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Secondary Structure of the Ribosomal Internal Transcribed Spacer (ITS) Region of Hypsizygus marmoreus (느티만가닥버섯의 ITS (internal transcribed spacer) 영역의 2차구조 분석)

  • Woo, Ju-Ri;Yoon, Hyeokjun;You, Young-Hyun;Lee, Chang-Yun;Kong, Won-Sik;Kim, Jong-Guk
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1260-1266
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    • 2013
  • The ribosomal DNA (rDNA) clusters of Hypsizygus marmoreus 3-10 and H. marmoreus 1-1 were analyzed in this study. The small subunit (SSU) and intergenic spacer 2 (IGS 2) was partially sequenced. The internal transcribed spacer 1 (ITS 1), 5.8S, internal transcribed spacer 2 (ITS 2), large subunit (LSU), intergenic spacer 1 (IGS 1), and 5S were completely sequenced. The rDNA clusters of H. marmoreus 3-10 and H. marmoreus 1-1 were 7,049 bp in length. The sequence of SSU rDNA, which corresponded to 18S rDNA, was 1,796 bp in length, and the sequence of LSU rDNA, which corresponded to 28S rDNA, was 3,348 bp in length. The ITS region that variable region and IGS region that non-transcribed spacer was 462 bp and 1,290 bp in length. The sequence of 5.8S rDNA and 5S rDNA was 153 bp and 43 bp in length, respectively. The 17 bp of the rDNA cluster in the H. marmoreus 3-10 strain was different to that in the H. marmoreus 1-1 strain, with 2 bp in the SSU, 3 bp in the ITS, 9 bp in the LSU, and 3 bp in the IGS. The analysis of the secondary structure revealed that the ITS regions of H. marmoreus 3-10 and H. marmoreus 1-1 have five stem-loop structures. Interestingly, among these structures, one different nucleotide sequence resulted in a different secondary structure in stem-loop V.

Meat Species Identification using Loop-mediated Isothermal Amplification Assay Targeting Species-specific Mitochondrial DNA

  • Cho, Ae-Ri;Dong, Hee-Jin;Cho, Seongbeom
    • Food Science of Animal Resources
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    • v.34 no.6
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    • pp.799-807
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    • 2014
  • Meat source fraud and adulteration scandals have led to consumer demands for accurate meat identification methods. Nucleotide amplification assays have been proposed as an alternative method to protein-based assays for meat identification. In this study, we designed Loop-mediated isothermal amplification (LAMP) assays targeting species-specific mitochondrial DNA to identify and discriminate eight meat species; cattle, pig, horse, goat, sheep, chicken, duck, and turkey. The LAMP primer sets were designed and the target genes were discriminated according to their unique annealing temperature generated by annealing curve analysis. Their unique annealing temperatures were found to be $85.56{\pm}0.07^{\circ}C$ for cattle, $84.96{\pm}0.08^{\circ}C$ for pig, and $85.99{\pm}0.05^{\circ}C$ for horse in the BSE-LAMP set (Bos taurus, Sus scrofa domesticus and Equus caballus); $84.91{\pm}0.11^{\circ}C$ for goat and $83.90{\pm}0.11^{\circ}C$ for sheep in the CO-LAMP set (Capra hircus and Ovis aries); and $86.31{\pm}0.23^{\circ}C$ for chicken, $88.66{\pm}0.12^{\circ}C$ for duck, and $84.49{\pm}0.08^{\circ}C$ for turkey in the GAM-LAMP set (Gallus gallus, Anas platyrhynchos and Meleagris gallopavo). No cross-reactivity was observed in each set. The limits of detection (LODs) of the LAMP assays in raw and cooked meat were determined from $10pg/{\mu}L$ to $100fg/{\mu}L$ levels, and LODs in raw and cooked meat admixtures were determined from 0.01% to 0.0001% levels. The assays were performed within 30 min and showed greater sensitivity than that of the PCR assays. These novel LAMP assays provide a simple, rapid, accurate, and sensitive technology for discrimination of eight meat species.

Inhibition of DNA-dependent Protein Kinase by Blocking Interaction between Ku Complex and Catalytic Subunit of DNA-dependent Protein Kinase

  • Kim, Chung-Hui;Cuong, Dang-Van;Kim, Jong-Su;Kim, Na-Ri;Kim, Eui-Yong;Han, Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.1
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    • pp.9-14
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    • 2003
  • Recent studies indicated that cancer cells become resistant to ionizing radiation (IR) and chemotherapy drugs by enhanced DNA repair of the lesions. Therefore, it is expected to increase the killing of cancer cells and reduce drug resistance by inhibiting DNA repair pathways that tumor cells rely on to escape chemotherapy. There are a number of key human DNA repair pathways which depend on multimeric polypeptide activities. For example, Ku heterodimer regulatory DNA binding subunits (Ku70/Ku80) on binding to double strand DNA breaks (DSBs) are able to interact with 470-kDa DNA-dependent protein kinase catalytic subunit (DNA-PKcs), and are essential for DNA-dependent protein kinase (DNA-PK) activity. It has been known that DNA-PK is an important factor for DNA repair and also is a sensor-transmitting damage signal to downstream targets, leading to cell cycles arrest. Our ultimate goal is to develop a treatment of breast tumors by targeting proteins involved in damage-signaling pathway and/or DNA repair. This would greatly facilitate tumor cell cytotoxic activity and programmed cell death through DNA damaging drug treatment. Therefore, we designed a domain of Ku80 mutants that binds to Ku70 but not DNA end binding activity and used the peptide in co-therapy strategy to see whether the targeted inhibition of DNA-PK activity sensitized breast cancer cells to irradiation or chemotherapy drug. We observed that the synthesized peptide (HNI-38) prevented DNA-PKcs from binding to Ku70/Ku80, thus resulting in inactivation of DNA-PK activity. Consequently, the peptide treated cells exhibited poor to no DNA repair, and became highly sensitive to IR or chemotherapy drugs, and the growth of breast cancer cells was inhibited. Additionally, the results obtained in the present study also support the physiological role of resistance of cancer cells to IR or chemotherapy.

Characteristics of Enterotoxigenic genes and Antibiotic Susceptibility of Staphylococcus aureus Isolated from Tomato Farms in Western Gyeongnam (서부 경남지역 토마토 농장에서 분리된 Staphylococcus aureus의 내독소 유전자의 특성과 항생제 감수성)

  • Kim, Jin-Soo;Lee, Jin-Ha;Kim, Ji-Hun;Choi, Ju-Mi;Kim, Se-Ri;Ha, Sang-Do;Kim, Keun-Sung;Lee, Kyu-Ho;Kim, Min-Gon;Kim, Kwang-Yup;Kim, Cheol-Ho;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.38 no.2
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    • pp.295-303
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    • 2006
  • To provide microbial information for the safety of agricultural production, the presence of enterotoxin genes and antibiotic susceptibility of 14 isolated Staphylococcus aureus (11.7%) strains were investigated using PCR-based methods and disk diffusion method, respectively. Among enterotoxin-encoding genes, sea was detected from two isolates (14.3%), sea and sed genes were co-detected from three isolates (21.4%), and sea, sed, and see genes in seven isolates (50.0%), whereas seb, sec, and tsst were not detected in any isolate. Nine (64.3%), eight (57.1%), six (42.9%), two (14.3%), and one (7.2%) isolates were resistant to penicillin, novobiocin, amphicillin, erythromycin and oxacillin, and doxycycline and kanamycin, respectively. Methicilline-resistant S. aureus was found in roller of B farm and in hydroponic solution of D farm.

A Study on Perception of Shippers about the Service Quality of Logistics Center in Port Distripark (항만배후단지 물류센터 서비스품질에 대한 화주 인식 연구)

  • Kweon, Jung-Dae;Kwon, Kyu-Ri;Kim, Yul-Seong
    • Journal of Navigation and Port Research
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    • v.41 no.6
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    • pp.415-422
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    • 2017
  • In a situation where there is excessive competition among logistics centers due to the low price of logistics centers for attracting freight volume, the conditions provided by the logistics centers are similar. Therefore, in order to determine the logistics center, we want to find out what kind of differentiated service the shippers desire, as well as what level of service they want in addition to al ow price. There are currently no studies about the service quality of logistics centers. The components of the service quality of the logistics center were extracted by applying them to the logistics center based on the existing service quality theory. Factor analysis revealed five dimensions of service quality: tangibility, reliability, empathy, assurance, and know-how. It was found that service quality had a statistically significant influence on customer satisfaction, from the investigation of the causal effect relationship. In addition, 'Know-How' among these factors has more influence on customer satisfaction, so it is important to accumulate differentiated 'Know-How' only in logistics centers.

Quantitative Analysis of Milk-Derived microRNAs and Microbiota during the Manufacturing and Ripening of Soft Cheese

  • Oh, Sangnam;Park, Mi-Ri;Ryu, Sangdon;Maburutse, Brighton E.;Kim, Ji-Uk;Kim, Younghoon
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1566-1575
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    • 2017
  • MicroRNAs (miRNAs) are abundant in bovine milk and milk derived from other livestock, and they have functional roles in infants and in the secretion process of mammary glands. However, few studies have evaluated miRNAs in dairy processes, such as during cheese making and ripening. Thus, we investigated the characteristics of milk-derived miRNAs during the manufacturing and ripening of Camembert cheese as well as the microbiota present using the quantitative reverse transcription polymer chain reaction (RT-qPCR) and 16S rRNA pyrosequencing, respectively. Pyrosequencing showed that the cheese microbiota changed dramatically during cheese processing, including during the pasteurization, starter culture, and ripening stages. Our results indicated that the RNA contents per $200mg/200{\mu}l$ of the sample increased significantly during cheese-making and ripening. The inner cheese fractions had higher RNA contents than the surfaces after 12 and 22 days of ripening in a time-dependent manner (21.9 and 13.2 times higher in the inner and surface fractions than raw milk, respectively). We performed a comparative analysis of the miRNAs in each fraction by RT-qPCR. Large amounts of miRNAs (miR-93, miR-106a, miR-130, miR-155, miR-181a, and miR-223) correlated with immune responses and mammary glands were present in aged cheese, with the exception of miR-223, which was not present on the surface. Considerable amounts of miRNAs were also detected in whey, which is usually disposed of during the cheese-making process. Unexpectedly, there were no significant correlations between immune-related miRNAs and the microbial populations during cheese processing. Taken together, these results show that various functional miRNAs are present in cheese during its manufacture and that they are dramatically increased in amount in ripened Camembert cheese, with differences according to depth.

Prevalence of major enteric pathogens in different feeding groups of pig in Korean pig farms (국내 양돈장의 사육구간별 주요 소화기질병 원인체 유병율 조사)

  • Jung, Youn-Soo;Park, Yu-Ri;Kang, Dae-Young;Han, Do-Hyun;Yoon, Duhak;Jung, Byeong-Yeal;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.39 no.4
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    • pp.211-219
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    • 2016
  • For determining the prevalence of major enteric pathogens, clinical examination and etiological diagnosis were carried out on 75 Korean pig farms. Enteric disease-suspected signs were observed in 90.7% of the farms and the incidence and severity were higher in younger age groups of the pigs. Five of seven pathogens were detected in 375 fecal samples collected from the 75 farms, and the farm-level prevalence of porcine rotavirus group A (PoRVA), pathogenic Escherichia (E.) coli, Lawsonia (L.) intracelluraris, Salmonella spp., and Brachyspira (B.) hyodysenteriae was 54.7%, 54.7%, 16.0%, 10.7% and 2.7%, respectively. PoRVA was extensively infected in suckling and weaning pig groups. The prevalence of pathogenic E. coli was highest in suckling period, and after the period, it exhibited a tendency to decrease. Salmonella spp. and L. intracelluraris were detected in all feeding groups of pigs in a ratio of 1.3~6.7%. B. hyodysenteriae was detected in 1.3~2.7% of growing and fattening pig groups but not detected in suckling and weaning pig groups. At least one or more pathogens were detected in 30.1% of 375 fecal samples. Among these, 25.0% or 5.1% of cases were single or mixed infection. Enteric disease signs of the pigs were significantly co-related with the detection of PoRVA, pathogenic E. coli or Salmonella spp. (P<0.01) but not with L. intracelluraris or B. hyodysenteriae (P>0.05). Conclusively, it will be expected that these data obtained in this study are very useful for subsequent studies and prevention strategies for swine enteric disease in Korean pig farms.

Isolation and Yield Enhancement of Primo Vessels Inside of Rabbit Lymph Vessels by Using Sound Wave Vibration (음파진동을 이용한 토끼의 림프관내 프리모관 분리와 수득률 향상 특성 연구)

  • Heo, June-Yi;Chung, Ji-Hwan;Choi, Dong-Hee;Lee, Hye-Rie;Noh, Young-Il;Han, Moon-Young;Jeong, Yeon-Chul;Lee, Jae-Yeon;Seo, Kyeong-Ju;Park, Ji-Su;Kim, Nu-Ri;Lee, Kyu-Hwan;Bae, Yu-Mi;Lee, Eun-Sae;Lee, Sang-Suk
    • Korean Journal of Acupuncture
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    • v.30 no.2
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    • pp.122-130
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    • 2013
  • Objectives : The fact that Primo vessel as a new circulatory system exists in the lymphatic vessels of the rabbits which were treated with sound wave vibration therapy is confirmed with the anatomy of rabbit. Isolated Primo vessel is investigated, particularly focused on morphological features. Methods : Before the anatomy, a rabbit in a cage box was laid on the sound wave vibrating apparatus and then is applied by various frequencies and intensities for 30 min with music that a rabbit likes. Results : Isolation and observation of Primo vessel was easier when the lymphatic circular system was applied at a certain frequency and intensity of 7 Hz and 50. The probability of observation for Primo vessel enhanced to 90%. Conclusions : The sound wave therapy by a vibration apparatus is considered necessary as major process to facilitate the reproducible isolation and observation of Primo vessel.

Monitoring of nervous necrosis virus in fertilized eggs of walleye pollock (Gadus chalcogrammus) (명태(Gadus chalcogrammus) 수정란에서 신경괴사증바이러스(nervous necrosis virus) 모니터링)

  • Nam, U-Hwa;Lee, Jong-Hyuk;Kim, Mi-Ri;Jang, Su-Rim;Yoon, Do-Hyun;Seo, Joo-Young;Kwon, O-Nam;Kim, Jeong-Ho
    • Journal of fish pathology
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    • v.31 no.1
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    • pp.9-13
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    • 2018
  • We previously monitored nervous necrosis virus (NNV) in brain samples of artificially produced walleye Pollock (Gadus chalcogrammus) seedlings, with a low prevalence (1.8%, 1/55) but no clinical symptoms. Given that this virus is considered one of the most serious viral threats for almost all marine aquaculture fish species and characterized by both vertical and horizontal transmission, it would be interesting to monitor NNV in the fertilized eggs as well. We collected fertilized walleye pollock eggs from the farms located in Goseong during January to March, 2017. Approximately 50 mg of eggs were periodically taken from 4 each different batches, and 37 different pooled sample sets in total were made during sampling period. RNA was extracted from the eggs by using Trizol and cDNA was synthesized for RT-PCR for detecting NNV. Primers and PCR conditions are the same as previously described. As a result, NNV was not detected from any of the sample sets by one step PCR (0%, 0/37), suggesting NNV may not be a threat in walleye pollock aquaculture in Korea at present time. However, continuous monitoring for NNV should be conducted because introducing a new species into aquaculture industry involves potentials of disease outbreak and NNV is already known to cause outbreaks in gadoid fishes.

Antimetastatic Effects of Crude Polysaccharide Isolated from Polygonati Rhizoma on 4T1 Breast Cancer Cells by Activation of Innate Immune System (황정(黃精)으로부터 유래한 조다당류의 선천면역 활성에 의한 유방암 세포주 전이 억제 효과)

  • Ji, Hae-Ri;Hwang, Deok-Sang;Lee, Chang-Hoon;Jang, Jun-Bok;Lee, Jin-Moo
    • The Journal of Korean Obstetrics and Gynecology
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    • v.32 no.4
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    • pp.1-13
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    • 2019
  • Objective: This study is aimed to investigate the anti-tumor metastasis by innate immunomodulating effects of crude polysaccharide isolated from Polygonati Rhizoma (CP-PR) on 4T1 breast cancer cells. Methods: CP-PR was isolated from Polygonati Rhizoma. Antimetastatic experiments were conducted in vivo mouse model by using 4T1 breast cancer cells. The cell viability of CP-PR was tested with normal spleen and 4T1 breast cancer cells. To observe the activation of macrophages with/without 4T1 breast cancer cells, production of tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$), interleukin-6 (IL-6), IL-10 and IL-12 were measured with enzyme-linked immunosorbent assay (ELISA), respectively. In addition, the lysis of YAC-1 cells and the production of granzymes were measured to observe the activation of natural killer (NK) cell. Results: Intravenous administration of CP-PR significantly inhibited metastasis of 4T1 breast cancer cells. In an in vitro cytotoxicity analysis, CP-PR affected the growth of normal spleen and 4T1 breast cancer cells above specific concentration. The production of $TNF-{\alpha}$, IL-6, IL-10 and IL-12 were significantly increased in macrophages with CP-PR. As compared with control, CP-PR showed significantly higher production of $TNF-{\alpha}$, IL-10 and IL-12 in macrophages co-cultured with 4T1 breast cancer cells. The lysis of YAC-1 cells and the production of granzymes were significantly up regulated by CP-PR. Conclusion: CP-PR appears to have considerable activity on the anti-metastasis by activation of innate immune system.