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  1. Article ; Online: Molecular Mechanisms of Lipopolysaccharide (LPS) Induced Inflammation in an Immortalized Ovine Luteal Endothelial Cell Line (OLENDO)

    Aykut Gram / Mariusz P. Kowalewski

    Veterinary Sciences, Vol 9, Iss 99, p

    2022  Volume 99

    Abstract: Escherichia coli ( E. coli ) is the most common Gram-negative bacterium causing infection of the uterus or mammary gland and is one of the major causes of infertility in livestock. In those animals affected by E. coli driven LPS-mediated infections, ... ...

    Abstract Escherichia coli ( E. coli ) is the most common Gram-negative bacterium causing infection of the uterus or mammary gland and is one of the major causes of infertility in livestock. In those animals affected by E. coli driven LPS-mediated infections, fertility problems occur in part due to disrupted follicular and luteal functionality. However, the molecular mechanisms by which LPS induces inflammation, and specifically, the role of LPS in the disruption of capillary morphogenesis and endothelial barrier function remain unclear. Here, we hypothesized that LPS may lead to alterations in luteal angiogenesis and vascular function by inducing inflammatory reactions in endothelial cells. Accordingly, OLENDO cells were treated with LPS followed by evaluation of the expression of selected representative proinflammatory cytokines: NF-kB , IL6 , IL8 , TNFα , and ICAM 1 . While TNFα was not affected by treatment with LPS, transcripts of NF-kB , IL6 , and IL8 were affected in a dosage-dependent manner. Additionally, the activity of TLR2 and TLR4 was blocked, resulting in suppression of the LPS-induced expression of ICAM 1 , NF-kB , IL6 , and IL8 . Inhibition of the PKA or MAPK/ERK pathways suppressed the LPS-stimulated expression of NF-kB , IL6 , and IL8 , whereas blocking the PKC pathway had the opposite effect. Furthermore, LPS-induced phosphorylation of Erk1 and Erk2 was inhibited when the TLR4 or MAPK/ERK pathways were blocked. Finally, LPS seems to induce inflammatory processes in OLENDO cells via TLR2 and TLR4, utilizing different signaling pathways.
    Keywords toll-like receptor ; ovary ; interleukins ; PKA ; PKC ; MAPKs ; Veterinary medicine ; SF600-1100
    Subject code 630
    Language English
    Publishing date 2022-02-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article: The role of hypoxia and HIF1α in the regulation of STAR-mediated steroidogenesis in granulosa cells

    Kowalewski, Mariusz Pawel / Alois Boos / Aykut Gram

    Molecular and Cellular Endocrinology. 2015 Feb. 05, v. 401

    2015  

    Abstract: The adaptive responses to hypoxia are mediated by hypoxia-inducible factor 1 alpha (HIF1α). Its role, however, in regulating steroidogenesis remains poorly understood. We examined the role of hypoxia and HIF1α in regulating steroid acute regulatory ... ...

    Abstract The adaptive responses to hypoxia are mediated by hypoxia-inducible factor 1 alpha (HIF1α). Its role, however, in regulating steroidogenesis remains poorly understood. We examined the role of hypoxia and HIF1α in regulating steroid acute regulatory protein (STAR) expression and steroidogenesis in immortalized (KK1) mouse granulosa cells under progressively lowering O2 concentrations (20%, 15%, 10%, 5%, 1%). Basal and dbcAMP-stimulated progesterone synthesis was decreased under severe hypoxia (1% and 5% O2). The partial hypoxia revealed opposing effects, with a significant increase in steroidogenic response at 10% O2 in dbcAMP-treated cells: Star-promoter activity, mRNA and protein expression were increased. The hypoxia-stimulated STAR expression was PKA-dependent. Binding of HIF1α to the Star-promoter was potentiated under partial hypoxia. Inhibition of the transcriptional activity or expression of HIF1α suppressed STAR-expression. HIF1α appears to be a positive regulator of basal and stimulated STAR-expression, which under partial hypoxia is capable of increasing the steroidogenic capacity of granulosa cells.
    Keywords granulosa cells ; hypoxia ; hypoxia-inducible factor 1 ; messenger RNA ; mice ; oxygen ; progesterone ; protein synthesis ; regulatory proteins ; steroidogenesis ; transcription (genetics)
    Language English
    Dates of publication 2015-0205
    Size p. 35-44.
    Publishing place Elsevier Ireland Ltd
    Document type Article
    ZDB-ID 187438-x
    ISSN 1872-8057 ; 0303-7207
    ISSN (online) 1872-8057
    ISSN 0303-7207
    DOI 10.1016/j.mce.2014.11.023
    Database NAL-Catalogue (AGRICOLA)

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1127: Show issues Location:
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  3. Article: Prostaglandin-mediated effects in early canine corpus luteum: In vivo effects on vascular and immune factors

    Tavares Pereira, Miguel / Aykut Gram / Renata Nowaczyk / Alois Boos / Bernd Hoffmann / Tomasz Janowski / Mariusz P. Kowalewski

    Reproductive Biology. 2019 Mar., v. 19, no. 1

    2019  

    Abstract: Prostaglandins (PGs) are important regulators of the early corpus luteum (CL) in the dog. Whereas, initially, CL is gonadotropin independent, in the second half of its lifespan, hypophyseal support is required. The transition period is marked by ... ...

    Abstract Prostaglandins (PGs) are important regulators of the early corpus luteum (CL) in the dog. Whereas, initially, CL is gonadotropin independent, in the second half of its lifespan, hypophyseal support is required. The transition period is marked by decreased availability of PGs, in particular of PGE2. We previously reported that inhibition of COX2/PTGS2 in vivo suppressed luteal production of PGE2, lowered circulating progesterone and negatively affected luteal development. Therefore, bitches were treated with a COX2-specific blocker, firocoxib, for 5, 10, 20 and 30 days after ovulation, leading to suppression of the steroidogenic machinery. Control groups received a placebo for the same periods. Considering the wide range of possible modulatory roles of PGs shown in different organ systems, this follow-up project aimed to understand further possible PG-mediated effects in early canine CL. Thirty-four (34) factors related predominantly to vascularization and immune response were screened (mRNAs and proteins) on samples from the above described in vivo study. Most of the effects were observed during the transitional period (days 20 and 30). The inhibition of COX2 diminished the expression of angiopoietin family members ANGPT1, -2, Tie1 and -2 receptors. The expression of endothelin (ET)-1 was increased. Concerning the immune system, increased expression of the pro-inflammatory cytokines, IL1β, IL6 and IL12a, and elevated expression levels of CD4, was observed. Cumulatively, besides its involvement in regulating steroidogenesis, our results indicate a broader role of PGs in the canine CL, including modulation of angiogenesis, vascular stabilization and local immunomodulation. Possible cross-species translational effects are strongly implied.
    Keywords angiogenesis ; bitches ; corpus luteum ; immune response ; immune system ; immunomodulation ; in vivo studies ; interleukin-12 ; interleukin-1beta ; interleukin-6 ; longevity ; messenger RNA ; ovulation ; placebos ; progesterone ; prostaglandins ; receptors ; steroidogenesis
    Language English
    Dates of publication 2019-03
    Size p. 100-111.
    Publishing place Elsevier B.V.
    Document type Article
    ZDB-ID 2189316-0
    ISSN 1642-431X
    ISSN 1642-431X
    DOI 10.1016/j.repbio.2019.02.001
    Database NAL-Catalogue (AGRICOLA)

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 6095: Show issues Location:
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  4. Article: Cells expressing CD4, CD8, MHCII and endoglin in the canine corpus luteum of pregnancy, and prepartum activation of the luteal TNFα system

    Nowaczyk, Renata M / Alois Boos / Aykut Gram / Ewelina Jursza-Piotrowska / Mariusz P. Kowalewski / Marta J. Siemieniuch

    Theriogenology. 2017 Aug., v. 98

    2017  

    Abstract: In the dog, knowledge about involvement of the immune system in controlling luteal function is restricted to observations showing a time-dependent invasion of immune cells into the corpus luteum (CL) of non-pregnant bitches. Therefore, this study ... ...

    Abstract In the dog, knowledge about involvement of the immune system in controlling luteal function is restricted to observations showing a time-dependent invasion of immune cells into the corpus luteum (CL) of non-pregnant bitches. Therefore, this study investigated the presence of CD4-, CD8-, MHCII- and endoglin-expressing cells in CL collected throughout pregnancy from pre-implantation until prepartum luteolysis. Immunohistochemistry and semi-quantitative RT-PCR were applied. The time-dependent expression of CD4, CD8 and endoglin was more strongly related to formation of the CL, whereas MHCII was induced during luteolysis. Next, the luteal expression of TNFα and its receptors, TNFR1 and TNFR2, was analyzed in non-pregnant dogs between days 5–65 after ovulation and during pregnancy. Moreover, the effects of progesterone withdrawal were investigated in mid-pregnant dogs treated with an antigestagen aglepristone. The TNFα system was induced in the early CL of non-pregnant dogs. In pregnant dogs, expression of TNFα did not vary much, contrasting with increased expression of both receptors in the post-implantation period and significantly decreased expression at mid-gestation; prepartum luteolysis was characterized by increased TNFR2 expression. Apart from the downregulated expression of TNFR1, the changes observed following antigestagen treatment resembled those observed during normal prepartum luteolysis. A modulatory function of the TNFα system during formation of the canine CL is suggested, possibly related to the strong accompanying vascularization and luteal infiltration with activated macrophages. Contrasting with the slow luteal regression in non-pregnant dogs, in pregnant animals the upregulation of TNFR2 expression during prepartum luteolysis implies functional involvement of the TNFα system during that time.
    Keywords bitches ; corpus luteum ; immunohistochemistry ; luteolysis ; macrophages ; ovulation ; pregnancy ; progesterone ; receptors ; reverse transcriptase polymerase chain reaction ; tumor necrosis factor-alpha
    Language English
    Dates of publication 2017-08
    Size p. 123-132.
    Publishing place Elsevier Inc.
    Document type Article
    ZDB-ID 189232-0
    ISSN 1879-3231 ; 0093-691X
    ISSN (online) 1879-3231
    ISSN 0093-691X
    DOI 10.1016/j.theriogenology.2017.05.003
    Database NAL-Catalogue (AGRICOLA)

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 3929: Show issues Location:
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