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Extracellular vesicles-coupled miRNAs from oviduct and uterus modulate signaling pathways related to lipid metabolism and bovine early embryo development

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Journal of Animal Science and Biotechnology 2024年第4期15卷 1528-1549页

作者： Rosane mazzarella Karina Canon‑Beltran Yulia N.Cajas meriem Hamdi encina m.gonzalez Juliano C.da Silveira Claudia L.V.Leal D.Rizos Department of Animal Reproduction INIA-CSICMadridSpain Department of Biochemistry and molecular Biology Veterinary FacultyComplutense University of Madrid(UCM)MadridSpain Department Agrarian Production Technical University of MadridUPMMadridSpain Departamento de Ciencias Biologicas Universidad Tecnica Particular de LojaUTPLLojaEcuador Department of Anatomy and Embryology FV-UCMMadridSpain Department of Veterinary medicine FZEA-USPPirassunungaBrazil

Background Extracellular vesicles(EVs) present in oviductal(OF) and uterine fluid(UF) have been shown to enhance bovine embryo quality during in vitro culture by reducing lipid contents and modulating lipid metabolism-related genes(LmGs), while also influencing cell proliferation, suggesting their involvement on the regulation of different biological pathways. The regulation of signaling pathways related to cell differentiation, proliferation, and metabo-lism is crucial for early embryo development and can determine the success or failure of the pregnancy. Bioactive molecules within EVs in maternal reproductive fluids, such as micro RNAs(miRNAs), may contribute to this regulatory process as they modulate gene expression through post-transcriptional *** This study evaluated miRNA cargo in OF-EVs from the early luteal phase and UF-EVs from the mid-luteal phase, coinciding with embryo transit within oviduct and uterus in vivo, and its possible influence on LmGs and sign-aling pathways crucial for early embryo development. A total of 333 miRNAs were detected, with 11 exclusive to OF, 59 to UF, and 263 were common between both groups. From the 20 differentially expressed miRNAs, 19 up-regulated in UF-EVs(bta-miR-134, bta-miR-151-3p, bta-miR-155, bta-miR-188, bta-miR-181b, bta-miR-181d, bta-miR-224, bta-miR-23b-3p, bta-miR-24-3p, bta-miR-27a-3p, bta-miR-29a, bta-miR-324, bta-miR-326, bta-miR-345-3p, bta-miR-410, bta-miR-652, bta-miR-677, bta-miR-873 and bta-miR-708) and one(bta-miR-148b) in OF-EVs. These miRNAs were predicted to modulate several pathways such as Wnt, Hippo, mAPK, and lipid metabolism and degradation. Differ-ences in miRNAs found in OF-EVs from the early luteal phase and UF-EVs from mid-luteal phase may reflect differ-ent environments to meet the changing needs of the embryo. Additionally, miRNAs may be involved, particularly in the uterus, in the regulation of embryo lipid metabolism, immune system, and *** Our study su

关键词： Embryo-maternal interaction Epigenetic Exosomes mammalian Preimplantation Reproductive fluids

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Extracellular vesicles from oviductal and uterine fluids supplementation in sequential in vitro culture improves bovine embryo quality

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Journal of Animal Science and Biotechnology 2023年第1期14卷 114-133页

作者： Cláudia Lima Verde Leal Karina Canón-Beltrán Yulia N.Cajas meriem Hamdi Aracelli Yaryes maría Gemma millán de la Blanca Paula Beltrán-Brena Rosane mazzarella Juliano Coelho da Silveira Alfonso Gutiérrez-Adán encina m gonzález Dimitrios Rizos Department of Animal Reproduction National Center Institute for Agriculture and Food Research and Technology(CSIC-INIA)28040 MadridSpain. Departamento de medicina Veterinaria Faculdade de Zootecnia e Engenharia de AlimentosUniversidade de Sao Paulo(FZEA-USP)PirassunungaBrazil. Facultad de Ciencias Agrarias y Ambientales Programa de Medicina VeterinariaFundacion Universitaria Juan de CastellanosTunjaColombia. Laboratorio de Biotecnologia de la Reproduccion Animal Facultad de Ciencias AgropecuariasUniversidad de Cuenca(UC)EC010205 CuencaEcuador. Department of Anatomy and Embryology Veterinary Faculty-Universidad Complutense de Madrid(UCM)MadridSpain.

Background:In vitro production of bovine embryos is a well-established technology,but the in vitro culture(IVC)system still warrants improvements,especially regarding embryo *** study aimed to evaluate the effect of extracellular vesicles(EVs)isolated from oviductal(OF)and uterine fluid(UF)in sequential IVC on the development and quality of bovine *** were cultured in SOF supplemented with either BSA or EVs-depleted fetal calf serum(dFCS)in the presence(BSA-EV and dFCS-EV)or absence of EVs from OF(D1 to D4)and UF(D5 to D8),mimicking in vivo *** from oviducts(early luteal phase)and uterine horns(mid-luteal phase)from slaughtered heifers were isolated by size exclusion *** rate was recorded on days 7-8 and their quality was assessed based on lipid contents,mitochondrial activity and total cell numbers,as well as survival rate after *** mRNA abundance for lipid metabolism-related transcripts and levels of phosphorylated hormonesensitive lipase(pHSL)proteins were also ***,the expression levels of 383 miRNA in OF-and UF-EVs were assessed by ***:Blastocyst yield was lower(Pments compared with dFCS *** rates after vitrification/warming were improved in dFCS-EVs(Pmber in dFCS-EV and BSA-EV compared with respective controls(dFCS and BSA),while lipid content was decreased in dFCSEV(Pmitochondrial activity did not change(P>0.05).Lipid metabolism transcripts were affected by EVs and showed interaction with type of protein source in medium(PPARGC1B,LDLR,CD36,FASN and PNPLA2,PmiRNA were differentially expressed between OF-and UF-EVs and only bta-miR-148b was increased in OF-EVs(Pmimicking physiological conditions using EVs from OF and UF in sequential IVC does not affect embryo development but improves blast

关键词： Cattle Cryopreservation Embryo development Exosomes Lipid metabolism miRNAs Oviduct Uterus

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Extracellular vesicles-coupled miRNAs from oviduct and uterus modulate signaling pathways related to lipid metabolism and bovine early embryo development.

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畜牧与生物技术杂志(英文版) 2024年第4期 1528页

作者： Rosane mazzarella KarinaCañón-Beltrán Yulia N.Cajas meriem Hamdi encina m.gonzález Juliano C.da Silveira Claudia L.V.Leal D.Rizos

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Extracellular vesicles from oviductal and uterine fluids supplementation in sequential in vitro culture improves bovine embryo quality

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畜牧与生物技术杂志(英文版) 2023年第1期

作者： Cláudia Lima Verde Leal Karina Ca?ón-Beltrán Yulia N.Cajas meriem Hamdi Aracelli Yaryes maría Gemma millán de la Blanca Paula Beltrán-Bre?a Rosane mazzarella Juliano Coelho da Silveira Alfonso Gutiérrez-Adán encina m gonzález Dimitrios Rizos

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Plasticity of Xyloglucan Composition in Bean （Phaseolus vulgaris）-Cultured Cells during Habituation and Dehabituation to Lethal Concentrations of Dichlobenil

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molecular Plant 2010年第3期3卷 603-609页

作者： A. Alonso-Simon L. Neumetzler R Garcia-Angulo A.E. encina J.L. Acebes J.m. Alvarez T. Hayashi Area de Fisiologia Vegetal Universidad de Leon E-24071 Leon Spain max Planck Institute of molecular Plant Physiology 14476 Golm Germany Kyoto University RISH Gokasho Uji Kyoto 611-0011 Japan

Bean cells that have been habituated to grow in a lethal concentration （12 μm） of 2,6-dichlorobenzonitrile （dichlobenil or DCB, a cellulose biosynthesis inhibitor） are known to have decreased cellulose content in their cell walls. Xyloglucan, which is bound to cellulose and together with it forms the main loading network of plant cell walls, has also been described to decrease in habituated cells, but whether the change on cellulose affects the xyloglucan structure besides its abundance has not been analyzed. Fragmentation analysis with xyloglucan-specific endoglucanase （XEG） and endocellulase revealed that habituation to DCB caused a change in the fine structure of xyloglucan, namely a decrease in fucosyl residues attached to the galactosyl-xylosyl residues along the glucan backbone. After the removal of herbicide from the medium （dehabituated cells）, xyloglucan recovered its fucosyl residues. In addition, some cello-oligosaccharides could be detected only in habituated cells＇ xyloglucan digested by XEG and endocellulase, corresponding to a glucan co- valently bound or co-precipitated with the hemicelluloses. These results show that structural flexibility of cell walls relies in part on the plasticity of xyloglucan composition and opens up new perspectives to further research in this field.

关键词： Bean 2,6-dichlorobenzonitrile habituation and dehabituation OLImP (Oligosaccharides mass Profiling) xyloglucan.

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Phenolic metabolism and molecular mass distribution of polysaccharides in cellulose-deficient maize cells

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Journal of Integrative Plant Biology 2017年第7期59卷 475-495页

作者： maría de Castro Romina martínez-Rubio José L.Acebes Antonio encina Stephen C.Fry Penélope García-Angulo área de Fisiología Vegetal.Dpto.Ingenieríay Ciencias Agrarias.Facultad de Biologíay Ciencias Ambientales.Universidad de León The Edinburgh Cell Wall Group Institute of Molecular Plant Sciences School of Biological Sciences Daniel Rutherford BuildingThe King's Buildings

As a consequence of the habituation to low levels of dichlobenil （DCB）, cultured maize cells presented an altered hemicellulose cell fate with a lower proportion of strongly wall-bound hemicelluloses and an increase in soluble extracellular polymers released into the culture medium. The aim of this study was to investigate the relative molecular mass distributions of polysaccharides as well as phenolic metabolism in cells habituated to low levels of DCB （1.5 μm）. Generally, cell wall bound hemicelluloses and sloughed polymers from habituated cells were more homogeneously sized and had a lower weight-average relative molecular mass. In addition, polysaccharides underwent massive cross-linking after being secreted into the cell wall, but this cross-linking was less pronounced in habituated cells than in non-habituated ones. However, when relativized, ferulic acid and p-coumaric acid contents were higher in this habituated cell line. Feasibly, cells habituated to low levels of DCB synthesized molecules with a lower weight-average relative molecular mass, although cross-linked, as a part of their strategy to compensate for the lack of cellulose.

关键词： Phenolic metabolism and molecular mass distribution of polysaccharides in cellulose-deficient maize cells were for

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The biosynthesis and wall-binding of hemicelluloses in cellulose-deficient maize cells:An example of metabolic plasticity

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Journal of Integrative Plant Biology 2015年第4期57卷 373-387页

作者： maría de Castro Janice G.miller José Luis Acebes Antonio encina Penélope García-Angulo Stephen C.Fry The Edinburgh Cell Wall Group Institute of Molecular Plant SciencesSchool of Biological SciencesDaniel Rutherford BuildingThe King's Buildings Laboratory of Plant Physiology Faculty of Biological and Environmental SciencesUniversity of Léon

Cell-suspension cultures（Zea mays L.,Black mexican sweet corn） habituated to 2,6-dichlorobenzonitrile（DCB） survive with reduced cellulose owing to hemicellulose network modification. We aimed to de fine the hemicellulose metabolism modifications in DCB-habituated maize cells showing a mild reduction in cellulose at different stages in the culture cycle. Using pulse-chase radiolabeling, we fed habituated and non-habituated cultures with [3H]arabinose,and traced the distribution of 3H-pentose residues between xylans, xyloglucans and other polymers in several cellular compartments for 5 h. Habituated cells were slower taking up exogenous [3H]arabinose. Tritium was incorporated into polysaccharide-bound arabinose and xylose residues, but habituated cells diverted a higher proportion of their new [3H]xylose residues into（hetero） xylans at the expense of xyloglucan synthesis. During logarithmic growth, habituated cells showed slower vesicular traf ficking of polymers,especially xylans. moreover, habituated cells showed a decrease in the strong wall-binding of all pentose-containing polysaccharides studied; correspondingly, especially in log phase cultures, habituation increased the proportion of 3H-hemicelluloses（[3H]xylans and [3H]xyloglucan） sloughed into the medium. These findings could be related to the cel walls’ cellulose-deficiency, and consequent reduction in binding sites for hemicelluloses; the data could also re fl ect the habituated cells’ reduced capacity to integrate arabinox ylans by extra-protoplasmic phenolic cross-linking, as well as xyloglucans, during wall assembly.

关键词： Arabinoxylan cell wall cross-linking 2,6-dichlorobenzonitrile Hemicellulose biosynthesis xyloglucan

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Ectopic lignification in primary cellulose-deficient cell walls of maize cell suspension cultures

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Journal of Integrative Plant Biology 2015年第4期57卷 357-372页

作者： Hugo mélida Asier Largo-Gosens Esther Novo-Uzal Rogelio Santiago Federico Pomar Pedro García Penélope García-Angulo José Luis Acebes Jesús álvarez Antonio encina Plant Physiology Laboratory Faculty of Biological and Environmental SciencesUniversity of León Centre for Plant Biotechnology and Genomics (CBGP) Politechnical University of Madrid Department of Plant Biology University of Murcia Plant Biology and Soil Sciences Department Faculty of BiologyUniversity of Vigo Environmental Agrobiology Soil and Plant Quality (University of Vigo)Associated Unit to Biological Mission of Galicia (CSIC) Department of Animal Biology Plant Biology and EcologyUniversity of A Coru?a Department of molecular Biology(área de Genética) Faculty of Biological and Environmental SciencesUniversity of León

maize （Zea mays L.） suspension-cultured cells with up to 7o% less cellulose were obtained by stepwise habituation to dichlobenil （DCB）, a cellulose biosynthesis inhibitor. Cellulose deficiency was accompanied by marked changes in cell wall matrix polysaccharides and phenolics as revealed by Fourier transform infrared （FTIR） spectroscopy. Cell wall compositional analysis indicated that the cellulosedeficient cell walls showed an enhancement of highly branched and cross-linked arabinoxylans, as well as an increased content in ferulic acid, diferulates and p-coumaric acid, and the presence of a polymer that stained positive for phloroglucinol. In accordance with this, cellulose-deficient cell walls showed a fivefold increase in Klason-type lignin. Thioacidolysis/GC-mS analysis of cellulose-deficient cell walls indicated the presence of a lignin-like polymer with a Syringyl/Guaiacyl ratio of 1.45, which differed from the sensu stricto stress-related lignin that arose in response to shortterm DCB-treatments. Gene expression analysis of these ceils indicated an overexpression of genes specific for the biosynthesis of monolignol units of lignin. A study of stress signaling pathways revealed an overexpression of some of the iasmonate signaling pathway genes, which might trigger ectopic lignification in response to cell wall integrity disruptions. In summary, the structural plasticity of primary cell walls is proven, since a lignification process is possible in response to cellulose impoverishment.

关键词： Cellulose DCB dichlobenil ectopic lignin maize

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Bispecific sigma-1 receptor antagonism and mu-opioid receptor partial agonism:WLB-73502,an analgesic with improved efficacy and safety profile compared to strong opioids

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Acta Pharmaceutica Sinica B 2023年第1期13卷 82-99页

作者： Alba Vidal-Torres Begoña Fernández-Pastor mónica García Eva Ayet Anna Cabot Javier Burgueño Xavier monroy Bertrand Aubel Xavier Codony Luz Romero Rosalía Pascual maria Teresa Serafini Gregorio encina Carmen Almansa Daniel Zamanillo manuel merlos Josémiguel Vela WeLab Barcelona Parc Cientific de BarcelonaBarcelona 08028Spain

Opioids are the most effective painkillers,but their benefit-risk balance often hinder their therapeutic ***-73502 is a dual,bispecific compound that binds sigma-1(S1R)and mu-opioid(mOR)***-73502 is an antagonist at the *** behaved as a partial mOR agonist at the G-protein pathway and produced no/unsignificantβ-arrestin-2 recruitment,thus demonstrating low intrinsic efficacy on mOR at both signalling *** its partial mOR agonism,WLB-73502exerted full antinociceptive efficacy,with potency superior to morphine and similar to oxycodone against nociceptive,inflammatory and osteoarthritis pain,and superior to both morphine and oxycodone against neuropathic ***-73502 crosses the blood-brain barrier and binds brain S1R and mOR to an extent consistent with its antinociceptive *** to morphine and oxycodone,tolerance to its antinociceptive effect did not develop after repeated 4-week ***,contrary to opioid comparators,WLB-73502 did not inhibit gastrointestinal transit or respiratory function in rats at doses inducing full efficacy,and it was devoid of proemetic effect(retching and vomiting)in ferrets at potentially effective ***-73502 benefits from its bivalent S1R antagonist and partial mOR agonist nature to provide an improved antinociceptive and safety profile respect to strong opioid therapy.

关键词： WLB-73502 Sigma-1 receptor mu-opioid receptor Biased agonist Analgesic activity Chronic pain Neuropathic pain Safety

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The role of hypoxia-induced factor 1α in breast cancer

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Journal of Cancer metastasis and Treatment 2019年第6期4卷 44-54页

作者： Carlos Henrique F.Peiró Jéssica A.encina matheus m.Perez Glauco S.A.Aquino Glaucia L.Veiga Fernando Fonseca Beatriz C.A.Alves Laboratório de Análises Clínicas da Faculdade e medicina do ABC Santo AndréSP CEP 09060-650Brazil Instituto de Ciências Farmacêuticas Universidade Federal de São PauloDiademaSP CEP 09972-270Brazil

Breast cancer usually grows very quickly,becoming insensitive to blood flow in nearby veins;because of that,inside solid tumors it's possible to find a hypoxic environment,in other words,an environment where oxygen is less *** feature of cancer is its angiogenesis rate,because of the high energy demand,new blood vessels must be produced to take nutrients inside the solid tumor *** with normal blood flow bringing the cancer oxygen and nutrients,its cells favor hypoxia,in an event known as Warburg *** to the Warburg Effect,cells,even with normal oxygen rates,prefer to use fermentation instead of the citric acid cycle to produce *** the cancer to operate normally in hypoxia,a transcription factor family is activated,known as hypoxia-induced factors(HIF),composed of a HIF-1βand a HIF-1α*** HIF-1αis expressed during hypoxia,it is a great target for treatments and a breast cancer *** of the role of HIF-1αin cancer and the high incidence of breast cancer worlwide,this review was performed in order to bring the most recent results concerning the role HIF-1αcan exert in breast cancer development and progression.

关键词： Breast neoplasms hypoxia-induced factor 1 HIF-1α

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