• Title/Summary/Keyword: m/RT

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Geographical origin discrimination of Korean variety, Angelica gigas Nakai by using TDU (Thermal Desorption Unit)-GC/MS (TDU-GC/MS를 이용한 한국품종 참당귀 국산 판별 기법)

  • Lee, Mi-Na;Kim, Yoon-Suk;Kim, Won-Il;Kim, Jeong-Gyu;Kwon, Oh-Kyung
    • Journal of Applied Biological Chemistry
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    • v.63 no.1
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    • pp.29-33
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    • 2020
  • The objective of this study was to develop a fast and accurate method of variety discrimination and geographical discrimination origin of Korean angelica (Korean variety, Angelica gigas Nakai) by using TDU-GC/MS. Two peaks of decursin and decursinol, which are coumarin derivatives were identified in the range of Total Ion Chromatogram (TIC) RT 26.9-27.2 of the Korean angelica by GC/MS analysis at the time of condensation in a refrigerated condensation system after thermal desorption of sample extracts. In case of Chinese angelica (Chinese variety), ligustilide peak was detected at the RT 17.2. In order to investigate the difference of volatile components according to the geographical origin of Korean variety, the mass spectra were measured by TDU-GC/MS at the range of m/z 40-400 amu. The TIC of domestic cultivation and Chinese cultivation of the Korean variety, Angelica gigas Nakai showed the same tendency as a whole. However, in partial scans of TIC, two peaks detected at 15.54 and 16.05 of RT showed different peak patterns between Korean angelica (Korean variety) cultivated in Korea and in China. The ratio of Peak A (RT 15.54) and B (RT 16.05) was 0.0-0.2 for domestic cultivation and 0.5-2.8 for Chinese cultivation, confirming the possibility of discriminating origin by comparing the TIC peak pattern of TDU-GC/MS.

Early Efficacy of Taxotere and Cisplatin Chemo-Radiotherapy for Advanced Cervical Cancer

  • Ke, Qing-Hua;Zhou, Shi-Qiong;Du, Wei;Lei, Yong;Huang, Min;Luo, Fei;Yang, Ji-Yuan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.2
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    • pp.617-619
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    • 2012
  • The aim of this study was to investigate the early outcome of the taxotere and cisplatin chemoradiotherapy for advanced cervical cancer. Fifty-six cases (FIGO II b to IVa) were divided randomly into two groups: radiotherapy alone (28 cases) and radiation plus chemotherapy (TP) group. There was no difference in radiotherapy between the two groups. The RT+C cases who received TP regimen during the radiation, and DDP once weekly injection of vain, according to 20$mg/m^2$ and taxotere once weekly iv according to 35 $mg/m^2$. These regimens were given for 4~5weeks, and some medicines to control vomiting were available for the RT+C cases. The two groups received an oral medicine MA 160mg every day during the treatment. Regarding early outcome, the complete remission rate was 64.3% and partial remission rate was 35.7% in RT+C. The complete remission rate was 32.1% and partial remission rate was 39.3% in RT. The total response rate and complete remission in the RT+C group were higher than that in the RT group. We conclude that taxotere and cisplatin chemoradiotherapy can improve the early outcome of the advanced cervical cancer, the adverse effects being endurable.

Rapid and Sensitive Detection of Lettuce Necrotic Yellows Virus and Cucumber Mosaic Virus Infecting Lettuce (Lactuca sativa L.) by Reverse Transcription Loop-Mediated Isothermal Amplification

  • Zhang, Yubao;Xie, Zhongkui;Fletcher, John D;Wang, Yajun;Wang, Ruoyu;Guo, Zhihong;He, Yuhui
    • The Plant Pathology Journal
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    • v.36 no.1
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    • pp.76-86
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    • 2020
  • Cucumber mosaic virus (CMV) is damaging to the growth and quality of lettuce crops in Lanzhou, China. Recently, however, for the first time an isolate of lettuce necrotic yellows virus (LNYV) has been detected in lettuce crops in China, and there is concern that this virus may also pose a threat to lettuce production in China. Consequently, there is a need to develop a rapid and efficient detection method to accurately identify LNYV and CMV infections and help limit their spread. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed to detect the nucleoprotein (N) and coat protein (CP) genes of LNYV and CMV, respectively. RT-LAMP amplification products were visually assessed in reaction tubes separately using green fluorescence and gel electrophoresis. The assays successfully detected both viruses in infected plants without cross reactivity recorded from either CMV or LNYV or four other related plant viruses. Optimum LAMP reactions were conducted in betaine-free media with 6 mM Mg2+ at 65℃ for LNYV and 60℃ for 60 min for CMV, respectively. The detection limit was 3.5 pg/ml and 20 fg/ml using RT-LAMP for LNYV and CMV plasmids, respectively. Detection sensitivity for both RT-LAMP assays was greater by a factor of 100 compared to the conventional reverse transcription polymerase chain reaction assays. This rapid, specific, and sensitive technique should be more widely applied due to its low cost and minimal equipment requirements.

Comparison of clinical diagnostic performance between commercial RRT-LAMP and RT-qPCR assays for SARS-CoV-2 detection

  • Kim, Hye-Ryung;Park, Jonghyun;Han, Hyung-Soo;Kim, Yu-Kyung;Jeon, Hyo-Sung;Park, Seung-Chun;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.44 no.3
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    • pp.163-168
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    • 2021
  • The rapid and reliable detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) plays a key role in isolating infected patients and preventing further viral transmission. In this study, we evaluated the clinical diagnostic performances of a commercial real-time reverse transcription loop-mediated isothermal amplification (RRT-LAMP) assay (Isopollo® COVID-2 assay, M-monitor, Daegu, Korea) using eighty COVID-19 suspected clinical samples and compared these with the results of a commercial real-time reverse transcription polymerase chain reaction (RT-qPCR) assay (AllplexTM 2019-nCoV rRT-QPCR Assay, SeeGene, Seoul, Korea). The results of the RRT-LAMP assay targeting the N or RdRp gene of SARS-CoV-2 showed perfect agreement with the RT-qPCR assay results in terms of detection. Furthermore, the RRT-LAMP assay was completed in just within a 20-min reaction time, which is significantly faster than about the 2 h currently required for the RT-qPCR assay, thus enabling prompt decision making regarding the isolation of infected patients. The RRT-LAMP assay will be a valuable tool for rapid, sensitive, and specific detection of SARS-CoV-2 in human or unexpected animal clinical cases.

A Study on performance of geothermal heatpump using domestic supply water source and geothermal source during winter (상수도열원과 지열원을 이용한 동절기 지열히트펌프 성능평가에 대한 연구)

  • Lee, Byoungdoo;Lee, Sejin;Lee, Daewoo
    • 한국신재생에너지학회:학술대회논문집
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    • 2011.05a
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    • pp.194.1-194.1
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    • 2011
  • 본 연구에서는 지열히트펌프 시스템의 열원으로써 지열이외에 건축물의 미활용 에너지라고 할 수 있는 상수도의 에너지를 활용하여 지중열교환기의 천공길이를 줄이는 것이 주요 목적이며, 또한 건물의 미활용에너지를 냉난방에너지원으로써 이용 가능한 것을 보여주는 것에 있다. 실험은 4인 가족기준으로 3RT 용량의 히트펌프를 설치하고 인당 평균 177 liter/day 기준으로 하루에 약 710 liter/day의 물을 사용하는 것으로 가정하였다(환경부 2007년 상수도 통계값). 시간당 가정내에서 사용하는 물량은 일정하지 않아 일일 8시간 사용하는 것으로 하여 약 1.5 LPM 으로 실험하였다. 저수조의 크기 및 지열 히트펌프의 열원으로써 사용가능한 열량을 계산하기 위해 CFD 시물레이션을 수행하였다. CFD의 결과 상수도를 급수하기 위한 저수조의 크기는 $2m^3$로 결정하였으며 이때 열원으로써 사용가능한 열량은 약 0.7RT였다. 48시간의 실험기간 동안 저수조를 통해 얻은 열원은 0.6RT 였으며 100m의 지중열교환기를 통해 얻은 열원은 2RT 였다. 히트펌프 자체의 난방 COP는 평균 4.2를 나타내었으며 펌프등의 소비전력을 포함한 System COP는 4.0 나타내었다. 이번 연구를 통해 건물의 미활용에너지인 저수조의 물을 이용하여 지열히트펌프의 열원으로써 이용 가능하며 기존의 지열히트펌프 시스템대비 천공길이 단축, 시공비 저감이 가능한 것을 볼 수 있었다.

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Effects of Melatonin on Gene Expression of IVM/IVF Porcine Embryos

  • Jang, H.Y.;Kong, H.S.;Choi, K.D.;Jeon, G.J.;Yang, B.K.;Lee, C.K.;Lee, H.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.1
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    • pp.17-21
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    • 2005
  • The effect of melatonin on in vitro embryo development and the expression of antioxidant enzyme gene in preimplantation porcine embryos was determined by modified semi-quantitative single cell RT-PCR. Porcine embryos derived from in vitro maturation /in vitro fertilization were cultured in 5% $CO_2$ and 20% $O_2$ at $37^{\circ}C$ in NCSU23 medium. Melatonin was added to medium at concentration of 1nM, 5 nM, and 10 nM. When treated with 1nM (39.0%) of melatonin, the developmental rate of embryos beyond the morula stage were higher than that of control group (31.0%) (p<0.05). Number of inner cell mass and tropectoderm cell in control (23.0${\pm}$0.5 and 17.3${\pm}$0.8), 1 nM (23.6${\pm}$0.6 and 19.0${\pm}$0.5), and 5 nM (23.3${\pm}$1.1 and 16.3${\pm}$0.8) treated with melatonin were higher than in 10 nM (20.0${\pm}$0.5 and 13.3${\pm}$0.8) treated with melatonin (p<0.05). To develop an mRNA phenotypic map for the expression of catalase, bax and caspase-3, single cell RT-PCR analysis were carried out in porcine IVM/IVF embryo. Catalase was detected in 0, 1 and 5 nM supplemented with melatonin, but bax and caspase-3 were detected in 10 nM treated with melatonin.

Enhancing value of quality assurance rounds in improving radiotherapy management: a retrospective analysis from King Hussein Cancer Center in Jordan

  • Khader, Jamal K.;Al-Mousa, Abdelatif M.;Mohamad, Issa A.;Abuhijlih, Ramiz A.;Al-Khatib, Sondos A.;Alnsour, Anoud Z.;Asha, Wafa A.;Ramahi, Shada W.;Hosni, Ali A.;Abuhijla, Fawzi J.
    • Radiation Oncology Journal
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    • v.37 no.1
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    • pp.60-65
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    • 2019
  • Purpose: The quality assurance (QA) chart rounds are multidisciplinary meetings to review radiation therapy (RT) treatment plans. This study focus on describing the changes in RT management based on QA round reviews in a single institution. Materials and Methods: After 9 full years of implementation, a retrospective review of all patients whose charts passed through departmental QA chart rounds from 2007 to 2015. The reviewed cases were presented for RT plan review; subcategorized based on decision in QA rounds into: approved, minor modifications or major modifications. Major modification defined as any substantial change which required patient re-simulation or re-planning prior to commencement of RT. Minor modification included treatment plan changes which didn't necessarily require RT re-planning. Results: Overall 7,149 RT treatment plans for different anatomical sites were reviewed at QA rounds. From these treatment plans, 6,654 (93%) were approved, 144 (2%) required minor modifications, while 351 (5%) required major modifications. Major modification included changes in: selected RT dose (96/351, 27%), target volume definition (127/351, 36%), organs-at-risk contouring (10/351, 3%), dose volume objectives/constraints criteria (90/351, 26%), and intent of treatment (28/351, 8%). The RT plans which required major modification according to the tumor subtype were as follows: head and neck (104/904, 12%), thoracic (12/199, 6%), gastrointestinal (33/687,5%), skin (5/106, 5%), genitourinary (16/359, 4%), breast (104/2387, 4%), central nervous system (36/846, 4%), sarcoma (11/277, 4%), pediatric (7/251, 3%), lymphoma (10/423, 2%), gynecological tumors (2/359, 1%), and others (11/351, 3%). Conclusion: Multi-disciplinary standardized QA chart rounds provide a comprehensive and an influential method on RT plans and/or treatment decisions.

Effect of Radiation on mRNA Expression of Ceruloplasmin Gene (방사선 조사에 따른 U-937 세포의 Ceruloplasmin 유전자에서 mRNA 발현 변화)

  • 오연경;임희영;김종수;윤충효;김인규;윤병수
    • Toxicological Research
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    • v.20 no.1
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    • pp.31-36
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    • 2004
  • Against environmental stress, ceruloplasmin which is a plasma protein, are believed to play central roles in antioxidant- or peroxidase-activity in blood stream to remove free radicals, which may be caused by exposing of $\gamma$-irradiation. In human U-937 cells exposed to $\gamma$-irradiation, the levels of mRNA in ceruloplasmin gene were measured on 0, 4, 12, 24 hr after exposing by using comparative RT-PCR (Reverse transcriptase-polymerase chain reaction) which was achieved to compare with house keeping genes such as $\beta$-actin and hprt. After $\gamma$-irradiation of 100 rads or 200 rads, the total quantities of RNA were increased as dose and time dependent manner. On the contrary, the variation of mRNA expression in ceruloplasmin was not found until 4 hr after irradiation. After 12 hr and 24 hr of irradiation, the levels of mRNA in ceruloplasmin were significantly increased as dose and time dependent manner than un-exposed cells.

Cloning, Expression and Hormonal Regulation of Steroidogenic Acute Regulatory Protein Gene in Buffalo Ovary

  • Malhotra, Nupur;Singh, Dheer;Sharma, M.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.2
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    • pp.184-193
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    • 2007
  • In mammalian ovary, steroidogenic acute regulatory (StAR) protein mediates the true rate-limiting step of transport of cholesterol from outer to inner mitochondrial membrane. Appropriate expression of StAR gene represents an indispensable component of steroidogenesis and its regulation has been found to be species specific. However, limited information is available regarding StAR gene expression during estrous cycle in buffalo ovary. In the present study, expression, localization and hormonal regulation of StAR mRNA were analyzed by semi-quantitative RT-PCR in buffalo ovary and partial cDNA was cloned. Total RNA was isolated from whole follicles of different sizes, granulosa cells from different size follicles and postovulatory structures like corpus luteum and Corpus albicans. Semi-quantitative RT-PCR analyses showed StAR mRNA expression in the postovulatory structure, corpus luteum. No StAR mRNA was detected in total RNA isolated from whole follicles of different size including the preovulatory follicle (>9 mm in diameter). However, granulosa cells isolated from preovulatory follicles showed the moderate expression of StAR mRNA. To assess the hormonal regulation of StAR mRNA, primary culture of buffalo granulosa cells were treated with FSH (100 ng/ml) alone or along with IGF-I (100 ng/ml) for 12 to 18 h. The abundance of StAR mRNA increased in cells treated with FSH alone or FSH with IGF-I. However, effect of FSH with IGF-I on mRNA expression was found highly significant (p<0.01). In conclusion, differential expression of StAR messages was observed during estrous cycle in buffalo ovary. Also, there was a synergistic action of IGF-I on FSH stimulation of StAR gene.

A Simple Method for Elimination of False Positive Results in RT-PCR

  • Martel, Fatima;Grundemann, Dirk;Schomig, Edgar
    • BMB Reports
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    • v.35 no.2
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    • pp.248-250
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    • 2002
  • Discrimination between the amplification of mRNA and contaminating genomic DNA is a common problem when performing a reverse transcriptase-polymerase chain reaction (RT-PCR). Even after treatment of the samples with DNAse, it is possible that negative controls (samples in which no reverse transcriptase was added) will give positive results. This indicates that there was amplification of DNA, which was not generated during the reverse transcriptase step. The possibility exists that Taq DNA polymerase acts as a reverse transcriptase, generating cDNA from RNA during the PCR step. In order to test this hypothesis, we incubated samples with a DNAse-free RNAse after the cDNA synthesis. Comparison of the results that were obtained from these samples (incubated with or without DNAse-free RNAse) confirms that the reverse transcriptase activity of Taq DNA polymerase I is a possible source of false positive results when performing RT-PCR from intronless genes. Moreover, we describe here a simple and rapid method to overcome the false positive results that originate by this activity of Taq polymerase.