Influence of antiphospholipid antibodies on the monocyte inflammatory and adhesion profile

Antifosfolipidni sindrom je sistemska avtoimunska bolezen, za katero so značilne žilne tromboze in/ali zapleti v nosečnosti ob stalni prisotnosti antifosfolipidnih protiteles. Antifosfolipidna protitelesa se preko interakcij s trombociti, endotelnimi celicami in monociti vpletajo v patogenezo tromboze pri antifosfolipidnem sindromu. Posledica tovrstnih interakcij so spremembe v fenotipu tarčnih celic, ki se kaže v povečanem izražanju protrombotičnih molekul. Pri monocitih pride do povečanega izražanja vnetnih citokinov in tkivnega faktorja, pri endotelnih celicah pa se poleg tega poveča tudi izražanje adhezijskih molekul, kar vzpodbudi interakcije endotelnih celic z monociti. Zelo malo študij se osredotoča na vprašanje, ali antifosfolipidna protitelesa vzpodbujajo interakcije monocitov z endotelijem tudi preko vpliva na adhezijske molekule izražene na monocitih. Namen naše raziskave je zato bil, da poleg vpliva antifosfolipidnih protiteles na izražanje vnetnih citokinov in tkivnega faktorja, raziščemo še vpliv antifosfolipidnih protiteles na izražanje adhezijskih molekul na monocitih. Tako celice THP1 kot tudi iz krvi izolirane monocite smo stimulirali s frakcijami IgG bolnikov z antifosfolipidnim sindromom ob sočasni prisotnosti lipopolisaharida in dodatku protrombina. Najprej smo s poskusi na celicah THP1 preko merjenja količine izločenega tumor nekrotičnega dejavnika α v celično okolje opredelili ustrezne pogoje stimulacije pri kateri pride do aktivacije celic. Kasneje smo, ob uporabi opredeljenih pogojev stimulacije, ovrednotili še količine drugih izločenih vnetnih citokinov (tumor nekrotični dejavnik α, interlevkin 6) v okolje celic THP1 in monocitov. Ob enem smo na nivoju mRNA merili spremembe v izražanju genov za vnetne citokine (tumor nekrotični dejavnik α, interlevkin 6, 1β, 8) ter gena za tkivni faktor v celicah THP1. Izražanje adhezijskih molekul (CD49d, CD11b, CD62L) na površini monocitov smo določili s pretočno citometrijo. Naši rezultati kažejo, da antifosfolipidna protiteles ob prisotnosti lipopolisaharida aktivirajo monocitne celice, ob čemer pride do povečanega izražanja vnetnih citokinov, tkivnega faktorja in do spremenjenega izražanja adhezijskih molekul. V celicah THP1 smo izmerili povečanje izražanja genov za tumor nekrotični dejavnik α in interlevkine 6, 1β, 8. Rezultate o povečanem izražanju tumor nekrotičnega dejavnika α in interlevkina 6 smo dodatno dokazali tudi na proteinskem nivoju, ko smo njune povišane vrednosti izmerili v celičnem okolju celic THP1 in monocitov. V celicah THP1 smo izmerili tudi povečano izražanje gena za tkivni faktor. Na monocitih smo izmerili spremenjen profil adhezijskih molekul, in sicer povečano izražanje CD49d in CD62L ter zmanjšano izražanje CD11b.Antiphospholipid syndrome is a systemic autoimmune disease characterized by vascular thrombosis and/or complications in pregnancy in the constant presence of antiphospholipid antibodies. Antiphospholipid antibodies are involved in the pathogenesis of thrombosis in antiphospholipid syndrome through interactions with platelets, endothelial cells and monocytes. Such interactions result in changes in the target cell phenotype, which turns into prothrombotic. In monocytes and endothelial cells, these changes show as increased expression of inflammatory cytokines and tissue factor. In endothelial cells, the expression of adhesion molecules is also increased, which promotes interactions of endothelial cells with monocytes. Very few studies focused on the question whether these interactions are also promoted trough increased expression of adhesion molecules on the monocytes. The purpose of our study was therefore that in addition to the effect of antiphospholipid antibodies on the expression of inflammatory cytokines and tissue factor, we also investigate the effect of antiphospholipid antibodies on the expression of adhesion molecules on monocytes. THP1 cells as well as monocytes isolated from blood were stimulated with IgG fractions of patients with antiphospholipid syndrome in the presence of lipopolysaccharide and the addition of prothrombin. First, we determined the appropriate stimulation conditions by which the activation of cells is achieved through measuring the amount of tumor necrotic factor α secreted into the cell environment. Later, using the defined stimulation conditions, the effects of antiphospholipid antibodies were evaluated. We measured secreted proinflammatory cytokines (tumor necrotic factor α, interleukin 6) by THP1 cells and monocytes, changes in the expression of genes for proinflammatory cytokines (tumor necrotic factor α, interleukin 6, 1β, 8) and tissue factor in THP1 cells and changes in the expressions of adhesion molecules (CD49d, CD11b, CD62L) on the surface of monocytes by flow cytometry. Our results show that antiphospholipid antibodies in the presence of lipopolysaccharide activate monocytic cells, resulting in the increased expression of inflammatory cytokines, tissue factor, and altered expression of adhesion molecules. In THP1 cells, increased gene expression of tumor necrotic factor α and interleukin 6, 1β, 8 was measured. In addition, we measured increased secretion of tumor necrotic factor α and interleukin 6 into the cell environment of THP1 cells and monocytes. In THP1 cells we also measured increased gene expression of tissue factor. An altered profile of adhesion molecules was measured on monocytes, namely increased expression of CD49d and CD62L and decreased expression of CD11b

Occupational lifting as risk factor for pregnant woman and fetus

Uvod: V diplomskem delu smo predstavili vpliv dvigovanja bremen na delovnem mestu in vpliv na žensko ter plod. Izpostavljenost dvigovanju težkih bremen poveča tveganje za neugodne izide nosečnosti. Privede lahko do sprememb v rasti in razvoju ploda, pelvične in/ali medenične bolečine, spontanega splava, prezgodnjega poroda, mrtvorojenosti. Delodajalec je dolžen ukrepati takoj, ko ga delavka obvesti o nosečnosti. Namen: Namen diplomskega dela je raziskati poznavanje posledic dvigovanja bremen med ženskami. Zanima nas tudi odziv delodajalca ter poznavanje pravic. Metode dela: V teoretičnem delu je bila uporabljena deskriptivna metoda dela s pregledom domače in tuje strokovne in znanstvene literature v knjigah, zbornikih in člankih. Literaturo smo iskali v spletnih podatkovnih bazah Medline, PubMed, CINAHL, Proquest, ScienceDirect, Google učenjak. V drugem delu diplomskega dela je bila uporabljena kvantitativna metoda z vprašalnikom, ki smo ga posredovali s pomočjo spletne strani 1ka. Posredovali smo jo v Facebook skupine z naslovom Nosečka forum in Mamice Poljanske doline. Vprašalnik je bil namenjen ženskam v zadnjem trimesečju nosečnosti ali mamicam, ki so rodile v preteklem letu. Vprašalnik je bil sestavljen iz dvajsetih vprašanj, ki smo jih postavili na podlagi pregleda literature. Rezultati: Vse ženske v naši raziskavi (N=216), ki so bile med nosečnostjo zaposlene, so svojega delodajalca o nosečnosti obvestile. Večina jih je mnenja, da se je delodajalec na novico odzval pozitivno in da je skrbel za njihovo ter otrokovo varnost. Skoraj vse so bile pred in med nosečnostjo izpostavljene vsaj enemu dejavniku tveganja za njeno in otrokovo zdravje. Delodajalci in nadrejeni so nosečnico s pravicami seznanili le v skupno 15 %. Dve tretjini jih med nosečnostjo ni bilo v bolniškem staležu. 72 % jih med nosečnostjo ni imelo težav. Večina jih ni nikoli izračunala mase enkratnega bremena in vsote bremen, ki jih dvignejo dnevno. Skoraj vse dvigujejo bremena tudi v svojem prostem času. Razprava in zaključek: Informacije o pravicah so od delodajalcev in nadrejenih pridobile le v redkih primerih. Ženske se teže dvignjenih bremen ne zavedajo, saj se le redko vprašajo, kakšno težo vsakodnevno dvigujejo. Babica lahko tekom prenatalnega obdobja vpliva na samo področje dvigovanja bremen na delovnem mestu. Ženske bi lahko v šolah za starše in dispanzerjih poučevali o dovoljeni teži in načinu dvigovanja.Introduction: In the literature review we researched the impact of lifting heavy loads on the workplace and the impact of lifting on the woman and fetus. With heavy lifting the chances of negative outcomes on pregnancy increases. Lifting can lead to changes in fetal growth and development, pelvic and/or pelvic pain, miscarriage, premature birth, stillbirth. The employer is obliged to act as soon as the worker informs him of the pregnancy. Purpose: The purpose of this paper is to investigate knowledge of the consequences of lifting heavy loads among pregnant women. We are also interested in an employer response and knowledge of rights during pregnancy. Methods: In theoretical part we used a descriptive method with a review of domestic and foreign, professional and scientific literature in books, proceedings and articles. We searched for the literature in databases as Medline, PubMed, CINAHL, Proquest, ScienceDirect, Google Scholar. In the second part of the diploma we used the quantitative method. We used the questionnaire, which was submitted through the website 1ka. The questionnair was sent to Facebook groups Nosečka forum and Mamice Poljanske doline. It was meant for women in last trimester of pregnancy and for those who gave birth in the past year. The questionnaire consisted of twenty questions that were asked based on the literature review. Results: All women in our study (N=216) who were employed during pregnancy told about pregnancy to their employer. In most cases employer responded positively to the news and that it took care of her and her child\u27s safety. Almost all of them were exposed to at least one risk factor before and during pregnancy. Only 15% of pregnant women have been informed of their rights by employers and superiors. Two thirds of them were not on sick leave during pregnancy. 72% had no helath problems during pregnancy. Most have never calculated the mass of a one-time load and the sum of the loads they lift on a daily basis. Almost all of them lift loads even in their spare time. Discussion and conclusion: In most cases, employers responded positively to the news about preganncy. Women believe that both her and her child\u27s safety have been taken care of during pregnancy. Women have rarely obtained information about rights from employers and superiors. Women are not aware of the weight of the lifted loads, as they rarely imagine the weight they lift on a daily basis. Midwife can have influence to lifting on work during preganancy. This things can be tought during schools for parents and doctors visits

Antiphospholipid Antibody Syndrome-Associated Increased Surface Expression of VLA4 Integrin on Human Monocytes

Antiphospholipid syndrome (APS) is a systemic autoimmune disorder characterized by thrombosis and/or obstetric complications in the presence of antiphospholipid antibodies (aPL). Catastrophic APS (CAPS) is the most severe form of the disease, in which microvascular thromboses develop rapidly, leading to multiorgan failure. Monocytes, along with endothelial cells, are critical players in the pathogenesis of APS. Recruitment of these cells to the site of injury/inflammation involves a series of events, including capture, rolling, adhesion enhancement, and transmigration, which are controlled by surface adhesion molecules. The aim of our study was to investigate the surface adhesion profile of monocytes from APS patients and monocytes stimulated in vitro with aPL from a CAPS patient. The surface expression of the adhesion molecules LFA1, L-selectin, MAC1, PSGL1, and VLA4 was analyzed by flow cytometry. To our knowledge, this preliminary study was the first to show that VLA4 was significantly increased on the surface of monocytes from APS patients. Moreover, in vitro stimulations mimicking CAPS showed an even greater increase in VLA4. Our data suggest that the surface adhesion profile on monocytes is altered in APS and CAPS and may be involved in the thrombotic pathophysiology of the disease by enhancing monocyte adhesion