• Title/Summary/Keyword: Cell-based assay

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Effects of the cis-Acting Element in the 3' End of Porcine $\beta$-Casein Gene on the Expression in Mammary Epithelial Cells (돼지 $\beta$-Casein 유전자의 3' 말단 부위의 cis-Acting Element가 유선 상피 세포내의 발현에 미치는 영향)

  • Lee, Hwi-Cheul;Kim, Byoung-Ju;Byun, Sung-June;Lee, Seung-Hoon;Kim, Min-Ji;Chung, Hee Kyoung;Lee, Hyun-Gi;Jo, Su-Jin;Chang, Won-Kyong;Park, Jin-Ki;Lee, Poong-Yeon
    • Reproductive and Developmental Biology
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    • v.32 no.3
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    • pp.153-158
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    • 2008
  • Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.

Diagnostic Value of Serum Procalcitonin in Febrile Infants Under 6 Months of Age for the Detection of Bacterial Infections (발열이 있는 6개월 미만의 영아에서 세균성 감염에 대한 procalcitonin의 진단적 가치)

  • Kim, Nam Hyo;Kim, Ji Hee;Lee, Taek Jin
    • Pediatric Infection and Vaccine
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    • v.16 no.2
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    • pp.142-149
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    • 2009
  • Purpose : The aim of this study was to determine the diagnostic value of serum procalcitonin (PCT) compared with that of C-reactive protein (CRP) and the total white blood cell count (WBC) in predicting bacterial infections in febrile infants<6 months of age. Methods : A prospective study was performed with infants <6 months of age who were admitted to the Department of Pediatrics with a fever of uncertain source between July and September 2008. Spinal taps were performed according to clinical symptoms and physical examination. Serum PCT levels were measured using an enzyme-linked fluorescent assay. Results : Seventy-one infants (mean age, 2.62 months) were studied. Twenty-six infants (36.6%) had urinary tract infections (UTIs), and 22 infants (31.0%) had viral meningitis. The remaining infants had acute pharyngitis (n=1), herpangina (n=1), upper respiratory tract infections (n=7), acute bronchiolitis (n=8), acute gastroenteritis (n=4), and bacteremia (n=2). The median WBC and CRP levels were significantly higher in infants with UTIs than in infants with viral meningitis. However, there were no differences in the median PCT levels between the groups (0.14 ng/mL vs. 0.11 ng/mL, P=0.419). The area under the receiver operating characteristic curve was 0.792 (95% CI, 0.65-0.896) for WBC, 0.77 (95% CI, 0.626-0.879) for CRP, and 0.568 (95% CI, 0.417-0.710) for PCT. An elevated WBC count (>11,920/${\mu}L$) and an increased CRP level (>1.06mg/dL) were significant predictors of UTIs based on multiple logistic regression analysis. Conclusion : Serum PCT concentrations should be interpreted with caution in infants <6 months of age with a fever of uncertain source.

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Association of Genetic Polymorphism of Glutathione S-transferase M1, T1 and N-acetyltransferase 1 with Lung Cancer (폐암발생과 Glutathione S-transferase M1, T1 및 N-acetyltransferase 1의 유전적 다형성과의 연관성에 관한 연구)

  • Lee, Seung-Joon;Park, Gye-Young;Oh, Yeon-Mok;Kang, Dae-Hee;Cho, Soo-Hun;Kim, Soo-Ung;Yoo, Chul-Gyu;Lee, Chun-Taeck;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.4
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    • pp.471-477
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    • 1999
  • Background : Smoking and high-risk occupation have been known to be the risk factors of lung cancer. The carcinogen-metabolizing enzymes in human body such as glutathione S-transferase M1, T1 and N-acetyltransferase 1 have also been regarded as risk factors in many cancers, because the activities of those enzymes play a role in metabolizing the carcinogen. A case-control study was conducted to evaluate the genetic polymorphism of GSTM1, T1 and NAT1 in lung carcinogenesis in Korean men. Methods : The histologically proven lung cancer cases were recruited from Seoul National University Hospital. The patients of more than 40-year-old with the nonmalignant urinary tract diseases were recruited as controls from the same hospitals. The informations of demographical characteristics and smoking were obtained by interview or chart review and the genetic polymorphisms of GSTM1, T1 and NAT1 were determined by PCR-based assay. The statistical analyses were performed by linear logistic regression. Results : The number of case-control was 118 and 150, respectively. The smoking history was significantly higher in the lung cancer patients than the controls. The prevalence of GSTM1 null-type was statistically higher(OR=2.25 ; 95% CI=1.12-4.51) in squamous cell carcinoma than other genotypes, but other histologic types were not The prevalence of GSTT1 null-type were not statistically higher than other genotypes in all histologic types. The fast acetylator of NAT1 was more prevalent than normal(OR=2.13 ; 95% CI=1.04-4.40) in all lung cancer patients. Conclusion : The null-type of GSTM1 and fast acetylator of NAT1 are associated with development of lung cancer in Korean men.

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Effects of Quinone Reductase Induction and Cytotoxicity of the Angelica radix Extracts (당귀 추출성분의 세포독성 효과와 Quinone Reductase 유도활성 효과)

  • 배송자;한은주;노승배
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.1
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    • pp.147-152
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    • 2000
  • Various lines of evidence suggest that dietary components protect the initiation of carcinogenesis. In this study, the ethanol extracts (AGE) and the methanol and hexane partition layers (AGEM, AGEH) of the Angelica radix were screened for their cytotoxic effects using the MTT assay on HepG2, HeLa, MCF7 and SW626 cells and for their ability to induce quinone reductase (QR) in HepG2 cells. AGEM and AGEH of the Angelica radix showed the strongest cytotoxic effects on HepG2 and HeLa cells. Cell growth was inhibited by 99.8% and 99.8% on HepG2 cells and 99.3% and 99.4% on HeLa cells, at dose of $100\;\mu\textrm{g}/ml$ of AGEM and AGEH extracts respectively. AGE and AGEH significantly induced QR activities in the HepG2 cells. The QR activities of HepG2 cells grown in the presence of AGE, AGEH, and AGEM at the concentration of $50\;\mu\textrm{g}/mL$ were 313.5, 273.3 and 133.3 nmol/min/mg protein, respectively. Therefore, based on these studies, Angelica radix may be developed into a potentially useful cancer chemopreventive agent.

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LP-M, a Novel Butanol-Extracts Isolated from Liriope platyphylla, could Induce the Neuronal Cell Survival and Neuritic Outgrowth in Hippocampus of Mice through Akt/ERK Activation on NGF Signal Pathway (맥문동(Liriope platyphylla)의 새로운 부탄올 추출물인 LP-M이 Akt/ERK NGF receptor signaling pathway를 통해 뇌조직에서 신경세포의 생존과 성장에 미치는 영향에 관한 연구)

  • Nam, So-He;Choi, Sun-Il;Goo, Jun-Seo;Kim, Ji-Eun;Lee, Yoen-Kyung;Hwang, In-Sik;Lee, Hye-Ryun;Lee, Young-Ju;Lee, Hong-Gu;Choi, Young-Whan;Hwang, Dae-Youn
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1234-1243
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    • 2011
  • Liriope platyphylla has been used in oriental medicine as an effective medical plant to improve symptoms of cough, sputum production, neurodegenerative disorders, obesity and diabetes for long time. In order to investigate the effects of novel extracts on nerve growth factors (NGF)-stimulated neuritic outgrowth, the alteration of NGF expression and NGF receptor signaling pathway were detected in neuroblastoma cells and C57BL/6 mice. Of a total of 13 novel extracts, 4 extracts (LP-E, LP-M, LP-M50, LP2E17PJ) showed high viability on MTT assay. Also, all of these extracts induced NGF secretion and NGF mRNA expression in neuroblastoma cells. However, the NGF-induced neuritic outgrowth from PC12 cells was only stimulated by LP-E, LP-M and LP-M50. Furthermore, we selected LP-M as a best candidate, based on method and amounts of extraction, in order to verify its effect in mice. C57BL/6 mice were treated with 50 mg/kg of LP-M for 2 weeks and the effects on NGF regulation were analyzed with various methods. The expression of NGF mRNA was significantly increased in LP-M treated mice compared to vehicle treated mice. Also, the signaling pathway of p75NTR was inhibited in the cortex by LP-M treatment, with no change in the hippocampus of brain. However, the signaling pathway of TrkA was dramatically activated in only hippocampus via LP-M treatment. Therefore, these results suggest that the novel four extracts of L. platyphylla may contribute to the regulation of NGF expression and secretion in neuronal cells. LP-M was especially considered to be an excellent candidate for a neurodegenerative disease-therapeutic drug.

Antimutagenicity and Immuno Activity of Extracts from Epimedium koreanum Nakai Containing Different Icariin Quantity (Icariin 함량에 따른 삼지구엽초 추출물의 항돌연변이 및 면역활성)

  • Park, Myoung-Su;Kim, Seo-Jin;Forghani, Fereidoun;Rahman, S.M.E.;Eo, Ji-Hyun;Eun, Jong-Bang;Oh, Deog-Hwan
    • Food Science and Preservation
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    • v.18 no.6
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    • pp.938-945
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    • 2011
  • Epimedium koreanum Nakai is a wild herb commonly consumed in South Korea due to its beneficial health effects. In the present study, the antimutagenic and immunoactivities of extracts from E. koreanum Nakai containing different icariin quantities were investigated for food use. In the Ames test, both the water and ethanol extracts were found not to have a mutagenic effect on Salmonella typhimurium TA98 and TA100, respectively. The E. koreanum Nakai extracts showed over 80 and 90% antimutagenic effects on benzo(${\alpha}$)pyrene (B(a)P) in S. typhimurium TA98 and TA100, respectively. Moreover, all the extracts showed over 70% antimutagenicity on S. typhimurium TA98 and TA100 against 4-nitroquinoline-1-oxide (4NQO). The E. koreanum Nakai extract with ethanol showed strong antimutagenic activity, higher than that of the water extract and the sequenced KE9412, KE9408, and KE9405. In the immunomodulating activity test, the effect of E. koreanum Nakai on the B (Rhamos) and T (Jukat) cells were investigated. The immunoactivity results showed that the growth and viability of the B and T cells increased and were activated more in KE9405 (1.8 times), KE9408 (1.6 times), and KE9412 (1.32 times) in the water extracts, and least in KE9412 (1.74 times), KE9408 (1.52 times), and KE9405 (1.4 times) in the ethanol extracts. In the case of both the water and ethanol extracts ($500{\mu}g/mL$) from E. koreanum Nakai, the highest cell number of the human B (Rhamos) and T (Jukat) cells was observed on day 4 in KE9405 and KE9412, and on day 5 in KE9408. Based on the obtained results, the development of E. koreanum Nakai as a food material is recommended.

Biochemical Assessment of Deer Velvet Antler Extract and its Cytotoxic Effect including Acute Oral Toxicity using an ICR Mice Model (ICR 마우스 모델을 이용한 녹용 추출물의 생화학적 평가 및 급성 경구 독성을 포함한 세포 독성 효과)

  • Ramakrishna Chilakala;Hyeon Jeong Moon;Hwan Lee;Dong-Sung Lee;Sun Hee Cheong
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.430-441
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    • 2023
  • Velvet antler is widely used as a traditional medicine, and numerous studies have demonstrated its tremendous nutritional and medicinal values including immunity-enhancing effects. This study aimed to investigate different deer velvet extracts (Sample 1: raw extract, Sample 2: dried extract, and Sample 3: freeze-dried extract) for proximate composition, uronic acid, sulfated glycosaminoglycan, sialic acid, collagen levels, and chemical components using ultra-performance liquid chromatography-quadrupole-time-of-light mass spectrometry. In addition, we evaluated the cytotoxic effect of the deer velvet extracts on BV2 microglia, HT22 hippocampal cells, HaCaT keratinocytes, and RAW264.7 macrophages using the cell viability MTT assay. Furthermore, we evaluated acute toxicity of the deer velvet extracts at different doses (0, 500, 1000, and 2000 mg/kg) administered orally to both male and female ICR mice for 14 d (five mice per group). After treatment, we evaluated general toxicity, survival rate, body weight changes, mortality, clinical signs, and necropsy findings in the experimental mice based on OECD guidelines. The results suggested that in vitro treatment with the evaluated extracts had no cytotoxic effect in HaCaT keratinocytes cells, whereas Sample-2 had a cytotoxic effect at 500 and 1000 ㎍/mL on HT22 hippocampal cells and RAW264.7 macrophages. Sample 3 was also cytotoxic at concentrations of 500 and 1000 ㎍/mL to RAW264.7 and BV2 microglial cells. However, the mice treated in vivo with the velvet extracts at doses of 500-2000 mg/kg BW showed no clinical signs, mortality, or necropsy findings, indicating that the LD50 is higher than this dosage. These findings indicate that there were no toxicological abnormalities connected with the deer velvet extract treatment in mice. However, further human and animal studies are needed before sufficient safety information is available to justify its use in humans.