Lab Life ****************************************************************************************** * Lab Life ****************************************************************************************** As a Pharmacy student at my home university, during my Erasmus traineeship I have been sta of Hradec Králové, where Charles University has its Pharmacy faculty. Here, I have been wo department of Biological and Medical Sciences, specifically with a group of scientists who endothelial dysfunction. The main aim of this research is to establish whether the up-regulation of certain protein dysfunction are causative factors or whether they are simply markers of endothelial dysfun the proteins are proven to be causative of endothelial dysfunction there is a possibility development to prevent endothelial dysfunction from occurring. We have been looking specif soluble endoglin, which is a plasma protein that’s highly expressed in vascular endothelia levels of soluble endoglin are found in many conditions, such as hypertension, diabetes me hypercholesterolemia, which we know are linked to endothelial dysfunction, although the ex endoglin is not fully understood in these instances. Therefore, the department hypothesise levels of soluble endoglin, along with a high fat diet, may affect aortic endothelium in v To try and confirm the hypothesis, throughout my stay here, I have had the chance to try v that are used but have mostly been focussing on Western Blotting. Western Blotting is a me distinguish protein expression with the possibility of being able to quantify changes in p also. Prior to Western Blotting, electrophoresis has to be carried out to separate protein molecular weights. Ultimately each ‘blot’ represents a specific protein from a different s these blots can be analysed by intensity (a darker blot represents up-regulation of the pr usually expect a difference in expression of inflammatory mediators, such as COX and iNOS, samples, indicating whether inflammation is present or not.  I have also had the chance to see an ELISA and immunofluorescence tests which identify pro ELISA works based on specific binding, which produces a colorimetric result. In the instan an ELISA, it produced a yellow colour at the end point of the reaction. The intensity of t measured which directly correlates to the amount of protein present. As a large number of tested in one go, the intensity of one sample can be compared to the other samples present hand, immunofluorescence works by binding of an antibody, with a fluorescent marker attach protein. This can then be examined under a microscope and can be used to locate where in a is expressed. In our case it is useful to know whether various proteins are expressed dire endothelium or inside the atherosclerotic plaque itself. It has been very interesting for me to see and try Western Blotting, along with the other we learnt about them in our lectures and many of us wondered when we might possibly ever u certainly useful to have the background knowledge to the techniques allowing a deeper unde what was actually happening. Furthermore, I had no idea Western Blotting usually takes aro complete – our lecturers had made it sound like it could be done in half an hour! My time here has also taught me science doesn’t always go to plan – even when you follow t to the letter. For instance, getting a good result in Western Blotting is often very trick are so many steps it can be very hard to identify what went wrong and how to correct it. T particularly disheartening after two days hard work, especially if you need the results ur say ‘there’s no such thing as a bad result’ but in Western Blotting that could certainly b you’re in the dark room and nothing is showing up on your membranes!  Nevertheless, I feel has given me a realistic insight into laboratory work, when things don’t always go right t perseverance is often required. Overall, gaining some experience in a laboratory environment has been really valuable for something I’ve seen much of before. Not only has it allowed me to consider whether this mi I would like to pursue in the future but it has also given me an understanding to the wide pharmacy and what a career in pharmacy can involve, whilst relating what we have learnt in reality. Fiona Smith is a second year student from Cardiff Unive Faculty of Pharmacy at the Hradec Králové campus of Cha interests include music, travelling and dance.