Macrophage migration inhibitory factor is involved in endovascular trophoblast cell function in vitro

Macrophage migration inhibitory factor (MIF) is a multifunctional cytokine abundantly present at the feto-maternal interface proposed to play a role in establishment of pregnancy. We have previously shown that pharmacological inhibition of enzymatic activity of MIF decreases extravillous trophoblast invasion and migration in vitro. This study aimed to further elucidate potential role of endogenous trophoblast MIF, and to assess its importance for endovascular trophoblast cell function in particular. Attenuation of MIF by siRNA reduced HTR-8/SVneo cell invasion through Matrigel (59 % of control), expression of integrin α1 (86 % of control) and levels of MMP2 and MMP9 (87 % and 57 % of control, respectively). MIF specific siRNA reduced the ability of HTR-8/SVneo to differentiate in to endothelial-like phenotype, as determined by Matrigel tube formation assay. The total tube length was decreased to 68.6 %, while the number of branching points was reduced to 57.8 % of control. HTR-8/SVneo cell capacity to integrate into HUVEC monolayers was reduced by knock-down of MIF. This could be partly caused by reduced N-cadherin expression to 63 % of control, which decreased with knock-down of MIF, as the expression of this protein was recently shown essential for trophoblast-endothelial interaction. These novel findings indicate a novel role for trophoblast MIF in spiral artery remodeling process

Supplementary data for article: Stanković, N.; Šenerović, L.; Bojic-Trbojevic, Z.; Vuckovic, I.; Vicovac, L.; Vasiljevic, B.; Nikodinović-Runić, J. Didehydroroflamycoin Pentaene Macrolide Family from Streptomyces Durmitorensis MS405(T): Production Optimization and Antimicrobial Activity. Journal of Applied Microbiology 2013, 115 (6), 1297–1306. https://doi.org/10.1111/jam.12326

Supporting information for: [https://doi.org/10.1111/jam.12326]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1438]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3486

Supplementary data for article: Stanković, N.; Šenerović, L.; Bojic-Trbojevic, Z.; Vuckovic, I.; Vicovac, L.; Vasiljevic, B.; Nikodinović-Runić, J. Didehydroroflamycoin Pentaene Macrolide Family from Streptomyces Durmitorensis MS405(T): Production Optimization and Antimicrobial Activity. Journal of Applied Microbiology 2013, 115 (6), 1297–1306. https://doi.org/10.1111/jam.12326

Supporting information for: [https://doi.org/10.1111/jam.12326]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1438]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3486

Expression of prolactin receptors in the human extra villous cytotrophoblast cell line HTR-8/SVNEO

Prolactin is a pituitary hormone that is widely produced at extrapitutitary locations. One of the first described sites of its production is the decidualized endometrium, suggesting that it could be potentially significant for local processes such as immunomodulation, embryo implantation, and survival. In this study, expression of prolactin receptor protein is investigated by immunocytochemistry and Western blot in the HTR-8/SVneo cell line, often used as a model of first-tri­mester trophoblast. The obtained data enabled us to identify both long and intermediate forms of prolactin receptors, which were shown to be significant in PRL signaling in other tissues

Monoclonal antibody 26 cross-reactive with beta(2)-glycoprotein I affects human trophoblast invasion in vitro

Objective: Monoclonal antibody 26 (MAb 26) raised against tetanus toxoid has documented cross-reactivity with beta(2)-glycoprotein I. Passive introduction of this antibody in mice results in an antiphospholipid syndrome-like condition. We investigated the effects of MAb 26 on first trimester human trophoblast in vitro. Study design: Binding of MAb 26 to placental tissue trophoblast, isolated cytotrophoblast and HTR-8/SVneo cells was analyzed by immunohisto(cyto)chemistry. Possible effects on cell invasion in vitro were assessed by Matrigel assay. Effects on cell viability were assessed by MU test. A possibility that MAb 26 induces change in levels of effector molecules important for cell invasion was investigated. Integrin subunits alpha(1), alpha(5) and beta(1), and galectin-1, were analyzed by qPCR and Western blot. Metalloproteinases -2 and -9 were assessed by gelatin zymography. Results: Immunohisto(cyto)chemistry showed binding of MAb 26 to placental tissue trophoblast, isolated cytotrophoblast and HTR-8/SVneo cells. The antibody had a significant inhibitory effect on cell invasion by both isolated cytotrophoblast and HTR-8/SVneo. The antibody induced significant decrease in protein levels of metalloproteinases, integrin subunit alpha(1) and galectin-1. Cell viability was not affected. Conclusion: MAb 26 reduces trophoblast invasion in vitro through decreased levels of metalloproteinases-2 and -9, integrin alpha(1) and galectin-1

Didehydroroflamycoin pentaene macrolide family from Streptomyces durmitorensis MS405(T): production optimization and antimicrobial activity

AimsThe aim of this study was to improve production of pentaene 32,33-didehydroroflamycoin (DDHR) in Streptomyces durmitorensis MS405 strain to obtain quantities sufficient for in depth analysis of antimicrobial properties. Methods and ResultsThrough classical medium optimization conditions for stable growth, DDHR production within 7days of incubation was established. Yields of 215mgl(-1) were achieved in shake flask experiments in complex medium with mannitol as the primary carbon source. DDHR had poor antibacterial activity with minimal inhibitory concentrations (MIC) of 400gml(-1) for Staphylococcus aureus and Bacillus subtilis, while MIC of 70gml(-1) was determined for Candida albicans. Using flow cytometry and fluorescent microscopy, it was demonstrated that DDHR induced membrane damage in C.albicans followed by cell death. Combination studies with known antifungal nystatin showed that DDHR is a promising agent for the development of novel antimycotic treatments potentially less toxic for human cells. ConclusionsPentaene didehydroroflamycoin has no antibacterial activity but can be further developed for the application in antifungal therapy. Significance and Impact of the StudyThis study is the first report on the stable and production in high yields of a novel pentaene family that acts on Candida cell membranes and can be used in combination with known antifungals. Polyenes are still antifungal antibiotics of choice, and therefore, isolation and production of new lead structures are highly significant.Free full text: [https://doi.org/10.1111/jam.12326]Peer-reviewed manuscript: [http://cherry.chem.bg.ac.rs/handle/123456789/3486]Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3487