• Title/Summary/Keyword: Animal disease model

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The Immunosuppressive Potential of Cholesterol Sulfate Through T Cell Microvilli Disruption

  • Jeong-Su Park;Ik-Joo Chung;Hye-Ran Kim;Chang-Duk Jun
    • IMMUNE NETWORK
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    • v.23 no.3
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    • pp.29.1-29.23
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    • 2023
  • Cholesterol (CL) is required for various biomolecular production processes, including those of cell membrane components. Therefore, to meet these needs, CL is converted into various derivatives. Among these derivatives is cholesterol sulfate (CS), a naturally produced CL derivative by the sulfotransferase family 2B1 (SULT2B1), which is widely present in human plasma. CS is involved in cell membrane stabilization, blood clotting, keratinocyte differentiation, and TCR nanocluster deformation. This study shows that treatment of T cells with CS resulted in the decreased surface expression of some surface T-cell proteins and reduced IL-2 release. Furthermore, T cells treated with CS significantly reduced lipid raft contents and membrane CLs. Surprisingly, using the electron microscope, we also observed that CS led to the disruption of T-cell microvilli, releasing small microvilli particles containing TCRs and other microvillar proteins. However, in vivo, T cells with CS showed aberrant migration to high endothelial venules and limited infiltrating splenic T-cell zones compared with the untreated T cells. Additionally, we observed significant alleviation of atopic dermatitis in mice injected with CS in the animal model. Based on these results, we conclude that CS is an immunosuppressive natural lipid that impairs TCR signaling by disrupting microvillar function in T cells, suggesting its usefulness as a therapeutic agent for alleviating T-cell-mediated hypersensitivity and a potential target for treating autoimmune diseases.

Pharmacokinetics of oxolinic acid in cultured olive flounder Paralichthys olivaceus by oral administration, injection and dipping (Oxolinic acid의 경구투여, 주사 및 약욕에 따른 넙치, Paralichthys olivaceus 체내 약물동태학적 특성)

  • Jung, Sung-Hee;Choi, Dong-Lim;Kim, Jin-Woo;Jo, Mi-Ra;Jee, Bo-Young;Seo, Jung-Soo
    • Journal of fish pathology
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    • v.22 no.2
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    • pp.125-135
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    • 2009
  • The pharmacokinetic properties of oxolinic acid (OA) were studied after oral administration, intraperitoneal injection and dipping to cultured olive flounder, Paralichthys olivaceus (average 90 g, $23{\pm}1{^{\circ}C}$). Plasma samples were taken at 3, 5, 10, 15, 24, 30, 48, 96 and 144 h post-dose. In oral dosage at 15, 30 and 60 ㎎/㎏, the peak plasma concentrations of OA, which attained at 10~15 h post-dose, were 1.92, 2.45 and 3.72 $\mu{g}/m\ell$, respectively. In intraperitoneal injection with 10 and 20 ㎎/㎏, the peak plasma concentrations of OA, which attained at 10 h post-dose, were 4.1 and 4.8 $\mu{g}/m\ell$, respectively. In dipping in 30 and 50 ppm for 1 h, peak concentrations were observed at 5 h and 30 h post-dose, were 0.22 and 0.38 $\mu{g}/m\ell$, respectively. The kinetic profile of absorption, distribution and elimination of OA in plasma were analyzed fitting to a one-compartment model by WinNonlin program. Calculated parameters for a single oral dosage of 15, 30 and 60 ㎎/㎏, respectively, were: AUC (the area under the concentration-time curve)=70.93, 120.0 and 141.86 $\mu{g}$ $h/m\ell$ $T_{max}$ (time for maximum concentration)=16.22, 20.39 and 17.33 h; $C_{max}$ (maximum concentration)=���D1.61, 2.40 and 3.01 $\mu{g}/m\ell$. Following intraperitoneal injection of 10 and 20 ㎎/㎏, these parameters were AUC=184.7 and 315.92 $\mu{g}$ $h/m\ell$ $T_{max}$=5.91 and 6.26 h; $C_{max}$=4.19 and 4.45 $\mu{g}/m\ell$. Following dipping at 30 and 50 ppm, these parameters were AUC=17.58 and 21.69 $\mu{g}$ $h/m\ell$ $T_{max}$=19.08 and 31.43 h; $C_{max}$x=0.22 and 0.25 $\mu{g}/m\ell$.

Hypotriglyceridemic effects of brown seaweed consumption via regulation of bile acid excretion and hepatic lipogenesis in high fat diet-induced obese mice

  • Han, A-Reum;Kim, Jae-Hoon;Kim, Eunyoung;Cui, Jiamei;Chai, In-Suk;Zhang, Guiguo;Lee, Yunkyoung
    • Nutrition Research and Practice
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    • v.14 no.6
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    • pp.580-592
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    • 2020
  • BACKGROUND/OBJECTIVES: The present study aimed to further investigate the potential health beneficial effects of long-term seaweed supplementation on lipid metabolism and hepatic functions in DIO mice. MATERIALS/METHODS: Four brown seaweeds (Undaria pinnatifida [UP], Laminaria japonica [LJ], Sargassum fulvellum [SF], or Hizikia fusiforme [HF]) were added to a high fat diet (HFD) at a 5% ratio and supplemented to C57BL/6N mice for 16 weeks. Triglycerides (TGs) and total cholesterol (TC) in the liver, feces, and plasma were measured. Fecal bile acid (BA) levels in feces were monitored. Hepatic insulin signaling- and lipogenesis-related proteins were evaluated by Western blot analysis. RESULTS: Fasting blood glucose levels were significantly reduced in the LJ, SF, and HF groups compared to the HFD group by the end of 16-week feeding period. Plasma TG levels and hepatic lipid accumulation were significantly reduced in all 4 seaweed supplemented groups, whereas plasma TC levels were only suppressed in the UP and HF groups compared to the HFD group. Fecal BA levels were significantly elevated by UP, LJ, and SF supplementation compared to HFD feeding only. Lastly, regarding hepatic insulin signaling-related proteins, phosphorylation of 5'-AMP-activated protein kinase was significantly up-regulated by all 4 types of seaweed, whereas phosphorylation of protein kinase B was up-regulated only in the SF and HF groups. Lipogenesis-related proteins in the liver were effectively down-regulated by HF supplementation in DIO mice. CONCLUSIONS: Brown seaweed consumption showed hypotriglyceridemic effects in the prolonged DIO mouse model. Specifically, combinatory regulation of BA excretion and lipogenesis-related proteins in the liver by seaweed supplementation contributed to the reduction of plasma and hepatic TG levels, which inhibited hyperglycemia in DIO mice. Thus, the discrepant and species-specific functions of brown seaweeds provide novel insights for the selection of future targets for therapeutic agents.

Tumorigenic Effects of Endocrine-Disrupting Chemicals are Alleviated by Licorice (Glycyrrhiza glabra) Root Extract through Suppression of AhR Expression in Mammalian Cells

  • Chu, Xiao Ting;Cruz, Joseph Dela;Hwang, Seong Gu;Hong, Heeok
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.13
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    • pp.5117-5121
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    • 2014
  • Endocrine-disrupting chemicals (EDCs) have been reported to interfere with estrogen signaling. Exposure to these chemicals decreases the immune response and causes a wide range of diseases in animals and humans. Recently, many studies showed that licorice (Glycyrrhiza glabra) root extract (LRE) commonly called "gamcho" in Korea exhibits antioxidative, chemoprotective, and detoxifying properties. This study aimed to investigate the mechanism of action of LRE and to determine if and how LRE can alleviate the toxicity of EDCs. LRE was prepared by vacuum evaporation and freeze-drying after homogenization of licorice root powder that was soaked in 80% ethanol for 72 h. We used 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) as an EDC, which is known to induce tumors or cancers; MCF-7 breast cancer cells were used as a tumorigenic model. These were treated with TCDD and various concentrations of LRE (0, 50, 100, 200, $400{\mu}g/mL$) for 24, 48, and 72 h. As a result, TCDD stimulated MCF-7 cell proliferation, but LRE significantly inhibited TCDD-induced MCF-7 cell proliferation in a dose- and time-dependent manner. Expression of TCDD toxicity-related genes, i.e., aryl hydrocarbon receptor (AhR), AhR nuclear translocator, and cytochrome P450 1A1, were subsequently down-regulated by LRE in a dose-dependent manner. Analysis of cell cycle distribution after treatment of MCF-7 cells with TCDD and various concentrations of LRE showed that LRE inhibited the proliferation of MCF-7 cells via G2/M phase arrest. Reverse transcription-polymerase chain reaction and Western blot analyses also revealed that LRE dose-dependently increased the expression of the tumor suppressor genes p53 and p27 and down-regulated the expression of cell cycle-related genes. These data suggest that LRE can mitigate the tumorigenic effects of TCDD in breast cancer cells by suppression of AhR expression and cell cycle arrest. Thus, LRE can be used as a potential toxicity-alleviating agent against EDC-mediated disease.

Upregulation of aquaporin 2 and aquaporin 4 in the water-deprived mongolian gerbil (Meriones unguiculatus) kidney (절수시 Mongolian gerbil (Meriones unguiculatus) 콩팥의 Aquaporin 2, Aquaporin 4 발현변화)

  • Song, Ji-Hyun;Kwon, Jin-Seuk;Kim, Yong-Hwan;Park, Yong-Deok;Han, Ki-Hwan;Ryu, Si-Yun;Jung, Ju-Young
    • Korean Journal of Veterinary Research
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    • v.47 no.4
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    • pp.363-370
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    • 2007
  • Mongolian gerbil (Meriones unguiculatus) has been as an model animal for studing the neurological disease such as stroke and epilepsy because of the congenital incompleteries in Willis circle, as well as the investigation of water metabolism because of the long time-survival in the condition of water-deprived desert condition, compared with other species animal. Aquaporin 2 (AQP2) expressed at the surface of principal cells in collecting duct results from an equilibrium between the AQP2 in intracellular vesicles and the AQP2 on the plasma membrane. Aquaporin 4 (AQP4), which is expressed in cell in a wide range of organ, is also present in the collecting duct principal cells where this is abundant in the basolateral plasma membranes and represent potential exit pathways from the cell for water entering via AQP2. In this research, we divide 3 groups of which each group include the 5 animals. In the study of 7 or 14 days water restricted condition, we investigated the AQP2 and AQP4 by using a quantitative immunohistochemistry in the kidney. The results obtained in this study were summarized as followings. AQP2 is abundant in the apical plasma membrane and apical vesicles in the collecting duct principal cell and at rare abundance in connecting tubules. In the water-deprived Mongolian gerbil kidney, expression of AQP2 was continuosly increased in the cortical collecting duct and inner medullary collecting duct. This increase was both the apical region and cytoplasm. AQP4 is mainly expressed in the inner medulla, although some expression is also noted in the more proximal segment. In the water-deprived Mongolian gerbil kidney, AQP4 was also increased in the inner medullary collecting duct. Immunoactivity was increased in entire inner medullary collecting duct and newly detected in cytoplasm of principal cell. These findings suggest that increased levels of AQP2 and AQP4 in the cortical and inner medulalry collecting duct may play a important role for maintain fluid balance in the water-deprived kidney.

A Study on Estimation of Factors Affecting Duration of Milk Flow and Milk Flow Rate and Their Relationships with Milk Yields of Dairy Cattle (유우의 착유 지속시간과 유속에 미치는 요인 및 산유량과의 상관관계 추정에 관한 연구)

  • Cho, Y.M.;Park, B.H.;Ahn, B.S.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.517-524
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    • 2004
  • The objective of this study was to investigate the environmental effects on duration time of milk flow, peak milk flow, and average milk flow in teats, and to estimate their relationships with milk yields in morning and evening milking, and to provide fimdamental information for the further study on their relationships with clinical mastitis and somatic cell in milk. A total of 6,768 milking records were studied in 72 Holstein cows. The influences of season, parity, lactation month, and milking interval on characteristics of milk flow considering in linear model were significant(p<.05). Duration of milk flow was longest at milking in fall, past first parity and second month of lactation, and with milking interval over than 13.5hrs. Average milk flow rate and peak milk flow rate were highest at milking in summer, past first parity and 8${\sim}$10 months of lactation, and with milking interval over than 13.5hrs. Milk flow rate was positively correlated to milk yield, and negatively correlated to the duration of milk flow. However duration milk flow was positively correlated to the milk yield with high level of correlation coefficient(+.60). For the establishment of optimum selection criteria on these traits, other aspects such as the udder health, disease and respective economic weights of milk flow characteristics in this study must be considered to develop the indices.

Advancement and Application of Somatic Cell Nuclear Transfer Technique in Dog

  • Oh, H.J.;Hong, S.G.;Park, J.E.;Kim, M.J.;Gomez, M.N.;Kim, M.K.;Kang, J.T.;Kim, J.E.;Jang, G.;Lee, B.C.
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2009.02a
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    • pp.49-57
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    • 2009
  • The cloning of canids was succeeded in 2005, several years after the birth of Dolly the sheep and also after the cloning of numerous other laboratory and farm animal species. The delay of successful somatic cell nuclear transfer (SCNT)was due to the unique reproductive characteristics of the female dogin comparison to other domestic mammals, such as ovulation of immature canine oocyte and a requirement of 25 days for the completion of meiosis within the oviduct (Holst & Phemister, 1971). When the technology for the recovery of in vivo matured oocyte was established, the application of cloning also became possible and cloned dog offspring were obtained. This report summarizes the progress of technical procedures that are required for cloning canids and the application of this technique. The first cloned dog, Snuppy, was achieved using an in vivo-matured oocyte which was enucleated and transferred with an adult skin cell of male Afghan hound. After establishment of a criterion of well-matured oocyte for the improvement of SCNT efficiency, we obtained three cloned female Afghan hound and a toy poodle cloned from 14 year-old aged Poodle using SCNT through this factor. To date, cloned dogs appeared to be normal and those that have reached puberty have been confirmed to be fertile. Through application of canine SCNT technique, first, we demonstrated that SNCT is useful for conserving the breed of endangered animal from extinction through cloning of endangered gray wolves using inter-species SCNT and keeping the pure pedigree through the cloning of Sapsaree, a Korean natural monument. Secondly, we showed possibility of human disease model cloned dog and transgenic cloned dog production through cloning of red fluorescent protein expressing dog. Finally, SCNT can be used for the propagation of valuable genotypes for making elite seed stock and pet dog. In summary, dog cloning is a reproducible technique that offers the opportunity to preserve valuable genetics and a potential step towards the production of gene targeted transgenic cloned dogs for the study of human diseases.

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Effects of Bia-hwan (Féiér-wán) on the Ovariectomized Rat Model of Osteoporosis (비아환(肥兒丸)이 난소적출로 유발된 흰쥐의 골다공증 모델에 미치는 영향)

  • Cho, Chang-Young;Kim, Eun-Young;Kim, Dong Hee;Kim, MinBeom;Kim, Sang-bae;Yang, KyuJin;Sohn, Youngjoo;Jung, Hyuk-Sang
    • Journal of Korean Medicine Rehabilitation
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    • v.27 no.2
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    • pp.19-27
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    • 2017
  • Objectives Osteoporosis is a common disease in adults with prevalence rate of more than 40%, and occurs more to female than to male. The purpose of this study is to identify the effect of Bia-hwan ($F{\acute{e}}i{\acute{e}}r$-$w{\acute{a}}n$) on the osteoporosis of ovariectomized rats. Methods 24 female rats were randomly assigned to a SHAM group, a control group, and a BAH group. Ovaries of the control group and the BAH group were extracted. After than, general animal feed were given to the SHAM group and the control group, animal feed contained BAH were given to the BAH group through mouths of them. After 8 weeks, rats were sacrificed and their weight, calcium, phosphorus, estradiol, total-cholesterol, triglyceride, ALP, albumin, AST, ALT, the weight of femur and tibia ash per body ratio, both area and thickness of trabecular bone, the area of osteoblast and the number of osteoclast were measured. Results The level of calcium, phosphorus, AST in the serum of the BAH group increased significantly compared to the control group. There was no significant difference between the control group and the BAH group in the level of estradiol, total-cholesterol, triglyceride, ALP, albumin, and ALT in the serum. There was no significant difference between the control group and the BAH group in the weight of femur and tibia ash per body ratio. The thickness of trabecular of the BAH group increased significantly compared to the control group, but there was no significant difference in the trabecular area. The area of osteoblast and the number of osteoclast of the BAH group decreased significantly compared to the control group. Conclusions From the results of the above study, oral intake of BAH can inhibit the decrease of calcium, phosphorus in blood and prevent the thinning process of the trabecular by suppressing activation of osteoclast. Thus, BAH may have effects on the treatment and prevention of osteoporosis.

A brief method for preparation of gintonin-enriched fraction from ginseng

  • Choi, Sun-Hye;Jung, Seok-Won;Kim, Hyun-Sook;Kim, Hyeon-Joong;Lee, Byung-Hwan;Kim, Joon Yong;Kim, Jung-Hyun;Hwang, Sung Hee;Rhim, Hyewon;Kim, Hyoung-Chun;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.39 no.4
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    • pp.398-405
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    • 2015
  • Background: Ginseng has been used as a tonic for invigoration of the human body. In a previous report, we identified a novel candidate responsible for the tonic role of ginseng, designated gintonin. Gintonin induces $[Ca^{2+}]_i$ transient in animal cells via lysophosphatidic acid receptor activation. Gintonin-mediated $[Ca^{2+}]_i$ transient is linked to anti-Alzheimer's activity in transgenic Alzheimer's disease animal model. The previous method for gintonin preparation included multiple steps. The aim of this study is to develop a simple method of gintonin fraction with a high yield. Methods: We developed a brief method to obtain gintonin using ethanol and water. We extracted ginseng with fermentation ethanol and fractionated the extract with water to obtain water-soluble and water-insoluble fractions. The water-insoluble precipitate, rather than the water-soluble supernatant, induced a large $[Ca^{2+}]_i$ transient in primary astrocytes. We designated this fraction as gintonin-enriched fraction (GEF). Results: The yield of GEF was approximately 6-fold higher than that obtained in the previous gintonin preparation method. The apparent molecular weight of GEF, determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was equivalent to that obtained in the previous gintonin preparation method. GEF induced $[Ca^{2+}]_i$ transient in cortical astrocytes. The effective dose (ED50) was $0.3{\pm}0.09{\mu}g/mL$. GEF used the same signal transduction pathway as gintonin during $[Ca^{2+}]_i$ transient induction in mouse cortical astrocytes. Conclusion: Because GEF can be prepared through water precipitation of ginseng ethanol extract and is easily reproducible with high yield, it could be commercially utilized for the development of gintoninderived functional health food and natural medicine.

Utilizing cell-free DNA to validate targeted disruption of MYO7A in rhesus macaque pre-implantation embryos

  • Junghyun Ryu;Fernanda C. Burch;Emily Mishler;Martha Neuringer;Jon D. Hennebold;Carol Hanna
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.4
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    • pp.292-297
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    • 2022
  • Direct injection of CRISPR/Cas9 into zygotes enables the production of genetically modified nonhuman primates (NHPs) essential for modeling specific human diseases, such as Usher syndrome, and for developing novel therapeutic strategies. Usher syndrome is a rare genetic disease that causes loss of hearing, retinal degeneration, and problems with balance, and is attributed to a mutation in MYO7A, a gene that encodes an uncommon myosin motor protein expressed in the inner ear and retinal photoreceptors. To produce an Usher syndrome type 1B (USH1B) rhesus macaque model, we disrupted the MYO7A gene in developing zygotes. Identification of appropriately edited MYO7A embryos for knockout embryo transfer requires sequence analysis of material recovered from a trophectoderm (TE) cell biopsy. However, the TE biopsy procedure is labor intensive and could adversely impact embryo development. Recent studies have reported using cell-free DNA (cfDNA) from embryo culture media to detect aneuploid embryos in human in vitro fertilization (IVF) clinics. The cfDNA is released from the embryo during cell division or cell death, suggesting that cfDNA may be a viable resource for sequence analysis. Moreover, cfDNA collection is not invasive to the embryo and does not require special tools or expertise. We hypothesized that selection of appropriate edited embryos could be performed by analyzing cfDNA for MYO7A editing in embryo culture medium, and that this method would be advantageous for the subsequent generation of genetically modified NHPs. The purpose of this experiment is to determine whether cfDNA can be used to identify the target gene mutation of CRISPR/Cas9 injected embryos. In this study, we were able to obtain and utilize cfDNA to confirm the mutagenesis of MYO7A, but the method will require further optimization to obtain better accuracy before it can replace the TE biopsy approach.