• 제목/요약/키워드: single PCR

검색결과 1,036건 처리시간 0.038초

겹삼잎국화 에탄올 추출물의 지방세포 분화 억제 효과 (Inhibitory Effect of the Ethanol Extract of Rudbeckia laciniata var. hortensis Bailey on Adipocyte Differentiation in 3T3-L1 Cells)

  • 남건희;위지향;김상용;백지영;김영민
    • 생명과학회지
    • /
    • 제29권10호
    • /
    • pp.1152-1158
    • /
    • 2019
  • 겹삼잎국화는 한국에서 복통, 위염에 민간요법으로 사용되는 약초로 알려져있다. 하지만, 겹삼잎국화를 민간요법으로 사용되고 있으나 이 약초에 대한 연구는 거의 수행되어지지 않았다. 본 연구에서는 겹삼잎국화의 의학적 효과를 증명하기 위해 항비만 관련 연구를 수행하였다. 70% 에탄올을 이용한 겹삼잎국화 추출물의 항 비만 효과에 대한 연구를 위해 3T3-L1 지방전구세포를 사용하였으며, 지방전구세포가 지방세포로 성숙하는 것을 방지하고 이에 관련된 단백질 인자의 발현을 확인하기 위해 Oil Red O 염색법과 웨스턴 블롯, PCR을 통한 mRNA 분석이 수행되었다. DM (인슐린 칵테일)을 처리하여 지방전구세포의 성숙을 유도한 후, 겹삼잎국화 에탄올 추출물을 처리한 결과, $100{\mu}g/ml$ 농도에서 지방세포 분화 억제 효과와 세포 내 트리글리세리드(TG)의 수준이 현저하게 감소한 것을 확인하였다. 또한, 지방전구세포의 성숙에 관여하는 여러가지 단백질 등의 인자 발현율 또한 확인하였다. 겹삼잎국화 에탄올 추출물로 인해 $PPAR{\gamma}$, $C/EBP{\alpha}$, LPL, FAS와 같은 지방 세포의 성숙을 유도하는 인자가 조절되었으며, Acetyl-CoA carboxylase, AMP-activated protein kinase (AMPK) 등과 같은 단백질의 발현도 억제하여 분자적 기전 또한 규명하였다. 결론적으로, 겹삼잎국화 에탄올 추출물은 지방전구세포의 활성에 영향을 끼치지 않으며 지방세포로의 성숙을 유도하는 인자 및 단백질의 발현을 선택적으로 조절하여 항 비만 효과를 갖는 것으로 확인되었다.

국내 주택에서 물 피해 유형에 따른 부유곰팡이 농도 수준 평가 및 동정 분석 (Assessment of the level and identification of airborne molds by the type of water damage in housing in Korea)

  • 이주영;황은설;이정섭;권명희;정현미;서성철
    • 실내환경 및 냄새 학회지
    • /
    • 제17권4호
    • /
    • pp.355-361
    • /
    • 2018
  • Mold grows more easily when humidity is higher in indoor spaces, and as such is found more often on wetted areas in housing such as walls, toilets, kitchens, and poorly managed spaces. However, there have been few studies that have specifically assessed the level of mold in the indoor spaces of water-damaged housing in the Republic of Korea. We investigated the levels of airborne mold according to the characteristics of water damage types and explored the correlation between the distribution of mold genera and the characteristics of households. Samplings were performed from January 2016 to June 2018 in 97 housing units with water leakage or condensation, or a history of flooding, and in 61 general housing units in the metropolitan and Busan area, respectively. Airborne mold was collected on MEA (Malt extract agar) at flow rate of 100 L/min for 1 min. After collection, the samples were incubated at $25^{\circ}C$ for 120 hours. The cultured samples were counted and corrected using a positive hole conversion table. The samples were then analyzed by single colony culture, DNA extraction, gene amplification, and sequencing. By type of housing, concentrations of airborne mold were highest in flooded housing, followed by water-leaked or highly condensed housings, and then general housing. In more than 50% of water-damaged housing, the level of airborne mold exceeded the guideline of Korea's Ministry of Environment ($500CFU/m^3$). Of particular concern was the fact that the I/O ratio of water-damaged housing was greater than 1, which could indicate that mold damage may occur indoors. The distribution patterns of the fungal species were as follows: Penicillium spp., Cladosporium spp. (14%), Aspergillus spp. (13%) and Alternaria spp. (3%), but significant differences of their levels in indoor spaces were not found. Our findings indicate that high levels of mold damage were found in housing with water damage, and Aspergillus flavus and Penicillium brevicompactum were more dominant in housing with high water activity. Comprehensive management of flooded or water-damaged housing is necessary to reduce fungal exposure.

Monitoring Culicine Mosquitoes (Diptera: Culicidae) as a Vector of Flavivirus in Incheon Metropolitan City and Hwaseong-Si, Gyeonggi-Do, Korea, during 2019

  • Bahk, Young Yil;Park, Seo Hye;Kim-Jeon, Myung-Deok;Oh, Sung-Suck;Jung, Haneul;Jun, Hojong;Kim, Kyung-Ae;Park, Jong Myong;Ahn, Seong Kyu;Lee, Jinyoung;Choi, Eun-Jeong;Moon, Bag-Sou;Gong, Young Woo;Kwon, Mun Ju;Kim, Tong-Soo
    • Parasites, Hosts and Diseases
    • /
    • 제58권5호
    • /
    • pp.551-558
    • /
    • 2020
  • The flaviviruses are small single-stranded RNA viruses that are typically transmitted by mosquitoes or tick vectors and are etiological agents of acute zoonotic infections. The viruses are found around the world and account for significant cases of human diseases. We investigated population of culicine mosquitoes in central region of Korean Peninsula, Incheon Metropolitan City and Hwaseong-si. Aedes vexans nipponii was the most frequently collected mosquitoes (56.5%), followed by Ochlerotatus dorsalis (23.6%), Anopheles spp. (10.9%), and Culex pipiens complex (5.9%). In rural regions of Hwaseong, Aedes vexans nipponii was the highest population (62.9%), followed by Ochlerotatus dorsalis (23.9%) and Anopheles spp. (12.0%). In another rural region of Incheon (habitat of migratory birds), Culex pipiens complex was the highest population (31.4%), followed by Ochlerotatus dorsalis (30.5%), and Aedes vexans vexans (27.5%). Culex pipiens complex was the predominant species in the urban region (84.7%). Culicine mosquitoes were identified at the species level, pooled up to 30 mosquitoes each, and tested for flaviviral RNA using the SYBR Green-based RT-PCR and confirmed by cDNA sequencing. Three of the assayed 2,683 pools (989 pools without Anopheles spp.) were positive for Culex flaviviruses, an insect-specific virus, from Culex pipiens pallens collected at the habitats for migratory birds in Incheon. The maximum likelihood estimation (the estimated number) for Culex pipiens pallens positive for Culex flavivirus was 25. Although viruses responsible for mosquito-borne diseases were not identified, we encourage intensified monitoring and long-term surveillance of both vector and viruses in the interest of global public health.

국화왜화바이로드 저항성 국화 유전자원 선발 (Identification of Chrysanthemum Genetic Resources Resistant to Chrysanthemum Stunt Viroid (CSVd))

  • 박상근;최성열;곽용범
    • 현장농수산연구지
    • /
    • 제22권1호
    • /
    • pp.131-142
    • /
    • 2020
  • 전 세계적으로 국화에 큰 피해를 야기하는 국화왜화바이로이는 단일나선형의 감염성 RNA 병원체로, 1997년 국내에서 처음 발견된 이후 전국적으로 빠르게 전파되고 있다. 그러나 이러한 심각성 에도 불구하고, 국화왜화바이로이드는 재배적, 화학적 방제가 불가능하기 때문에 예방 이외에는 방제할 수 있는 방법이 거의 없는 실정이다. 따라서 국화 재배에 있어 국화왜화바이로이드병을 방제하기 위해서는 저항성 품종의 육성이 절실하다. 이에 본 연구에서는 국화왜화바이로이드 저항성 품종 육성을 위한 기반 구축을 위해 유전자원을 선발하고자 하였다. 스프레이 167 품종과 스탠다드 25품종 등 총 192 품종을 공시하여 접목즙액접종법을 통해 병 저항성 검정을 실시한 결과, 대부분의 품종에서 초장이 감소하는 전형적인 왜화 증상을 나타냈으며, 화경이 감소하고 화색이 퇴화되는 증상을 보였다. 그러나 일부 품종에서는 이와 같은 병징이 나타나지 않거나 지연되는 경향을 보였다. 192 품종 가운데 5% 이하의 초장 감소율을 보인 'Chiwerel'과 'Jeongheungdaesin'은 저항성, 20% 미만의 초장감소율을 보인 'Inga' 등 6 품종은 중도저항성, 그리고 그 외 184품종은 감수성으로 평가되었다. 저항성 발현이 안정적으로 나타나는 'Chiwerel' 등의 품종들은 국화왜화바이로이드병 저항성 품종을 육종하는 교배모본으로 활용될 수 있을 것으로 기대된다.

광화학적 뇌경색 백서 모델에서 황련의 항염증 및 운동기능 회복에 미치는 효과 (Effects of Coptidis Rhizoma on the Anti-inflammation and Motor Recovery in Photothrombotic Brain Infarction Model in Rats)

  • 이수경;이인;신선호;김은영;신병철
    • 대한본초학회지
    • /
    • 제24권1호
    • /
    • pp.179-189
    • /
    • 2009
  • Objectives : Coptidis Rhizoma (Coptis japonica MAKINO; CR) is a well known crude drug as antimicrobial, antibacterial, anti-inflammatory, antioxidant activity. However, there is no study of the effect of CR on brain infarction and it's mechanism. The aim of this study was to investigate the effects on ischemic stroke induced by photothrombotic infarction by evaluating the functional & neuronal recovery after brain infarction. Materials & Methods : Male Sprague-Dawley rats (250-300 g) were induced photothrombotic brain infarction on sensorimotor cortex, and brain infarction volume by image J software (NIH, USA) after Nissl stain, also single pellet reaching task as a functional motor recovery were observed. After orally pretreated by CR (500 mg/kg) or normal saline as a sham control before 7 days from the time of photothrombotic infarction, rats were sacrificed. After then we analysed anti-inflammatory cytokines (TNF-$\alpha$, IL-6, IL-1$\beta$), by RT-PCR and ELISA method, and immunohistochemistry (GFAP, connexin-43) as a marker of neural plasticity. Results : CR (100, 250, 500 mg/kg) decreased the infarction volume dose-dependently, however the effect of 500mg/kg of CR (CR 500) showed the best (P=0.051). Also, CR 500 decreased the infarction volume time-dependently, the most effective time was 3-7 days after stroke. Photothrombosis increased inflammatory cytokines after infarction, CR 500 suppressed significantly mRNA expression of IL-1$\beta$, IL-6 and TNF-$\alpha$. In serum, CR 500 decreased the amount of IL-1$\beta$, 12h, 24h and 48h respectively (p < 0.05), also decreased that of IL-6 and TNF-$\alpha$, 12h respectively (p < 0.05) after infarction. The more astrocytes were observed and neural plasticity was facilitated in the rat brain of CR 500 than that of sham control in immunohistochemistry. Conclusions : This results suggest that CR decrease infarction volume and improve functional motor recovery in acute stage in photothrombotic ischemic infarction model in the mechanism of anti-inflammation and promoting neural plasticity.

국내 잎들깨에서 발생한 잠두위조바이러스2의 특성 구명 (Characterization of broad bean wilt virus 2 isolated from Perilla frutescens in Korea)

  • 김현선;변희성;최유지;최현용;서장균;최홍수;이봉춘;김미경;곽해련
    • 환경생물
    • /
    • 제41권1호
    • /
    • pp.1-13
    • /
    • 2023
  • 잠두위조바이러스2 (BBWV2)는 경제적으로 중요한 원예, 관상작물에 심각한 피해를 주는 바이러스 중의 하나다. BBWV2의 넓은 기주 범위, 매개충 방제의 어려움 등 효과적인 치료제가 없기 때문에, 병을 예방하거나 저항성 품종의 개발 등을 위해서는 BBWV2의 새로운 분리주들의 특성 조사와 연구가 필요하다. 본 연구에서는 2019년 금산지역의 잎들깨 재배농가에서 분리된 BBWV2 분리주(BBWV2-GS-PF)의 유전학적·생물학적 특성을 구명하고 기존에 보고된 분리주들과의 특성을 비교하였다. 순수 분리된 BBWV2-GS-PF는 기존 분리주와 달리 들깨에서 심한 모자이크 증상과 고추에서 원형 반점 증상을 보였다. BBWV2-GS-PF의 유전자 계통분석 결과, 기존에 알려진 2개의 그룹과 약 76~80%의 비교적 낮은 유전자 상동성을 보이며 계통이 분화된 것을 확인할 수 있었다.

Synergistic Effect of Hydrogen and 5-Aza on Myogenic Differentiation through the p38 MAPK Signaling Pathway in Adipose-Derived Mesenchymal Stem Cells

  • Wenyong Fei;Erkai Pang;Lei Hou;Jihang Dai;Mingsheng Liu;Xuanqi Wang;Bin Xie;Jingcheng Wang
    • International Journal of Stem Cells
    • /
    • 제16권1호
    • /
    • pp.78-92
    • /
    • 2023
  • Background and Objectives: This study aims to clarify the systems underlying regulation and regulatory roles of hydrogen combined with 5-Aza in the myogenic differentiation of adipose mesenchymal stem cells (ADSCs). Methods and Results: In this study, ADSCs acted as an in vitro myogenic differentiating mode. First, the Alamar blue Staining and mitochondrial tracer technique were used to verify whether hydrogen combined with 5-Aza could promote cell proliferation. In addition, this study assessed myogenic differentiating markers (e.g., Myogenin, Mhc and Myod protein expressions) based on the Western blotting assay, analysis on cellular morphological characteristics (e.g., Myotube number, length, diameter and maturation index), RT-PCR (Myod, Myogenin and Mhc mRNA expression) and Immunofluorescence analysis (Desmin, Myosin and 𝛽-actin protein expression). Finally, to verify the mechanism of myogenic differentiation of hydrogen-bound 5-Aza, we performed bioinformatics analysis and Western blot to detect the expression of p-P38 protein. Hydrogen combined with 5-Aza significantly enhanced the proliferation and myogenic differentiation of ADSCs in vitro by increasing the number of single-cell mitochondria and upregulating the expression of myogenic biomarkers such as Myod, Mhc and myotube formation. The expressions of p-P38 was up-regulated by hydrogen combined with 5-Aza. The differentiating ability was suppressed when the cells were cultivated in combination with SB203580 (p38 MAPK signal pathway inhibitor). Conclusions: Hydrogen alleviates the cytotoxicity of 5-Aza and synergistically promotes the myogenic differentiation capacity of adipose stem cells via the p38 MAPK pathway. Thus, the mentioned results present insights into myogenic differentiation and are likely to generate one potential alternative strategy for skeletal muscle related diseases.

Development and Validation of an SNP Marker for Identifying Xanthomonas oryzae pv. oryzae Thai Isolates That Break xa5-Mediated Bacterial Blight Resistance in Rice

  • Tebogo Balone;Ananda Nuryadi Pratama;Werapat Chansongkram;Thanita Boonsrangsom;Kawee Sujipuli;Kumrop Ratanasut
    • The Plant Pathology Journal
    • /
    • 제40권5호
    • /
    • pp.451-462
    • /
    • 2024
  • Xanthomonas oryzae pv. oryzae (Xoo) is a pathogenic bacterium responsible for bacterial blight (BB) disease in rice, primarily mediated by the interaction between the plant and pathogen. The virulence mechanism involves the activation of the Sugars Will Eventually be Exported Transporter (SWEET) gene family in rice by transcription activator-like effectors derived from Xoo. The BB resistance gene xa5 has been identified as one of the most effective genes against Thai Xoo isolates, but xa5-mediated resistance-breaking Xoo strains have emerged. This study aimed to develop a single nucleotide polymorphism (SNP) marker for precise identification of xa5-mediated resistance-breaking Xoo. Comparative genomics of Thai Xoo isolates Xoo16PK001 and Xoo16PK002, which were incompatible and compatible with rice variety IRBB5 carrying xa5, respectively, identified eight SNP positions for the development of an SNP marker. The SNP marker XooE6 yields a specific 1,143 bp PCR product unique to Xoo16PK002. Screening 61 Thai isolates using XooE6 identified two positives: Xoo20PL010 and Xoo20UT002. Inoculation tests on rice varieties IRBB5 and IRBB13 demonstrated compatibility with IRBB5 and incompatibility with IRBB13, which bears Xa5 and xa13. Xoo16PK001 (XooE6-negative) showed different virulence. Inoculation on IRBB21 harboring Xa5, Xa13, and Xa21 resulted in partial resistance to both XooE6-positive and -negative strains. XooE6-positive strains up-regulated SWEET11 and suppressed SWEET14 in IRBB5, while Xoo16PK001 slightly induced SWEET11 but activated SWEET14 in IRBB13. This highlights the potential of XooE6 to identify xa5-mediated resistance-breaking Xoo strains and elucidate their pathogenic mechanisms through the upregulation of SWEET11.

Promoter Polymorphisms of ST3GAL4 and ST6GAL1 Genes and Associations with Risk of Premalignant and Malignant Lesions of the Cervix

  • de los Angeles Rivera-Juarez, Maria;Rosas-Murrieta, Nora Hilda;Mendieta-Carmona, Victoriano;Hernandez-Pacheco, Raquel Esneidy;Zamora-Ginez, Irma;Rodea-Avila, Carlos;Apresa-Garcia, Teresa;Garay-Villar, Onix;Aguilar-Lemarroy, Adriana;Jave-Suarez, Luis Felipe;Diaz-Orea, Maria Alicia;Milflores-Flores, Lorena;Reyes-Salinas, Juan Salvador;Ceja-Utrera, Francisco Javier;Vazquez-Zamora, Victor Javier;Vargas-Maldonado, Tomas;Reyes-Carmona, Sandra;Sosa-Jurado, Francisca;Santos-Lopez, Gerardo;Reyes-Leyva, Julio;Vallejo-Ruiz, Veronica
    • Asian Pacific Journal of Cancer Prevention
    • /
    • 제15권3호
    • /
    • pp.1181-1186
    • /
    • 2014
  • Sialyltransferase gene expression is altered in several cancers, including examples in the cervix. Transcriptional regulation of the responsible genes depends on different promoters. We aimed to determine the association of single-nucleotide polymorphisms in the B3 promoter of the ST3GAL4 gene and the P1 promoter of the ST6GAL1 gene with cervical premalignant lesions or cervical cancer. A blood sample and/or cervical scrapes were obtained from 104 women with normal cytology, 154 with premalignant lesions and 100 with cervical cancer. We also included 119 blood samples of random donors. The polymorphisms were identified by sequencing from PCR products. For the B3 promoter, a fragment of 506 bp (from nucleotide -408 to +98) was analyzed, and for the P1 promoter a 490 bp (-326 to +164) fragment. The polymorphism analysis showed that at SNP rs10893506, genotypes CC and CT of the ST3GAL4 B3 promoter were associated with the presence of premalignant lesions (OR=2.89; 95%CI 1.72-4.85) and cervical cancer (OR=2.23; 95%CI 1.27-3.91). We detected only one allele of each polymorphism in the ST6GAL1 P1 promoter. We did not detect any genetic variability in the P1 promoter region in our study population. Our results suggest that the rs10893506 polymorphism -22C/T may increase susceptibility to premalignant and malignant lesions of the cervix.

닭의 품종 간 스트레스 반응 정도 비교 분석 (Comparison of Stress Response between Korean Native Chickens and Single Comb White Leghorns subjected to a High Stocking Density)

  • 손시환;조은정;박단비;장인석;문양수
    • 한국가금학회지
    • /
    • 제41권2호
    • /
    • pp.115-125
    • /
    • 2014
  • 본 연구에서는 닭의 품종에 따른 개체의 스트레스 반응정도를 알아보고자 한국 재래닭과 단관 백색 레그혼종을 공시하고, 고밀도 사육에 따른 생리적 스트레스 표지 값을 비교 분석하였다. 스트레스 반응 정도는 혈액과 각 조직별 세포들에 대한 텔로미어 함량, DNA 손상율 및 열 스트레스 단백질(HSPs) 유전자의 발현율을 분석하고 비교하였다. 텔로미어 함량 및 감축율은 양적 형광 접합 보인법으로 분석하였고, DNA 손상율은 Comet assay로 분석하였다. 열 스트레스 단백질 유전자 발현율은 HSP70, HSP90-${\alpha}$, HSP90-${\beta}$ 및 HMGCR을 표적으로 하여 real-time PCR로 분석하였다. 분석 결과, 한국 재래닭과 단관 백색 레그혼 간 품종에 따른 체중, 증체량, 텔로미어 감축율 및 DNA 손상율의 차이는 없는 것으로 나타났다. 그러나 고밀도 사육과 같은 스트레스 사양 관리는 품종에 상관없이 닭의 성장을 저해하고, 텔로미어 감축 및 DNA 손상을 촉진시키는 것으로 나타났다. 한편, HMGCR을 제외한 HSPs 유전자 발현율의 경우, 밀사 사육에 따른 요인뿐만 아니라, 품종 간에도 유의적 차이를 보였다. HSPs의 분석에 따른 스트레스 정도는 단관 백색 레그혼종이 한국 재래닭에 비해 보다 민감하게 반응하는 것으로 나타났다. 따라서 품종과 관계없이 닭에 있어 고밀도 사육이 강한 환경적 스트레스 요인임을 시사하고, 품종 간 스트레스의 반응 정도는 레그혼종이 한국 재래닭에 비해 다소 높은 것으로 사료된다.