• Title/Summary/Keyword: Differential Expression

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Differential Proteome Expression of In vitro Proliferating Bovine Satellite Cells from Longissimus Dorsi, Deep Pectoral and Semitendinosus Muscle Depots in Response to Hormone Deprivation and Addition

  • Rajesh, Ramanna Valmiki;Kim, Seong-Kon;Park, Mi-Rim;Park, Min-Ah;Jang, Eun-Joung;Hong, Seung-Gu;Chang, Jong-Soo;Yoon, Du-Hak;Kim, Tae-Hun;Lee, Hyun-Jeong
    • Journal of Animal Science and Technology
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    • v.51 no.6
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    • pp.459-470
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    • 2009
  • The aim of this study was to analyze the proteome of proliferating bovine satellite cells from longissimus dorsi, deep pectoral and semitendinosus muscle depots which had been subjected to hormonal deprivation or addition in culture. For hormone deprivation or addition studies, the cells were either grown in 10% charcoal-dextran stripped fetal bovine serum (CD-FBS) or in 10% FBS supplemented medium. Further to analyze the effect of insulin like growth factor (IGF-1) and testosterone (TS), the cells were grown in 10% CD-FBS containing IGF-1 (10 ng/ml) or TS (10 nM). Results have shown that hormone deprivation had a negative impact on proliferation of the cells from each of the muscle depots. In case of IGF-1 and TS addition, the proliferation levels were low compared with that of the cells grown in 10% FBS. Hence, to gain the insights of the proteins that are involved in such divergent levels of proliferation, the proteome of such satellite cells proliferating under the above mentioned conditions were analyzed using 2D-DIGE and MALDI-ToF/ToF. Thirteen proteins during hormone deprivation and nine proteins from hormone addition were found to be differentially expressed in all the cultures of the cells from the three depots. Moreover, the results highlighted in this study offer a role for each differentially expressed protein with respect to its effect on positive or negative regulation of cell proliferation.

Stem Cells and Cell-Cell Communication in the Understanding of the Role of Diet and Nutrients in Human Diseases

  • Trosko James E.
    • Journal of Food Hygiene and Safety
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    • v.22 no.1
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    • pp.1-14
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    • 2007
  • The term, "food safety", has traditionally been viewed as a practical science aimed at assuring the prevention acute illnesses caused by biological microorganisms, and only to a minor extent, chronic diseases cause by chronic low level exposures to natural and synthetic chemicals or pollutants. "food safety" meant to prevent microbiological agents/toxins in/on foods, due to contamination any where from "farm to Fork", from causing acute health effects, especially to the young, immune-compromised, genetically-predisposed and elderly. However, today a broader view must also include the fact that diet, perse (nutrients, vitamins/minerals, calories), as well as low level toxins and pollutant or supplemented synthetic chemicals, can alter gene expressions of stem/progenitor/terminally-differentiated cells, leading to chronic inflammation and other mal-functions that could lead to diseases such as cancer, diabetes, atherogenesis and possibly reproductive and neurological disorders. Understanding of the mechanisms by which natural or synthetic chemical toxins/toxicants, in/on food, interact with the pathogenesis of acute and chronic diseases, should lead to a "systems" approach to "food safety". Clearly, the interactions of diet/food with the genetic background, gender, and developmental state of the individual, together with (a) interactions of other endogenous/exogenous chemicals/drugs; (b) the specific biology of the cells being affected; (c) the mechanisms by which the presence or absence of toxins/toxicants and nutrients work to cause toxicities; and (d) how those mechanisms affect the pathogenesis of acute and/or chronic diseases, must be integrated into a "system" approach. Mechanisms of how toxins/toxicants cause cellular toxicities, such as mutagenesis; cytotoxicity and altered gene expression, must take into account (a) irreversible or reversal changes caused by these toxins or toxicants; (b)concepts of thresholds or no-thresholds of action; and (c) concepts of differential effects on stem cells, progenitor cells and terminally differentiated cells in different organs. This brief Commentary tries to illustrate this complex interaction between what is on/in foods with one disease, namely cancer. Since the understanding of cancer, while still incomplete, can shed light on the multiple ways that toxins/toxicants, as well as dietary modulation of nutrients/vitamins/metals/ calories, can either enhance or reduce the risk to cancer. In particular, diets that alter the embryo-fetal micro-environment might dramatically alter disease formation later in life. In effect "food safety" can not be assessed without understanding how food could be 'toxic', or how that mechanism of toxicity interacts with the pathogenesis of any disease.

Identification of Differentially Expressed Radiation-induced Genes in Cervix Carcinoma Cells Using Suppression Subtractive Hybridization (자궁경부암세포에서 방사선조사시 차등 발현되는 유전자 동정)

  • Kim Jun-Sang;Lee Young-Sook;Lee Jeung Hoon;Lee Woong-Hee;Seo Eun Young;Cho Moon-June
    • Radiation Oncology Journal
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    • v.23 no.1
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    • pp.43-50
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    • 2005
  • Purpose : A number of genes and their products are Induced early or late following exposure of cells to ionizing radiation. These radiation-Induced genes have various effects on irradiated cells and tissues. Suppression subtractive hybridization (SSH) based on PCR was used to Identify the differentially expressed genes by radiation in cervix carcinoma cells. Materials and Methods : Total RNA and poly $(A)^+$ mRNA were Isolated from Irradiated and non-irradiated HeLa cells. Forward- and reverse-subtracted cDNA libraries were constructed using SSH. Eighty-eight clones of each were used to randomly select differentially expressed genes using reverse Northern blotting (dot blot analysis). Northern blotting was used to verify the screened genes. Results : Of the 17t clones, 10 genes in the forward-subtracted library and 9 genes In the reverse-subtracted library were identified as differentially expressed radiation-induced genes by PCR-select differential screening. Three clones from the forward-subtracted library were confirmed by Northern blotting, and showed increased expression in a dose-dependent manner, including a telomerase catalytic subunit and sodium channel-like protein gene, and an ESTs (expressed sequence tags) gene. Conclusion : We Identified differentially expressed radiation-induced genes with low-abundance genes with SSH, but further characterization of theses genes are necessary to clarify the biological functions of them.

Kernel Integration Scheme for 2D Linear Elastic Direct Boundary Element Method Using the Subparametric Element (저매개변수 요소를 사용한 2차원 선형탄성 직접 경계요소법의 Kernel 적분법)

  • Jo, Jun-Hyung;Park, Yeongmog;Woo, Kwang-Sung
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.25 no.5
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    • pp.413-420
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    • 2012
  • In this study, the Kernel integration scheme for 2D linear elastic direct boundary element method has been discussed on the basis of subparametric element. Usually, the isoparametric based boundary element uses same polynomial order in the both basis function and mapping function. On the other hand, the order of mapping function is lower than the order of basis function to define displacement field when the subparametric concept is used. While the logarithmic numerical integration is generally used to calculate Kernel integration as well as Cauchy principal value approach, new formulation has been derived to improve the accuracy of numerical solution by algebraic modification. The subparametric based direct boundary element has been applied to 2D elliptical partial differential equation, especially for plane stress/strain problems, to demonstrate whether the proposed algebraic expression for integration of singular Kernel function is robust and accurate. The problems including cantilever beam and square plate with a cutout have been tested since those are typical examples of simple connected and multi connected region cases. It is noted that the number of DOFs has been drastically reduced to keep same degree of accuracy in comparison with the conventional isoparametric based BEM. It is expected that the subparametric based BEM associated with singular Kernel function integration scheme may be extended to not only subparametric high order boundary element but also subparametric high order dual boundary element.

Inflammatory Mediators Modulate NK Cell-stimulating Activity of Dendritic Cells by Inducing Development of Polarized Effector Function

  • Kim, Kwang-Dong;Choi, Seung-Chul;Lee, Eun-Sil;Kim, Ae-Yung;Lim, Jong-Seok
    • IMMUNE NETWORK
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    • v.7 no.3
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    • pp.133-140
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    • 2007
  • Background: It is well established that cross talk between natural killer (NK) cells and myeloid dendritic cells (DC) leads to NK cell activation and DC maturation. In the present study, we investigated whether type 1-polarized DC (DC1) matured in the presence of IFN-${\gamma}$ and type 2-polarized DC (DC2) matured in the presence of PGE2 can differentially activate NK cells. Methods: In order to generate DC, plastic adherent monocytes were cultured in RPMI 1640 containing GM-CSF and IL-4. At day 6, maturation was induced by culturing the cells for 2 days with cytokines or PGE2 in the presence or absence of LPS. Each population of DC was cocultured with NK cells for 24 h. The antigen expression on DC was analyzed by flow cytometry and cytokine production in culture supernatant was measured by ELISA or a bioassay for TNF-${\alpha}$ determination. NK cell-mediated lysis was determined using a standard 4h chromium release assay. Results: DC2, unlike DC1, had weak, if any, ability to induce NK cell activation as measured by IFN-${\gamma}$ production and cytolytic activity. DC2 were weakly stimulated by activated NK cells compared to DC1. In addition, IFN-${\gamma}$-primed mature DC appeared to be most resistant to active NK cell-mediated lysis even at a high NK cell/DC ratio. On the other hand, PGE2-primed DC were less resistant to feedback regulation by NK cells than IFN-${\gamma}$-primed mature DC. Finally, we showed that the differential effect of two types of DC population on NK cell activity is not due to differences in their ability to form conjugates with NK cells. Conclusion: These results suggest that different combinations of inflammatory mediators differentially affect the effector function of DC and, as a result, the function of NK cells, eventually leading to distinct levels of activation in adaptive immunity.

Comparison of Skin Injury Induced by β- and γ-irradiation in the Minipig Model

  • Kim, Joong-Sun;Jang, Hyosun;Bae, Min-Ji;Shim, Sehwan;Jang, Won-Seok;Lee, Sun-Joo;Park, Sunhoo;Lee, Seung-Sook
    • Journal of Radiation Protection and Research
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    • v.42 no.4
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    • pp.189-196
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    • 2017
  • Background: The effects of radiation on tissues vary depending on the radiation type. In this study, a minipig model was used to compare the effects of ${\beta}$-rays from $^{166}Ho$ and ${\gamma}$-rays from $^{60}Co$ on the skin. Materials and Methods: In this study, the detrimental effects of ${\beta}$- and ${\gamma}$-irradiation on the skin were assessed in minipigs. The histopathological changes in the skin from 1 to 12 weeks after exposure to 50 Gy of either ${\beta}$- (using $^{166}Ho$ patches) or ${\gamma}$- (using $^{60}Co$) irradiation were assessed. Results and Discussion: The skin irradiated by ${\beta}$-rays was shown to exhibit more severe skin injury than that irradiated by ${\gamma}$-rays at 1-3 weeks post-exposure; however, while the skin lesions caused by ${\beta}$-rays recovered after 8 weeks, the ${\gamma}$-irradiated skin lesions were not repaired after this time. The observed histopathological changes corresponded with gross appearance scores. Seven days post-irradiation, apoptotic cells in the basal layer were detected more frequently in ${\beta}$-irradiated skin than in ${\gamma}$-irradiated skin. The basal cell density and skin thickness gradually decreased until 4 weeks after ${\gamma}$- and ${\beta}$- irradiation. In ${\beta}$-irradiated skin lesions, and the density and thickness increased sharply back to control levels by 6-9 weeks. However, this was not the case in ${\gamma}$-irradiated skin lesions. In ${\gamma}$-irradiated skin, cyclooxygenase-2 (COX-2) was shown to be expressed in the epidermis, endothelial cells of vessels, and fibroblasts, while ${\beta}$-irradiated lesions exhibited COX-2 expression that was mostly limited to the epidermis. Conclusion: In this study, ${\beta}$-rays were shown to induce more severe skin injury than ${\gamma}$-rays; however, the ${\beta}$-rays-induced injury was largely repaired over time, while the ${\gamma}$-rays-induced injury was not repaired and instead progressed to necrosis. These findings reveal the differential effects of ${\gamma}$- and ${\beta}$-irradiation on skin and demonstrate the use of minipigs as a beneficial experimental model for studying irradiation-induced skin damage.

Development of Target-Controlled Infusion System in Plasma Concentration. PART1 : Establishment of Pharmacokinetic Model and Verification (혈중 목표 농도 자동 조절기(TCI) 개발 PART1 : 약동학적 모델의 수립과 검증)

  • 안재목;길호영
    • Journal of Biomedical Engineering Research
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    • v.23 no.5
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    • pp.341-349
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    • 2002
  • The target controlled infusion(TCI) pump system is a logical approach to the development of improved administration techniques of an intravenous anaesthetic agent. The principle of TCI system is based on an understanding of the pharmacokinetic properties, three or four compartment model. The TCI system is optimal and flexible control of the plasma drug concentration. But the clinical goal is always to achieve a therapeutic drug effect, not a therapeutic concentration. So we developed the algorithm to target the concentration at the site of drug effect rather than the concentration in the plasma. If impulse drug is inputted into body, the decline of plasma concentration with time is shown, resulting in the expression of the differential equation. Therefore, we must reformulate our three-compartment model as four-compartment model with the effect compartment. And we tested plasma targeting and effect targeting algorithm by computer simulation using four-compartment model. So we developed the TCI capable of applying all intravenous drugs by adjusting individual pharmacokinetic parameters independently.

Evaluation of Biological Effects of Low Concentrations of Mercury Chloride (II) and Ionizing Radiation in the Prepubertal Male Rats (미성숙 웅성 흰쥐를 이용한 이온화 방사선 조사 및 저농도 염화수은(II)의 음용에 따른 위해성 비교 평가)

  • Kim Ji Hyang;Kim Jin Kyu;Yoon Yong Dal
    • Korean Journal of Environmental Biology
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    • v.22 no.3
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    • pp.411-418
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    • 2004
  • Mercury, one of the most diffused and hazardous organ-specific environmental contaminants, exists in a wide variety of physical and chemical states. The murcury with the nature which evaporates easily can cause an acute or chronic mercury poisoning to workers at mercury-handling workplaces. Although many studies indicate that mercury induces a deleterious damage, little has been reported from the investigations of mercury effects at surrounding levels in living things. The purpose of this study was to evaluate the biological effects of mercury chloride and ionizing radiation. Prepubertal male F344 rats were administered mercury chloride in drinking water throughout the experimental period or were given wholebody irradiation with a dose of 6.5 Gy. The amount changed of body weight during the experimental period showed a 4.9% rise in the mercury-treated group and 14.4% decline in the irradiated group compared with the level of the control group. The results of hematological analysis (red blood cells, white blood cells, hemoglobin, and hematocrit) indicated the differential effects of mercury chloride and ionizing radiation. However the concentration of cortisol as assessed by radioimmunoassay increased in both of the groups. Relative expressions of mRNA related to mitochondrion-mediated apoptosis were investigated using semiquantitative reverse transcription polymerase chain reaction on gonad and urinary organs of the experimental groups. While the expression of Bcl-2 mRNA exhibited different patterns depending on the organs or the experimental groups, both of the experimental groups showed a conspicuous expressions of Bax mRNA. In conclusion, the target organ of mercury chloride seems to be a urinary organ and the pattern of damage induced by mercury chloride differs from that by ionizing radiation.

Preparation and Release Behavior of Ipriflavone-Loaded PLGA Microsphere for Tissue Engineered Bone (이프리플라본을 함유한 생분해성 PLGA 미립구의 제조 및 조직공학적 골재생을 위한 영향평가)

  • So, Jung-Won;Jang, Ji-Wook;Kim, Soon-Hee;Kim, Geun-Ah;Choi, Jin-Hee;Rhee, John-M.;Son, Young-Suk;Min, Byoung-Hyun;Khang, Gil-Son
    • Polymer(Korea)
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    • v.33 no.1
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    • pp.26-32
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    • 2009
  • The aim of this research was to prepare microparticulate systems based on poly (lactide-co-glycolide)(PLGA) for the local release of ipriflavone in order to reduce bone loss. We developed the IP loaded PLGA microspheres using relatively simple oil-in-water(O/W) solvent evaporation method. HPLC was used to perform the in vitro release test of IP and morphology of cell attached on the micro-spheres was investigated using SEM. Cytotoxicity was assayed by cell counting kit-8 (CCK-8) test. Osteogenic differential cells were analyzed by ALP activity. Through RT-PCR analysis, we observed osteocalcin, ALP, and Type I collagen mRNA expression. The release of IP in vitro was more prolonged over 42 days and IP/PLGA microspheres showed the improvement on the cell proliferation, ALP activity and RT-PCR comparing with control (only PLGA). This initial research will be used to direct future work involved in developing this composite injectable bone tissue engineering system.

A study of PCNA Expression in Gastric Adenoma and Adenocarcinoma (위선종 및 위선암종에 있어서 PCNA 발현 양상에 관한 연구)

  • Kim, Mi-Jin;Choi, Won-Hee;Lee, Tae-Sook
    • Journal of Yeungnam Medical Science
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    • v.12 no.1
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    • pp.1-9
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    • 1995
  • A monoclonal antibody to PCNA, which can be used on routinely processed tissue, was applied to 25 cases of gastric adenomas and 64 cases of gastric adenocarcinomas in order to diffentiate adenoma and adenocarcinoma and also to evaluate the prognostic value in adenocarcinoma. The results were summerized as follows: The peNA labelling index was $29.14{\pm}12.77%$ in control, $44.09{\pm}17.11%$ in adenoma and $80.15{\pm}10.69$ in adenocarcinoma, resulting in significant increase in adenocarcinoma compared to adenoma. In adenocarcinoma, no significant correlation was observed between PCNA labelling index and histologic grade, and there was increased tendency of PCNA labelling index in proportion to depth of invasion without statistical significance. The PCNA index was significantly increased in advanced adenocarcinoma compared to early gastric carcinoma, and also in positive nodal metastasis group than in negative group. From above results, the PCNA stain will be able to provide a helpful method for the differential diagnosis between gastric adenoma and adenocarcinoma, and could be a useful prognostic factor in adenocarcinoma if other factors are considered together.

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