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Lactogenic hormones alter cellular and extracellular microRNA expression in bovine mammary epithelial cell culture

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Journal of Animal Science and Biotechnology 2016年第4期7卷 504-513页

作者： Susumu Muroya Tatsuro Hagi Ataru Kimura hisashi aso Masatoshi Matsuzaki Masaru Nomura Animal Products Research Division NARO Institute of Livestock and Grassland Science Faculty of Agriculture and Life Science Hirosaki University International Education and Research Center for Food and Agricultural Immunology(CFAI) Graduate School of Agricultural ScienceTohoku University

Background： Bovine milk contains not only a variety of nutritional ingredients but also microRNAs （miRNAs） that are thought to be secreted by the bovine mammary epithelial cells （BMECs）. The objective of this study was to elucidate the production of milk-related miRNAs in BMECs under the influence of lactogenic hormones. Results： According to a microarray result of milk exosomal miRNAs prior to cellular analyses, a total of 257 miRNAs were detected in a Holstein cow milk. Of these, 18 major miRNAs of interest in the milk were selected for an expression analysis in BMEC culture that was treated with or without dexamethasone, insulin, and prolactin （DIP） to induce a lactogenic differentiation. Quantitative polymerase chain reaction （qPCR） results showed that the expressions of miR-21-Sp （P = 0.005）, miR-26a （P = 0.016）, and miR-320a （P = 0.011） were lower in the DIP-treated cells than in the untreated cells. In contrast, the expression of miR-339a （P-- 0.017） in the cell culture medium were lower in the DiP-treated culture than in the untreated culture. Intriguingly, the miR-148a expression in cell culture medium was elevated by DIP treatment of BMEC culture （P = 0.018）. The medium-to-cell expression ratios of miR- 103 （P = 0.025）, miR-148a （P 〈 0.001）, and miR-223 （P = 0.013） were elevated in the DIP-treated BMECs, suggesting that the lactogenic differentiation-induced secretion of these three miRNAs in BMECs. A bioinformatic analysis showed that the miRNAs down-regulated in the BMECs were associated with the suppression of genes related to transcriptional regulation, protein phosphorylation, and tube development. Conclusion： The results suggest that the miRNAs changed by lactogenic hormones are associated with milk protein synthesis, and mammary gland development and maturation. The elevated miR-148a level in DIP-treated BMECs may be associated with its increase in milk during the lactation period of cows.

关键词： Bovine mammary epithelial cell Lactogenic differentiation microRNA Milk Secretion

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The TLR Expression Pattern on Monocyte-Derived Macrophages for Lipopolysaccharid Stimulation of Calves

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Agricultural Sciences in China 2009年第7期8卷 864-871页

作者： GUO Yi-jie ZHAO Guo-Qi HUO Yong-jiu Sachi Tanaka hisashi aso Takahiro Yamaguchi Laboratory of Animal Nutrition Graduate School of Animal Science and Technology Yangzhou University Yangzhou 225009 P.R. China Laboratory of Functional Morphology Graduate School of Agricultural Science Tohoku University 1-1 Tsutsumidori-Amamiyamachi Aobaku 981-8555 Sendai Japan

In this paper, toll-like receptor expression pattern in monocytes-derived macrophages by lipopolysaccharid （LPS） stimulation was examined. Jugular venous blood samples from 4 Japanese calves were obtained and the peripheral blood mononuclear cells （PBMC） were isolated. The PBMC were cultured for 7 d so as to collect monocytes-derived macrophages in Repcell. The PBMC were stimulated by LPS for 24 h and the mRNA expression pattern of TLR and cytokines in monocytes-derived macrophages （Mod-Mφ） was analyzed. Results showed that LPS stimulation of Mod-Mφ could increase the mRNA levels of the genes of TNF-α, IL-6, and IL-8. In addition, the mRNA levels of the genes of TNF-α and IL-6 in the group of LPS stimulation were most significantly （P 〈 0.01） higher than those in control group and the mRNA levels of TLR1, 3, 5, 8, and 10 were significantly （P 〈 0.05） decreased after LPS stimulation. There was no difference in the mRNA expressions of TLR2, 4, 6, and 7 between the groups of the control and LPS stimulation. Besides, expression of TLR9 was not found. It suggested that monocytes-derived macrophages could respond to LPS and they might take an important role in the innate immunity. The important function of the cells might contribute to better disease treatment.

关键词： TLR bovine macrophage LPS

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TLRS EXPRESSION PATTERN IN MONOCYTES AND MONOCYTE DERIVED MACROPHAGES OF CALVES

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TLRS EXPRESSION PATTERN IN MONOCYTES AND MONOCYTE DERIVED MA...

Evaluation of the Immunoregulatory Capacities of Feed Microbial Materials in Porcine Intestinal Immune and Epithelial Cells

中国畜牧兽医学会动物营养学分会第八届全国会员代表大会暨...

作者： Tanaka Sachi Watanabe Kouichi Ohwada Shyuichi aso hisashi Yamaguchi Takahiro College of of life science ohoku UniversityJapan College of of life science Tohoku UniversityJapan

＜正＞Monocytes and macropHages have numerous important functions on the immune defense system,because they phagocytose foreign antigens,regulate immunocompetent cells and produce several kinds of *** innate immunity that is crucial for more rapid defense mechanisms is initiated by the recognition of pathogens on germ-line encoded pattern recognition *** recognition is mainly attributed to the family of Toll-like re-

关键词： Expression Monocytes Monocyte Calves

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Open Journal of Veterinary Medicine 2014年第3期4卷 15-28页

作者： Naosuke Kumagae Julio Villena Yohsuke Tomosada Hisakazu Kobayashi Paulraj Kanmani hisashi aso Takashi Sasaki Motohiko Yoshida Hiroshi Tanabe Isao Shibata Tadao Saito Haruki Kitazawa Cell Biology Laboratory Graduate School of Agricultural Science Tohoku University Sendai Japan Food and Feed Immunology Group Tohoku University Sendai Japan Food and Feed Immunology Group Tohoku University Sendai Japan Japan Society for the Promotion of Science (JSPS) Tokyo Japan Food and Feed Immunology Group Tohoku University Sendai Japan Laboratory of Immunobiotechnology Reference Centre for Lactobacilli (CERELA-CONICET) Tucuman Argentina Food and Feed Immunology Group Tohoku University Sendai Japan Scientific Feed Laboratory Co. Ltd. Tokyo Japan Scientific Feed Laboratory Co. Ltd. Tokyo Japan

The establishment of drug-free feeding systems has been required for secure and healthy livestock production. Although functional feed materials containing microorganisms as alternatives to enhance intestinal immunity are expected to be beneficial for reducing diarrhoea caused by pathogens in weaned piglets, the effects of such materials on porcine intestinal cells have not been investigated in detail. Therefore, this work evaluated the immunoregulatory functions of microbial feed materials in porcine intestinal immune and epithelial cells. Porcine immune cells isolated from Peyer’s patches and mesenteric lymph nodes were stimulated with six different feed materials containing microorganisms, and evaluated for lymphocyte mitogenicity and cytokine inductions. In addition, porcine intestinal epithelial cells were stimulated with the materials before treatment with heat-killed enterotoxigenic Escherichia coli (ETEC), and analyzed for the proinflammatory cytokine expressions. The material containing Bifidobacterium thermophilum significantly augmented lymphocytes’ mitogenicity and also induced a high expression of IL-2, IL-6 and IFN-γ in immune cells, and inhibited ETEC-induced overexpression of IL-6 and IL-8 via regulation of Toll-like receptor signaling. These results suggest that this feed material stimulates intestinal epithelial and immune cells to exert immunoregulation, suggesting that this feed is expected to contribute to promoting the health of piglets without using antimicrobial feed materials.

关键词： Feed Microbial Material Immunoregulatory Effect Porcine Intestinal Epitheliocytes Porcine Immune Cells

Bifidobacteria Upregulate Expression of Toll-Like Receptor Negative Regulators Counteracting Enterotoxigenic <i>Escherichia coli</i>Mediated Inflammation in Bovine Intestinal Epitheliocytes

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Open Journal of Veterinary Medicine 2013年第2期3卷 143-155页

作者： Kozue Murata Julio Villena Yohsuke Tomosada Risa Hara Eriko Chiba Tomoyuki Shimazu hisashi aso Yoshihito Suda Noriyuki Iwabuchi Jin-Zhong Xiao Tadao Saito Haruki Kitazawa Food and Feed Immunology Group Laboratory of Animal Products Chemistry Graduate School of Agricultural Science Tohoku University Sendai Japan

We previously established a bovine intestinal epithelial cell line (BIE cells) and showed that BIE cells are useful in vitro model system for the study of interactions between pathogenic and beneficial microorganisms and bovine intestinal epithelial cells (IECs). In the present study, we aimed to select potential immunomodulatory bifidobacteria that may be used to beneficially modulate the inflammatory response in bovine IECs. We also aimed to gain insight into the molecular mechanisms involved in the anti-inflammatory effect of bifidobacteria by evaluating the role of Toll-like receptor (TLR)-2 and TLR negative regulators in the regulation of proinflammatory cytokines production and MAPK, NF-κB and PI3K pathways activation in BIE cells. Five bifidobacteria strains were evaluated in this study and according to their capacity to modulate the inflammatory response of BIE cells. Despite the unique effect of each strain, four common points were found when comparing the effect of the high and moderate anti-inflammatory strains: 1) Upregulation of TLR negative regulators and the intensity of that upregulation was related to the different immunomodulatory capacity of each bifidobacteria strain;2) The balance between MAPK activation and MKP-1 upregulation affected the anti-inflammatory effect of bifidobacteria in BIE cells;3) The inhibition of PI3K pathway was related to the anti-inflammatory effect of bifidobacteria;4) The immunoregulatory effect of bifidobacteria in BIE cells is partially dependent on TLR2. This study shows that BIE cells can be used for the selection of immunoregulatory bifidobacteria and for studying the mechanisms involved in the protective activity of immunobiotics against TLR4-induced inflammatory damage. In addition, we have demonstrated that the anti-inflammatory effect of bifidobacteria was achieved by a complex interaction of multiple TLRs negative regulators as well as the inhibition/activation of multiple signaling pathways.

关键词： Bovine Intestinal Epithelial Cells Bifidobacteria ETEC TLRs Negative Regulators

Engendered nanoparticles for treatment of brain tumors

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Oncology Research 2025年第1期33卷 15-26页

作者： SOROUSH SOLEYMANI MOHAMMAD DOROUDIAN MAHDIEH SOEZI ALI BELADI KIARASH ASGARI aso MOBARAKSHAHI ARYANA AGHAEIPOUR RONAN MACLOUGHLIN Department of Cell and Molecular Biology School of Biological SciencesUniversity of LeicesterLeicesterLE17RHUK Department of Cell and Molecular Biology Faculty of Biological SciencesKharazmi UniversityTehran15719-14911Iran Cancer Epidemiology Research Center Aja University of Medical SciencesTehran14117-18541Iran Medical Biotechnology Research Center Aja University of Medical SciencesTehran14117-18541Iran Department of Life Sciences Faculty of Natural SciencesImperial College LondonLondonSW72AZUK School of Pharmacy and Biomolecular Sciences Royal College of Surgeons in Ireland(RCSI)DublinD02 YN77Ireland Research and Development Science and Emerging TechnologiesAerogen Ltd.Galway Business ParkGalwayH91 HE94Ireland School of Pharmacy and Pharmaceutical Sciences Trinity CollegeDublinD02 PN40Ireland

Brain metastasis and primary glioblastoma multiforme represent the most common and lethal malignant brain *** median survival time is typically less than a year after *** of the major challenges in treating these cancers is the efficiency of the transport of drugs to the central nervous *** blood-brain barrier is cooperating with advanced stages of *** blood-brain barrier poses a significant challenge to delivering systemic medications to brain *** delivery systems have emerged as promising tools for effectively crossing this ***,the development of smart nanoparticles brings new hope for cancer diagnosis and *** nanoparticles improve drug delivery efficiency,allowing for the creation of targeted and stimuli-responsive delivery *** review highlights recent advancements in nanoparticle and smart nanoparticle technologies for brain cancer treatment,exploring the range of nanoparticles under development,their applications,targeting strategies,and the latest progress in enhancing transport across the blood-brain *** also addresses the ongoing challenges and potential benefits of these innovative approaches.

关键词： Nanoparticles Smart nanoparticles Stimuli responsive nanoparticles Targeted nanoparticles Blood-brain barrier(BBB) Brain cancer

Prion Protein Binds to Aldolase A Produced by Bovine Intestinal M Cells

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Open Journal of Veterinary Medicine 2015年第3期5卷 43-60页

作者： Yuya Nagasawa Yu Takahashi Wataru Itani Hitoshi Watanabe Yusuke Hidaka Shotaro Morita Kei Suzuki Kouichi Watanabe Shyuichi Ohwada Haruki Kitazawa Morikazu Imamura Takashi Yokoyama Motohiro Horiuchi Suehiro Sakaguchi Shirou Mohri Michael T. Rose Tomonori Nochi hisashi aso Cellular Biology Laboratory Tohoku University Sendai Japan Food Immunology Group Tohoku University Sendai Japan Prion Disease Research Center National Institute of Animal Health Tsukuba Japan Laboratory of Veterinary Hygiene Hokkaido University Sapporo Japan Institute for Enzyme Research The University of Tokushima Tokushima Japan Institute of Biological Environmental and Rural Sciences Aberystwyth University Aberystwyth UK

Microfold (M) cells are a kind of intestinal epithelial cell in the follicle-associated epithelium (FAE) of Peyer’s patches. They can transport antigens and microorganisms to lymphoid tissues. Bovine spongiform encephalopathy (BSE) is a fatal neurodegenerative disorder in cattle. It is linked to variant Creutzfeldt-Jakob disease in humans. Although it is thought that M cells transport the BSE agent, the exact mechanism by which it crosses the intestinal barrier is not clear. We have bovine intestinal epithelial cell line (BIE cells), which can differentiate into the M cell type in vitro after stimulation, and which is able to transport the BSE agent. We show here that M cells are able to incorporate large numbers of PrP coated magnetic particles into intracellular vesicles, which we collected. The results of 2-DE show a specific protein associated with the PrP-coated particles, compared with non-coated particles. This protein was identified as aldolase A, a glycolytic pathway enzyme, using LC-MS/MS analysis. Aldolase A was synthesized and secreted by BIE cells, and increased during M cell differentiation. In the villi of the bovine intestine, aldolase A was detected on the surface of the epithelium and in the mucus droplet of goblet cells. In the FAE of bovine jejunal and ileal Peyer’s patches, aldolase A was localized on the surface and the apical part of the M cells. The binding of rbPrP to aldolase A was clearly detected and inhibited by pre-treatment of anti-aldolase A antibody. Aldolase A was co-stained with incorporated PrPSc in M-BIE cells. These results suggest that bovine M cells and goblet cells synthesize aldolase A, and that aldolase A may have the ability to bind PrP and associate with PrP in cellular vesicles. Therefore, aldolase A-positive M cells may play a key role in the invasion of BSE into the body.

关键词： Peyer’s Patch M Cell BIE Cells Aldolase A PrP Binding Protein

Phylogenetic relationships among Hystrix species and related species based on expressed sequence tag-polymerase chain reaction

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Journal of Systematics and Evolution 2011年第1期49卷 65-71页

作者： hisashi TSUJIMOTO Laboratory of Plant Genetics and Breeding Science Faculty of Agriculture Tottori University

Twelve species, including three Hystrix species, five Leymus species, Hordeum bogdanii, Pseudoroegneria spicata, Psathyrostachys huashanica, and Roegneria ciliaris, were used for expressed sequence tag-polymerase chain reaction (EST-PCR) assay. A total of 125 products were amplified by 72 sets of EST-PCR markers developed in barley, among which 106 (84.8%) products were found to be polymorphic. Each EST-PCR marker produced 0–6 polymorphic bands, with an average of 1.47. The relationship between H. duthiei ssp. duthiei and H. duthiei ssp. longearistata is close, but they are remote to H. patula. Hystrix duthiei ssp. duthiei and H. duthiei ssp. longearistata were clustered with Leymus species and Psathyrostachys huashanica, which suggested that they have close genetic relationships. The results of EST-PCR analysis are basically comparable with those obtained from studies on cytology, which indicated that EST-PCR can be used to assess the genetic relationships among the perennial species in Triticeae.

关键词： cluster analysis expressed sequence tag Hystrix molecular marker phylogenetic relationship.

Physical mapping of repetitive sequences and genome analysis in six Elymus species by in situ hybridization

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Journal of Systematics and Evolution 2011年第4期49卷 347-352页

作者： hisashi TSUJIMOTO Faculty of Agriculture Tottori University

Genome constitution and genetic relationships between six Elymus species were assessed by physical mapping of different repetitive sequences using a technique of sequential fluorescence in situ hybridization and genomic in situ *** six Elymus species are all naturally growing species in northwest China,namely,***,***,***,***,***,and *** StStHH genome consti- tution was revealed for *** and StStHHYY for the remainder *** chromosome could be clearly characterized by physical mapping with 18S-26S rDNA,5S rDNA,Afa-family,and AAG repeats,and be allocated to a certain genome by genomic in situ *** 5S rDNA sites,each in the H and St genomes,and three 18S-26S rDNA sites,two in the St genome and one in the Y genome,were uncovered in most of the *** strong Afa-family hybridization signals discriminated the H genome from the St and Y *** H and Y genome carried more AAG repeats than St.A common non-Robertsonian reciprocal translocation between the H and Y genomes was revealed in ***,***,*** and *** of molecular karyotypes strongly suggests that they can be classified into three groups,namely,***,***,and others.

关键词： Elymus fluorescence in situ hybridization repetitive sequences.