• Title/Summary/Keyword: quantitative PCR (qPCR)

검색결과 381건 처리시간 0.026초

Four Novel Synthetic Tryptamine Analogs Induce Head-Twitch Responses and Increase 5-HTR2a in the Prefrontal Cortex in Mice

  • Abiero, Arvie;Ryu, In Soo;Botanas, Chrislean Jun;Custodio, Raly James Perez;Sayson, Leandro Val;Kim, Mikyung;Lee, Hyun Jun;Kim, Hee Jin;Seo, Joung-Wook;Cho, Min Chang;Lee, Kun Won;Yoo, Sung Yeun;Jang, Choon-Gon;Lee, Yong Sup;Cheong, Jae Hoon
    • Biomolecules & Therapeutics
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    • 제28권1호
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    • pp.83-91
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    • 2020
  • Tryptamines are monoamine alkaloids with hallucinogenic properties and are widely abused worldwide. To hasten the regulations of novel substances and predict their abuse potential, we designed and synthesized four novel synthetic tryptamine analogs: Pyrrolidino tryptamine hydrochloride (PYT HCl), Piperidino tryptamine hydrochloride (PIT HCl), N,N-dibutyl tryptamine hydrochloride (DBT HCl), and 2-Methyl tryptamine hydrochloride (2-MT HCl). Then, we evaluated their rewarding and reinforcing effects using the conditioned place preference (CPP) and self-administration (SA) paradigms. We conducted an open field test (OFT) to determine the effects of the novel compounds on locomotor activity. A head-twitch response (HTR) was also performed to characterize their hallucinogenic properties. Lastly, we examined the effects of the compounds on 5-HTR1a and 5-HTR2a in the prefrontal cortex using a quantitative real-time polymerase chain reaction (qRT-PCR) assay. None of the compounds induced CPP in mice or initiated SA in rats. PYT HCl and PIT HCl reduced the locomotor activity and elevated the 5-HTR1a mRNA levels in mice. Acute and repeated treatment with the novel tryptamines elicited HTR in mice. Furthermore, a drug challenge involving a 7-day abstinence from drug use produced higher HTR than acute and repeated treatments. Both the acute treatment and drug challenge increased the 5-HTR2a mRNA levels. Ketanserin blocked the induced HTR. Taken together, the findings suggest that PYT HCl, PIT HCl, DBT HCl, and 2-MT HCl produce hallucinogenic effects via 5-HTR2a stimulation, but may have low abuse potential.

Exosomal miR-181b-5p Downregulation in Ascites Serves as a Potential Diagnostic Biomarker for Gastric Cancer-associated Malignant Ascites

  • Yun, Jieun;Han, Sang-Bae;Kim, Hong Jun;Go, Se-il;Lee, Won Sup;Bae, Woo Kyun;Cho, Sang-Hee;Song, Eun-Kee;Lee, Ok-Jun;Kim, Hee Kyung;Yang, Yaewon;Kwon, Jihyun;Chae, Hee Bok;Lee, Ki Hyeong;Han, Hye Sook
    • Journal of Gastric Cancer
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    • 제19권3호
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    • pp.301-314
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    • 2019
  • Purpose: Peritoneal carcinomatosis in gastric cancer (GC) patients results in extremely poor prognosis. Malignant ascites samples are the most appropriate biological material to use to evaluate biomarkers for peritoneal carcinomatosis. This study identified exosomal MicroRNAs (miRNAs) differently expressed between benign liver cirrhosis-associated ascites (LC-ascites) and malignant gastric cancer-associated ascites (GC-ascites), and validated their role as diagnostic biomarkers for GC-ascites. Materials and Methods: Total RNA was extracted from exosomes isolated from 165 ascites samples (73 LC-ascites and 92 GC-ascites). Initially, microarrays were used to screen the expression levels of 2,006 miRNAs in the discovery cohort (n=22). Subsequently, quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) analyses were performed to validate the expression levels of selected exosomal miRNAs in the training (n=70) and validation (n=73) cohorts. Furthermore, carcinoembryonic antigen (CEA) levels were determined in ascites samples. Results: The miR-574-3p, miR-181b-5p, miR-4481, and miR-181d were significantly downregulated in the GC-ascites samples compared to the LC-ascites samples, and miR-181b-5p showed the best diagnostic performance for GC-ascites (area under the curve [AUC]=0.798 and 0.846 for the training and validation cohorts, respectively). The diagnostic performance of CEA for GC-ascites was improved by the combined analysis of miR-181b-5p and CEA (AUC=0.981 and 0.946 for the training and validation cohorts, respectively). Conclusions: We identified exosomal miRNAs capable of distinguishing between non-malignant and GC-ascites, showing that the combined use of miR-181b-5p and CEA could improve diagnosis.

Spatial and seasonal distributions of the phototrophic dinoflagellate Biecheleriopsis adriatica (Suessiaceae) in Korea: quantification using qPCR

  • Kang, Hee Chang;Jeong, Hae Jin;Ok, Jin Hee;You, Ji Hyun;Jang, Se Hyeon;Lee, Sung Yeon;Lee, Kyung Ha;Park, Jae Yeon;Rho, Jung-Rae
    • ALGAE
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    • 제34권2호
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    • pp.111-126
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    • 2019
  • The phototrophic dinoflagellate Biecheleriopsis adriatica is a small suessioid species characterized by a fragile thin wall. Although the morphology of this dinoflagellate is well established, there is currently little information available on its distribution and the environmental factors that influence this distribution. Thus, to investigate the spatial and seasonal distributions of the vegetative cells of B. adriatica in Korean waters, surface water samples were collected on a seasonal basis from 28 stations in the East, West, and South Sea of Korea and Jeju Island from April 2015 to October 2018, and abundances of the vegetative cells of B. adriatica were quantified using quantitative real-time polymerase chain reactions, for which we developed the species-specific primer and probe set. Simultaneously, major environmental parameters, including temperature, salinity, nutrient concentrations, and dissolved oxygen concentrations were measured. The vegetative cells of B. adriatica were detected at 20 of the 28 sampling stations: 19 stations in summer and 6 in autumn, although from no stations in either spring or winter. The ranges of water temperature and salinity at sites where this species was detected were $17.7-26.4^{\circ}C$ and 9.9-34.3, respectively, whereas those of nitrate and phosphate concentrations were not detectable-96.2 and $0.18-2.66{\mu}M$, respectively. Thus, the sites at which this species is found are characterized by a narrow range of temperature, but wide ranges of salinity and concentrations of nitrate and phosphate. The highest abundance of the vegetative cells of B. adriatica was $41.7cells\;mL^{-1}$, which was recorded in Jinhae Bay in July 2018. In Jinhae Bay, the abundance of vegetative cells was significantly positively correlated with the concentration of nitrate, but was negatively correlated with salinity. On the basis of these findings, it appears that the abundance of B. adriatica vegetative cells shows strong seasonality, and in Jinhae Bay, could be affected by the concentrations of nitrate.

First report of the photosynthetic dinoflagellate Heterocapsa minima in the Pacific Ocean: morphological and genetic characterizations and the nationwide distribution in Korea

  • Lee, Sung Yeon;Jeong, Hae Jin;Kwon, Ji Eun;You, Ji Hyun;Kim, So Jin;Ok, Jin Hee;Kang, Hee Chang;Park, Jae Yeon
    • ALGAE
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    • 제34권1호
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    • pp.7-21
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    • 2019
  • The genus Heterocapsa is one of the major dinoflagellate groups, with some of its species having worldwide distributions. However, prior to the present study, the phototrophic species Heterocapsa minima has been reported only from the northeast Atlantic Ocean. Recently, H. minima was found in the Korean waters, and a clonal culture was established. This culture was used to examine the morphology of the Korean strain H. minima HMMJ1604 through light and scanning electron microscopy, as well as for its genetic characterization. Furthermore, to determine the nationwide distribution of H. minima in Korea, its abundance was quantified in the waters of 28 stations in all four seasons in 2016-2018 using the quantitative real-time polymerase chain reaction method. The overall morphology of H. minima HMMJ1604 was very similar to that of the Irish strain H. minima JK2. However, the Korean strain had five pores around the pore plate, whereas the Irish strain had six pores. When properly aligned, the sequences of the large subunit and internal transcribed spacer regions of the ribosomal DNA of the Korean strain were identical to those of the Irish strain. This species was detected in the waters of 26 out of 28 stations, but its abundance was greater than $1.0cells\;mL^{-1}$ at 8 stations. The highest abundance of H. minima was $44.4cells\;mL^{-1}$. Although this species was found in all seasons, its abundance was greater than $1.0cells\;mL^{-1}$ when the water temperature and salinity were $10.9-25.0^{\circ}C$ and 17.5-34.1, respectively. To the best knowledge, the present study reported for the first time that H. minima lives in the Pacific Ocean and is widely distributed in the Korean waters.

계대 배양 속도가 다른 과잉치 치수유래 줄기세포 간 유전자 발현 특성 (Gene Expression of Supernumerary Dental Pulp Related to the Subculture Speed: A Pilot Study)

  • 이유경;김종수;신지선;김종빈
    • 대한소아치과학회지
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    • 제46권2호
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    • pp.219-225
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    • 2019
  • 이 연구의 목적은 과잉치 치수 유래 줄기세포의 계대 배양 속도에 대한 상아모세포 연관 유전자의 발현을 비교하는 것이다. 줄기세포는 다른 여러 형태의 세포로 분화할 수 있는 미 분화된 세포이다. 이는 환경이나 특정 자극에 의해 세포 분열이 일어나며 근육이나 골 같은 특정 장기의 조직으로 분화할 수 있다. 20명의 어린이에서 발거한 과잉치에서 과잉치 치수 유래 줄기세포가 얻어졌다. 10계대까지 배양하는 동안 가장 빠른 속도로 계대 배양된 세포와 가장 느린 속도로 계대 배양된 세포 각 3계대와 10계대 세포를 얻어 실험을 진행하였다. 각 세포는 분화제를 처리한 군과 처리하지 않은 군으로 나누었다. 이 실험에서 발현도를 살펴본 유전자는 Osteonectin (ONT), Osteocalcin (OCN), Alkaline Phosphatase (ALP), Dentin matrix acidic phosphoprotein 1 (DMP-1), Dentin sialophosphoprotein (DSPP)이다. 분화가 된 세포가 전반적으로 더 높은 유전자 발현도를 보였으며, 미분화 세포는 10계대에서, 분화된 세포는 3계대에서 더 높은 유전자 발현도를 보였다. 빠른 계대 배양 속도를 보인 세포가 OCN과 DSPP를 제외하고 상대적으로 더 낮은 유전자 발현도를 보였다.

어린 돌기해삼 Apostichopus japonicus (Echinodermata; Holothuroidea) 방류 방법에 따른 열충격단백질90 유전자의 발현 분석 (Heat Shock Protein 90 Gene Expression in Juvenile Sea Cucumber Apostichopus japonicus (Echinodermata; Holothuroidea) according to Releasing Methods)

  • 이동한;이승헌;정동빈;손영창
    • 한국해양생명과학회지
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    • 제7권1호
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    • pp.29-36
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    • 2022
  • 돌기해삼 Apostichopus japonicus는 주요 양식 대상 무척추동물로서 우리나라 연안 해역에 서식하고 있다. 본 연구는 방류 방법에 따른 단기간의 생리학적 스트레스 정도를 평가하기 위하여 heat shock protein 90 (HSP90) 유전자의 발현 변화를 실시간 정량적 중합효소연쇄반응법으로 조사하였다. 어린 돌기해삼을 비닐봉지에 산소 포장하여 30분간 수송하거나 방류 해역의 간조기에 1시간 공기 중에 노출된 실험군의 HSP90 유전자 발현은 대조군의 HSP90 유전자 발현에 비하여 통계학적으로 유의미하게 증가하였다(수송 후 실험군 p=0.001; 간조기 실험군 p=0.032). 어린 돌기해삼을 방류 후 6시간까지 분석한 결과, 선상에서 씨뿌림 방식으로 방류된 6시간째의 개체 및 호스를 통과하여 수중으로 방류된 2~6시간째의 HSP90 유전자 발현율은 대조군에 비하여 약간 감소하는 경향을 보였다(씨뿌림 실험군 p=0.069; 호스 방류군 p=0.093). 한편, 잠수부에 의해 수중에서 방류된 어린 돌기해삼은 방류 후 시간이 경과할수록 HSP90 유전자 발현율은 증가하는 패턴이 관찰되었다(p=0.061). 이상의 결과는 방류된 어린 돌기해삼의 단기간 스트레스 반응 연구와 효과적인 방류 방법의 개발에 HSP90 유전자 발현이 유용하게 사용될 수 있음을 시사한다.

Ginsenoside Rg1 alleviates Aβ deposition by inhibiting NADPH oxidase 2 activation in APP/PS1 mice

  • Zhang, Han;Su, Yong;Sun, Zhenghao;Chen, Ming;Han, Yuli;Li, Yan;Dong, Xianan;Ding, Shixin;Fang, Zhirui;Li, Weiping;Li, Weizu
    • Journal of Ginseng Research
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    • 제45권6호
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    • pp.665-675
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    • 2021
  • Background: Ginsenoside Rg1 (Rg1), an active ingredient in ginseng, may be a potential agent for the treatment of Alzheimer's disease (AD). However, the protective effect of Rg1 on neurodegeneration in AD and its mechanism of action are still incompletely understood. Methods: Wild type (WT) and APP/PS1 AD mice, from 6 to 9 months old, were used in the experiment. The open field test (OFT) and Morris water maze (MWM) were used to detect behavioral changes. Neuronal damage was assessed by hematoxylin and eosin (H&E) and Nissl staining. Immunofluorescence, western blotting, and quantitative real-time polymerase chain reaction (q-PCR) were used to examine postsynaptic density 95 (PSD95) expression, amyloid beta (Aβ) deposition, Tau and phosphorylated Tau (p-Tau) expression, reactive oxygen species (ROS) production, and NAPDH oxidase 2 (NOX2) expression. Results: Rg1 treatment for 12 weeks significantly ameliorated cognitive impairments and neuronal damage and decreased the p-Tau level, amyloid precursor protein (APP) expression, and Aβ generation in APP/PS1 mice. Meanwhile, Rg1 treatment significantly decreased the ROS level and NOX2 expression in the hippocampus and cortex of APP/PS1 mice. Conclusions: Rg1 alleviates cognitive impairments, neuronal damage, and reduce Aβ deposition by inhibiting NOX2 activation in APP/PS1 mice.

Comparative metabolomic analysis in horses and functional analysis of branched chain (alpha) keto acid dehydrogenase complex in equine myoblasts under exercise stress

  • Jeong-Woong, Park;Kyoung Hwan, Kim;Sujung, Kim;Jae-rung, So;Byung-Wook, Cho;Ki-Duk, Song
    • Journal of Animal Science and Technology
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    • 제64권4호
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    • pp.800-811
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    • 2022
  • The integration of metabolomics and transcriptomics may elucidate the correlation between the genotypic and phenotypic patterns in organisms. In equine physiology, various metabolite levels vary during exercise, which may be correlated with a modified gene expression pattern of related genes. Integrated metabolomic and transcriptomic studies in horses have not been conducted to date. The objective of this study was to detect the effect of moderate exercise on the metabolomic and transcriptomic levels in horses. In this study, using nuclear magnetic resonance (NMR) spectroscopy, we analyzed the concentrations of metabolites in muscle and plasma; we also determined the gene expression patterns of branched chain (alpha) keto acid dehydrogenase kinase complex (BCKDK), which encodes the key regulatory enzymes in branched-chain amino acid (BCAA) catabolism, in two breeds of horses, Thoroughbred and Jeju, at different time intervals. The concentrations of metabolites in muscle and plasma were measured by 1H NMR (nuclear magnetic resonance) spectroscopy, and the relative metabolite levels before and after exercise in the two samples were compared. Subsequently, multivariate data analysis based on the metabolic profiles was performed using orthogonal partial least square discriminant analysis (OPLS-DA), and variable important plots and t-test were used for basic statistical analysis. The stress-induced expression patterns of BCKDK genes in horse muscle-derived cells were examined using quantitative reverse transcription polymerase chain reaction (qPCR) to gain insight into the role of transcript in response to exercise stress. In this study, we found higher concentrations of aspartate, leucine, isoleucine, and lysine in the skeletal muscle of Jeju horses than in Thoroughbred horses. In plasma, compared with Jeju horses, Thoroughbred horses had higher levels of alanine and methionine before exercise; whereas post-exercise, lysine levels were increased. Gene expression analysis revealed a decreased expression level of BCKDK in the post-exercise period in Thoroughbred horses.

Bisphenol 구조 유사체가 기수산 물벼룩 Ecdysteroid 경로에 미치는 영향 (Time-dependent Effects of Bisphenol Analogs on Ecdysteroid Pathway Related Genes in the Brackish Water Flea Diaphanosoma celebensis)

  • 인소연;이영미
    • 한국해양생명과학회지
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    • 제6권2호
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    • pp.73-79
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    • 2021
  • 비스페놀A(BPA)는 대표적인 내분비계 교란물질로 광범위한 사용으로 인해 환경 내에서 지속적으로 검출됨에 따라 인간을 비롯한 다양한 생물에서 성장, 발생, 생식 등에 유해한 영향을 미치는 것으로 알려져 있다. 따라서 BPA를 대체하기 위한 구조 유사체들이 개발되어 널리 사용되고 있으나 이러한 대체제들이 내분비계 교란 작용을 갖는지에 대한 연구가 필요하다. 본 연구에서는 BPA와 그 구조 유사체인 BPS와 BPF에 노출시킨 기수산 물벼룩 Diaphanosoma celebensis에서 탈피과정에 관여하는 ecdysteroid 합성(nvd, cyp314a1), receptors (EcRA, EcRB, USP, ERR), 그리고 하위 경로에 있는 유전자(HR3, E75, Vtg, VtgR)의 시간 별 발현 변화를 조사하였다. nvd와 cyp314a1 유전자의 발현은 BPA 보다 BPF에서 6시간 일찍 발현이 증가하는 양상을 보인 반면, BPS의 경우에는 이들 유전자의 발현이 24시간 내내 감소하는 양상을 보였다. BPA와 BPF 노출 시 EcR 유전자들의 발현 양상도 이와 유사한 경향을 보였다. ERR 유전자의 발현은 BPF와 BPS에서 BPA 보다 6시간 일찍 발현이 증가하는 양상을 보였고, HR3, E75, VtgR의 유전자 발현도 노출군에서 시간 차이는 있지만 유의하게 증가하는 양상을 보였다. 반면 Vtg는 24시간 이내에서는 크게 증가하지는 않았다. 이러한 결과는 BPA 뿐 아니라 BPF와 BPS도 탈피에 관여하는 호르몬의 합성 및 조절 경로의 유전자의 발현을 조절할 수 있으며, 서로 다른 기전으로 기수산 물벼룩의 내분비계를 교란시킬 수 있는 능력을 갖는다고 볼 수 있다. 본 연구는 비스페놀 구조 유사체가 기수산 물벼룩의 탈피과정에 관여하는 분자 경로 어떻게 영향을 미치는지를 이해하는데 도움이 될 것이다.

Green perilla leaf extract ameliorates long-term oxidative stress induced by a high-fat diet in aging mice

  • Edward, Olivet Chiamaka;Thomas, Shalom Sara;Cha, Kyung-Ok;Jung, Hyun-Ah;Han, Anna;Cha, Youn-Soo
    • Nutrition Research and Practice
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    • 제16권5호
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    • pp.549-564
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    • 2022
  • BACKGROUND/OBJECTIVES: Oxidative stress is caused by an imbalance between harmful free radicals and antioxidants. Long-term oxidative stress can lead to an "exhausted" status of antioxidant defense system triggering development of metabolic syndrome and chronic inflammation. Green perilla (Perilla frutescens) is commonly used in Asian cuisines and traditional medicine in southeast Asia. Green perilla possesses numerous beneficial effects including anti-inflammatory and antioxidant functions. To investigate the potentials of green perilla leaf extract (PE) on oxidative stress, we induced oxidative stress by high-fat diet (HFD) in aging mice. MATERIALS/METHODS: C57BL/6J male mice were fed HFD continuously for 53 weeks. Then, mice were divided into three groups for 12 weeks: a normal diet fed reference group (NDcon), high-fat diet fed group (HDcon), and high-fat diet PE treated group (HDPE, 400 mg/kg of body weight). Biochemical analyses of serum and liver tissues were performed to assess metabolic and inflammatory damage and oxidative status. Hepatic gene expression of oxidative stress and inflammation related enzymes were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: PE improved hepatopathology. PE also improved the lipid profiles and antioxidant enzymes, including hepatic glutathione peroxidase (GPx) and superoxide dismutase (SOD) and catalase (CAT) in serum and liver. Hepatic gene expressions of antioxidant and anti-inflammatory related enzymes, such as SOD-1, CAT, interleukin 4 (IL-4) and nuclear factor erythroid 2-related factor (Nrf2) were significantly enhanced by PE. PE also reduced the levels of hydrogen peroxide (H2O2) and malondialdehyde (MDA) in the serum and liver; moreover, PE suppressed hepatic gene expression involved in pro-inflammatory response; Cyclooxygenase-2 (COX-2), nitric oxide synthase (NOS), interleukin 1 beta (IL-1β), and interleukin 6 (IL-6). CONCLUSIONS: This research opens opportunities for further investigations of PE as a functional food and possible anti-aging agent due to its attenuative effects against oxidative stress, resulting from HFD and aging in the future.