepresiv, k němuž v posledních desetiletích celosvětově dochází, je zapotřebí chápat jako výsledek řady faktorů, mezi nimiž stěžejní roli hraje právě změna v chápání určitých emočních stavů, dříve považovaných za přirozené, jakožto symptomů psychické poruchy. Kromě obecné analýzy činitelů, jež spotřebu těchto léků podporují, pak práce obsahuje rovněž vlastní případovou studii jednoho z nich, jímž je způsob, jakým o antidepresivech pojednávají média. Zaměřuji se zde na novinové články, publikované mezi lety 1996 a 2016 v rámci českého celostátního tisku a pokouším se ukázat, jak i přes svou relativní neutralitu diskuze, které se v nich o problematice antidepresiv vedou, svým akcentováním medikalizovaného rámce pohledu na psychické problémy ve výsledku napomáhají vytvářet prostředí, které jejich užívání podporuje.The term medicalization refers to the process through which a previously nonmedical issue newly becomes conceived of as a medical one. The aim of this thesis is to show how the utilization of this concept can help lead to a better understanding of the contemporary phenomenon of mass antidepressant consumption. I attempt to demonstrate here that the rise in antidepressant use, which has been occurring worldwide in the last few decades, must be understood as the result of a host of factors, among which a key role is played by changes in the conception of certain emotional states, previously understood as normal, as symptoms of mental illness. Aside from a general analysis of the factors that contribute to the consumption of these medications, the thesis also contains my own case study of one of them, namely the ways in which antidepressants are portrayed by the media. I focus on articles, published between the years 1996 and 2016 in Czech national newspapers and attempt to show how discussions about antidepressants, despite their relative neutrality, by accenting a medicalized frame of understanding mental problems help to create an environment, which supports their consumption.Department of SociologyKatedra sociologieFilozofická fakultaFaculty of Art

Dundonald Links Golf Club, Skotsko

Zadání bakalářské práce reaguje na absenci reprezentativních prostorů a funkčního zázemí na golfovém hřišti Dundonald Links, které se nachází na západním pobřeží skotské správní oblasti Severní Ayshire. Hlavním cílem je navrhnout objekt, který bude kapacitně a esteticky disponovat prostory k pořádání nejvýznamnějších golfových turnajů. V současnosti je vybudované samotné golfové hřiště, u kterého je provizorně řešena klubovna. Návrh by měl respektovat potřeby provozu golfové klubovny: zázemí pro golfový klub (šatny, proshop, administrativa, aj.), hotelová část a restaurační část vč. technického zázemí a odpovídající veřejný prostor v dané lokalitě.Architectural proposal reacts to absence of representative spaces and functional background on golf course Dundonald Links that is located  on the west coast of the Scottish council area North Ayrshire. The main purpose is to design an object that is capacity and aesthetics wise appropriate for holding major golf events. Currently there has been built golf course itself with tentative lounge. The proposal should respect operational needs of the lounge: background for the golf club (dressing rooms, proshop, administration etc.), hotel and restaurant area with technical background and appropriate public area

All clinically-relevant blood components transmit prion disease following a single blood transfusion: a sheep model of vCJD

Variant CJD (vCJD) is an incurable, infectious human disease, likely arising from the consumption of BSE-contaminated meat products. Whilst the epidemic appears to be waning, there is much concern that vCJD infection may be perpetuated in humans by the transfusion of contaminated blood products. Since 2004, several cases of transfusion-associated vCJD transmission have been reported and linked to blood collected from pre-clinically affected donors. Using an animal model in which the disease manifested resembles that of humans affected with vCJD, we examined which blood components used in human medicine are likely to pose the greatest risk of transmitting vCJD via transfusion. We collected two full units of blood from BSE-infected donor animals during the pre-clinical phase of infection. Using methods employed by transfusion services we prepared red cell concentrates, plasma and platelets units (including leucoreduced equivalents). Following transfusion, we showed that all components contain sufficient levels of infectivity to cause disease following only a single transfusion and also that leucoreduction did not prevent disease transmission. These data suggest that all blood components are vectors for prion disease transmission, and highlight the importance of multiple control measures to minimise the risk of human to human transmission of vCJD by blood transfusion

Prion protein-specific antibodies that detect multiple TSE agents with high sensitivity

This paper describes the generation, characterisation and potential applications of a panel of novel anti-prion protein monoclonal antibodies (mAbs). The mAbs were generated by immunising PRNP null mice, using a variety of regimes, with a truncated form of recombinant ovine prion protein spanning residues 94–233. Epitopes of specific antibodies were mapped using solid-phase Pepscan analysis and clustered to four distinct regions within the PrP molecule. We have demonstrated the utility of these antibodies by use of Western blotting and immunohistochemistry in tissues from a range of different species affected by transmissible spongiform encephalopathy (TSE). In comparative tests against extensively-used and widely-published, commercially available antibodies, similar or improved results can be obtained using these new mAbs, specifically in terms of sensitivity of detection. Since many of these antibodies recognise native PrPC, they could also be applied to a broad range of immunoassays such as flow cytometry, DELFIA analysis or immunoprecipitation. We are using these reagents to increase our understanding of TSE pathogenesis and for use in potential diagnostic screening assays