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Mohamed Samy Yousef, Mohamed Ali Marey, Nina Hambruch, Hiroyuki Hayakawa, Takashi Shimizu, Hassan Ali Hussien, Abdel-Razek Khalifa Abdel-Razek, Christiane Pfarrer, Akio Miyamoto
Sperm are allogenic to the female genital tract; however, oviducts provide optimal conditions for survival and capacitation of these non-self cells until fertilization. Recently, we showed that oviduct-conditioned media and prostaglandin E2 (PGE2) suppress sperm phagocytosis by polymorphonuclear neutrophils (PMNs) under physiological conditions. We hypothesized that sperm binding to bovine oviduct epithelial cells (BOECs) could change the local innate immunity via PGE2. As the first step to obtain basic information, sub-confluent BOEC monolayers were co-cultured with swim-up sperm for 2 h...
2016: PloS One
Hamideh Yadegari, Arijit Biswas, Mohammad Suhail Akhter, Julia Driesen, Vytautas Ivaskevicius, Natascha Marquardt, Johannes Oldenburg
Disease-associated silent mutations are considered to affect the accurate pre-mRNA splicing either by influencing regulatory elements leading to exon skipping or by creating a new cryptic splice site (ss). The present study describes a new molecular pathological mechanism by which a silent mutation inhibits splicing and leads to intron retention. We identified a heterozygous silent mutation c.7464C>T in exon 44 of the von Willebrand factor (VWF) gene in a family with type 1 von Willebrand disease (VWD). In vivo and ex vivo transcript analysis revealed an aberrantly spliced transcript with intron 44 retained in the mRNA, implying disruption of the first catalytic step of splicing at 5' ss...
August 19, 2016: Blood
Sadjad Danesh Mesgaran, Jutta Sharbati, Ralf Einspanier, Christoph Gabler
BACKGROUND: The mammalian oviduct provides the optimal environment for gamete maturation including sperm capacitation, fertilization, and development of the early embryo. Various cell culture models for primary bovine oviductal epithelial cells (BOEC) were established to reveal such physiological events. The aim of this study was to evaluate 17 candidate mRNA expression patterns in oviductal epithelial cells (1) in transition from in vivo cells to in vitro cells; (2) during three consecutive cell culture passages; (3) affected by the impact of LOW or HIGH glucose content media; and (4) influenced by different phases of the estrous cycle in vivo and in vitro...
2016: Reproductive Biology and Endocrinology: RB&E
M A Marey, M S Yousef, R Kowsar, N Hambruch, T Shimizu, C Pfarrer, A Miyamoto
The local immune system in the oviduct has a unique ability to deal with pathogens, allogeneic spermatozoa, and the semi-allogeneic embryo. To achieve this, it seems likely that the oviduct possesses an efficient and strictly controlled immune system that maintains optimal conditions for fertilization and early embryo development. The presence of a proper sperm and/or embryo-oviduct interaction begs the question of whether the local immune system in the oviduct exerts beneficial or deleterious effects on sperm and early embryo; support or attack?...
July 2016: Domestic Animal Endocrinology
Lindsey Hawke, Mackenzie L Bowman, Man-Chiu Poon, Mary-Frances Scully, Georges-Etienne Rivard, Paula D James
Approximately 10% of von Willebrand factor (VWF) gene mutations are thought to alter messenger RNA (mRNA) splicing through disruption of consensus splice sites. This mechanism is likely underrecognized and affected by mutations outside consensus splice sites. During VWF synthesis, splicing abnormalities lead to qualitative defects or quantitative deficiencies in VWF. This study investigated the pathologic mechanism acting in 3 von Willebrand disease (VWD) families with putative splicing mutations using patient-derived blood outgrowth endothelial cells (BOECs) and a heterologous human embryonic kidney (HEK 293(T)) cell model...
July 28, 2016: Blood
Damien G Noone, Magdalena Riedl, Fred G Pluthero, Mackenzie L Bowman, M Kathryn Liszewski, Lily Lu, Yi Quan, Steve Balgobin, Reinhard Schneppenheim, Sonja Schneppenheim, Ulrich Budde, Paula James, John P Atkinson, Nades Palaniyar, Walter H A Kahr, Christoph Licht
Atypical hemolytic uremic syndrome and thrombotic thrombocytopenic purpura have traditionally been considered separate entities. Defects in the regulation of the complement alternative pathway occur in atypical hemolytic uremic syndrome, and defects in the cleavage of von Willebrand factor (VWF)-multimers arise in thrombotic thrombocytopenic purpura. However, recent studies suggest that both entities are related as defects in the disease-causing pathways overlap or show functional interactions. Here we investigate the possible functional link of VWF-multimers and the complement system on endothelial cells...
July 2016: Kidney International
R Kowsar, M A Marey, T Shimizu, A Miyamoto
Excess dietary protein intake in early lactation dairy cows resulting in blood urea nitrogen of greater than 19 to 20mg/dL is associated with decreased fertility. Little is known about the local interference of urea in the normal immunological environment of the oviduct that provides optimal conditions for early reproductive events. A bovine oviduct epithelial cell (BOEC) culture was used to determine how urea influences immune environment. The BOEC monolayer was supplemented with low (20mg/dL) and high (40mg/dL) concentrations of urea together with ovarian steroids, estradiol (1ng/mL) and progesterone (1ng/mL), and LH (10ng/mL) at concentrations observed during the preovulatory period...
July 2016: Journal of Dairy Science
Dieter Dauwe, Beatriz Pelacho, Arief Wibowo, Ann-Sophie Walravens, Kristoff Verdonck, Hilde Gillijns, Ellen Caluwe, Peter Pokreisz, Nick van Gastel, Geert Carmeliet, Maarten Depypere, Frederik Maes, Nina Vanden Driessche, Walter Droogne, Johan Van Cleemput, Johan Vanhaecke, Felipe Prosper, Catherine Verfaillie, Aernout Luttun, Stefan Janssens
BACKGROUND: Blood outgrowth endothelial cells (BOECs) mediate therapeutic neovascularization in experimental models, but outgrowth characteristics and functionality of BOECs from patients with ischemic cardiomyopathy (ICMP) are unknown. We compared outgrowth efficiency and in vitro and in vivo functionality of BOECs derived from ICMP with BOECs from age-matched (ACON) and healthy young (CON) controls. METHODS AND RESULTS: We isolated 3.6±0.6 BOEC colonies/100×10(6) mononuclear cells (MNCs) from 60-mL blood samples of ICMP patients (n=45; age: 66±1 years; LVEF: 31±2%) versus 3...
April 2016: Journal of the American Heart Association
Ricaurte Lopera-Vásquez, Meriem Hamdi, Beatriz Fernandez-Fuertes, Verónica Maillo, Paula Beltrán-Breña, Alexandra Calle, Alberto Redruello, Soraya López-Martín, Alfonso Gutierrez-Adán, María Yañez-Mó, Miguel Ángel Ramirez, Dimitrios Rizos
To evaluate the effect of conditioned media (CM) and Extracellular Vesicles (EVs) derived from bovine oviduct epithelial cell (BOEC) lines on the developmental capacity of bovine zygotes and the quality of embryos produced in vitro, presumptive zygotes were cultured under specific conditions. In experiment 1, zygotes were cultured either on monolayers from BOEC extended culture (E), together with fresh BOEC suspension cells, or with BOEC-CM from fresh or E-monolayers. In experiment 2, EVs were isolated from BOEC-CM and characterized (150-200 nm) by Nanosight® and electron microscopy...
2016: PloS One
Mohamed Ali Marey, Mohamed Samy Yousef, Jinghui Liu, Kazuhiro Morita, Motoki Sasaki, Hiroyuki Hayakawa, Takashi Shimizu, Ibrahim I Elshahawy, Akio Miyamoto
The oviduct is an active contractile tube that provides the proper environment for sperm transport, capacitation and survival. Oviductal contractions are regulated by autocrine/paracrine secretion of several factors, such as prostaglandins (PGs) and endothelin-1 (EDN-1). We have previously shown that during the preovulatory stage, sperm are exposed to polymorphonuclear neutrophils (PMNs) in the bovine oviduct, and the bovine oviduct epithelial cells (BOECs) secrete molecules including PGE2 that suppress sperm phagocytosis by PMNs in vitro...
April 22, 2016: Journal of Reproduction and Development
B Schmaltz-Panneau, Y Locatelli, S Uzbekova, C Perreau, P Mermillod
There are convincing arguments to suggest that the success of early reproductive events is reliant on a satisfactory dialogue between gametes-embryo and the oviduct epithelium. The aim of this study was to develop and characterize an in vitro model to study these interactions. Cattle zygotes produced in vitro were cultured in either SOF or TCM-199 in the presence or absence of bovine oviduct cell monolayers (BOEC), under 20% or 5% O2 . The embryonic development rate and its quality (cell numbers, cryosurvival) were evaluated, as were the BOEC contents in 11 candidate transcripts (real-time PCR) at different time points...
October 2015: Reproduction in Domestic Animals, Zuchthygiene
D J Groeneveld, T van Bekkum, R J Dirven, J-W Wang, J Voorberg, P H Reitsma, J Eikenboom
BACKGROUND: Endothelial von Willebrand factor (VWF) inhibits angiogenesis. Accordingly, blood outgrowth endothelial cells (BOECs) isolated from von Willebrand disease (VWD) patients showed enhanced in vitro angiogenesis when compared with healthy control BOECs. Characterization of the angiogenic response of VWD BOECs is limited and differences between the different types of VWD have not been investigated in detail. OBJECTIVES: The aim of this study was to further explore the potential pathogenic effect of VWF mutations on angiogenesis...
October 2015: Journal of Thrombosis and Haemostasis: JTH
Albert C Sek, Zhihui Xie, Kaoru Terai, Lauren M Long, Celeste Nelson, Arkadiusz Z Dudek, Kirk M Druey
Idiopathic systemic capillary leak syndrome (SCLS) is a rare and potentially fatal vascular disorder characterized by reversible bouts of hypotension and edema resulting from fluid and solute escape into soft tissues. Although spikes in permeability-inducing factors have been linked to acute SCLS flares, whether or not they act on an inherently dysfunctional endothelium is unknown. To assess the contribution of endothelial-intrinsic mechanisms in SCLS, we derived blood-outgrowth endothelial cells (BOEC) from patients and healthy controls and examined gene expression patterns...
2015: PloS One
Faruk Incecik, Ozlem M Herguner, Sakir Altunbasak
BACKGROUND: The early and late benign occipital epilepsies of childhood (BOEC) are described as two discrete electro-clinical syndromes, eponymously known as Panayiotopoulos and Gastaut syndromes. The purpose of this study was to identify predictors of failure to respond to the initial antiepileptic drug (AED). MATERIALS AND METHODS: A total of 42 children with BOEC were enrolled. Predictive factors were analyzed by survival methods. RESULTS: Among the 42, 25 patients (59...
July 2015: Journal of Neurosciences in Rural Practice
L Jordaens, M Arias-Alvarez, I Pintelon, S Thys, S Valckx, Y Dezhkam, P E J Bols, J L M R Leroy
Elevated non-esterified fatty acids (NEFAs) have been recognized as an important link between lipolytic metabolic conditions and impaired fertility in high-yielding dairy cows. However, NEFA effects on the oviductal micro-environment currently remain unknown. We hypothesize that elevated NEFAs may contribute to the complex pathology of subfertility by exerting a negative effect on bovine oviductal epithelial cell (BOEC) physiology. Therefore, the objectives of this study were to elucidate direct NEFA effects on BOEC physiology in three different in vitro cell culture systems...
October 1, 2015: Theriogenology
E Martínez-León, C Osycka-Salut, J Signorelli, P Pozo, B Pérez, M Kong, P Morales, S Pérez-Martínez, E S Díaz
STUDY QUESTION: Does fibronectin (Fn) stimulate the sperm capacitation process in humans? SUMMARY ANSWER: Fibronectin stimulates human sperm capacitation. WHAT IS KNOWN ALREADY: Capacitation is a process that occurs in the oviduct. It has been suggested that some molecules present in the oviductal fluid and cells as well as proteins present in the cumulus oophorus could be involved in the modulation of sperm function and their acquisition of fertilizing capacity...
September 2015: Human Reproduction
Sally Ibrahim, Dessie Salilew-Wondim, Franca Rings, Michael Hoelker, Christiane Neuhoff, Ernst Tholen, Christian Looft, Karl Schellander, Dawit Tesfaye
In the present study, we used an in vitro model to investigate the response of the oviduct with respect to inflammatory mediators and their regulatory microRNAs in case of bacterial infection and subsequent association with embryo survival. For this, we conducted two experiments. In the first experiment, cultured primary bovine oviductal cells (BOEC) were challenged with lipopolysaccharide (LPS) for 24h and the temporal expression pattern of inflammatory mediators and their regulatory microRNAs were measured at 0, 3, 6, 12, 24 and 48h after LPS treatment...
2015: PloS One
Jie Yin, Zhen-Ni Ma, Xiao-Juan Zhao, Chang-Geng Ruan
Compared with endothelial progenitor cells, outgrowth endothelial cells (BOECs) from peripheral blood are rich in protein for blood angiogenesis and cell adhesion, similar to mature endothelial cells in biological characteristics. Moreover, they are now replacing human umbilical vein endothelial cells for the latter's limited life span and drift of phenotype, and might become a new tool for exploring the vascular abnormalities. This study was aimed to establish the protocol of producing BOECs, and then analyze the cell phenotype and function of BOECs...
December 2014: Zhongguo Shi Yan Xue Ye Xue za Zhi
L Rapala, R R Starzynski, P Z Trzeciak, S Dabrowski, A M Duszewska
Elevated temperatures have a negative impact on bovine reproduction. One of its effects is an increased concentration of reactive oxygen species (ROS) which may lead to female infertility. Oxidative stress impairs oocyte maturation, fertilization, and embryo development, and it also influences the reproductive tract. One of the defence mechanisms against the increase of ROS is the synthesis of antioxidants. Thus, the aim of this study was to analyse the expression of antioxidant enzymes (superoxide dismutase 1, SOD1; catalase, CAT; and glutathione peroxidase 1, GPX1) in bovine oviduct epithelial cells (BOEC) cultured with or without embryos at elevated temperatures...
December 2014: Reproduction, Fertility, and Development
E Corbin, A Cordova, J Grosbois, P Mermillod
Previous experiments demonstrated that co-culture of bovine embryos with bovine oviducal epithelial cells (BOEC) improved blastocyst rate and quality (Cordova et al. 2014). However, the use of primary cell support for improving embryo development in vitro may introduce a higher variability of the results between different BOEC batches used, as well as sanitary risks. The use of well-controlled large batches of frozen BOEC may help to solve these problems. Therefore, the aim of the present study was to characterise the survival and functionality of frozen-thawed BOEC...
December 2014: Reproduction, Fertility, and Development
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