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Optimized Immunohistochemical Analysis of Cerebellar Purkinje Cells Using a Specific Biomarker, Calbindin D28k

  • Kim, Byung-Joo;Lee, So-Yeon;Kim, Hyung-Woo;Park, Eun-Jung;Kim, Jun;Kim, Sang-Jeong;So, In-Suk;Jeon, Ju-Hong
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.5
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    • pp.373-378
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    • 2009
  • Cerebellar Purkinje cells (PCs) play a crucial role in motor functions and their progressive degeneration is closely associated with spinocerebellar ataxias. Although immunohistochemical (IHC) analysis can provide a valuable tool for understanding the pathophysiology of PC disorders, the method validation of IHC analysis with cerebellar tissue specimens is unclear. Here we present an optimized and validated IHC method using antibodies to calbindin D28k, a specific PC marker in the cerebellum. To achieve the desired sensitivity, specificity, and reproducibility, we modified IHC analysis procedures for cerebellar tissues. We found that the sensitivity of staining varies depending on the commercial source of primary antibody. In addition, we showed that a biotin-free signal amplification method using a horseradish peroxidase polymer-conjugated secondary antibody increases both the sensitivity and specificity of ICH analysis. Furthermore, we demonstrated that dye filtration using a $0.22\;{\mu}m$ filter eliminates or minimizes nonspecific staining while preserving the analytical sensitivity. These results suggest that our protocol can be adapted for future investigations aiming to understand the pathophysiology of cerebellar PC disorders and to evaluate the efficacy of therapeutic strategies for treating' these diseases.

Functional Characterization of the C-Terminus of YhaV in the Escherichia coli PrlF-YhaV Toxin-Antitoxin System

  • Choi, Wonho;Yoon, Min-Ho;Park, Jung-Ho
    • Journal of Microbiology and Biotechnology
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    • v.28 no.6
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    • pp.987-996
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    • 2018
  • Bacterial programmed cell death is regulated by the toxin-antitoxin (TA) system. YhaV (toxin) and Pr1F (antitoxin) have been recently identified as a type II TA system in Escherichia coli. YhaV homologs have conserved active residues within the C-terminus, and to characterize the function of this region, we purified native YhaV protein (without denaturing) and constructed YhaV proteins of varying lengths. Here, we report a new low-temperature method of purifying native YhaV, which is notable given the existing challenges of purifying this highly toxic protein. The secondary structures and thermostability of the purified native protein were characterized and no significant structural destruction was observed, suggesting that the observed inhibition of cell growth in vivo was not the result of structural protein damage. However, it has been reported that excessive levels of protein expression may result in protein misfolding and changes in cell growth and mRNA stability. To exclude this possibility, we used an [$^{35}S$]-methionine prokaryotic cell-free protein synthesis system in vitro in the presence of purified YhaV, and two C-terminal truncated forms of this protein (YhaV-L and YhaV-S). Our results suggest that the YhaV C-terminal region is essential for mRNA interferase activity, and the W143 or H154 residues may play an analogous role to Y87 of RelE.

Clinical sign and transmission of foot-and-mouth disease in deer, Review (사슴에서의 구제역 증상과 전파 가능성)

  • Park, Jong-Hyeon;Lee, Kwang-Nyeong;Kim, Su-Mi;Ko, Young-Joon;Lee, Hyang-Sim;Cho, In-Soo
    • Korean Journal of Veterinary Service
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    • v.33 no.2
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    • pp.97-103
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    • 2010
  • Foot-and-mouth disease (FMD) commonly infects cloven-hoofed livestock animals such as cattle, pig, sheep, and goat and its clinical signs are well-known. Besides livestock, FMD can be transmitted among cloven-hoofed animals in the wild. FMD mostly affects livestock animals in farms, but, wild animals are likely to play a pivotal role in spreading the disease due to their way of free living. In the case of deer, the clinical signs of FMD vary widely from subclinical to severe infections. Thus, in some deer species, it may be hard to verify clinical signs of FMD. A deer may carry the virus up to 11 weeks after exposure, shedding the virus during the period. However, deer is not considered as a typical host for persistent infection like buffalo, cattle or sheep. In Korea, small-scale livestock farms which have less than 10 animals make up 63.6% of the entire livestock farms. Considering raising environment in deer farms, it is assumed that the risk of virus excretion and consequent transmission of FMD among deers is relatively lower than other cloven-hoofed animals. However, Sika deer and Elk which are typical deer species in Korea would manifest mild to subclinical symptoms upon FMD infection. Therefore, laboratory testing is necessary to confirm FMD in these animals because of difficulty in verifying clinical signs and the risk of virus shedding during inapparent infection.

Lesson Plan Development Based on Marjorie M. Brown’s Home Economics Curriculum Paradiam in Area of Human Development and Family Relationship (Morjorie M. Brown의 가정과 교과과정 모형에 근거한 ‘인간발달과 가족관계’ 영역의 학습지도안 개발)

  • 장혜경;유태명
    • Journal of Korean Home Economics Education Association
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    • v.6 no.2
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    • pp.109-131
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    • 1994
  • This study developed and proposed a lesson plan for the area of “Human development and family relationship”in secondary home economics curriculum. In current educational practice in secondary home economics curriculum, the employed goal and the nature of home economics are neither stated specifically nor closely interrelated. This problem has led us not being able to achieve pursuing goal of home economics education and to employ appropriate teaching methods. In view of this situation, the purpose of this study was to propose a lesson plan reflecting the contents and teaching methods to overcome difficulties and inefficiency in the current practice of secondary home economics education. The theoretical background of the lesson plan was based on the Marjorie M. Brown’s philosophy of home economics which can be summerized as follows: 1. The goal of home economics is to help individuals and families to become autonomous in their own family life and to be more capable of interacting with others in behalf of well-being of the family and of social evolution toward a free society. 2. The characteristics of home economics is conceptualized as practical science and personal service profession. 3. the nature of problems which home economics should deal with is perennial and practial. 4. Practial reasoning composed of historical understanding, critical action, and reflective thinking is appropriate for the previously mentioned goal and characteristics of home economics and for dealing with perennial practial problems. The lesson plan was proposed for one of perennial problem, “How should we understand, create, and maintain our family culture\ulcorner”by employing practial reasoning and teaching models such as “role play”and “classroom meeting.”

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Tungsten Recovery from Tungsten Carbide by Alkali Melt followed by Water Leaching (알칼리 용융 및 수 침출을 이용한 탄화텅스텐으로부터 텅스텐 회수)

  • Kim, Byoungjin;Kim, Suyun;Lee, Jaeryeong
    • Resources Recycling
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    • v.26 no.6
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    • pp.91-96
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    • 2017
  • Tungsten (W) recovery from tungsten carbide (WC) was researched by alkali melt followed by water leaching. The experiments of alkali melt were carried out with the change of the sort of alkali material, heating temperature, and the heating duration. Water leaching of W was performed in the fixed conditions ($25^{\circ}C$, 2 hr., slurry density: 10 g/L). From the mixture of WC and sodium nitrate ($NaNO_3$) in the molar ratio of 1:2, treated at $400^{\circ}C$ for 6 hours, only 63.3% of W might be leached by water leaching. With the increase of sodium hydroxide (NaOH) as a melting additive, the leachability increased. Finally it reached to 97.8 % with the melted mixture of ($WC:NaNO_3:NaOH$) in the ratio of (1:2:2). This imply that NaOH may play a role as a reaction catalyst by lowering Gibb's free energy for alkali melt reaction for WC.

Cell-Free miR-27a, a Potential Diagnostic and Prognostic Biomarker for Gastric Cancer

  • Park, Jong-Lyul;Kim, Mirang;Song, Kyu-Sang;Kim, Seon-Young;Kim, Yong Sung
    • Genomics & Informatics
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    • v.13 no.3
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    • pp.70-75
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    • 2015
  • MicroRNAs (miRNAs) have been demonstrated to play an important role in carcinogenesis. Previous studies revealed that miRNAs are present in human plasma in a remarkably stable form that is protected from endogenous RNase activity. In this study, we measured the plasma expression levels of three miRNAs (miR-21, miR-27a, and miR-155) to investigate the usefulness of miRNAs for gastric cancer detection. We initially examined plasma miRNA expression levels in a screening cohort consisting of 15 patients with gastric cancer and 15 healthy controls from Korean population, using TaqMan quantitative real-time polymerase chain reaction. We observed that the expression level of miR-27a was significantly higher in patients with gastric cancer than in healthy controls, whereas the miR-21 and miR-155a expression levels were not significantly higher in the patients with gastric cancer. Therefore, we further validated the miR-27a expression level in 73 paired gastric cancer tissues and in a validation plasma cohort from 35 patients with gastric cancer and 35 healthy controls. In both the gastric cancer tissues and the validation plasma cohort, the miR-27a expression levels were significantly higher in patients with gastric cancer. Receiver-operator characteristic (ROC) analysis of the validation cohort, revealed an area under the ROC curve value of 0.70 with 75% sensitivity and 56% specificity in discriminating gastric cancer. Thus, the miR-27a expression level in plasma could be a useful biomarker for the diagnosis and/or prognosis of gastric cancer.

Insulin sensitivity improvement of fermented Korean Red Ginseng (Panax ginseng) mediated by insulin resistance hallmarks in old-aged ob/ob mice

  • Cheon, Jeong-Mu;Kim, Dae-Ik;Kim, Kil-Soo
    • Journal of Ginseng Research
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    • v.39 no.4
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    • pp.331-337
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    • 2015
  • Background: The biological actions of various ginseng extracts have been studied for treating obesity and diabetes mellitus. However, few studies have evaluated the effects of fermented Korean Red Ginseng (Panax ginseng Meyer) on metabolic syndrome. The present study evaluated the antiobesity and antidiabetic effects of fermented red ginseng (FRG) on old-aged, obese, leptin-deficient (B6.V-Lepob, "ob/ob") mice. Methods: The animals were divided into three groups and given water containing 0%, 0.5%, and 1.0% FRG for 16 wk. The effect of FRG on ob/ob mice was determined by measuring changes in body weight, levels of blood glucose, serum contents of triglycerides, total cholesterol and free fatty acids, messenger RNA (mRNA) expressions of key factors associated with insulin action, such as insulin receptor (IR), lipoprotein lipase (LPL), glucose transporter 1 and 4 (GLUT1 and GLUT4), peroxisome proliferators-activated receptor gamma ($PPAR-{\gamma}$), and phosphoenolpyruvate carboxykinase (PEPCK) in the liver and in muscle, and histology of the liver and pancreas. Results: FRG-treated mice had decreased body weight and blood glucose levels compared with control ob/ob mice. However, anti-obesity effect of FRG was not evident rather than hypoglycemic effect in old aged ob/ob mice. The hyperlipidemia in control group was attenuated in FRG-treated ob/ob mice. The mRNA expressions of IR, LPL, GLUT1, GLUT4, $PPAR-{\gamma}$, and PEPCK in the liver and in muscle were increased in the FRG-treated groups compared with the control group. Conclusion: These results suggest that FRG may play a vital role in improving insulin sensitivity relative to reducing body weight in old-aged ob/ob mice.

A Study on Taxation Electronic Commerce (전자상거래의 조세정책에 관한 연구)

  • Kim, Ju-Taek
    • Korean Business Review
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    • v.15
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    • pp.59-78
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    • 2002
  • It is only 9 to 10 years since the use of internet became commercial. But electronic commerce is now a familiar form of commercial transaction. Though the ratio of electronic commerce compared to total commercial transaction is still low, it is certain that electronic commerce will be the major form of transactions in the future. In July 1, 1997, United States made a public "A Framework for Global Electronic Commerce" to set up Internet to be a duty free zone and declared to play the trigger role in getting the international discussion of "A Framework for Global Electronic Commerce". In July 8, 1997, EU also adopted "Bonn Declaration"-minimizing government regulations and imposing no new tax on electronic commerce. Focusing the international trend, we need to develop the tax policy that is suitable for our country situation as soon as we can. To be able to do that, we must make an endless effort like selecting professionals, making new administrative part and supporting the research.

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Effect of Ginseng on Calretinin Expression in Mouse Hippocampus Following Exposure to 835 MHz Radiofrequency

  • Aryal, Bijay;Maskey, Dhiraj;Kim, Myeung-Ju;Yang, Jae-Won;Kim, Hyung-Gun
    • Journal of Ginseng Research
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    • v.35 no.2
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    • pp.138-148
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    • 2011
  • Exponential rise in the use of mobile communication devices has generated health concerns due to radiofrequency (RF) exposure due to its close proximity to the head. Calcium binding proteins like calretinin regulate the levels of calcium ($Ca^{2+}$) which plays an important role in biological systems. Ginseng is known for maintaining equilibrium in the human body and may play a beneficial radioprotectant role against electromagnetic field (EMF) exposure. In the present study, we evaluated the radioprotective effects of red ginseng (RG) extract in a mouse model. Calretinin (CR) expression was measured using a free-floating immunohistochemical method in the hippocampus of mice after 835 MHz EMF exposure for 5 h/d for 5 d at specific absorption rate=1.6 W/kg for the different experimental groups. The control animals were treated with NaCl while the experimental animals received 10 mg/kg ginseng, or 30 mg/kg; EMF exposed mice were also treated with NaCl, 10 mg/kg ginseng (E10), or 30 mg/kg (E30). Decreases in CR immunoreactivity (IR) along with loss of CA1 and CA3 interneurons and infragranular cells were observed in the ENaCl group while such losses were not observed in the E10 and E30 groups. CR IR significantly increased in the RG-treated group compared to control and EMF-exposed groups treated with NaCl. The study demonstrates that RG extract can serve as a radioprotective agent that maintains $Ca^{2+}$ homeostasis and prevents neuronal loss in the brain hippocampal region caused by RF exposure.

Radioprotective Potential of Panax ginseng: Current Status and Future Prospectives (고려인삼의 방사선 방어효과에 대한 연구현황과 전망)

  • Nam, Ki-Yeul;Park, Jong-Dae;Choi, Jae-Eul
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.4
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    • pp.287-299
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    • 2011
  • Pharmacological effects of Panax ginseng have been demonstrated in cardiovascular system, endocrine secretion and immune system, together with antitumor, anti-stress and anti-oxidant activities. Modern scientific data show protective effect of ginseng against bone marrow cell death, increased survival rate of experimental animals, recovery of hematopoietic injury, immunopotentiation, reduction of damaged intestinal epithelial cells, inhibition of mutagenesis and effective protection against testicular damages, caused by radiation exposure. And also, ginseng acts in indirect fashion to protect radical processes by inhibition of initiation of free radical processes and thus reduces the radiation damages. The research has made much progress, but still insufficient to fully uncover the action mechanism of ginseng components on the molecule level. This review provides the usefulness of natural product, showing no toxic effects, as an radioprotective agent. Furthermore, the further clinical trials on radioprotection of ginseng need to be highly done to clarify its scientific application. The effective components of ginseng has been known as ginsenosides. Considering that each of these ginsenosides has pharmacological effect, it seems likely that non-saponin components might have radioprotective effects superior to those of ginsenosides, suggesting its active ingredients to be non-saponin series. These results also show that the combined effects of saponin and non-saponin components play an important role in the radioprotective effects of ginseng.