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Prof. Graca Ferreira Dias
Department of Morphology and Function, CIISA- Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, 1649-004 Lisbon, Portugal

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0 Cytokines
0 Endocrinology
0 Epigenetics
0 Fibrosis
0 Reproduction

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mare
endometrium
Corpus luteum
endometrosis
Cytokines
Fibrosis
Oviduct
Reproduction
Epigenetics

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Journal article
Published: 19 May 2021 in International Journal of Molecular Sciences
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Endometrosis is a reproductive pathology that is responsible for mare infertility. Our recent studies have focused on the involvement of neutrophil extracellular traps enzymes, such as elastase (ELA), in the development of equine endometrosis. Noscapine (NOSC) is an alkaloid derived from poppy opium with anticough, antistroke, anticancer, and antifibrotic properties. The present work investigates the putative inhibitory in vitro effect of NOSC on collagen type I alpha 2 chain (COL1A2) mRNA and COL1 protein relative abundance induced by ELA in endometrial explants of mares in the follicular or mid-luteal phases at 24 or 48 h of treatment. The COL1A2 mRNA was evaluated by qPCR and COL1 protein relative abundance by Western blot. In equine endometrial explants, ELA increased COL 1 expression, while NOSC inhibited it at both estrous cycle phases and treatment times. These findings contribute to the future development of new endometrosis treatment approaches. Noscapine could be a drug capable of preventing collagen synthesis in mare’s endometrium and facilitate the therapeutic approach.

ACS Style

Ana Amaral; Carina Fernandes; Anna Szóstek-Mioduchowska; Maria Rebordão; Dariusz Skarzynski; Graça Ferreira-Dias. Noscapine Acts as a Protease Inhibitor of In Vitro Elastase-Induced Collagen Deposition in Equine Endometrium. International Journal of Molecular Sciences 2021, 22, 5333 .

AMA Style

Ana Amaral, Carina Fernandes, Anna Szóstek-Mioduchowska, Maria Rebordão, Dariusz Skarzynski, Graça Ferreira-Dias. Noscapine Acts as a Protease Inhibitor of In Vitro Elastase-Induced Collagen Deposition in Equine Endometrium. International Journal of Molecular Sciences. 2021; 22 (10):5333.

Chicago/Turabian Style

Ana Amaral; Carina Fernandes; Anna Szóstek-Mioduchowska; Maria Rebordão; Dariusz Skarzynski; Graça Ferreira-Dias. 2021. "Noscapine Acts as a Protease Inhibitor of In Vitro Elastase-Induced Collagen Deposition in Equine Endometrium." International Journal of Molecular Sciences 22, no. 10: 5333.

Journal article
Published: 12 April 2021 in Animals
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The oviduct presents the ideal conditions for fertilization and early embryonic development. In this study, (i) vascularization pattern; (ii) microvascular density; (iii) transcripts of angiogenic factors (FGF1, FGF2, VEGF) and their receptors—FGFR1, FGFR2, KDR, respectively, and (iv) the relative protein abundance of those receptors were assessed in cyclic mares’ oviducts. The oviductal artery, arterioles and their ramifications, viewed by means of vascular injection-corrosion, differed in the infundibulum, ampulla and isthmus. The isthmus, immunostained with CD31, presented the largest vascular area and the highest number of vascular structures in the follicular phase. Transcripts (qPCR) and relative protein abundance (Western blot) of angiogenic factors fibroblast growth factor 1 (FGF1) and 2 (FGF2) and vascular endothelial growth factor (VEGF), and their respective receptors (FGFR1, FGFR2, VEGFR2 = KDR), were present in all oviduct portions throughout the estrous cycle. Upregulation of the transcripts of angiogenic receptors FGF1 and FGFR1 in the ampulla and isthmus and of FGF2 and KDR in the isthmus were noted. Furthermore, in the isthmus, the relative protein abundance of FGFR1 and KDR was the highest. This study shows that the equine oviduct presents differences in microvascular density in its three portions. The angiogenic factors VEGF, FGF1, FGF2 and their respective receptors are expressed in all studied regions of the mare oviduct, in agreement with microvascular patterns.

ACS Style

Pedro Pinto-Bravo; Maria Rebordão; Ana Amaral; Carina Fernandes; António Galvão; Elisabete Silva; Pedro Pessa-Santos; Graça Alexandre-Pires; Rosário Roberto da Costa; Dariusz Skarzynski; Graça Ferreira-Dias. Microvascularization and Expression of Fibroblast Growth Factor and Vascular Endothelial Growth Factor and Their Receptors in the Mare Oviduct. Animals 2021, 11, 1099 .

AMA Style

Pedro Pinto-Bravo, Maria Rebordão, Ana Amaral, Carina Fernandes, António Galvão, Elisabete Silva, Pedro Pessa-Santos, Graça Alexandre-Pires, Rosário Roberto da Costa, Dariusz Skarzynski, Graça Ferreira-Dias. Microvascularization and Expression of Fibroblast Growth Factor and Vascular Endothelial Growth Factor and Their Receptors in the Mare Oviduct. Animals. 2021; 11 (4):1099.

Chicago/Turabian Style

Pedro Pinto-Bravo; Maria Rebordão; Ana Amaral; Carina Fernandes; António Galvão; Elisabete Silva; Pedro Pessa-Santos; Graça Alexandre-Pires; Rosário Roberto da Costa; Dariusz Skarzynski; Graça Ferreira-Dias. 2021. "Microvascularization and Expression of Fibroblast Growth Factor and Vascular Endothelial Growth Factor and Their Receptors in the Mare Oviduct." Animals 11, no. 4: 1099.

Journal article
Published: 17 February 2021 in Biology
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Seminal plasma (SP) in the female genital tract induces changes that affect multiple reproductive processes. One of the active components in SP is the transforming growth factor β1 (TGF-β1), which has major roles in embryo development and pregnancy. Embryo transfer (ET) technology is welcomed by the pig industry provided that embryo quality at embryo collection as well as the fertility and prolificacy of the recipients after the ET is increased. This study evaluated different intrauterine infusion treatments at estrus (40 mL of SP, TGF-β1 cytokine in the extender, or the extender alone (control)) by mimicking an ET scenario in so-called “donor” (inseminated) and “recipient” (uninseminated) sows. On day 6 (day 0—onset of estrus), all “donors” were laparotomized to determine their pregnancy status (presence and developmental stage of the embryos). In addition, endometrial explants were collected from pregnant “donors” and cyclic “recipients,” incubated for 24 h, and analyzed for cytokine production. SP infusions (unlike TGF-β1 infusions) positively influenced the developmental stage of day 6 embryos. Infusion treatments differentially influenced the endometrial cytokine production, mainly in donors. We concluded that SP infusions prior to AI not only impacted the porcine preimplantation embryo development but also influenced the endometrial cytokine production six days after treatment, both in donors and recipients.

ACS Style

Cristina Martinez; Josep Cambra; Xiomara Lucas; Graça Ferreira-Dias; Heriberto Rodriguez-Martinez; Maria Gil; Emilio Martinez; Cristina Cuello; Inmaculada Parrilla. Intrauterine Infusion of TGF-β1 Prior to Insemination, Alike Seminal Plasma, Influences Endometrial Cytokine Responses but Does Not Impact the Timing of the Progression of Pre-Implantation Pig Embryo Development. Biology 2021, 10, 159 .

AMA Style

Cristina Martinez, Josep Cambra, Xiomara Lucas, Graça Ferreira-Dias, Heriberto Rodriguez-Martinez, Maria Gil, Emilio Martinez, Cristina Cuello, Inmaculada Parrilla. Intrauterine Infusion of TGF-β1 Prior to Insemination, Alike Seminal Plasma, Influences Endometrial Cytokine Responses but Does Not Impact the Timing of the Progression of Pre-Implantation Pig Embryo Development. Biology. 2021; 10 (2):159.

Chicago/Turabian Style

Cristina Martinez; Josep Cambra; Xiomara Lucas; Graça Ferreira-Dias; Heriberto Rodriguez-Martinez; Maria Gil; Emilio Martinez; Cristina Cuello; Inmaculada Parrilla. 2021. "Intrauterine Infusion of TGF-β1 Prior to Insemination, Alike Seminal Plasma, Influences Endometrial Cytokine Responses but Does Not Impact the Timing of the Progression of Pre-Implantation Pig Embryo Development." Biology 10, no. 2: 159.

Journal article
Published: 16 January 2021 in Animals
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Neutrophils can originate neutrophil extracellular traps (NETs). Myeloperoxidase (MPO) is a peroxidase found in NETs associated to equine endometrosis and can be inhibited by 4-aminobenzoic acid hydrazide (ABAH). Metallopeptidases (MMPs) participate in extracellular matrix stability and fibrosis development. The objectives of this in vitro work were to investigate, in explants of mare’s endometrium, (i) the ABAH capacity to inhibit MPO-induced collagen type I (COL1) expression; and (ii) the action of MPO and ABAH on the expression and gelatinolytic activity of MMP-2/-9. Explants retrieved from the endometrium of mares in follicular or mid-luteal phases were treated with MPO, ABAH, or their combination, for 24 or 48 h. The qPCR analysis measured the transcription of COL1A2, MMP2, and MMP9. Western blot and zymography were performed to evaluate COL1 protein relative abundance and gelatinolytic activity of MMP-2/-9, respectively. Myeloperoxidase elevated COL1 relative protein abundance at both treatment times in follicular phase (p < 0.05). The capacity of ABAH to inhibit MPO-induced COL1 was detected in follicular phase at 48 h (p < 0.05). The gelatinolytic activity of activated MMP-2 augmented in mid-luteal phase at 24 h after MPO treatment, but it was reduced with MPO+ABAH treatment. The activity of MMP-9 active form augmented in MPO-treated explants. However, this effect was inhibited by ABAH in the follicular phase at 48 h (p < 0.05). By inhibiting the pro-fibrotic effects of MPO, it might be possible to reduce the development of endometrosis. Metallopeptidase-2 might be involved in an acute response to MPO in the mid-luteal phase, while MMP-9 might be implicated in a prolonged exposition to MPO in the follicular phase.

ACS Style

Ana Amaral; Carina Fernandes; Maria Rebordão; Anna Szóstek-Mioduchowska; Karolina Lukasik; Pedro Pinto-Bravo; Luís Telo da Gama; Dariusz Jan Skarzynski; Graça Ferreira-Dias. Myeloperoxidase Inhibition Decreases the Expression of Collagen and Metallopeptidase in Mare Endometria under In Vitro Conditions. Animals 2021, 11, 208 .

AMA Style

Ana Amaral, Carina Fernandes, Maria Rebordão, Anna Szóstek-Mioduchowska, Karolina Lukasik, Pedro Pinto-Bravo, Luís Telo da Gama, Dariusz Jan Skarzynski, Graça Ferreira-Dias. Myeloperoxidase Inhibition Decreases the Expression of Collagen and Metallopeptidase in Mare Endometria under In Vitro Conditions. Animals. 2021; 11 (1):208.

Chicago/Turabian Style

Ana Amaral; Carina Fernandes; Maria Rebordão; Anna Szóstek-Mioduchowska; Karolina Lukasik; Pedro Pinto-Bravo; Luís Telo da Gama; Dariusz Jan Skarzynski; Graça Ferreira-Dias. 2021. "Myeloperoxidase Inhibition Decreases the Expression of Collagen and Metallopeptidase in Mare Endometria under In Vitro Conditions." Animals 11, no. 1: 208.

Conference paper
Published: 22 November 2020 in Advances in Animal Health, Medicine and Production
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Mare endometrial function involves innumerous mediators as hormones (progesterone, estradiol, oxytocin, prostaglandins), cytokines, nitric oxide, enzymes and others. When this complex interaction is disrupted reproductive function is impaired. While some mares carry on a pregnancy to term, others develop recurrent post-breeding endometritis and chronic degenerative fibrosis (endometrosis), leading to infertility. Endometrosis is also linked to oviduct fibrosis, which exacerbates infertility. A thorough etiology, diagnosis and pathogenesis of equine endometrosis is still incomplete. In the uterus, equine neutrophils may release neutrophil extracellular traps (NETs). Besides holding bacteria in loco and kill them, they have a deleterious effect by inducing collagen formation. Our in vitro studies have shown that NETs persistence might be linked to endometrosis, due to NETs proteases action, mainly elastase and cathepsin G. These NETs proteases increase PGF2α synthesis or PGF2α receptor transcripts. Also impaired PGE2 or PGE2 receptor 2 transcripts are associated to higher collagen type I production, characteristic of fibrosis. In order to fight NETs fibrotic effect, the use of an elastase inhibitor stimulated the in vitro production of anti-fibrotic PGE2 and inhibited the pro-fibrotic PGF2α. Since these are promising results, further studies should be performed to reduce the establishment of mare endometrial fibrosis in susceptible mares.

ACS Style

G. Ferreira-Dias; M. R. Rebordão; A. M. Galvão; R. Roberto-Da-Costa; A. Amaral; C. Fernandes; P. Pinto-Bravo; S. Morazzo; G. Alexandre-Pires; K. Lukasik; A. Szóstek-Mioduchowska; D. J. Skarzynski. What Goes Wrong from a Mare Healthy Endometrium to Endometrosis? Advances in Animal Health, Medicine and Production 2020, 528 -540.

AMA Style

G. Ferreira-Dias, M. R. Rebordão, A. M. Galvão, R. Roberto-Da-Costa, A. Amaral, C. Fernandes, P. Pinto-Bravo, S. Morazzo, G. Alexandre-Pires, K. Lukasik, A. Szóstek-Mioduchowska, D. J. Skarzynski. What Goes Wrong from a Mare Healthy Endometrium to Endometrosis? Advances in Animal Health, Medicine and Production. 2020; ():528-540.

Chicago/Turabian Style

G. Ferreira-Dias; M. R. Rebordão; A. M. Galvão; R. Roberto-Da-Costa; A. Amaral; C. Fernandes; P. Pinto-Bravo; S. Morazzo; G. Alexandre-Pires; K. Lukasik; A. Szóstek-Mioduchowska; D. J. Skarzynski. 2020. "What Goes Wrong from a Mare Healthy Endometrium to Endometrosis?" Advances in Animal Health, Medicine and Production , no. : 528-540.

Journal article
Published: 04 November 2020 in Animals
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In the context of porcine embryo transfer (ET) technology, understanding the tightly regulated local uterine immune environment is crucial to achieve an adequate interaction between the transferred embryos and the receiving endometrium. However, information is limited on the uterine immune status of cyclic-recipient sows when receiving embryos during ET. The present study postulated that the anti- and proinflammatory cytokine profile 6 days after the onset of estrus differs between endometria from uninseminated cyclic sows and blastocyst-bearing sows. On Day 6 of the cycle, endometrial explants were collected from sows inseminated or not inseminated during the postweaning estrus and cultured for 22 h. The culture medium was then analyzed for the contents of a total of 16 cytokines using Luminex MAP® technology. The results showed important differences in the endometrial production of most cytokines between the sow categories, with a predominant anti-inflammatory environment displayed by the blastocyst-bearing endometria. These findings suggest that sperm, seminal plasma (SP) and/or early embryos modify the uterine environment by inducing an immune-tolerant cytokine profile already visible at Day 6. Whether the SP or some of its active components may help to develop strategies to maximize the reproductive performance of recipients after ET needs further investigation.

ACS Style

Inmaculada Parrilla; Cristina Martinez; Josep Cambra; Xiomara Lucas; Graça Ferreira-Dias; Heriberto Rodriguez-Martinez; Cristina Cuello; Maria Gil; Emilio Martinez. Blastocyst-Bearing Sows Display a Dominant Anti-Inflammatory Cytokine Profile Compared to Cyclic Sows at Day 6 of the Cycle. Animals 2020, 10, 2028 .

AMA Style

Inmaculada Parrilla, Cristina Martinez, Josep Cambra, Xiomara Lucas, Graça Ferreira-Dias, Heriberto Rodriguez-Martinez, Cristina Cuello, Maria Gil, Emilio Martinez. Blastocyst-Bearing Sows Display a Dominant Anti-Inflammatory Cytokine Profile Compared to Cyclic Sows at Day 6 of the Cycle. Animals. 2020; 10 (11):2028.

Chicago/Turabian Style

Inmaculada Parrilla; Cristina Martinez; Josep Cambra; Xiomara Lucas; Graça Ferreira-Dias; Heriberto Rodriguez-Martinez; Cristina Cuello; Maria Gil; Emilio Martinez. 2020. "Blastocyst-Bearing Sows Display a Dominant Anti-Inflammatory Cytokine Profile Compared to Cyclic Sows at Day 6 of the Cycle." Animals 10, no. 11: 2028.

Journal article
Published: 19 June 2020 in Biomolecules
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The seminal plasma (SP) modulates the female reproductive immune environment after mating, and microRNAs (miRNAs) could participate in the process. Considering that the boar ejaculate is built by fractions differing in SP-composition, this study evaluated whether exposure of mucosal explants of the sow internal genital tract (uterus, utero-tubal junction and isthmus) to different SP-fractions changed the profile of explant-secreted miRNAs. Mucosal explants retrieved from oestrus sows (n = 3) were in vitro exposed to: Medium 199 (M199, Control) or M199 supplemented (1:40 v/v) with SP from the sperm-rich fraction (SRF), the post-SRF or the entire recomposed ejaculate, for 16 h. After, the explants were cultured in M199 for 24 h to finally collect the media for miRNA analyses using GeneChip miRNA 4.0 Array (Affymetrix). Fifteen differentially expressed (False Discovery Rate (FDR) < 0.05 and Fold-change ≥ 2) miRNAs (11 down- versus 4 up-regulated) were identified (the most in the media of uterine explants incubated with SP from post-SRF). Bioinformatics analysis identified that predicted target genes of dysregulated miRNAs, mainly miR-34b, miR-205, miR-4776-3p and miR-574-5p, were involved in functions and pathways related to immune response. In conclusion, SP is able to elicit changes in the miRNAs profile secreted by female genital tract, ultimately depending SP-composition.

ACS Style

Isabel Barranco; Lorena Padilla; Cristina A. Martinez; Manuel Alvarez-Rodriguez; Inmaculada Parrilla; Xiomara Lucas; Graça Ferreira-Dias; Marc Yeste; Heriberto Rodriguez-Martinez; Jordi Roca. Seminal Plasma Modulates miRNA Expression by Sow Genital Tract Lining Explants. Biomolecules 2020, 10, 933 .

AMA Style

Isabel Barranco, Lorena Padilla, Cristina A. Martinez, Manuel Alvarez-Rodriguez, Inmaculada Parrilla, Xiomara Lucas, Graça Ferreira-Dias, Marc Yeste, Heriberto Rodriguez-Martinez, Jordi Roca. Seminal Plasma Modulates miRNA Expression by Sow Genital Tract Lining Explants. Biomolecules. 2020; 10 (6):933.

Chicago/Turabian Style

Isabel Barranco; Lorena Padilla; Cristina A. Martinez; Manuel Alvarez-Rodriguez; Inmaculada Parrilla; Xiomara Lucas; Graça Ferreira-Dias; Marc Yeste; Heriberto Rodriguez-Martinez; Jordi Roca. 2020. "Seminal Plasma Modulates miRNA Expression by Sow Genital Tract Lining Explants." Biomolecules 10, no. 6: 933.

Journal article
Published: 16 May 2020 in Animals
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Neutrophil extracellular traps (NETs) fight endometritis, and elastase (ELA), a protease found in NETs, might induce collagen type I (COL1) accumulation in equine endometrium. Metallopeptidases (MMPs) are involved in extracellular matrix balance. The aim was to evaluate the effects of ELA and sivelestat (selective elastase inhibitor) on MMP-2 and MMP-9 expression and gelatinolytic activity, as well as the potential inhibitory effect of sivelestat on ELA-induced COL1 in equine endometrium. Endometrial explants from follicular (FP) and mid-luteal (MLP) phases were treated for 24 or 48 h with ELA, sivelestat, and their combination. Transcripts of COL1A2, MMP2, and MMP9 were evaluated by qPCR; COL1 protein relative abundance by Western blot, and MMP-2 and MMP-9 gelatinolytic activity by zymography. In response to ELA treatment, there was an increase in MMP2 mRNA transcription (24 h) in active MMP-2 (48 h), both in FP, and in MMP9 transcripts in FP (48 h) and MLP (24 h) (p < 0.05). Sivelestat inhibited ELA-induced COL1A2 transcripts in FP (24 h) and MLP (24 h, 48 h) (p < 0.05). The sivelestat inhibitory effect was detected in MMP9 transcripts in FP at 48 h (p < 0.05), but proteases activity was unchanged. Thus, MMP-2 and MMP-9 might be implicated in endometrium fibrotic response to ELA. In mare endometrium, sivelestat may decrease ELA-induced COL1 deposition and hinder endometrosis development.

ACS Style

Ana Amaral; Carina Fernandes; Maria Rosa Rebordão; Anna Szóstek-Mioduchowska; Karolina Łukasik; Barbara Gawronska-Kozak; Luís Telo Da Gama; Dariusz J. Skarzynski; Graça Ferreira-Dias. The In Vitro Inhibitory Effect of Sivelestat on Elastase Induced Collagen and Metallopeptidase Expression in Equine Endometrium. Animals 2020, 10, 863 .

AMA Style

Ana Amaral, Carina Fernandes, Maria Rosa Rebordão, Anna Szóstek-Mioduchowska, Karolina Łukasik, Barbara Gawronska-Kozak, Luís Telo Da Gama, Dariusz J. Skarzynski, Graça Ferreira-Dias. The In Vitro Inhibitory Effect of Sivelestat on Elastase Induced Collagen and Metallopeptidase Expression in Equine Endometrium. Animals. 2020; 10 (5):863.

Chicago/Turabian Style

Ana Amaral; Carina Fernandes; Maria Rosa Rebordão; Anna Szóstek-Mioduchowska; Karolina Łukasik; Barbara Gawronska-Kozak; Luís Telo Da Gama; Dariusz J. Skarzynski; Graça Ferreira-Dias. 2020. "The In Vitro Inhibitory Effect of Sivelestat on Elastase Induced Collagen and Metallopeptidase Expression in Equine Endometrium." Animals 10, no. 5: 863.

Journal article
Published: 29 January 2020 in Theriogenology
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Equine endometrosis is a multifactorial chronic degenerative condition, considered to be one of a major causes of equine infertility. The formation of periglandular fibrosis seems to be linked to chronic inflammation of the mare endometrium in a paracrine way and in a response to numerous forms of inflammatory stimuli elicit the net deposition of extracellular matrix (ECM) around the endometrial glands and stroma. We hypothesized some of these stimuli, such as monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6), and hyaluronan synthases (HASs), may share the nuclear factor-κB (NF-κB) dependent activation pathway. This study aimed to determine whether mRNA expression of MCP-1, IL-6, HASs, and proteins of canonical (RelA/NK-κβ1) and noncanonical (NK-κβ2) signaling pathways for NF-kB would change in subsequent categories of endometrosis during the estrous cycle. The expression of selected genes was established in mare endometrium (n = 80; Kenney and Doig categories I, IIA, IIB, III), obtained in the follicular phase (FLP) and mid-luteal phase (MLP). The high expression of RelA mRNA was observed in III, whereas of NK-κβ1 and NK-κβ2 also in IIA, and IIA and IIB, respectively. The expression of MCP-1 mRNA occurred constantly, regardless of the category, whereas IL-6 mRNA was low in IIA, IIB, and III. The expression of HAS 1 was high in IIA and HAS 3 in IIA, IIB, and III. All those changes were observed in FLP, but not MLP. Our results suggest that NF-κB may be involved in progression of the chronic degenerative condition of the mare endometrium, on both canonical and noncanonical pathways. The most important changes in target genes expression were observed only in FLP, which may suggest the hormone-dependent activation of the NF-κB-dependent fibrosis pathway.

ACS Style

Malgorzata Domino; Tomasz Jasinski; Ewa Kautz; Edyta Juszczuk-Kubiak; Graça Maria Leitão Ferreira Dias; Romuald Zabielski; Maria Sady; Zdzislaw Gajewski. Expression of genes involved in the NF-κB-dependent pathway of the fibrosis in the mare endometrium. Theriogenology 2020, 147, 18 -24.

AMA Style

Malgorzata Domino, Tomasz Jasinski, Ewa Kautz, Edyta Juszczuk-Kubiak, Graça Maria Leitão Ferreira Dias, Romuald Zabielski, Maria Sady, Zdzislaw Gajewski. Expression of genes involved in the NF-κB-dependent pathway of the fibrosis in the mare endometrium. Theriogenology. 2020; 147 ():18-24.

Chicago/Turabian Style

Malgorzata Domino; Tomasz Jasinski; Ewa Kautz; Edyta Juszczuk-Kubiak; Graça Maria Leitão Ferreira Dias; Romuald Zabielski; Maria Sady; Zdzislaw Gajewski. 2020. "Expression of genes involved in the NF-κB-dependent pathway of the fibrosis in the mare endometrium." Theriogenology 147, no. : 18-24.

Journal article
Published: 13 January 2020 in Theriogenology
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The innate and adaptive immune mechanisms are key components of regulation of reproductive physiological function and uterine disorders in equine uterus. The predominant immunological response in equine endometrium, characterized by an innate immune response, occurs under estrogens influence, in the follicular phase. Although, the increase in immune-related genes in equine endometrium during estrus has been suggested to play a role in uterine clearance after mating, immune cells and their product, i.e. cytokines play also mandatory role in the luteal development and maintenance, regression of equine corpus luteum, as well as in early pregnancy. Innate immune response is nonspecific and acts as the first line of defense against pathogens, foreign stimuli that include constituents of seminal fluid and local infections (endometritis). It has been recently established that a phagocytosis-independent mechanism to restrain bacteria, by means of neutrophil extracellular traps (NETs) formation, is involved in pathogenesis of in mare endometrial fibrosis (endometrosis). Moreover, persistent macrophages and mast cell activation could also have pro-fibrotic roles by secreting great amounts of pro-fibrotic factors and lead to fibrosis. This review will highlight the involvement of immune key components of the innate and adaptive immune system and their products in equine uterus and their contribution to reproductive physiological function and uterine disorders.

ACS Style

Dariusz J. Skarzynski; Anna Szóstek-Mioduchowska; Maria Rosa Rebordão; Beenu Moza Jalali; Katarzyna K. Piotrowska-Tomala; Natalia Leciejewska; Miriam Łazarczyk; Graça Maria Leitão Ferreira Dias. Neutrophils, monocytes and other immune components in the equine endometrium: Friends or foes? Theriogenology 2020, 150, 150 -157.

AMA Style

Dariusz J. Skarzynski, Anna Szóstek-Mioduchowska, Maria Rosa Rebordão, Beenu Moza Jalali, Katarzyna K. Piotrowska-Tomala, Natalia Leciejewska, Miriam Łazarczyk, Graça Maria Leitão Ferreira Dias. Neutrophils, monocytes and other immune components in the equine endometrium: Friends or foes? Theriogenology. 2020; 150 ():150-157.

Chicago/Turabian Style

Dariusz J. Skarzynski; Anna Szóstek-Mioduchowska; Maria Rosa Rebordão; Beenu Moza Jalali; Katarzyna K. Piotrowska-Tomala; Natalia Leciejewska; Miriam Łazarczyk; Graça Maria Leitão Ferreira Dias. 2020. "Neutrophils, monocytes and other immune components in the equine endometrium: Friends or foes?" Theriogenology 150, no. : 150-157.

Original research article
Published: 18 December 2019 in Frontiers in Veterinary Science
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Background: Seminal plasma (SP) promotes sperm survival and fertilizing capacity, and potentially affects embryo development, presumably via specific signaling pathways to the internal female genital tract. Objectives: This study evaluated how heterologous SP, infused immediately before postcervical artificial insemination (AI) affected embryo development and the transcriptional pattern of the pig endometria containing embryos. Materials and Methods: Postweaning estrus sows (n = 34) received 40-mL intrauterine infusions of either heterologous pooled SP or Beltsville Thawing Solution (BTS; control) 30 min before AI of semen extended to 10% of homologous SP. Embryos (all sows) and endometrium samples (3 sows/group) were removed during laparotomy 6 days after the infusion of SP or BTS to morphologically evaluate the embryos to determine their developmental stage and to analyze the endometrial transcriptome using microarrays (PORGENE 1.0 ST GeneChip array, Affymetrix) followed by qPCR for further validation. Results: Embryo viability was equal between the groups (~93%), but embryo development was significantly (P < 0.05) more advanced in the SP-treated group compared to control. A total of 1,604 endometrium transcripts were differentially expressed in the SP group compared to the control group. An enrichment analysis showed an overrepresentation of genes and pathways associated with the immune response, cytokine signaling, cell cycle, cell adhesion, and hormone response, among others. Conclusions: SP infusions prior to AI positively impacted the preimplantation embryo development and altered the expression of the endometrial genes and pathways potentially involved in embryo development.

ACS Style

Cristina A Martinez; Josep M. Cambra; Inmaculada Parrilla; Jordi Roca; Graça Maria Leitão Ferreira Dias; Francisco J. Pallares; Xiomara Lucas; Juan M. Vazquez; Emilio A. Martinez; Maria A. Gil; Heriberto Rodriguez-Martinez; Cristina Cuello; Manuel Alvarez-Rodriguez. Seminal Plasma Modifies the Transcriptional Pattern of the Endometrium and Advances Embryo Development in Pigs. Frontiers in Veterinary Science 2019, 6, 465 .

AMA Style

Cristina A Martinez, Josep M. Cambra, Inmaculada Parrilla, Jordi Roca, Graça Maria Leitão Ferreira Dias, Francisco J. Pallares, Xiomara Lucas, Juan M. Vazquez, Emilio A. Martinez, Maria A. Gil, Heriberto Rodriguez-Martinez, Cristina Cuello, Manuel Alvarez-Rodriguez. Seminal Plasma Modifies the Transcriptional Pattern of the Endometrium and Advances Embryo Development in Pigs. Frontiers in Veterinary Science. 2019; 6 ():465.

Chicago/Turabian Style

Cristina A Martinez; Josep M. Cambra; Inmaculada Parrilla; Jordi Roca; Graça Maria Leitão Ferreira Dias; Francisco J. Pallares; Xiomara Lucas; Juan M. Vazquez; Emilio A. Martinez; Maria A. Gil; Heriberto Rodriguez-Martinez; Cristina Cuello; Manuel Alvarez-Rodriguez. 2019. "Seminal Plasma Modifies the Transcriptional Pattern of the Endometrium and Advances Embryo Development in Pigs." Frontiers in Veterinary Science 6, no. : 465.

Journal article
Published: 19 November 2019 in Theriogenology
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Pregnancy loss during the normal lifespan of endometrial cups (∼37-120-150 days of gestation) may affect a mare’s ability to conceive again in the same breeding season, as equine chorionic gonadotropin (eCG) secretion by retained endometrial cups can lead to abnormal ovulations and follicular growth. While intrauterine kerosene infusion has anecdotally been proposed as a treatment for endometrial cup retention, there are no controlled studies evaluating kerosene’s ability to enhance endometrial cup regression following abortion. The objectives of this study were to assess uterine response, systemic side effects, and efficacy of intrauterine kerosene infusions after abortion. We hypothesized that kerosene infusions would hasten regression of endometrial cups without detrimental effects on the endometrium and the mare’s general health. Twelve light-breed mares were enrolled in the study after an experimentally induced abortion with cloprostenol (n = 12) by 60 ± 2 days of gestation. Mares were randomly allocated to receive an intrauterine infusion with 500 mL of kerosene (n = 6) or 500 mL saline (n = 6) on days 21 and 35 after pregnancy termination. Uterine biopsies were collected at days 7, 21, 35, and 49 post-abortion to evaluate the degree of endometrial fibrosis with Picrosirius Red Stain and to be graded according to the Kenney & Doig 1986 classification. Furthermore, histomorphometry analysis of the endometrium lining, glandular epithelium and glandular density was performed. Endometrial lymphocyte B CD20+, lymphocyte T CD3+, and macrophage IBA-1+ cell populations were characterized by immunohistochemistry. Physical examinations, blood cell counts, and serum biochemistry were performed before, and for 2 days after each uterine infusion. Serum samples were collected for assessment of eCG concentrations. Continuous data were analyzed with MIXED procedure with repeated measures in SAS, categorical data with LOGISTIC procedure of SAS. Significance was set at p < 0.05. Kerosene infusion did not affect complete blood cell counts, serum chemistry parameters, or physical examinations. Concentrations of eCG decreased over time (p < 0.001), but there were no differences between groups or time by group interactions (p = 0.72). Histological evaluation of the uterus showed no signs of increased fibrosis or degeneration in the treatment group. In conclusion, while kerosene infusions did not appear to have detrimental effects on mare health, our findings suggest that the use of kerosene in the uterus does not enhance the regression of endometrial cups by 49 days post-abortion.

ACS Style

Giorgia Podico; Igor F. Canisso; Patrick J. Roady; Scott M. Austin; Mariano Carossino; Udeni Balasuriya; Robyn E. Ellerbrock; Fabio Lima; Graça Maria Leitão Ferreira Dias; Robert H. Douglas. Uterine responses and equine chorionic gonadotropin concentrations after two intrauterine infusions with kerosene post early fetal loss in mares. Theriogenology 2019, 147, 202 -210.

AMA Style

Giorgia Podico, Igor F. Canisso, Patrick J. Roady, Scott M. Austin, Mariano Carossino, Udeni Balasuriya, Robyn E. Ellerbrock, Fabio Lima, Graça Maria Leitão Ferreira Dias, Robert H. Douglas. Uterine responses and equine chorionic gonadotropin concentrations after two intrauterine infusions with kerosene post early fetal loss in mares. Theriogenology. 2019; 147 ():202-210.

Chicago/Turabian Style

Giorgia Podico; Igor F. Canisso; Patrick J. Roady; Scott M. Austin; Mariano Carossino; Udeni Balasuriya; Robyn E. Ellerbrock; Fabio Lima; Graça Maria Leitão Ferreira Dias; Robert H. Douglas. 2019. "Uterine responses and equine chorionic gonadotropin concentrations after two intrauterine infusions with kerosene post early fetal loss in mares." Theriogenology 147, no. : 202-210.

Original research article
Published: 01 October 2019 in Frontiers in Endocrinology
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The regulation of corpus luteus (CL) luteolysis is a complex process involving a myriad of factors. Previously, we have shown the involvement of Nodal in functional luteolysis in mares. Presently, we ask the extent of which Nodal mediation of luteolysis is done through regulation of angioregression. We demonstrated the interaction between Nodal and hypoxia-inducible factor 1 α (HIF1α) and thrombospondin 1/thrombospondin receptor (TSP1/CD36) systems, could mediate angioregression during luteolysis. First, we demonstrated the inhibitory effect of Nodal on the vascular marker platelet/endothelial cell adhesion molecule 1 (CD31). Also, treatment of mid CL explants with vascular endothelial growth factor A (VEGFA) showed a trend on activin-like kinase 7 (Alk7) protein inhibition. Next, Nodal was also shown to activate HIF1α and in vitro culture of mid CL explants under decreased oxygen level promoted Nodal expression and SMAD family member 3 (Smad3) phosphorylation. In another experiment, the crosstalk between Nodal and TSP1/CD36 was investigated. Indeed, Nodal increased the expression of the anti-angiogenic TSP1 and its receptor CD36 in mid CL explants. Finally, the supportive effect of prostaglandin F2α (PGF2α) on TSP1/CD36 was blocked by SB431542 (SB), a pharmacological inhibitor of Nodal signaling. Thus, we evidenced for the first time the in vitro interaction between Nodal and both HIF1α and TSP1 systems, two conserved pathways previously shown to be involved in vascular regression during luteolysis. Considering the given increased expression of Nodal in mid CL and its role on functional luteolysis, the current results suggest the additional involvement of Nodal in angioregression during luteolysis in the mare, particularly in the activation of HIF1α and TSP1/CD36.

ACS Style

Edyta Walewska; Karolina M. Wołodko; Dariusz Skarzynski; Graça Maria Leitão Ferreira Dias; António Galvão. The Interaction Between Nodal, Hypoxia-Inducible Factor 1 Alpha, and Thrombospondin 1 Promotes Luteolysis in Equine Corpus Luteum. Frontiers in Endocrinology 2019, 10, 1 .

AMA Style

Edyta Walewska, Karolina M. Wołodko, Dariusz Skarzynski, Graça Maria Leitão Ferreira Dias, António Galvão. The Interaction Between Nodal, Hypoxia-Inducible Factor 1 Alpha, and Thrombospondin 1 Promotes Luteolysis in Equine Corpus Luteum. Frontiers in Endocrinology. 2019; 10 ():1.

Chicago/Turabian Style

Edyta Walewska; Karolina M. Wołodko; Dariusz Skarzynski; Graça Maria Leitão Ferreira Dias; António Galvão. 2019. "The Interaction Between Nodal, Hypoxia-Inducible Factor 1 Alpha, and Thrombospondin 1 Promotes Luteolysis in Equine Corpus Luteum." Frontiers in Endocrinology 10, no. : 1.

Supplement article
Published: 11 September 2019 in Reproduction in Domestic Animals
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Inflammation and fibroproliferative diseases may be modulated by epigenetic changes. Therefore, we suggest that epigenetic mechanisms could be involved in equine endometrosis pathogenesis. DNA methylation is one of the methods to evaluate epigenetics, through the transcription of methyltransferases (DNMT1, DNMT3A, DNMT3B). The correlation between DNMTs and collagen (COL) transcripts was assessed for the different Kenney and Doig's (Current Therapy in Theriogenology. Philadelphia: WB Saunders; 1986) endometrium categories. Endometrial biopsies were randomly collected from cyclic mares. Histological classification (category I, n = 13; II A, n = 17; II B, n = 12; and III, n = 7) and evaluation of COL1A2, COL3A1 and DNMTs transcripts by qPCR, were performed. Data were analysed by one‐way analysis of variance (ANOVA), Kruskal–Wallis test and Pearson correlation. As mares aged, there was an increase in endometrium fibrosis (p < .01), and in DNMT1 mRNA (p < .001). Considering DNMT3B transcripts for each category, there was an increase with fibrosis (p < .05). No changes were observed for DNMT1 and DNMT3A transcripts. However, DNMT3A mRNA levels were the highest in all categories (p < .01). In category I endometrium, a positive correlation was observed for transcripts of all DNMTs in both COLs (p < .01). In category IIA, this correlation was also maintained for all DNMTs transcripts in COL1A2 (p < .05), but only for DNMT3B in COL3A1 (p < .05). In category IIB, there was a positive correlation between DNMT3B and COL3A1 (p < .05). In category III, a positive correlation was only observed between DNMT3B and COL3A1 (p < .05). Our results suggest that there is a disturbance in COLs and DNMTs correlation during fibrosis.

ACS Style

Joana Alpoim-Moreira; Carina Fernandes; Maria Rosa Rebordão; Ana Amaral; Pedro Pinto-Bravo; Miguel Bliebernicht; Dariusz J. Skarzynski; Graça Maria Leitão Ferreira Dias. Collagens and DNA methyltransferases in mare endometrosis. Reproduction in Domestic Animals 2019, 54, 46 -52.

AMA Style

Joana Alpoim-Moreira, Carina Fernandes, Maria Rosa Rebordão, Ana Amaral, Pedro Pinto-Bravo, Miguel Bliebernicht, Dariusz J. Skarzynski, Graça Maria Leitão Ferreira Dias. Collagens and DNA methyltransferases in mare endometrosis. Reproduction in Domestic Animals. 2019; 54 ():46-52.

Chicago/Turabian Style

Joana Alpoim-Moreira; Carina Fernandes; Maria Rosa Rebordão; Ana Amaral; Pedro Pinto-Bravo; Miguel Bliebernicht; Dariusz J. Skarzynski; Graça Maria Leitão Ferreira Dias. 2019. "Collagens and DNA methyltransferases in mare endometrosis." Reproduction in Domestic Animals 54, no. : 46-52.

Research article
Published: 17 July 2019 in PLoS ONE
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Development of a healthy musculoskeletal system is of high concern for horse breeders and users. A longitudinal field study was performed in order to: (i) evaluate growth patterns and long-term changes on bone quality, bone metabolism, growth factors and metabolic variables in the Lusitano horse; and (ii) retrospectively assess whether these changes were related with radiographic findings regarding osteochondrosis-like lesions (OC) at the onset of training. Thirty-four Lusitano foals born and raised at four stud-farms, were periodically weighed (BW), and measured (withers height-WH) from birth to 36 months of age. On the same days, blood samples were collected for determination of osteocalcin, bone alkaline phosphatase, insulin-like growth factor I (IGF-I), leptin, insulin, glucose, parathyroid hormone (PTH), calcium, phosphorus and magnesium plasma concentrations, and quantitative ultrasound measurements were performed on the right third metacarpal bone (McIII). At the end of the study horses underwent radiographic examination of the four fetlocks, hocks and stifles. According to their radiographic status (OC negative vs. OC positive), Richards growth function was adjusted to BW and WH data. Instantaneous BW and WH growth rates (BW IADG and WH IADG) were calculated for each foal, from the resolution of the first derivative of growth models for seven age-classes. The presence of radiographic findings compatible with OC at the onset of training was associated with changes in BW and WH growth rates. Positive horses presented higher BW IADG from six to 18 months of age and lower WH IADG before 45 days of age (P<0.001). Speed of sound measurements (SOS), bone markers, growth factors and other metabolic variables change markedly with age (P<0.01). OC positive horses tended to have lower SOS values at the lateral region of McIII, lower IGF-I, and higher insulin and PTH concentrations (P<0.1). This study provides indirect evidence that monitoring foals’ growth during the first year of life may be of assistance in managing the occurrence of OC. Further studies with a higher number of animals and a controlled feed intake should be pursued.

ACS Style

Maria J. Fradinho; Luísa Mateus; Nuno Bernardes; Rui J. B. Bessa; Rui M. Caldeira; Graça Ferreira-Dias. Growth patterns, metabolic indicators and osteoarticular status in the Lusitano horse: A longitudinal study. PLoS ONE 2019, 14, e0219900 .

AMA Style

Maria J. Fradinho, Luísa Mateus, Nuno Bernardes, Rui J. B. Bessa, Rui M. Caldeira, Graça Ferreira-Dias. Growth patterns, metabolic indicators and osteoarticular status in the Lusitano horse: A longitudinal study. PLoS ONE. 2019; 14 (7):e0219900.

Chicago/Turabian Style

Maria J. Fradinho; Luísa Mateus; Nuno Bernardes; Rui J. B. Bessa; Rui M. Caldeira; Graça Ferreira-Dias. 2019. "Growth patterns, metabolic indicators and osteoarticular status in the Lusitano horse: A longitudinal study." PLoS ONE 14, no. 7: e0219900.

Journal article
Published: 03 November 2018 in Domestic Animal Endocrinology
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Prostaglandin E2 (PGE2) has contradictory effects in many organs. It may have pro-inflammatory, anti-inflammatory or anti-fibrotic roles, depending on the type of receptors to which it binds. By signaling through its receptors EP2 and EP4, PGE2 mediates anti-inflammatory and anti-fibrotic actions. In spite of chronic endometrial fibrosis (endometrosis) being a major cause of mare infertility, its pathogenesis is not fully understood. We have shown that contact of mare endometrium in vitro with neutrophil extracellular traps (NETs) proteases favors endometrial collagen type I production. Therefore, we investigated the involvement of the PGE2 pathway in collagen deposition on mare endometrium, challenged in vitro with proteases present in NETs. Mare endometria (Kenney and Doig categories I/IIA and IIB/III), obtained in the follicular (FLP) and mid-luteal (MLP) phases, were incubated for 24h with components found in NETs (elastase, cathepsin-G, myeloperoxidase). Secretion of PGE2 and specific PGE synthase (PGES) and PGE2 receptors (EP2, EP4) transcripts were evaluated. Impaired PGE2 production and low EP2 transcripts depended on the endometrial category and estrous cycle phases. Impairment of PGE2 and/or EP2 might play a role in FLP (category IIB/III) and MLP (I/IIA) endometrial fibrogenesis, due to the suppression of its anti-fibrotic capacity. In conclusion, priming of the endometrium with endogenous ovarian steroids might inhibit the anti-fibrotic PGE2 pathway either in healthy or pathological tissues with collagen formation after NETs proteases action.

ACS Style

Maria Rosa Rebordão; Ana Amaral; Karolina Łukasik; Anna Szóstek-Mioduchowska; Pedro Pinto-Bravo; António Galvão; Dariusz J. Skarzynski; Graça Maria Leitão Ferreira Dias. Impairment of the antifibrotic prostaglandin E2 pathway may influence neutrophil extracellular traps–induced fibrosis in the mare endometrium. Domestic Animal Endocrinology 2018, 67, 1 -10.

AMA Style

Maria Rosa Rebordão, Ana Amaral, Karolina Łukasik, Anna Szóstek-Mioduchowska, Pedro Pinto-Bravo, António Galvão, Dariusz J. Skarzynski, Graça Maria Leitão Ferreira Dias. Impairment of the antifibrotic prostaglandin E2 pathway may influence neutrophil extracellular traps–induced fibrosis in the mare endometrium. Domestic Animal Endocrinology. 2018; 67 ():1-10.

Chicago/Turabian Style

Maria Rosa Rebordão; Ana Amaral; Karolina Łukasik; Anna Szóstek-Mioduchowska; Pedro Pinto-Bravo; António Galvão; Dariusz J. Skarzynski; Graça Maria Leitão Ferreira Dias. 2018. "Impairment of the antifibrotic prostaglandin E2 pathway may influence neutrophil extracellular traps–induced fibrosis in the mare endometrium." Domestic Animal Endocrinology 67, no. : 1-10.

Journal article
Published: 01 October 2018 in Cytokine
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In the present report we describe the involvement of transforming growth factor B1 (TGF) in functional regression and structural luteolysis in the mare. Firstly, TGF and its receptors activin-like kinase (ALK) 5 and TGF receptor 2 were identified in corpus luteum (CL) steroidogenic, endothelial and fibroblast-like cells. Also, TGF and ALK5 protein expression were shown to be increased in Mid-, and Late-CL (p < 0.05). Subsequently, using an in vitro model with Mid-CL cells, we studied the role of TGF on secretory activity and cell viability. Cell treatment with TGF decreased progesterone (P4) and prostaglandin (PG) E2 concentrations in culture media (p < 0.05), and downregulated mRNA and protein of StAR, CYP11A1, cPGES and mPGES1 (p < 0.05). Conversely, TGF augmented PGF2a concentration in culture media, through PTGS2 and PGFS gene expression activation (p < 0.05). When cells were incubated with PGF2a, both TGF and ALK5 were upregulated (p < 0.05). Additionally, treatment with the pharmacological inhibitor of ALK5, ALK4 and ALK7 - SB431542 (SB) attenuated PGF2a functional and structural luteolytic actions. Indeed, SB blocked: (i) PGF2a inhibitory effect on StAR, CYP11A1, 3BHSD and mPGES1; (ii) PGF2a auto-amplification signal via PTGS2 and PGFS expression (p < 0.05); (iii) the PGF2a-induced BAX and FASL expression (p < 0.05). Finally, TGF decreased cell viability (p < 0.05) and promoted caspase 3 activity (p = 0.08) and the expression of pro-apoptotic FASL and BAX (p < 0.05). Our results suggest that TGF supports functional regression and structural luteolysis, and also confirm the importance of ALK5, ALK4 and ALK7 activation during PGF2a mediated luteolysis in mares.

ACS Style

António Galvão; Karolina M. Wołodko; Maria Rosa Rebordão; Dariusz Skarzynski; Graça Maria Leitão Ferreira Dias. TGFB1 modulates in vitro secretory activity and viability of equine luteal cells. Cytokine 2018, 110, 316 -327.

AMA Style

António Galvão, Karolina M. Wołodko, Maria Rosa Rebordão, Dariusz Skarzynski, Graça Maria Leitão Ferreira Dias. TGFB1 modulates in vitro secretory activity and viability of equine luteal cells. Cytokine. 2018; 110 ():316-327.

Chicago/Turabian Style

António Galvão; Karolina M. Wołodko; Maria Rosa Rebordão; Dariusz Skarzynski; Graça Maria Leitão Ferreira Dias. 2018. "TGFB1 modulates in vitro secretory activity and viability of equine luteal cells." Cytokine 110, no. : 316-327.

Short communication
Published: 21 September 2018 in Reproduction in Domestic Animals
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We have shown that bacteria induce neutrophil extracellular traps (NETs) in mare endometrium. Besides killing pathogens, NETs may contribute for endometrosis (chronic endometrium fibrosis). Since elastase (ELA) is a NETs component that regulates fibrosis and prostaglandin (PG) output, the aim was to evaluate if inhibition of ELA would affect collagen 1 (COL1) transcription and PGs secretion by endometrium explants, in different estrous cycle phases. Follicular‐FP (n = 8) and mid luteal–MLP (n = 7) phases explants were cultured for 24–48 hr with medium alone (Control), ELA (0.5 μg/ml,1 μg/ml), sivelestat ‐ ELA inhibitor (INH,10 μg/ml), or ELA (0.5 μg/ml,1 μg/ml) + INH (10 μg/ml). COL1 gene transcription was done by qRT‐PCR and PGE2 and PGF2α determination in culture medium by EIA. In FP, at 24 hr, ELA0.5 increased COL1 transcription (p < 0.001) but its inhibition (ELA0.5 + INH10) decreased COL1 transcription (p < 0.01) and PGF2α production (p < 0.05). Also, ELA0.5 + INH10 or ELA1 + INH10 raised PGE2 production (p < 0.01). At 48 hr, ELA1 increased COL1 transcription (p < 0.01) and PGF2α production (p < 0.001), but its inhibition (ELA1 + INH10) decreased these actions (p < 0.01; p < 0.05, respectively). Besides, ELA1 + INH10 incubation increased PGE2 (p < 0.05). PGF2α also augmented with ELA0.5 (p < 0.001), but lowered with ELA0.5 + INH10 (p < 0.01). In MLP, ELA0.5 up‐regulated COL1 transcription (24 hr, p < 0.01; 48 hr, p < 0.001), but ELA0.5 + INH10 decreased it (24 hr, p < 0.05; 48 hr, p < 0.001). At 48 hr, incubation with ELA1 also increased COL1 transcription and PGF2α production (p < 0.05), but PGF2α production decreased with ELA1 + INH10 incubation (p < 0.05). PGE2 production was higher in ELA1 + INH10 incubation (p < 0.05). Therefore, ELA inhibition may reduce the establishment of mare endometrial fibrosis by stimulating the production of anti‐fibrotic PGE2 and inhibiting pro‐fibrotic PGF2α.

ACS Style

Ana Amaral; Carina Fernandes; Karolina Lukasik; Anna Szóstek-Mioduchowska; Agnieszka Baclawska; Maria Rosa Rebordão; Joana Aguiar-Silva; Pedro Pinto-Bravo; Dariusz J. Skarzynski; Graça Maria Leitão Ferreira Dias. Elastase inhibition affects collagen transcription and prostaglandin secretion in mare endometrium during the estrous cycle. Reproduction in Domestic Animals 2018, 53, 66 -69.

AMA Style

Ana Amaral, Carina Fernandes, Karolina Lukasik, Anna Szóstek-Mioduchowska, Agnieszka Baclawska, Maria Rosa Rebordão, Joana Aguiar-Silva, Pedro Pinto-Bravo, Dariusz J. Skarzynski, Graça Maria Leitão Ferreira Dias. Elastase inhibition affects collagen transcription and prostaglandin secretion in mare endometrium during the estrous cycle. Reproduction in Domestic Animals. 2018; 53 ():66-69.

Chicago/Turabian Style

Ana Amaral; Carina Fernandes; Karolina Lukasik; Anna Szóstek-Mioduchowska; Agnieszka Baclawska; Maria Rosa Rebordão; Joana Aguiar-Silva; Pedro Pinto-Bravo; Dariusz J. Skarzynski; Graça Maria Leitão Ferreira Dias. 2018. "Elastase inhibition affects collagen transcription and prostaglandin secretion in mare endometrium during the estrous cycle." Reproduction in Domestic Animals 53, no. : 66-69.

Journal article
Published: 01 May 2018 in Livestock Science
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ACS Style

M.J. Fradinho; Luisa Mateus; R.J.B. Bessa; R.M. Caldeira; Graça Maria Leitão Ferreira Dias. Age-related changes of bone ultrasound measurements and metabolic indicators in the young horse. Livestock Science 2018, 211, 104 -110.

AMA Style

M.J. Fradinho, Luisa Mateus, R.J.B. Bessa, R.M. Caldeira, Graça Maria Leitão Ferreira Dias. Age-related changes of bone ultrasound measurements and metabolic indicators in the young horse. Livestock Science. 2018; 211 ():104-110.

Chicago/Turabian Style

M.J. Fradinho; Luisa Mateus; R.J.B. Bessa; R.M. Caldeira; Graça Maria Leitão Ferreira Dias. 2018. "Age-related changes of bone ultrasound measurements and metabolic indicators in the young horse." Livestock Science 211, no. : 104-110.

Journal article
Published: 06 February 2018 in Theriogenology
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Neutrophil extracellular traps (NETs) are DNA complexes carrying nuclear and cytoplasmic proteins, such as elastase (ELA), cathepsin-G (CAT) and myeloperoxidase (MPO). Mare endometrosis is a chronic degenerative process characterized by excessive collagen in endometrium. While NETs fight bacteria that cause endometritis, they may trigger endometrial fibrogenesis. The aim was to evaluate the in vitro effect of some NETs components on mare endometrial fibrogenesis and determine its relationship with histopathology or estrous cycle. Endometrial explants were incubated with NETs components (ELA, CAT, MPO or oxytocin). Collagen type I (COL1) protein and type I and III (COL3) gene transcription were evaluated in follicular and mid-luteal phases endometria (Kenney and Doig type I/IIA and IIB/III). Increased COL1 occurred with all NETs proteins, although endometrial response to each NETs protease depended on estrous cycle and/or endometrial category. Since ELA enhanced COL1 production, NETs persistence might be linked to endometrosis. Estrous cycle influenced COL1 protein concentration and COL3 transcripts, suggesting that follicular phase may favor endometrial collagen production. However, luteal phase endometria with moderate or severe lesions may be also susceptible to fibrotic effects of NETs constituents. These data propose that NETs involvement in chronic endometritis in mares may act as putative endometrial fibrogenic mediators.

ACS Style

Maria Rosa Rebordão; Ana Amaral; Karolina Łukasik; Anna Szóstek-Mioduchowska; Pedro Pinto-Bravo; Antonio M. Galvao; Dariusz J. Skarzynski; Graça Ferreira-Dias. Constituents of neutrophil extracellular traps induce in vitro collagen formation in mare endometrium. Theriogenology 2018, 113, 8 -18.

AMA Style

Maria Rosa Rebordão, Ana Amaral, Karolina Łukasik, Anna Szóstek-Mioduchowska, Pedro Pinto-Bravo, Antonio M. Galvao, Dariusz J. Skarzynski, Graça Ferreira-Dias. Constituents of neutrophil extracellular traps induce in vitro collagen formation in mare endometrium. Theriogenology. 2018; 113 ():8-18.

Chicago/Turabian Style

Maria Rosa Rebordão; Ana Amaral; Karolina Łukasik; Anna Szóstek-Mioduchowska; Pedro Pinto-Bravo; Antonio M. Galvao; Dariusz J. Skarzynski; Graça Ferreira-Dias. 2018. "Constituents of neutrophil extracellular traps induce in vitro collagen formation in mare endometrium." Theriogenology 113, no. : 8-18.