• Title/Summary/Keyword: PCR condition

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A Survey on the Actual Condition of Products not Labeled with Allergens (알레르기 유발물질 미표시 제품 실태 조사)

  • Kim, Kyung-Seon;Song, Sung-Min;Kwon, Sung-Hee;Jang, Seung-Eun;Lee, Bo-Min;Kim, Meyong-Hee;Han, Young-Sun;Hur, Myung-Je;Kwon, Mun-Ju
    • Journal of Food Hygiene and Safety
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    • v.36 no.3
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    • pp.257-263
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    • 2021
  • For this survey, PCR (polymerase chain reaction) testing was conducted using 14 species-specific primers to monitor the labeling of allergy-causing substances in various foods. Sixty samples from stationary stores near elementary schools and imported confectionery shops were tested, including snacks, candies, and chocolate. Allergens of milk, wheat, eggs, tomatoes, almonds and peanuts were detected in 30 cases (50.0%). In addition, many products were detected as either containing unlabeled substances or not showing allergen-related information and labeling in Korean. In order to ensure that consumers are able to purchase products safely and securely, a system for thorough guidance and monitoring of allergen-related labeling by domestic manufacturing and processing companies and import-related companies is required.

Transforming Growth Factor-$\beta$ is a Possible Paracrine Mediator in the Human Endometrial Decidualization (인간자궁내막의 탈락막화 (Decudualization)에 있어서 TGF-$\beta$ (Transforming Growth Factor-$\beta$)의 역할)

  • Park, Dong-Wook;Choi, Dong-Soon;Kim, Mi-Ran;Hwang, Kyung-Joo;Jo, Mi-Yeong;Ahn, Seong-Hee;Min, Churl-K.;Ryu, Hee-Sug
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.1
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    • pp.65-75
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    • 2003
  • Objectives: To investigate the role of TGF (Transforming growth factor-$\beta$) involved in the paracrinic communication during decidualization between UEC (uterine epithelial cells) and USC (uterine stromal cells), we have employed a co-culture system composed of human endometrial epithelial and stromal cells in defined hormonal conditions. Design: In the co-culture, endometrial epithelial cells cultured in the matrigel-coated cell culture insert are seeded on top of the endometrial stromal cells cultured within a collagen gel. The co-culture was maintained for 48 hours under the following hormonal conditions: progesterone dominant condition (100 nM P4 and 1 nM E2) or estrogen-dominant condition (100 nM E2 and 1 nM P4). 10 ng/ ml HGF and/or 10 ng/ml TGF-$\beta$1 are added. Methods: RT-PCR is utilized to detect mRNAs quantitatively. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining are utilized to detect proteins in the tissue. Results: Prolactin mRNA is expressed in the co-cultured stromal cells under the progesterone dominant condition. TGF-$\beta$1 and its receptors are expressed in both the co-cultured epithelial and stromal cells irrespective of the steroid present, which is in contrast with no or negligible expression of TGF-$\beta$1 or its receptor in cells separately cultured. Both estrogen and progesterone significantly elevate the concentration of hepatocyte growth factor (HGF) in the conditioned medium of the co-culture with the value of 4, 325 pg/ml in E2-dominant and 2, 000 pg/ml in P4-dominant condition compare to 150 pg/ml in no hormone. In separately cultured stromal cells, administration of HGF induces the expression of TGF receptor 1 in both hormonal conditions, but induction of TGF receptor 2 is only manifest in the P4-dominant condition. Administration of TGF-$\beta$ and HGF directly induce the decidualization marker prolactin mRNA in separately cultured stromal cells. Conclusion: It is likely that steroid hormones induces prolactin mRNA indirectly by promoting the cell to cell communication between the stromal and the epithelial cells. TGF-$\beta$ and HGF are two possible paracrine mediators in the human endometrial decidualization.

Identification of Novel Salt Stress-responsive Genes Using the Activation Tagging System in Arabidopsis (애기장대에서 activation tagging system을 이용한 새로운 고염 스트레스 반응 유전자의 동정)

  • Seok, Hye-Yeon;Nguyen, Linh Vu;Bae, Hyoungjoon;Ha, Jimin;Kim, Ha Yeon;Lee, Sun-Young;Moon, Yong-Hwan
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1030-1041
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    • 2018
  • Abiotic stresses limit the growth and productivity of plants. Cellular adaptation to abiotic stresses requires coordinated regulation in gene expression directed by complex mechanisms. This study used the activation tagging system to identify novel salt stress-responsive genes. The study selected 9 activation tagging lines that showed salt stress-tolerant phenotypes during their germination stages. Thermal asymmetric interlaced-PCR (TAIL-PCR) was used to identify the T-DNA tagging sites on the Arabidopsis genome in selected activation tagging lines, including AT7508, AT7512, AT7527, AT7544, AT7548, and AT7556. RT-PCR analysis showed that ClpC2/HSP93-III (At3g48870), plant thionin family (At2g20605), anti-muellerian hormone type-2 receptor (At3g50685), vacuolar iron transporter family protein (At4g27870), and microtubule-associated protein (At5g16730) were activated in AT7508, AT7512, AT7527, AT7544, and AT7556, respectively. Interestingly, in AT7548, both the genes adjacent to the T-DNA insertion site were activated: Arabinogalactan protein 13 (AGP13) (At4g26320) and F-box/RNI-like/FBD-like domains-containing protein (At4g26340). All of the seven genes were newly identified as salt stress-responsive genes from this study. Among them, the expression of ClpC2/HSP93-III, AGP13, F-box/RNI-like/FBD-like domains-containing protein gene, and microtubule-associated protein gene were increased under salt-stress condition. In addition, AT7508, AT7527, and AT7544 were more tolerant to salt stress than wild type at seedling development stage, functionally validating the screening results of the activation tagging lines. Taken together, our results demonstrate that the activation tagging system is useful for identifying novel stress-responsive genes.

Identification of Equine Heat Shock Proteins Gene and Their mRNA Expression Analysis after Exercise (말의 열충격 단백질(heat shock proteins)의 특성 구명과 운동 후 유전자의 발현 분석)

  • Cho, Hyun-Woo;Park, Jeong-Woong;Choi, Jae-Young;Sivakumar, S.;Kim, Nam-Young;Shin, Teak-Soon;Cho, Seong-Keun;Kim, Byeong-Woo;Cho, Byung-Wook
    • Journal of Life Science
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    • v.24 no.2
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    • pp.105-111
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    • 2014
  • The purpose of this study was to characterize equine heat-shock protein (Hsp) genes and analyze their expression pattern in various horse tissues and blood leukocytes after exercise. In a previous study, RNA sequencing of blood and skeletal muscles of thoroughbreds before and after exercise was performed using differently expressed gene (DEG) analysis. Three Hsp genes (HspH1, Hsp90${\alpha}$ and Hsp70) were selected by DEG analysis and were found to be differentially expressed in either blood or muscle. To validate and extend previous observations on these genes, we performed RT-PCR analyses of horse tissue as well as real-time qPCR analyses of blood leukocytes after exercise. mRNA expression of these Hsp genes was found to be ubiquitous in the analyzed tissues (including thyroid, colon, skeletal muscle, cecum, kidney, spinal cord, heart, and lung). In addition, Hsp mRNA expression of these genes in extracted whole blood increased after 120 minutes of exercise compared to the baseline condition. These results are in agreement with the results of human and other experimental animals, suggesting that regulatory mechanisms that are responsible for upregulation of Hsp gene transcription may be conserved among species. Further investigations to correlate Hsp gene expression patterns with athletic performance or recovery processes after exercise are warranted.

A Study of Epstein-Barr Virus, and Human Leukocyte Antigen Typing in Children with Acute Infectious Mononucleosis (급성 전염성 단핵구증 환아에서 Epstein-Barr 바이러스의 감염형과 사람 백혈구 항원형 연구)

  • Hahn, Seung-Hoon;Shin, Wan-Shik;Han, Hoon;Kang, Jin-Han
    • Clinical and Experimental Pediatrics
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    • v.46 no.5
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    • pp.467-473
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    • 2003
  • Purpose : The Epstein-Barr virus(EBV), gamma herpesvirus, is an important pathogen that is widespread around the world. The EBV causes various diseases depending on the geographic location, and on the immunity or the premorbid condition of the person exposed to EBV. To evaluate EBV typing may be the most important step to figure out the pathogenesis of EBV associated diseases, and we need to re-evaluate the pathologic role of human leukocyte antigen(HLA) in developing Epstein- Barr virus associated acute infectious mononucleosis by using newly developed methods. Methods : This study included 24 children(age range : 6 to 13 years), serologically confirmed with acute infectious mononucleosis. The control group for the HLA type consisted of 200 age-matched healthy children. To classify HLA I, modified ARMs-PCR was used, while modified PCR-SSOP was utilized in typing of HLA II. Also, we performed EBV typing in study patients by using a one-step PCR. Results : The results of HLA types : In HLA class I, HLA-A24 was positive in 69 of 200 healthy children and positive in 14 of 24 patients in the study group(relative risk : 3.5724, chi-square; 5.26, P<0.05). In HLA class II, HLA-DRB1*07 was detected in 18 of 200 healthy children, and eight of 24 patients in the study group(relative risk; 506173, chi-square; 9.73, P<0.01). The results of EBV types : In the research group, 20(83.8%) of 24 patients were shedding type A virus, while 4(16.7%) were type B. Conclusion : We conclude that development of infectious mononucleosis may be associated with HLA types, and these results suggest that acute infectious mononucleosis could have hereditary traits. And we confirm that type A EBV is highly prevalent in patients with acute infectious mononucleosis in Korea. Also, our results suggest that further large scale studies, including adult groups, regarding the association between pathogenesis of EBV with HLA-DP or HLA-DQ will be warranted.

Embryo Transfer with Sex-Determined Hanwoo Embryos Produced by In-vitro Fertilization (성감별된 한우 체외수정란의 수정란 이식)

  • 김용준;이창민;정구남;이해리;조성우;김용수;신동수;홍유미;유일정
    • Journal of Embryo Transfer
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    • v.18 no.2
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    • pp.97-108
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    • 2003
  • In-vitro fertilized Hanwoo embryos were biopsied for sex determination by PCR. Biopsied embryos were incubated for 1∼2 hours for the recovery. Those sexed Hanwoo embryos were transferred to 49 Hanwoo and 16 Holstein recipients from February 2000 to February 2001. Of 65 recipients, 14 cows(12 Hanwoo and 2 Holstein) delivered the same offspring as sex-determined by PCR, therefore the conception rate was 21.5%. 1. Total 65 embryos(male 35, female 30) were transferred to recipients, and 14 calves (male 6, female 8) were delivered. In comparison between sex by PCR method and sex of calves born after embryo transfer, the accuracy of sex determination was 100.0%. 2. The conception rate after transfer with biopsied embryo between Hanwoo and Holstein was 24.5% and 12.5% 3. The conception rate after transfer with biopsied embryo between fresh and frozen-thawed embryos was 23.5% and 14.3%. 4. The conception rate according to the season when embryo was transferred was 11.8, 29.4, 23.5 and 20.0% for spring, summer, autumn and winter, respectively. 5. The conception rate according to embryo quality after biopsy was 41.7, 30.0 and 0.0% for excellent, good and fair quality. 6. The conception rate according to thickness of uterine horn was 71.4, 18.9, 11.8 and 0.0% for 0, +, ++ and +++ thickness. 7. The conception rate according to the site in the uterine hem where embryo was put was 30.0, 20.0 and 10.0% for cranial, mid, and caudal part of uterine horn. 8. The conception rate according to the quality of corpus luteum ipsilateral to the uterine horn where embryos was transferred was 41.2, 14.3 and 15.4% for excellent, good and fair quality. 9. The conception rate according to the time required for embryo transfer was 18.2, 30.0, 30.0, 0.0 and 25.0% for 10, 15, 20, 25 and 30 minutes. 10. The conception rate according to parity of recipients was 26.5, 19.1, 14.3 and 0.0% for the primiparous, the 2nd parous, the 3rd parous and the 4th parous recipients. These results indicated that fresh embryos are more demanded than frozen-thawed embryos for good conception rate in embryo transfer with biopsied-sexed embryo. Also, it was indicated that we should consider embryo-recovering condition, recipient's uterine thickness, transfer site in uterine horn, quality of corpus luteum, time required for transfer and parity of recipient to achieve good conception rate in ET with biopsied-sexed embryos.

Environmental factors Associated with Disease Development of Garlic White Rot Caused by Two Species of Sclerotium (온도와 토양습도가 마늘 흑색썩음균핵병 발생에 미치는 영향)

  • Kim Yong-Ki;Kwon Mi-Kyung;Shim Hong-Sik;Kim Tack-Soo;Yeh Wan-Hae;Cho Weon-Dae;Choi In-Hu;Lee Seong-Chan;Ko Sug-Ju;Lee Yong-Hwan;Lee Chan-Jung
    • Research in Plant Disease
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    • v.11 no.2
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    • pp.128-134
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    • 2005
  • This study was conducted to elucidate effect of environmental factors on the development of white rot. In order to identify the causal agents causing white rot of Allium crops, we compared DNA profiles of a representative isolate, Sclerotium cepivorum, introduced from foreign country with Korean isolates using UP-PCR. As a result, Sclerotium isolates forming round-shaped sclerotia were identified as Sclerotium cepivorum pertaining in UP-PCR b group and Sclerotium isolates farming anamorphic-shaped sclerotia presumed to be a novel species of Sclerotium based on DNA profiles of UP-PCR. There was a big difference in DNA band pattern between two species of Sclerotium isolated in Korea. Electron micrographs of scanning electron microscope and transmission electron microscope showed morphological differences in sclerotial surface structure and rind layers between two species of Sclerotium. There were more wrinkles and pore spaces on sclerotial surface of Sclerotium sp. forming anamorphic-shaped sclerotia than that of Sclerotium cepivorum forming round-shaped sclerotia. Both of two white rot pathogens grew well at the temperature range of $10-25^{\circ}C$ with optimal temperature of $20^{\circ}C$. Sclerotia of the two pathogens were well formed at $20^{\circ}C$ and well germinated at the temperature range of $20-24^{\circ}C$, Effect of pre-incubation of sclerotia on destruction of sclerotial dormancy of two pathogens was evaluated through storing sclerotia under different temperature condition. The sclerotia of the two pathogens showed an increased capacity to germinate on potato dextroise agar when the sclerotia were incubated for 7 days at $10^{\circ}C$ after pre-treatment at $35^{\circ}C$ for 7 days. At that time, germination rate of Sclerotium sp. and 5. cepivorum was $100\%\;and\;70\%$, respectively. Flooding period and treatment temperature had an effect on sclerotial survival rate of the two pathogens. As flooding period and treatment temperature increased, sclerotial germination rate of the two pathogens decreased. It was confirmed that soil humidity played an important role on development of white rot. It was the highest disease incidence of garlic white rot when garlic were sown at potted soils infested with the two pathogens and adjusted soil humidity to $15\%$ (field moisture capacity, about -300 mb). As soil humidity increase or decrease based on $15\%$ of soil humidity, disease incidence decreased move and more.

Role of Cordycepin and Adenosine on the Phenotypic Switch of Macrophages via Induced Anti-inflammatory Cytokines

  • Shin, Seul-Mee;Moon, Sun-Hee;Park, Yoon-Hee;Kwon, Jeong-Hak;Lee, Seung-Jeong;Lee, Chong-Kil;Cho, Kyung-Hae;Kim, Kyung-Jae
    • IMMUNE NETWORK
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    • v.9 no.6
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    • pp.255-264
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    • 2009
  • Background: Chronic low grade inflammation is closely linked to type II diabetes, obesity, and atherosclerosis. Macrophages play a key role in the regulation of pro- or anti-inflammatory actions at the lesion sites of disease. Components of cordyceps militaris, cordycepin and adenosine, have been used for the modulation of inflammatory diseases. The effects of cordycepin in the modulation of macrophages have yet to be be elucidated. We investigated the effects of cordycepin and adenosine on the morphological changes of macrophages under the inflammatory condition of LPS and an anti-inflammatory condition involving high concentrations of adenosine. Methods: We confirmed the mRNA levels of the M1/M2 cytokine genes through RT-PCR and morphological change. Results: LPS-activated macrophages returned to their inactivated original shape, i.e., they looked like naive macrophages, through the treatment with high concentrations of cordycepin ($40{\mu}g/ml$). LPS and adenosine activated macrophages also returned to their original inactivated shapes after cordycepin treatment; however, at relatively higher levels of cordycepin than adenosine. This change did not occur with relatively low concentrations of cordycepin. Adenosine down-regulated the gene expression of M1 cytokines (IL-$1{\beta}$, TNF-${\alpha}$) and chemokines (CX3CR1, RANTES), such as cordycepin. Additionally, M2 cytokines (IL-10, IL-1ra, TGF-${\beta}$) were up-regulated by both cordycepin and adenosine. Conclusion: Based on these observations, both cordycepin and adenosine regulated the phenotypic switch on macrophages and suggested that cordycepin and adenosine may potentially be used as immunomodulatory agents in the treatment of inflammatory disease.

Isolation, Identification, and Characterization of Aero-Adaptive Campylobacter jejuni

  • LEE YOUNG-DUCK;MOON BO-YOUN;CHOI JUNG-PIL;CHANG HAK-GIL;NOH BONG-SOO;PARK JONG-HYUN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.992-1000
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    • 2005
  • Campylobacter is one of the emerging foodborne pathogens, and its worldwide incidence rate is extremely high. This study was undertaken to isolate and identify Campylobacter strains from chicken carcasses in the local markets, and analyze their characteristics regarding oxygen tolerance. They were isolated after aerobic enrichment and identified by biochemical, physiological, and morphological characteristics, PCR, and 16S rDNA sequencing. Their oxygen tolerances were analyzed in terms of the cell surface hydrophobicity, cell fatty acid composition, and oxidoreductase. Five strains of C. jejuni were isolated and identified from 61 isolates from 50 chickens. Among them, C. jejuni IC21 grew well in Brucella broth and commercial milk under aerobic condition. However, in the aerobic exposure, the cell surface hydrophobicity of C. jejuni IC21 was almost the same as the other isolates, even though its morphology changed from the spiral-bacilli form into the coccoid form. Fatty acid analyses showed that all Campylobacter strains had a high composition of $C_{19:1}$, cyclopropane fatty acid, and that the amount of the other fatty acids were very similar between them. Interestingly, however, only oxidoreductase activities of C. jejuni IC21 increased highly under aerobic exposure even though its activities were almost the same as the other C. jejuni strains just after microaerobic culture. It had 11.8 times higher catalase activity, 4.4 times higher for SOD, and 2.0 times higher for NADH oxidase activities. Therefore, in the case of the aero-adaptive C. jejuni IC21, expression of oxidoreductase significantly increased under oxidative stressed condition, which might allow it to survive for a longer time and grow on food under aerobic exposure. Such new strain might be one of the explanations for the increase of campylobacteriosis.

Differentiation Inductions Altered Telomere Length and Telomerase Activity in Human Dental Pulp-Derived Mesenchymal Stem Cell

  • Lee, Hyeon-Jeong;Jeon, Ryoung-Hoon;Park, Byung-Joon;Jang, Si-Jung;Lee, Sung-Lim;Rho, Gyu-Jin;Kim, Seung-Joon;Lee, Won-Jae
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.2
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    • pp.93-99
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    • 2019
  • Telomeres are known as a specialized region in the end of chromosomes to protect DNA destruction, but their lengths are shortened by repetition of cell division. This telomere shortening can be preserved or be elongated by telomerase and TERT expression. Although a certain condition in the cells may affect to the cellular and molecular characteristics, the effect of differentiation induction to telomere length and telomerase activity in mesenchymal stem cells (MSCs) has been less studied. Therefore, the present study aimed to uncover periodical alterations of telomere length, telomerase activity and TERT expression in the dental pulp-derived MSCs (DP-MSCs) under condition of differentiation inductions into adipocytes and osteoblasts on a weekly basis up to 3 weeks. Shortening of telomere was significantly (p < 0.05) identified from early-middle stages of both differentiations in comparison with undifferentiated DP-MSCs by non-radioactive chemiluminescent assay and qRT-PCR method. Telomere length in undifferentiated DP-MSCs was 10.5 kb, but the late stage of differentiated DP-MSCs which can be regarded as the adult somatic cell exhibited 8.1-8.6 kb. Furthermore, the relative-quantitative telomerase repeat amplification protocol or western blotting presented significant (p < 0.05) decrease of telomerase activity since early stages of differentiations or TERT expression from middle stages of differentiations than undifferentiated state, respectively. Based on these results, it is supposed that shortened telomere length in differentiated DP-MSCs was remained along with prolonged differentiation durations, possibly due to weakened telomerase activity and TERT expression. We expect that the present study contributes on understanding differentiation mechanism of MSCs, and provides standardizing therapeutic strategies in clinical application of MSCs in the animal biotechnology.