• Title/Summary/Keyword: mouse 3T3-L1

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Micro-Spot Atmospheric Pressure Plasma Production for the Biomedical Applications

  • Hirata, T.;Tsutsui, C.;Yokoi, Y.;Sakatani, Y.;Mori, A.;Horii, A.;Yamamoto, T.;Taguchi, A.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.02a
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    • pp.44-45
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    • 2010
  • We are currently conducting studies on culturing and biocompatibility assessment of various cells such as neural stem cells and induced pluripotent stem cells(IPS cells) on carbon nanotube (CNT), on nerve regeneration electrodes, and on silicon wafers with a focus on developing nerve integrated CNT based bio devices for interfacing with living organisms, in order to develop brain-machine interfaces (BMI). In addition, we are carried out the chemical modification of carbon nanotube (mainly SWCNTs)-based bio-nanosensors by the plasma ion irradiation (plasma activation) method, and provide a characteristic evaluation of a bio-nanosensor using bovine serum albumin (BSA)/anti-BSA binding and oligonucleotide hybridization. On the other hand, the researches in the case of "novel plasma" have been widely conducted in the fields of chemistry, solid physics, and nanomaterial science. From the above-mentioned background, we are conducting basic experiments on direct irradiation of body tissues and cells using a micro-spot atmospheric pressure plasma source. The device is a coaxial structure having a tungsten wire installed inside a glass capillary, and a grounded ring electrode wrapped on the outside. The conditions of plasma generation are as follows: applied voltage: 5-9 kV, frequency: 1-3 kHz, helium (He) gas flow: 1-1.5 L/min, and plasma irradiation time: 1-300 sec. The experiment was conducted by preparing a culture medium containing mouse fibroblasts (NIH3T3) on a culture dish. A culture dish irradiated with plasma was introduced into a $CO_2$-incubator. The small animals used in the experiment involving plasma irradiation into living tissue were rat, rabbit, and pick and are deeply anesthetized with the gas anesthesia. According to the dependency of cell numbers against the plasma irradiation time, when only He gas was flowed, the growth of cells was inhibited as the floatation of cells caused by gas agitation inside the culture was promoted. On the other hand, there was no floatation of cells and healthy growth was observed when plasma was irradiated. Furthermore, in an experiment testing the effects of plasma irradiation on rats that were artificially given burn wounds, no evidence of electric shock injuries was found in the irradiated areas. In fact, the observed evidence of healing and improvements of the burn wounds suggested the presence of healing effects due to the growth factors in the tissues. Therefore, it appears that the interaction due to ion/radicalcollisions causes a substantial effect on the proliferation of growth factors such as epidermal growth factor (EGF), nerve growth factor (NGF), and transforming growth factor (TGF) that are present in the cells.

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Analysis of SNPs in Bovine CSRP3, APOBEC2 and Caveolin Gene Family (소의 CSRP3, APOBEC2, Caveolin 유전자들의 단일염기다형 분석)

  • Bhuiyan, M.S.A.;Yu, S.L.;Kim, K.S.;Yoon, D.;Park, E.W.;Jeon, J.T.;Lee, J.H.
    • Journal of Animal Science and Technology
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    • v.49 no.6
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    • pp.719-728
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    • 2007
  • The cysteine and glycine rich protein 3 (CSRP3), apolipoprotein B mRNA editing enzyme catalytic polypeptide‐like 2(APOBEC2) and caveolin (CAV) gene family(CAV1, CAV2, CAV3) have been reported to play important roles for carcass and meat quality traits in pig, mouse, human and cattle. As an initial step, we investigated SNPs in these 5 genes among eight different cattle breeds. Eighteen primer pairs were designed from bovine sequence data of NCBI database to amplify the partial gene fragments. Sequencing results revealed 9 SNPs in the coding regions of three caveolin genes, 1 SNP in CSRP3 and 3 SNPs in APOBEC2 gene. All the identified SNPs were confirmed by PCR-RFLP. Also, 9 more intronic SNPs were detected in these genes. However, all identified mutations in the coding region do not change amino acid sequence. Allelic distributions were significantly different for 5 SNPs in CAV2, CAV3, CSRP3 and APOBEC2 genes among the eight different breeds. These results gave some clues about the polymorphisms of these genes among the cattle breeds and will be useful for further searches for identifying association between these SNPs and meat quality traits in cattle.

The Effects of Progesterone on Cell Mediated Immunity to Trophoblast in Woman with Recurrent Pregnancy Loss (반복 유산환자의 말초혈액 단핵구와 태반항원을 체외 공동 배양시 세포 매개 면역반응에 프로게스테론이 미치는 영향)

  • Choi, Bum-Chae;Hill, Joseph A.
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.3
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    • pp.399-405
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    • 1997
  • Progesterone is necessary for successful pregnancy and had immunosuppressive properties. Peripheral blood mononuclear cells (PBMC) from many women with unexplained recurrent spontaneous abortion responded to trophoblast extract in vitro by prolifertion and releasing soluble, heat-labile factors that are toxic to mouse embryos (embryotoxic factors). Accumulating evidence suggests that T Helper (Th)-1 type immunity to trophoblast is correlated with embryotoxic factor production and is associated with pregnancy loss, while Th2-type immunity is associated with successful gestation. The objective of this study was to determine whether progesterone can inhibit Th1-type cytokine secretion (IFN-${\gamma}$, TNF-${\alpha}$) by trophoblast-activated peripheral blood mononuclear cells from 23 nonpregnant women (age 25-35) with unexplained recurrent abortion (median 5, range 3 to 15)who otherwise produce embryotoxic factors in response to trophoblast. We also determined whether progesterone affected Th2-type cytokines (IL-4, IL-10) in this system in vitro and if IL-10 (1,500 pg/mL) could inhibit Th1-type immunity to trophoblast. IFN-${\gamma}$ was detected in 17 of 23 (74%) trophoblast stimulated PBMC culture supernatants ($77.94{\pm}23.79$ pg/mL) containing embryotoxic activity. TNF-${\alpha}$ was detected in 19 (83%) of these same supernatants ($703.15{\pm}131.36$ pg/mL). In contrast, none of the supernatants contained detectable levels of IL-4 or IL-10. Progesterone ($10^{-5}$, $10^{-7}$, $10^{-9}$M) inhibited Th1-type immunity in a dose dependent manner, but had no effect on Th2-type cytokine secretion. The inhibitory effects of progesterone were abrogated with RU486, but did not affect Th2-type cytokine secretion in trophoblast-activated cell cultures. IL-10, like progesterone also inhibited Th1-type cytokine secretion but had no effect on Th2-type cytokines. These data suggest that therapies designed to suppress Th1-type cytokine secretion in women with recurrent abortion who have evidence of Th1-type immunity to trophoblast may be efficacious in preventing pregnancy loss and should be tested in appropriately designed clinical trials.

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Investigation of ${\alpha}$-Glucosidase Inhibitory Activity of Ethanolic Extracts from 19 Species of Marine Macroalgae in Korea

  • Jeong, So-Young;Qian, Zhong-Ji;Jin, Yeong-Jun;Kim, Gi-Ok;Yun, Pil-Yong;Cho, Tae-Oh
    • Natural Product Sciences
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    • v.18 no.2
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    • pp.130-136
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    • 2012
  • In the present work, we have collected 19 species of macroalgae (9 Phaeophta and 10 Rhodophyta) f rom all around of Korea: Dictyopteris divaricata, D. prolifera, Myelophycus cavus, Papenfussiella kuromo, Petalonia zosterifolia, Petrospongium rugosum, Rugulopteryx okamurae, Sargassum fulvellum, S. muticum, Callophyllis japonica, Gloiopeltis tenax, Gracilaria longissima, Gracilaria vermiculophylla, Grateloupia asiatica, Grateloupia lanceolata, Grateloupia sparsa, Grateloupia turuturu, Grateloupia sp, and Polyopes affinis. The macroalgal species were extracted by 70% ethanol (EtOH) for 24 h and evaluated its inhibitory effects on ${\alpha}$-glucosidase. Among ethanol extracts, Myelophycus cavus showed the most effectively inhibitory activity ($IC_{50}$, 2.17 ${\mu}g/ml$) against ${\alpha}$-glucosidase, followed by Sargassum fulvellum (<$IC_{50}$, 8.13 ${\mu}g/ml$), Dictyopteris prolifera ($IC_{50}$, 16.66 ${\mu}g/ml$), Rugulopteryx okamurae ($IC_{50}$, 50.63 ${\mu}g/ml$), and Petrospongium rugosum ($IC_{50}$, 101.62 ${\mu}g/ml$). Furthermore, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay showed no cytotoxicity on mouse pre-adipocytes cell line (3T3-L1). These results suggest that some edible macroalgae merit further evaluation for clinical usefulness as anti-diabetic functional foods.

Use of enzyme-linked immunosorbent assay (ELISA) for detection of toxoplasmosis in dogs (ELISA 법을 이용한 개 톡소플라즈마병의 조기진단에 관한 연구)

  • Suh, Myung-deuk;Joo, Hoo-don;Lee, Byung-hoon
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.491-500
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    • 1991
  • This study was conducted to detect the serum antibodies in the experimentally toxoplasma infected dogs and street dogs by use the of an enzyme-linked immunosorbent assay (ELISA). And this test was performed on the polystylene microplate by coating with the tachyzoites soluble antigen of T gondii (RH strain), incubated with sera diluted then, added with HPO-conjugated rabbit anti-dog IgG and o-phenylenediamine used as a substrate. Tachyzoites of T gondii harvested from mouse peritoneal cavity were purified by 30, 40 and 50% Percoll density gradient centrifugation and used as the source of antigen. The results obtained were summarized as follows; 1. The highest ratio of positive to negative (P/N ratio) was obtained at the level of $l{\mu}g/ml$ protein concentration of antigen with the 1/4000 dilution of the conjugate measured by checker-board titration. It was regarded as the optimum concentration of the antigen and conjugate. 2. Cut-off value in this IgG ELISA was 0.375 that was determined by mean absorbance (at 492nm) of IFA negative serum added with the dauble value of the standard deviation $(mean{\pm}2S.D.)$. 3. Serum ELISA IgG antibodies to T gondii in the exyerimentally infected dogs were detected firstly at the Week 3 after inoculation and the highest titer was recognized at the Week 4, 5 and 6 after inoculation. 4. Stability of the antigen absorbed in the microplates that were preserved at $4^{\circ}C$ and $-25^{\circ}C$ separately were prolonged up to 3 weeks and 10 weeks at $4^{\circ}C$ and $-25^{\circ}C$, respectively. However the reproducibility was not reliable after the preservation of 4 weeks and longer. 5. Positive rate of the specific antibodies in 312 test sera was 28.5% and there was no significant differences between the male (27.8%) and female (29.5%), respectively. 6. The IgG ELISA was proved to be a specific procedure for the detection of antibodies to canine toxoplasma infection and also evaluated as a screening test for the large scale of test samples in laboratory.

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Effects of Extracts from Sarcocarp, Peels, and Seeds of Avocado on Osteoblast Differentiation and Osteoclast Formation (아보카도 과육, 과피 및 씨 추출물이 조골세포 분화 및 파골세포 형성에 미치는 영향)

  • Kim, Mi-Jin;Im, Nam-Kyung;Yu, Mi-Hee;Kim, Hyun-Jeong;Lee, In-Seon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.7
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    • pp.919-927
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    • 2011
  • Avocado (Persea americana Mill., Family Lauraceae) is an important subtropical crop in the Americas where it has been cultivated for several thousand years. To investigate the bioactivities of avocado, which acts on bone formation, we prepared methanol extracts from the sarcocarp, peels, and seeds of avocado. The methanol extracts of peels and seeds showed higher bone-forming activity than avocado sarcocarp extracts accompanied by MC3T3-E1 osteoblast proliferation and alkaline phosphatase (ALP) activity. Additionally, the extracts of sarcocarp and peel from avocado also decreased tartrate-resistant acid phosphatase (TRAP) activity against differentiation of osteoclasts, derived from mouse bone marrow macrophages. The hexane fraction from avocado peels showed strong bone-forming activity accompanied by osteoblast proliferation and ALP activity (170.7${\pm}$8.4%), and the ethyl acetate fraction from avocado peel decreased TRAP activity (5.2${\pm}$0.3%) and differentiated osteoclasts at 50 ${\mu}g$/mL. Therefore, avocado is expected to be a natural source for developing medicinal agents to prevent bone-related diseases, such as osteoporosis, by increasing osteoblast differentiation and reducing osteoclast activity.

Effects of Combined Preparation (DWP715) Containing Alaska pollack Extract, Maltol, Ascorbic Acid and Nicotinamide on Decreasing of Blood Alcohol Concentration, Anti- fatigue and Anti-oxidation (북어엑스 및 말톨 함유 복합 조성물(DWP715)의 혈중 알콜농도 저하, 항피로 및 항산화 효과)

  • Cho, Jae-Youl;Kim, Ae-Ra;Yeon, Je-Duk;Lim, Seung-Wook;Lee, Jae-Hwi;Yoo, Eun-Sook;Yu, Young-Hyo;Park, Myung-Hwan
    • Korean Journal of Food Science and Technology
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    • v.29 no.1
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    • pp.167-172
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    • 1997
  • Effect of combined preparation (DWP715) containing Alaska pollack extract, maltol, ascorbic acid and nicotinamide on decreasing of blood alcohol was evaluated in human blood. Treatment of DWP715 prior to administration of 25% alcohol (100 mL) decreased alcohol concentration in blood and showed significant difference after 2 hours. The pharmacokinetic parameters such as area under the concentration-time curve (AUC), $C_{max},\;T_{max}\;and\;T_{1/2}$ were also decreased and delayed when compared with control values. Effects of DWP715 on anti-fatigue and anti-oxidation activities were also studied in the restraint stress model using various parameters (GOT, GPT, LDH values and organ weights) on mild condition and examined through the content of lipid peroxide induced by 2% $CCl_4$ in mouse livers. While GPT level, thymus and adrenal weight were not influenced by DWP715 dosing, LDH, GOT level and spleen weight used as a parameter against fatigue and stress states were recovered almost to the nomal level. Furthermore, lipid peroxidation due to $CCl_4$ was significantly inhibited by DWP715 treatment. These results suggest that DWP715 seems to metabolize the blood alcohol rapidly and to restore the damaged liver and fatigue conditions which was caused by alcohol metabolism to normal condition.

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Molecular Cloning and Characterization of Bovine HMGA1 Gene

  • Yu, S.L.;Chung, H.J.;Sang, B.C.;Bhuiyan, M.S.A.;Yoon, D.;Kim, K.S.;Jeon, J.T.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.11
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    • pp.1662-1669
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    • 2007
  • The high mobility group AT-hook1 (HMGA1) proteins are known to be related to the regulation of gene transcription, replication and promotion of metastatic progression in cancer cells. The loss of expression by disrupting the HMGA1 gene affects insulin signaling and causes diabetes in the mouse. Previously identified single nucleotide polymorphism (SNP) of HMGA1 was significantly associated with fat deposition traits in the pig. In this study, we identified 3,935 bp nucleotide sequences from exon 5 to exon 8 of the bovine HMGA1 gene and its mRNA expression was observed by quantitative real-time PCR. Six single nucleotide polymorphisms in the bovine HMGA1 gene were detected and the allele frequencies of these SNPs were investigated using the PCR-RFLP method in nine cattle breeds including Limousin, Simmental, Brown Swiss, Hereford, Angus, Charolais, Hanwoo, Brahman and Red Chittagong cattle. The map location showed that the bovine HMGA1 gene was also closely located with a previously identified meat quality QTL region indicating this gene is the most likely positional candidate for meat quality traits in cattle.

Effects of Mizoribine on MHC-Restricted Exogenous Antigen Presentation in Dendritic Cells

  • Song, Young-Cheon;Han, Shin-Ha;Kim, Hyun-Yul;Kim, Kwang-Hee;Kwon, Jeung-Hak;Lee, Sang-Jin;Ha, Nam-Joo;Lee, Young-Hee;Lee, Chong-Kil;Kim, Kyung-Jae
    • Archives of Pharmacal Research
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    • v.29 no.12
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    • pp.1147-1153
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    • 2006
  • Mizoribine (MZR) has been shown to possess immunosuppressive activity that selectively inhibits the proliferation of lymphocytes by interfering with inosine monophosphate dehydrogenase. The efficacy of MZR is not only in patients who have had renal transplantation, but also in patients with rheumatoid arthritis (RA), lupus nephritis, and primary nephritic syndrome. Because the exact mechanism of its immunosuppressive action is not clear, the object of this study was to examine the ability of MZR to regulate the antigen presenting cells (APCs), dendritic cells (DCs). In this work, we tested whether MZR ($1{\sim}10\;{\mu}g/mL$) could inhibit the cross-presentation of DCs. DC2.4 cells ($H-2K^{b}$) or bone marrow-derived DCs (BM-DCs) generated from BM cells of C57BL/6 mouse ($H-2K^{b}$) were cultured in the presence of MZR with OVA-microspheres, and the amount of OVA peptide-class I MHC complexes was measured by a T cell hybridoma, B3Z, that recognizes OVA (257-264 : SIINFEKL)-$H-2K^{b}$ complex and expresses-galactosidase. MZR profoundly inhibited the expression of SIINFEKL-$H-2K^{b}$ complexes. This inhibitory activity of MZR appeared to affect the phagocytic activity of DCs. MZR also decreased IL-2 production when we examined the effects of MZR on $CD4^{+}$ T cells. These results provide an understanding of the mechanism of immunosuppressive activity of MZR on the inhibition of MHC-restricted antigen presentation and phagocytic activity in relation to their actions on APCs.

Functional study of Villin 2 protein expressed in longissimus dorsi muscle of Korean native cattle in different growth stages

  • Jin, Yong-Cheng;Han, Jeng-A;Xu, Cheng-Xiong;Kang, Sang-Kee;Kim, Sang-Hun;Seo, Kang-Suk;Yoon, Du-Hak;Choi, Yun-Jaie;Lee, Hong-Gu
    • BMB Reports
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    • v.45 no.2
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    • pp.102-107
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    • 2012
  • The aim of this study was to investigate protein profiles related to the induction of adipogenesis within the bovine longissimus dorsi muscle (BLDM) by proteomic analysis. We analyzed BLDM proteins at different growth stages to clarify the physiological mechanisms of marbled muscle development in 20 head of Korean native cattle (11 month: 10 head, 17 month: 10 head). BLDM proteins were analyzed by two-dimensional electrophoresis and image analysis. Villin 2 was specifically identified by mass spectrometry and a protein search engine. Villin 2 protein expression in BLDM decreased during the fat development stage in test steers. In a Western blot cell culture study of spontaneously immortal bovine muscle fibroblasts, the abundance of Villin 2 was shown to be down-regulated during differentiation into muscle. In 3T3-L1 mouse embryonic fibroblasts, Villin 2 was decreased during differentiation into adipocytes. The results suggest that Villin 2 may be related to the induction of transdifferentiation and adipogenesis in bovine longissimus dorsi muscle.