PCSK9 inhibition alters the lipidome of plasma and lipoprotein fractions

Background and aims: While inhibition of proprotein convertase subtilisin/kexin type 9 (PCSK9) is known to result in dramatic lowering of LDL-cholesterol (LDL-C), it is poorly understood how it affects other lipid species and their metabolism. The aim of this study was to characterize the alterations in the lipidome of plasma and lipoprotein particles after administration of PCSK9 inhibiting antibody to patients with established coronary heart disease. Methods: Plasma samples were obtained from patients undergoing a randomized placebo-controlled phase II trial (EQUATOR) for the safe and effective use of RG7652, a fully human monoclonal antibody inhibiting PCSK9 function. Lipoprotein fractions were isolated by sequential density ultracentrifugation, and both plasma and major lipoprotein classes (VLDL-IDL, LDL, HDL) were subjected to mass spectrometric lipidomic profiling. Results: PCSK9 inhibition significantly decreased plasma levels of several lipid classes, including sphingolipids (dihydroceramides, glucosylceramides, sphingomyelins, ceramides), cholesteryl esters and free cholesterol. Previously established ceramide ratios predicting cardiovascular mortality, or inflammation related eicosanoid lipids, were not altered. RG7652 treatment also affected the overall and relative distribution of lipids in lipoprotein classes. An overall decrease of total lipid species was observed in LDL and VLDL thorn IDL particles, while HDL-associated phospholipids increased. Following the treatment, LDL displayed reduced lipid cargo, whereas relative lipid proportions of the VLDL thorn IDL particles were mostly unchanged, and there were relatively more lipids carried in the HDL particles. Conclusions: Administration of PCSK9 antibody significantly alters the lipid composition of plasma and lipoprotein particles. These changes further shed light on the link between anti-PCSK9 therapies and cardiovascular risk. (C) 2018 Elsevier B.V. All rights reserved.Peer reviewe

The ether lipid precursor hexadecylglycerol stimulates the release and changes the composition of exosomes derived from PC-3 cells

Exosomes are vesicles released by cells after fusion of multivesicular bodies with the plasma membrane. In this study, we have investigated whether ether lipids affect the release of exosomes in PC-3 cells. To increase the cellular levels of ether lipids, the ether lipid precursor hexadecylglycerol was added to cells. Lipidomic analysis showed that this compound was in fact able to double the cellular levels of ether lipids in these cells. Furthermore, increased levels of ether lipids were also found in exosomes released by cells containing high levels of these lipids. Interestingly, as measured by nanoparticle tracking analysis, cells containing high levels of ether lipids released more exosomes than control cells, and these exosomes were similar in size to control exosomes. Moreover, silver staining and Western blot analyses showed that the protein composition of exosomes released in the presence of hexadecylglycerol was changed; the levels of some proteins were increased, and the levels of others were reduced. In conclusion, this study clearly shows that an increase in cellular ether lipids is associated with changes in the release and composition of exosomes

Henkilöstön palkitseminen kuntien ruoka- ja siivouspalveluissa

Tämän opinnäytetyön tavoitteena oli hakea ja koota tietoa julkisella sektorilla käytössä olevista palkitsemismenetelmistä ja tarkastella niiden käyttömahdollisuuksia kuntien ruoka- ja siivouspalveluissa. Työn tarkoituksena oli selvittää ja analysoida käytössä olevien palkitsemismenetelmien vaikutus henkilöstön rekrytointiin, työmotivaatioon, työn tehostamiseen ja sairauspoissaoloihin. Ruoka- ja siivouspalvelujen asiantuntijapalveluita tuottava Damico Oy oli toimeksiantajana opinnäytetyölle. Opinnäytetyön tekijä aloitti palvelupäällikkönä Hollolan ruoka- ja siivouspalveluissa vuonna 2017. Palvelupäällikön aiempi lähes 15 vuoden esimieskokemus on hankittu saman kunnan palveluksessa erikoisalana ammattikeittiöosaaminen. Empiirisen tutkimuksen teoreettinen viitekehys koostui tiedonkeruusta koskien yksityisen ja julkisen sektorin palkitsemismenetelmiä. Opinnäytetyön tutkimusmenetelminä käytettiin kvalitatiivista avointa haastattelua ja kvantitatiivista kyselytutkimusta. Asiantuntijalausunnon kannustepalkkioista antoi Minna Dammert, Damico Oy:stä. Haastateltavina henkilöinä olivat Kuntatyönantajien johtava työmarkkina-asiamies ja Hollolan henkilöstöpäällikkö. Kyselytutkimukset kohdennettiin yksityisen ja julkisen sektorin ruoka- ja siivouspalvelujen vastuuhenkilöille sekä Hollolan ja kolmen muun kunnan ruoka- ja siivouspalvelujen henkilöstölle. Työlle asetettu tavoite ja tarkoitus toteutuivat teoreettisen viitekehyksen ja haastattelujen osalta hyvin. Kyselytutkimukseen vastanneiden määrä jäi tavoitteesta, mutta otanta oli riittävä vastausten analysoinnille ja palkitsemismenetelmien vaikuttavuuden arvioinnille. Tutkimustulokset osoittavat, että palkitsemisella voidaan vaikuttaa myönteisesti rekrytointeihin, työmotivaatioon, työn tehostamiseen ja sairauspoissaoloihin kuntien ruoka- ja siivouspalveluissa. Kunnissa käytetään erilaisia palkitsemismenetelmiä. Ohjeistus henkilöstön palkitsemiseen oli kuitenkin vaihtelevaa; osalta kunnista puuttuivat tavoitteet ja riittävät mittarit. Haastattelujen tuloksena esimiestaitojen tukeminen ja kehittäminen henkilöstöjohtamisessa ja palkitsemisen kokonaisuuden hallinnassa nousivat ykkösprioriteetiksi, jotka tulisi ratkaista työnantajan kanssa. Käytetyt tutkimusmenetelmät ja -tulokset koko laajuudessaan mahdollistavat monialaisen jatkotutkimuksen sekä palkitsemiselle että johtamistaitojen kehittämiselle.The goal of this bachelor’s thesis was to search and gather information about reward systems utilized by the public sector, and to examine the possibilities they could offer to catering and services in municipalities. The purpose of this thesis was to research and analyze the effect of already existing reward systems on recruiting, work motivation, work rationalization and sick leaves. Damico Oy was the commissioner of this thesis. The author of this thesis started as a Service Manager at catering and services of Hollola in 2017. The Service Manager had 15 years of prior experience working as a manager for the same municipality, specializing in professional institutional kitchens. The framework of the empirical research was composed of data acquisition about the reward systems of the private and public sectors. Research methods utilized in this thesis were qualitative interview and quantitative questionnaire. Expert opinion was provided by Minna Dammert, Damico Oy. The executive Labour Market Advisor and Personnel Manager of Hollola were interviewed for the research. The questionnaires targeted the leaders of catering and services in private and public sectors, and the employees in that field in municipalities. The goal of this thesis was achieved, and its purpose fulfilled as far as the theoretical framework and interviews are concerned. The questionnaire did not produce as many answers as was planned, but sampling was sufficient for analysis, which made it possible to have the effectivity of the reward systems assessed. The research shows that rewarding can have a positive effect on all studied fields in catering and services of municipalities. In the municipalities different kinds of reward systems were utilized. Instructions of personnel rewards varied; some of the municipalities were missing goals and sufficient indicators. Interviews showed that supporting and developing management skills, when it comes to personnel management and the rewarding procedures, should be prioritized and resolved in co-operation with the employer. The research methods used and the results in their entirety enable multi-disciplinary follow-up research of rewarding systems and to management skills progress

LC-ESI-MS/MS Analysis of arachidonic acid and its metabolites prostaglandins, HETEs, thromboxanes and leukotrienes in mammalian cells and tissues

Almost all mammalian cells produce eicosanoids and their oxidation products, which are synthesized rapidly by cyclooxygenase, lipoxygenase or epoxygenase reactions from arachidonic, homogammalinolenic, eicosapentanoic acids. They have physiological effects at trace concentrations, such as local mediation of inflammatory responses (1). Mass spectrometry coupled with electrospray ionization has been widely utilized for trace analysis of eicosanoids (2) combined with HPLC. We describe a LC-ESI-MS/MS method for analysis of arachidonic acid and its metabolites; prostaglandins, HETEs, thromboxanes and leukotrienes, within a single sample run. The method will enable targeted differential profiling and quantitative analysis of these compounds. The compounds of interest are extracted by liquid-liquid or solid phase extraction. The analytes are resolved by reversed phase chromatography, ionized by electrospray in negative ion mode and detected in MRM mode. Our aim is to measure at the order of 50 eicosanoid, HETE, and fatty acid molecular species in parallel at tissue level concentrations in order to evaluate active biochemical pathways in specific biological state. We will describe the analytical method, as well as demonstrate its utility on mouse model tissue samples related to studies of type 2 diabetes and metabolic syndrome