• Title/Summary/Keyword: Host system

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Knock-in of Enhanced Green Fluorescent Protein or/and Human Fibroblast Growth Factor 2 Gene into β-Casein Gene Locus in the Porcine Fibroblasts to Produce Therapeutic Protein

  • Lee, Sang Mi;Kim, Ji Woo;Jeong, Young-Hee;Kim, Se Eun;Kim, Yeong Ji;Moon, Seung Ju;Lee, Ji-Hye;Kim, Keun-Jung;Kim, Min-Kyu;Kang, Man-Jong
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.11
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    • pp.1644-1651
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    • 2014
  • Transgenic animals have become important tools for the production of therapeutic proteins in the domestic animal. Production efficiencies of transgenic animals by conventional methods as microinjection and retrovirus vector methods are low, and the foreign gene expression levels are also low because of their random integration in the host genome. In this study, we investigated the homologous recombination on the porcine ${\beta}$-casein gene locus using a knock-in vector for the ${\beta}$-casein gene locus. We developed the knock-in vector on the porcine ${\beta}$-casein gene locus and isolated knock-in fibroblast for nuclear transfer. The knock-in vector consisted of the neomycin resistance gene (neo) as a positive selectable marker gene, diphtheria toxin-A gene as negative selection marker, and 5' arm and 3' arm from the porcine ${\beta}$-casein gene. The secretion of enhanced green fluorescent protein (EGFP) was more easily detected in the cell culture media than it was by western blot analysis of cell extract of the HC11 mouse mammary epithelial cells transfected with EGFP knock-in vector. These results indicated that a knock-in system using ${\beta}$-casein gene induced high expression of transgene by the gene regulatory sequence of endogenous ${\beta}$-casein gene. These fibroblasts may be used to produce transgenic pigs for the production of therapeutic proteins via the mammary glands.

Syphacia obvelata: A New Hope to Induction of Intestinal Immunological Tolerance in C57BL/6 Mice

  • Taghipour, Niloofar;Mosaffa, Nariman;Rostami-Nejad, Mohammad;Homayoni, Mohamad Mohsen;Mortaz, Esmaeil;Aghdaei, Hamid Asadzadeh;Zali, Mohammad Reza
    • Parasites, Hosts and Diseases
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    • v.55 no.4
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    • pp.439-444
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    • 2017
  • The ability of nematodes to manipulate the immune system of their host towards a Th2 and T regulatory responses has been proposed to suppress the inflammatory response. Clinical trials have proposed a useful effect of helminth infections on improvement of inflammatory disorders. In this study, we investigated the immunomodulatory effect of Syphacia obvelata infection to induce intestinal tolerance in C57BL/6 mice. Mice were infected through the cagemates with self-infected BALB/c mice. Four weeks post-infection, expression levels of $IFN-{\gamma}$, $TNF-{\alpha}$, IL-17, and IL-10 were assessed in the supernatant of mesenteric lymph node (MLN) culture. $Foxp3^+Treg$ were measured in MLN cells by flow cytometry. In the S. obvelata-infected group, the percentage of Tregs ($5.2{\pm}0.4$) was significantly higher than the control ($3.6{\pm}0.5$) (P<0.05). The levels of IL-10 ($55.3{\pm}2.2$ vs $35.2{\pm}3.2$), IL-17 ($52.9{\pm}3.8$ vs $41{\pm}1.8$), $IFN-{\gamma}$ ($44.8{\pm}4.8$ vs $22.3{\pm}2.3$) and $TNF-{\alpha}$ ($71.1{\pm}5.8$ vs $60.1{\pm}3.3$) were significantly increased in infected mice compared to the control group (P<0.05). The above results showed the potential effects of S. obvelata to induce intestinal tolerance. Therefore, it seems that S. obvelata may increase the immunological suppressive function in the intestinal tract.

In Vivo Angiogenic Capacity of Stem Cells from Human Exfoliated Deciduous Teeth with Human Umbilical Vein Endothelial Cells

  • Kim, Ji-Hye;Kim, Gee-Hye;Kim, Jae-Won;Pyeon, Hee Jang;Lee, Jae Cheoun;Lee, Gene;Nam, Hyun
    • Molecules and Cells
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    • v.39 no.11
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    • pp.790-796
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    • 2016
  • Dental pulp is a highly vascularized tissue requiring adequate blood supply for successful regeneration. In this study, we investigated the functional role of stem cells from human exfoliated deciduous teeth (SHEDs) as a perivascular source for in vivo formation of vessel-like structures. Primarily isolated SHEDs showed mesenchymal stem cell (MSC)-like characteristics including the expression of surface antigens and in vitro osteogenic and adipogenic differentiation potentials. Moreover, SHEDs were positive for NG2, ${\alpha}$-smooth muscle actin (SMA), platelet-derived growth factor receptor beta ($PDGFR{\beta}$), and CD146 as pericyte markers. To prove feasibility of SHEDs as perivascular source, SHEDs were transplanted into immunodeficient mouse using Matrigel with or without human umbilical vein endothelial cells (HUVECs). Transplantation of SHEDs alone or HUVECs alone resulted in no formation of vessel-like structures with enough red blood cells. However, when SHEDs and HUVECs were transplanted together, extensive vessel-like structures were formed. The presence of murine erythrocytes within lumens suggested the formation of anastomoses between newly formed vessel-like structures in Matrigel plug and the host circulatory system. To understand underlying mechanisms of in vivo angiogenesis, the expression of angiogenic cytokine and chemokine, their receptors, and MMPs was compared between SHEDs and HUVECs. SHEDs showed higher expression of1VEGF, SDF-$1{\alpha}$, and $PDGFR{\beta}$ than HUVECs. On the contrary, HUVECs showed higher expression of VEGF receptors, CXCR4, and PDGF-BB than SHEDs. This differential expression pattern suggested reciprocal interactions between SHEDs and HUVECs and their involvement during in vivo angiogenesis. In conclusion, SHEDs could be a feasible source of perivascular cells for in vivo angiogenesis.

Association of Anxiety and Depressive Symptoms with Periodontal Disease in Korean Women (한국 여성의 불안 및 우울증상과 치주질환의 관련성)

  • Lee, Eun-Sun;Do, Kyung-Yi;Lee, Kang-Sook
    • Journal of dental hygiene science
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    • v.17 no.1
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    • pp.73-80
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    • 2017
  • Periodontal diseases occur from the interplay between increased bacterial response and the response of the host immune system over time. Anxiety and depression can impair immunological defense mechanisms, causing accumulation of periodontopathogens and thus exacerbating periodontal disease. We investigated the relationship of anxiety and depression to periodontal diseases in Korean women. In this study, 3,551 women aged ${\geq}19$ years were evaluated based on data from the first year (2010) of the Fifth Korea National Health and Nutrition Examination Survey. The analysis of the factors that caused periodontal diseases revealed that dental floss or interdental toothbrush nonuse behaviors have been shown to increase the risk of periodontal disease (odds ratio [OR], 1.49; 95% confidence interval [CI], 1.14~1.95). After adjusting for conditions such as age, marital status, income, educational level, economic activity, diabetes mellitus, smoking, drinking, and frequencies of toothbrushing and interdental cleaning, we found that anxiety and depression increased the risk of developing periodontal diseases (OR, 1.47; 95% CI, 1.04~2.09). People with anxiety and depression have a higher prevalence of periodontal diseases than people without anxiety and depression. Thus, periodic periodontal care and effective self-care education are needed to manage periodontal diseases.

Elemental Composition of Authigenic Siderites in the Early Holocene Coastal Sediments, Western Coast of Korea and Their Depositional Implication (한국 서해 초기현세 퇴적물중 자생 능철석의 원소 성분과 퇴적학적 의미)

  • Cho, J.W.;Lim, D.I.
    • Journal of the Korean earth science society
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    • v.23 no.8
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    • pp.697-706
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    • 2002
  • Authigenic siderite grains, ranging 100 to 250-${\mu}$m in diameter, are abundant in an about 8,600-year-old sediment layer in Namyang Bay, west coast of Korea. The siderites exhibit the aggregated spherulitic morphology with well-developed rhombs on the grain surfaces. They consist mostly of FeCO$_3$ (average, 65%) and MnCO$_3$ (average, 22%) with low Mg/Ca ratio (less than 0.4) in their bulk composition. A series of compositional ternary discrimination diagrams, together with high Mn and low Mg contents, show that only meteoric porewater was involved in siderite precipitation, assuming that depositional environment of host sediment is an organic-rich freshwater system. Considering a series of results such as radiocarbon age, authigenic Mn-rich siderite and lithological features, siderite-hosting sediment (unit Tl) is interpreted as freshwater swamp or bog deposition, infilling the topographic depressions that locally existed before the formation of mid-to-late Holocene tidal deposits. Center-to-margin compositional variation within individual grain is very systematic; Mn and Ca decrease towards the margin of a siderite grain, while Fe and Mg increase. It suggests that the spherulitic siderites were precipitated in this sedimentary layer in a series during the early diagenesis of MnOx-FeOx reduction under steady-state.

Growth and Development of the Academic Societies and Animal Production in Korea, China and Asia over the Last 50 Years

  • Han, In K.;Ha, Jong K.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.6
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    • pp.906-914
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    • 2009
  • The Korean Society of Animal Science (KSAS) was officially born on October 8, 1956 under the leadership of Professor Sang W. Yun of Seoul National University, Seoul, Korea a few years after the end of the Korean War. At that time, there were 0.9 million Korean native cattle, 1.3 million pigs and 8.9 million chickens in Korea. Per capita income for Korea (US$ 66) or China (US$ 59) was about 10% of Asian's average income (US$ 513) in 1956. Korea produced less than 0.2 million M/T of formula feed and consumed 6.1 kg/person/year of animal products. One could say that Korea was an example of an under-developed country in the world. Although the first issue of the Proceedings of the KSAS was published on October 28, 1958, regular quarterly journals of the KSAS were not published until March 1, 1969. Major activities other than publishing its journal were: holding an annual meeting and/or scientific forum at national or international level. The Asian-Australasian Association of Animal Production Societies (AAAP) was founded on September 1, 1980 at Kuala Lumpur, Malaysia with founding members from Australia, Indonesia, Japan, Korea, Malaysia, NZ, Philippines and Thailand. Thirteen AAAP Animal Science Congresses have been held in its 28 year history. Hosting countries were Malaysia (1980), Philippines (1982), Korea (1985), NZ (1987), Taiwan (1990), Thailand (1992), Indonesia (1994), Japan (1996), Australia (2000), India (2002), Malaysia (2004), Korea (2006) and Vietnam(2008). In 1988, significant progress of the AAAP was made by creating an official English journal of the AAAP entitled "Asian-Australasian Journal of Animal Sciences (AJAS)" under the initiative of the KSAS. This journal is now published monthly and distributed to more than 50 countries in the AAAP region and the world. It should be mentioned that the KSAS was able to successfully host the $3^{rd}$ AAAP Animal Science Congress in 1985 and the 12th in 2006, as well as the $8^{th}$ WCAP in 1998. During the last 50 years of KSAS history, per capita income of Korea increased to US$ 17,690 (268 fold), formula feed production increased to 15 million M/T (97 fold) and consumption of animal products increased to 105 kg/person/year (17 fold). Cattle, pig and chicken numbers also increased to 2.5 million (2.8 fold), 9 million (7.4 fold) and 119 million (13 fold). This trend was also found for China and Asia, even if the rate of growth was slightly lower than that of Korea. It is expected that a similar rate of growth in economics, animal numbers, formula feed production and animal protein intake will likely be achieved by other Asian countries in the $21^{st}$ century with somewhat lower rate of development than that of Korea.

Antitumor and immunomodulatory effects of a hot water extract of Phellinus baumii using Sarcoma-180 in the mouse (Pheflinus baumii 자실체 추출물의 Sarcoma-180 에 대한 항암 및 면역효과)

  • Ha, Hyo-Cheol;Kim, Hyun-Pyo;Sim, Ji Young;Chang, Yun-Hee;Kim, Hyun Su
    • Journal of Mushroom
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    • v.2 no.4
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    • pp.169-174
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    • 2004
  • A hot water extract(HWE-P4) was separated from the fruit bodies of PMO-P4, and its antitumor and immunomodulatory activities against sarcoma-180 in ICR mice were investigated. The internal transcribed spacer(ITS) regions from PMO-P4 were amplified using polymerase chain reaction(PCR) and sequenced. The results revealed that PMO-P4 was belong to the Phellinus baumii. When oral administration at the dose of 160mg/kg/day in the mice until the end of the experiment with 2 week's pre-feeding of the HWE-P4, the survival rate of the mice was 152% for 50days after the inoculation of sarcoma-180 and the suppression rate of the tumor growth was 35.3%(p<0.05) for 28 days after inoculation of sarcoma-180. The HWE-P4 increased 71.4% of the CD4/CD8 ratio and 5-fold of the expression of CD25(IL-2 receptor chain) compared with the control. From these results, the antitumor activity of HWE-P4 is exerted through its immunomodulating activity on the host's immune system.

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Immunomodulatory effect of bee pollen extract in macrophage cells (꿀벌 꽃가루 열수 추출물의 큰포식세포 면역활성 효과)

  • Kim, Yi-Eun;Cho, Eun-Ji;Byun, Eui-Hong
    • Korean Journal of Food Science and Technology
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    • v.50 no.4
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    • pp.437-443
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    • 2018
  • Activation of macrophages plays an important role in the host-immune system. In this study, we investigated the functional roles and related signaling mechanism of hot-water extracts of bee pollen (BPW) in RAW 264.7 macrophages. Since BPW did not exert cytotoxicity at concentrations ranging from 62.5 to $250{\mu}g/mL$ in macrophage cells, a concentration of $250{\mu}g/mL$ was used as the maximum dose of BPW throughout subsequent experiments. BPW increased inducible nitric oxide synthase-mediated nitric oxide production in a concentration-dependent manner. Additionally, BPW was found to induce macrophage activation by augmenting the expression of cell surface molecules (cluster of differentiation; CD80/86, and major histocompatibility complex; MHC class I/II) and production of pro-inflammatory cytokines (tumor necrosis $factor-{\alpha}$, interleukin-6, and $IL-1{\beta}$) through mitogen-activated protein kinase and nuclear $factor-{\kappa}B$ signaling pathways in RAW 264.7 macrophages. Taken together, our results indicate that BPW could potentially be used as an immunomodulatory agent.

Expression of Inulinase Gene by Yeast Constitutive Promoters (효모의 구성적 Promoter들에 의한 Inulinase 유전자의 발현)

  • 김연희;남수완
    • Journal of Life Science
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    • v.9 no.2
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    • pp.153-159
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    • 1999
  • To express constitutively the inulinase gene (INUl) of Kluyveromyces marxianus in Saccharomyces cerevisiae, three yeast promoters such as GAPDH, ADH1 and ENO1 were connected upstream of INUl. The resulting plasmids, pYIGP, pADHl-INU, and pENO-INU were introduced to S. cerevisiae SEY2102 host strain, respectively, and then each transformants were selected by staining of colonies on sucrose-agar plate. When the yeast transformants were cultivated on 2$\%$ dextrose media, the total expression levels of inulinase reached to 1.11 unit/mL, 0.88 unit/mL, and 0.69 unit/mL for respective GAPDH, ADH1, and ENO1 promoter systems. On 4% dextrose media, however, the inulinase activities were observed at 2.00 unit/mL for pYIGP, 0.71 unit/mL for pADH1-INU, and 1.40 unit/mL for pENO-INU. This result indicates that the constitutive expression of INUl was significantly affected by the initial concentration of dextrose and the promoter strength was in the order GAPDH, ENO1, and ADH1 promoter at high dextrose concentration. Taking into account the plasmid stability, however, it is suggested that the ENO1 promoter system is more suitable for the INU1 expression on high dextrose medium or in the fed-batch cultivation accumulating ethanol at high level.

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Electrochemical Characteristics of Zn and Si Ion-doped HA Films on Ti-6Al-4V by PEO Treatment

  • Lim, Sang-Gyu;Hwang, In-Jo;Choe, Han-Cheol
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2016.11a
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    • pp.199-199
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    • 2016
  • Commercially pure titanium (cp-Ti) and Ti alloys (typically Ti-6Al-4V) display excellent corrosion resistance and biocompatibility. Although the chemical composition and topography are considered important, the mechanical properties of the material and the loading conditions in the host have, conventionally. Ti and its alloys are not bioactive. Therefore, they do not chemically bond to the bone, whereas they physically bond with bone tissue. The electrochemical deposition process provides an effective surface for biocompatibility because large surface area can be served to cell proliferation. Electrochemical deposition method is an attractive technique for the deposition of hydroxyapatite (HAp). However, the adhesions of these coatings to the Ti surface needs to be improved for clinical used. Plasma electrolyte oxidation (PEO) enables control in the chemical com position, porous structure, and thickness of the $TiO_2$ layer on Ti surface. In addition, previous studies h ave concluded that the presence of $Ca^{+2}$ and ${PO_4}^{3-}$ ion coating on porous $TiO_2$ surface induced adhesion strength between HAp and Ti surface during electrochemical deposition. Silicon (Si) in particular has been found to be essential for normal bone and cartilage growth and development. Zinc (Zn) plays very important roles in bone formation and immune system regulation, and is also the most abundant trace element in bone. The objective of this work was to study electrochemical characteristcs of Zn and Si coating on Ti-6Al-4V by PEO treatment. The coating process involves two steps: 1) formation of porous $TiO_2$ on Ti-6Al-4V at high potential. A pulsed DC power supply was employed. 2) Electrochemical tests were carried out using potentiodynamic and AC impedance methoeds. The morphology, the chemical composition, and the micro-structure an alysis of the sample were examined using FE-SEM, EDS, and XRD. The enhancements of the HAp forming ability arise from $Si/Zn-TiO_2$ surface, which has formed the reduction of the Si/Zn ions. The promising results successfully demonstrate the immense potential of $Si/Zn-TiO_2$ coatings in dental and biomaterials applications.

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