• Title/Summary/Keyword: detection rate

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Validation of Stem-loop RT-qPCR Method on the Pharmacokinetic Analysis of siRNA Therapeutics (Stem-loop RT-qPCR 분석법을 이용한 siRNA 치료제의 생체시료 분석법 검증 및 약물 동태학적 분석)

  • Kim, Hye Jeong;Kim, Taek Min;Kim, Hong Joong;Jung, Hun Soon;Lee, Seung Ho
    • Journal of Life Science
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    • v.29 no.6
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    • pp.653-661
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    • 2019
  • The first small interfering RNA (siRNA) therapeutics have recently been approved by the Food and Drug Administration in the U.S., and the demand for a new RNA therapeutics bioanalysis method-which is essential for pharmacokinetics, including the absorption, distribution, metabolism, and excretion of siRNA therapeutics-is rapidly increasing. The stem-loop real-time qPCR (RT-qPCR) assay is a useful molecular technique for the identification and quantification of small RNA (e.g., micro RNA and siRNA) and can be applied for the bioanalysis of siRNA therapeutics. When the anti-HPV E6/E7 siRNA therapeutic was used in preclinical trials, the established stem-loop RT-qPCR assay was validated. The limit of detection was sensitive up to 10 fM and the lower limit of quantification up to 100 fM. In fact, the reliability of the established test method was further validated in three intra assays. Here, the correlation coefficient of $R^2$>0.99, the slope of -3.10 ~ -3.40, and the recovery rate within ${\pm}20%$ of the siRNA standard curve confirm its excellent robustness. Finally, the circulation profiles of siRNAs were demonstrated in rat serum, and the pharmacokinetic properties of the anti-HPV E6/E7 siRNA therapeutic were characterized using a stem-loop RT-qPCR assay. Therefore, the stemloop RT-qPCR assay enables accurate, precise, and sensitive siRNA duplex quantification and is suitable for the quantification of small RNA therapeutics using small volumes of biological samples.

INTENSIVE MONITORING SURVEY OF NEARBY GALAXIES (IMSNG)

  • Im, Myungshin;Choi, Changsu;Hwang, Sungyong;Lim, Gu;Kim, Joonho;Kim, Sophia;Paek, Gregory S.H.;Lee, Sang-Yun;Yoon, Sung-Chul;Jung, Hyunjin;Sung, Hyun-Il;Jeon, Yeong-beom;Ehgamberdiev, Shuhrat;Burhonov, Otabek;Milzaqulov, Davron;Parmonov, Omon;Lee, Sang Gak;Kang, Wonseok;Kim, Taewoo;Kwon, Sun-gill;Pak, Soojong;Ji, Tae-Geun;Lee, Hye-In;Park, Woojin;Ahn, Hojae;Byeon, Seoyeon;Han, Jimin;Gibson, Coyne;Wheeler, J. Craig;Kuehne, John;Johns-Krull, Chris;Marshall, Jennifer;Hyun, Minhee;Lee, Seong-Kook J.;Kim, Yongjung;Yoon, Yongmin;Paek, Insu;Shin, Suhyun;Taak, Yoon Chan;Kang, Juhyung;Choi, Seoyeon;Jeong, Mankeun;Jung, Moo-Keon;Kim, Hwara;Kim, Jisu;Lee, Dayae;Park, Bomi;Park, Keunwoo;O, Seong A
    • Journal of The Korean Astronomical Society
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    • v.52 no.1
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    • pp.11-21
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    • 2019
  • Intensive Monitoring Survey of Nearby Galaxies (IMSNG) is a high cadence observation program monitoring nearby galaxies with high probabilities of hosting supernovae (SNe). IMSNG aims to constrain the SN explosion mechanism by inferring sizes of SN progenitor systems through the detection of the shock-heated emission that lasts less than a few days after the SN explosion. To catch the signal, IMSNG utilizes a network of 0.5-m to 1-m class telescopes around the world and monitors the images of 60 nearby galaxies at distances D < 50 Mpc to a cadence as short as a few hours. The target galaxies are bright in near-ultraviolet (NUV) with $M_{NUV}$ < -18.4 AB mag and have high probabilities of hosting SNe ($0.06SN\;yr^{-1}$ per galaxy). With this strategy, we expect to detect the early light curves of 3.4 SNe per year to a depth of R ~ 19.5 mag, enabling us to detect the shock-heated emission from a progenitor star with a radius as small as $0.1R_{\odot}$. The accumulated data will be also useful for studying faint features around the target galaxies and other science projects. So far, 18 SNe have occurred in our target fields (16 in IMSNG galaxies) over 5 years, confirming our SN rate estimate of $0.06SN\;yr^{-1}$ per galaxy.

Efficient virus elimination for apple dwarfing rootstock M.9 and M.26 via thermotherapy, ribavirin and apical meristem culture (사과 왜성대목 M.9 및 M.26의 고온, ribavirin, 생장점 배양을 통한 바이러스 제거)

  • Kwon, Young Hee;Lee, Joung Kwan;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong;Huh, Yoon Sun;Park, Eui Kwang;Yoon, Yeo Joong
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.228-235
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    • 2019
  • Apple (Malus pumila) is one of the most economically important fruits in Korea. but virus infection has decreased the sustainable production of apples and caused serious problems such as yield loss and poor fruit quality. Virus or viroid infection including apple chlorotic leaf spot virus (ACLSV), apple stem pitting virus (ASPV), apple stem grooving virus (ASGV), apple mosaic virus (ApMV) and apple scar skin viroid (ASSVd) have been also reported in Korea. In many cases, as apple gets infected with virus and viroid with no specific symptoms, the damage and symptoms caused by the viruses are not detected. In our research, viruses in the rootstock were eliminated for a virus-free apple dwarfing rootstock of M.9 and M.26. The virus elimination methods were apical meristem culture, thermotherapy ($37^{\circ}C$, 6 weeks) and chemotherapy($Ribavirin^{(R)}$). The detection of apple viruses was accomplished by Enzyme-linked Immuno-Sorbent Assay (ELlSA) and reverse transcription-polymerase chain reaction (RT-PCR). RT- PCR method was 10 ~ 30% more sensitive than the ELISA method. The efficiency of virus elimination was enhanced in apical meristem culture method. The acquisition rate of virus-free apple dwarfing rootstocks was 30 ~ 40% higher in apical meristem culture. After the meristem culturing of M.9, the infection ratio of ACLSV, ASPV and ASGV was 45%, 60% and 50%, respectively. In the apple dwarfing rootstock of M.26, the infection ratio of ACLSV, ASPV and ASGV was 40%, 55% and 55%, respectively. Based on this study, the best method for the production of virus-free apple dwarfing rootstocks was the apical meristem culture.

A Case Study: Improvement of Wind Risk Prediction by Reclassifying the Detection Results (풍해 예측 결과 재분류를 통한 위험 감지확률의 개선 연구)

  • Kim, Soo-ock;Hwang, Kyu-Hong
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.23 no.3
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    • pp.149-155
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    • 2021
  • Early warning systems for weather risk management in the agricultural sector have been developed to predict potential wind damage to crops. These systems take into account the daily maximum wind speed to determine the critical wind speed that causes fruit drops and provide the weather risk information to farmers. In an effort to increase the accuracy of wind risk predictions, an artificial neural network for binary classification was implemented. In the present study, the daily wind speed and other weather data, which were measured at weather stations at sites of interest in Jeollabuk-do and Jeollanam-do as well as Gyeongsangbuk- do and part of Gyeongsangnam- do provinces in 2019, were used for training the neural network. These weather stations include 210 synoptic and automated weather stations operated by the Korean Meteorological Administration (KMA). The wind speed data collected at the same locations between January 1 and December 12, 2020 were used to validate the neural network model. The data collected from December 13, 2020 to February 18, 2021 were used to evaluate the wind risk prediction performance before and after the use of the artificial neural network. The critical wind speed of damage risk was determined to be 11 m/s, which is the wind speed reported to cause fruit drops and damages. Furthermore, the maximum wind speeds were expressed using Weibull distribution probability density function for warning of wind damage. It was found that the accuracy of wind damage risk prediction was improved from 65.36% to 93.62% after re-classification using the artificial neural network. Nevertheless, the error rate also increased from 13.46% to 37.64%, as well. It is likely that the machine learning approach used in the present study would benefit case studies where no prediction by risk warning systems becomes a relatively serious issue.

Anti-listeria Activity of Lactococcus lactis Strains Isolated from Kimchi and Characteristics of Partially Purified Bacteriocins (김치에서 분리한 Lactococcus lactis 균주의 항리스테리아 활성 및 부분 정제된 박테리오신의 특성)

  • Son, Na-Yeon;Kim, Tae-Woon;Yuk, Hyun-Gyun
    • Journal of Food Hygiene and Safety
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    • v.37 no.2
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    • pp.97-106
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    • 2022
  • Listeria monocytogenes (L. monocytogenes) is one of gram-positive foodborne pathogens with a very high fatality rate. Unlike most foodborne pathogens, L. monocytogenes is capable of growing at low temperatures, such as in refrigerated foods. Thus, various physical and chemical prevention methods are used in the manufacturing, processing and distribution of food. However, there are limitations to the methods such as possible changes to the food quality and the consumer awareness of synthetic preservatives. Thus, the aim of this study was to evaluate the anti-listeria activity of lactic acid bacteria (LAB) isolated from kimchi and characterize the bacteriocin produced by Lactococcuslactis which is one of isolated strains from kimchi. The analysis on the anti-listeria activity of a total of 36 species (Lactobacillus, Weissella, Lactobacillus, and Lactococcus) isolated from kimchi by the agar overlay method revealed that L. lactis NJ 1-10 and NJ 1-16 had the highest anti-listeria activity. For quantitatively analysis on the anti-listeria activity, NJ 1-10 and NJ 1-16 were co-cultured with L. monocytogenes in Brain Heat Infusion (BHI) broth, respectively. As a result, L. monocytogenes was reduced by 3.0 log CFU/mL in 20 h, lowering the number of bacteria to below the detection limit. Both LAB strains showed anti-listeria activity against 24 serotypes of L. monocytogenes, although the sizes of clear zone was slightly different. No clear zone was observed when the supernatants of both LAB cultures were treated with proteinase-K, indicating that their anti-listerial activities might be due to the production of bacteriocins. Heat stability of the partially purified bacteriocins of NJ 1-10 and NJ 1-16 was relatively stable at 60℃ and 80℃. Yet, their anti-listeria activities were completely lost by 60 min of treatment at 100℃ and 15 min of treatment at 121℃. The analysis on the pH stability showed that their anti-listeria activities were the most stable at pH 4.01, and decreased with the increasing pH value, yet, was not completely lost. Partially purified bacteriocins showed relatively stable anti-listeria activities in acetone, ethanol, and methanol, but their activities were reduced after chloroform treatment, yet was not completely lost. Conclusively, this study revealed that the bacteriocins produced by NJ 1-10 and NJ 1-16 effectively reduced L. monocytogenes, and that they were relatively stable against heat, pH, and organic solvents, therefore implying their potential as a natural antibacterial substance for controlling L. monocytogenes in food.

Effect of Gastric Cancer Screening on Patients with Gastric Cancer: A Nationwide Population-based Study (위암 환자에서 국가암검진의 효과)

  • Cho, Young Suk;Lee, Sang Hoon;So, Hyun Ju;Kim, Dong Wook;Choi, Yoon Jung;Jeon, Han Ho
    • Journal of Digestive Cancer Reports
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    • v.8 no.2
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    • pp.102-108
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    • 2020
  • Background: This study was performed to evaluate the effect of gastric cancer screening through analysis of screening-related data. Methods: We investigated claims data of gastric cancer from 2009 to 2015. We evaluated whether the screening was performed to prior to registration as patients with gastric cancer. The effect of gastric cancer screening was also analyzed by gender. Results: We collected total 196,293 patients with gastric cancer. 74% of them had previous experience of gastric cancer screening. In patients with screening, early gastric cancer was 33.4% and advanced gastric cancer was 17.3%. 22,548 (15.5%) patients were diagnosed with gastric cancer within 2 years after screening. In the case of patients without screening, early gastric cancer was 15.1% and advanced gastric cancer was 25.3%. In case of men, 76% of them confirmed gastric cancer through screening, and 70.2% of women confirmed the gastric cancer. In both men and women, the rate of early gastric cancer was higher among those with screening than those without screening. Conclusion: In this study, we were able to indirectly confirm the stage shift of gastric cancer screening. However, within 2 years after screening, not a few patients with gastric cancer were diagnosed. Therefore, more studies are warranted to in the future.

Validation of an Analytical Method for Deacetylasperulosidic acid, Total Sugar and Monosaccharide Analysis in Fermented Morinda citrifolia Polysaccharide Powder (발효노니 다당체 분말의 deacetylasperulosidic acid, 총당 및 단당류 분석법 검증)

  • Kwon, Heeyeon;Choi, Jisoo;Kim, Soojin;Kim, Eunmin;Uhm, Jihyun;Kim, Bokyung;Lee, Jaeyeon;Kim, Yongdeok
    • Journal of Food Hygiene and Safety
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    • v.37 no.4
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    • pp.216-224
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    • 2022
  • This study was aimed at validating the analysis methods for deacetylasperulosidic acid (DAA), total sugar, galacturonic acid, glucose, and galactose, which are the indicator components of fermented Morinda citrifolia polysaccharide extract (Vitalbos). We modified the previously reported methods for validating the analytical methods. The specificity, linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ) were measured using phenol-sulfuric acid method and high-performance liquid chromatography (HPLC). The retention time and spectrum of the standard solution of Vitalbos coincided, confirming the specificity. The calibration curve correlation coefficient (R2), of five indicator components, ranged from 0.9995-0.9998, indicating excellent linearity of 0.99 or more. The intra-day and inter-day precision range of the assay was 0.14-3.01%, indicating a precision of less than 5%. The recovery rate was in the range of 95.13-105.59%, presenting excellent accuracy. The LOD ranged from 0.39 to 0.84 ㎍/mL and the LOQ ranged from 1.18 to 2.55 ㎍/mL. Therefore, the analytical method was validated for DAA, total sugar, galacturonic acid, glucose, and galactose, in Vitalbos. The indicator component content in Vitalbos was determined using a validated method. The contents of DAA, total sugar, galacturonic acid, glucose, and galactose were 2.31±0.06, 475.92±5.95, 72.83±1.05, 71.63±2.44, and 67.30±2.31 mg/g of dry weight, respectively. These results suggest that the developed analytical method is efficient and could contribute to the quality control of Vitalbos, as a healthy functional food material.

Study on the Physiology of Optimal reproductive age in Korean Native Cattle (한우의 최적 번식적령기의 생리적 현상에 관한 연구)

  • Seong, H.H.;Lee, J.H.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.4 no.1
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    • pp.110-118
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    • 2002
  • This study was conducted to investigate the physiology of optimal reproductive age in Hanwoo for protection to decrease of reproductivity and improvement of production of offspring. Thirty two cows were devided into 4 groups of treatments : T1(12 months of age and 0.5kg daily gain), T2(12 months of age and 0.8kg daily gain), T3(15 months of age and 0.5kg daily gain) and T4(18 months of age and 0.5kg daily gain). The days of the first heat of treated cows were 263.3±6.4 days and average weight was 181.1±11.3kg. The conception rates of first insemination were 25%(T1) and 75%(T4), and the number of insemination of T3 and T4(both 1.5) was lower than those of T1 and T2(2.3 and 2.4), respectively. With regard to estrus return after the first parturition, T1, T2, T3 and T4 showed 66.2,76.7, 62.4 and 68.5 days, respectively, indicating the average days of estrus return was 65.7. Plasma progesterone(P4) concentration was nearly the same during the observation periods of treated cows and P4 began to be detected after 12months. Only 5(15.6%) out of 32cows showed normal estrus cycle and ovulation before 12 months. During the peri-parturition period, P4 concentration was rapidly decreased and there was no detection of P4 from parturition to 40 days after milking. P4 would be released again on 45 day after parturition. The results imply that the optimal reproductive age of Hanwoo heifers would be around at the 14 months of age, 110cm height and 265kg weight.

Occurrence of Viruses and Viroids in Chrysanthemum Plants (Dendranthema morifolium) Cultivated in Yesan-gun, Chungcheongnam-do in Korea (충남 예산 지역의 국화에서 바이러스 및 바이로이드 병들의 발생 현황)

  • Yoon Hyun, Bang;Eun Gyeong, Song;Younghye, Lee;Ki Hyun, Ryu
    • Research in Plant Disease
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    • v.28 no.4
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    • pp.237-244
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    • 2022
  • Chrysanthemum plants are one of the most economically important plants in South Korea. Both virus and viroid can cause diseases and economic damage to the plants. In this study, we investigated the detection of seven viruses and two viroids in 350 chrysanthemum plants cultivated in Yesan-gun, Chungcheongnam-do. Two viruses, chrysanthemum virus B (CVB) and tomato aspermy virus (TAV), and two viroids, chrysanthemum chlorotic mottle viroid (CChMVd) and chrysanthemum stunt viroid (CSVd), were detected in this study. The two viruses were detected in six samples and one sample, respectively. The two viroids were detected in 97 samples and 21 samples, respectively. The nucleotide sequences of the CVB-CN-Y, TAV-CN-Y, CChMVd-CN-Y, and CSVd-CN-Y obtained in this study showed 83.7-86.9%, 99.2-100.0%, 94.4-99.5%, and 95.7-99.7% identity, respectively, compared to their other strains/isolates. The CVB-CN-Y and TAV-CN-Y showed the greatest nucleotide sequence homology to CVB-GS1 and three TAV isolates (TAV-V, TAV-P, and TAV-ChJ), respectively. The CChMVd-CN-Y and CSVd-CN-Y showed the greatest nucleotide sequence homology to CChMVd-Horst and four CSVd isolates (Au1.1, K4pop, Sagae, and Tochigi), respectively. This study is the report on the infection rate of viruses and viroids in chrysanthemum plants cultivated in Yesan-gun in 2021.

Tumorigenesis after Injection of Lung Cancer Cell Line (SW-900 G IV) into the Pleural Cavity of Nude Mice (누드마우스의 흉강에 폐암세포주의 주입에 의한 종양형성과 HER2/neu와 TGF-${\beta}_1$의 발현)

  • Park, Eok-Sung;Kim, Song-Myung;Kim, Jong-In
    • Journal of Chest Surgery
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    • v.43 no.6
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    • pp.588-595
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    • 2010
  • Background: Base on types of tumor, the types of expressed tumor is diverse and the difference in its expression rate is even more various. Due to such reasons an animal model is absolutely needed for a clinical research of lung cancer. The author attempted oncogenesis by cultivating a cell line of non-small cell carcinoma and then injecting it inside thoracic cavities of nude mice. The author conducted quantitative analyses of HER2/neu tumor gene - an epidermal growth factor receptor (EGFR) related to lung cancer, and TGF-${\beta}_1$, which acts as a resistance to cell growth inhibition and malignant degeneration. In order to investigate achievability of the oncogenesis, histological changes and the expression of cancer gene in case of orthotopic lung cancer is necessary. Material and Method: Among 20 immunity-free male BALB/c, five nude mice were selected as the control group and rest as the experimental group. Their weights ranged from 20 to 25 gm (Orient, Japan). After injection of lung cancer line (SW900 G IV) into the pleural cavity of nude mice, They were raised at aseptic room for 8 weeks. HER2/neu was quantitatively analyzed by separating serum from gathered blood via chemiluminiscent immunoassay (CLIA), and immunosandwitch method was applied to quantitatively analyze TGF-${\beta}_1$. SPSS statistical program (SPSS Version 10.0, USA) was implemented for statistical analysis. Student T test was done, and cases in which p-value is less than 0.05 were considered significant. Result: Even after lung cancer was formed in the normal control group or after intentionally injected lung cancer cell line, no amplification of HER2/neu gene showed reaction. However, the exact quantity of TGF-${\beta}_1$ was $28,490{\pm}8,549pg/mL$, and the quantity in the group injected with lung cancer cell was $42,362{\pm}14,449pg/mL$, meaning 1.48 times highly Significant (p<0.483). It proved that HER2/neu gene TGF-${\beta}_1$ had no meaningful interconnection. Conclusion: TGF-${\beta}_1$ gene expressed approximately 1.48 times amplification in comparison to the control group. The amplification of TGF-${\beta}_1$ meant somatic recuperation inhibition mechanism due to carcinogenesis in nude mice was definitely working. It may be implemented as a quantitative analysis that allows early detection of lung cancer in human body.