• 제목/요약/키워드: Direct PCR

검색결과 410건 처리시간 0.026초

마우스에서 VEGF발현 Naked DNA 벡터인 pCK-VEGF의 약동력학 및 조직내 분포 (Pharmacokinetics and Biodistribution in Mice of pCK-VEGF Expressing Human Vascular Endothelial Growth Factor)

  • 도현미;고준일;이종진;손미원;조홍찬;김종묵;김병문;김선영
    • 약학회지
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    • 제45권1호
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    • pp.71-77
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    • 2001
  • We recently developed a high efficiency expression vectors pCK, which drives a high level of gene expression in the skeletal muscles of mice. In this study, we investigated the pharmacokinetics and biodistribution of pCK-VEGF expressing human VEGF165 after intravenous or intramuscular administration. The quantity of pCK-VEGF in the tissues of mice was measured by the PCR method which has a detection limit of approximately 1 pg of the exogenously added plasmid. In the case of intravenous administration, the half life of the pCK-VEGF plasmid in the bloodstream was 1.68 min. After intra-muscular administration, the half life of pCK-VEGF plasmid in the bloodstream was 6.78 min. At 90 min post-administration, 30% of the injected pCK-VEGF was found at the site of injection, where it persisted for up to 8 hours. Less than 1.6% of the injected pCK-VEGF plasmid DNA was detected in highly vascularized tissues such as the lung, kidney; and liver at 90 min post-administration, but the plasmid was undetectable at later time points. These results suggested that intramuscularly administrated pCK-VEGF persisted for longer periods of time in muscles than in other tissues and that direct intra-muscular injection of pCK-VEGF might be useful for local therapeutic angiogenesis.

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Macrophage의 IL-1$\beta$Gene Expression 대한 Lectin-conjugated Ellagitannin의 효과 (Effects of Lectin-conjugated Ellagitannin on the IL-1$\beta$, Gene Expression of Macrophage)

  • 김한준;김민수;이민원;최영욱;김하형;이도익
    • 약학회지
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    • 제46권3호
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    • pp.197-202
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    • 2002
  • Lectin-conjugated praecoxin A, which has developed as a missile antitumor drug, is the one that has conjugated with wheat germ agglutinin (WGA), a kind of carbohydrate-binding protein (lectin) especially bound to melanoma. Praecoxin A is a kind of tannin extracted and purified from plants. Beside this direct antitumor effect as tannins, we have examined an activation of macrophage by lectin-conjugated praecoxin A. We also confirmed the gene expression of IL-1 $\beta$, both in vitro and in vivo. We added 1, 10, 100 $\mu\textrm{g}$/mι of lectin-conjugated praecoxin A, 10 $\mu\textrm{g}$/mι of lectin, 10 $\mu\textrm{g}$/mι of praecoxin A to normal murine macrophage and analyzed the extracted total RNAs by RT-PCR after 4, 8, 12, 24 hours. Our data demonstrated that lectin-conjugated praecoxin A increased IL-1$\beta$, mRNA expression in a dose dependent manner. However, the effectiveness of lectin-conjugated praecoxin A was not superior to lectin and praecoxin A.

일본의 근적외선분광법에 대한 제약회사 응용 및 현황 (Application Study of Chemoinfometrical Near-Infrared Spectroscopic Method to Evaluate for Polymorphic Content of Pharmaceutical Powders)

  • Otsuka, Makoto
    • 한국근적외분광분석학회:학술대회논문집
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    • 한국근적외분광분석학회 2002년도 강연요지집
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    • pp.97-117
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    • 2002
  • A chemoinfometrical method for quantitative determination of crystal content of indomethacin (IMC) polymorphs based on fourie-transformed near-infrared (FT-NIR) spectroscopy was established. A direct comparison of the data with the ones collected from using the conventional powder X-ray diffraction method was performed. Pure $\alpha$ and ${\gamma}$ forms of IMC were prepared using published methods. Powder X-ray diffraction profiles and NIR spectra were recorded for six kinds of standard materials with various content of ${\gamma}$ form IMC. The principal component regression (PCR) analyses were performed based on normalized NIR spectra sets of standard samples of known content of IMC ${\gamma}$ form. A calibration equation was determined to minimize the root mean square error of the prediction. The predicted ${\gamma}$ form content values were reproducible and had a relatively small standard deviation. The values of ${\gamma}$ form content predicted by two methods were in close agreement. The results were indicated that NIR spectroscopy provides for an accurate quantitative analysis of crystallinity in polymorphs compared with the results obtained by conventional powder X-ray diffractometry.

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Analyze of Hepatitis A Virus Genotypes in Eastern Area of Jeonnam, Korea

  • Seo, Min-Young;Lee, Hyeok-Jae;Han, Weon-Dong
    • 대한의생명과학회지
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    • 제17권1호
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    • pp.27-37
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    • 2011
  • Hepatitis A virus (HAV) is a causative agent of triggering acute hepatitis which is transmitted by person-to-person contact and or fecal-oral route. In previous studies, most hepatitis A virus (HAV) isolates had been genotype IA in Korea. Recently, a small number of different genotypes were reported with an upsurge of acute hepatitis by HAV. Therefore, the distribution of HAV genotypes was investigated. RNA was extracted from anti-HAV IgM positive sera which were collected from February to August 2009, at a tertiary care hospital in eastern Jeonnam, Korea. Nested reverse transcription PCR and direct sequencing for VP1/P2A region of the HAV were performed. A total of 365 cases with suspected acute hepatitis were tested for anti-HAV IgM and positive results were obtained in 24 sera (9.0%), which were collected 2 to 15 days (median, 7 days) after the onset of symptoms. Of the 24 seropositive samples, 14 (58.3%) samples were positive for HAV RNA, among which 4 isolates (28.6%) were genotype IA and the other 10 (71.4%) were genotype IIIA. Both IA and IIIA genotypes were isolated from 5~6 neighboring administrative districts throughout the year without geographic or seasonal restrictions. HAV genotypes (IA and IIIA) were observed from the eastern Jeonnam for the studied.

Alum Directly Modulates Murine B Lymphocytes to Produce IgG1 Isotype

  • Jin, Bo-Ra;Kim, Sun-Jin;Lee, Jeong-Min;Kang, Seong-Ho;Han, Hye-Ju;Jang, Young-Saeng;Seo, Goo-Young;Kim, Pyeung-Hyeun
    • IMMUNE NETWORK
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    • 제13권1호
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    • pp.10-15
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    • 2013
  • Aluminum hydroxide (alum) is the most widely used adjuvant in human vaccines. Nevertheless, it is virtually unknown whether alum acts on B cells. In the present study, we explored the direct effect of alum on Ig expression by murine B cells in vitro. LPS-activated mouse spleen B cells were cultured with alum, and the level of isotype-specific Ig secretion, IgG1 secreting cell numbers, and Ig germ-line transcripts (GLT) were measured using ELISA, ELISPOT, and RT-PCR, respectively. Alum consistently enhanced total IgG1 production, numbers of IgG1 secreting cells, and $GLT{\gamma}1$ expression. These results demonstrate that alum can directly cause IgG1 isotype switching leading to IgG1 production.

A simple model for selection and rapid advancement of transgenic progeny in sorghum

  • Visarada, K.B.R.S.;Saikishore, N.;Kuriakose, S.V.;Rani, V. Shobha;Royer, M.;Rao, S.V.;Seetharama, N.
    • Plant Biotechnology Reports
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    • 제2권1호
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    • pp.47-58
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    • 2008
  • To select agronomically useful transgenic plants, a large number of transgenic events are initially produced, gene transfer confirmed, and advanced to obtain homozygous lines for testing in field trials. Direct in planta assays for identifying the transgene carriers in the segregating progeny are based on the activity of selectable marker gene and are easy, simple and inexpensive. For this purpose, expression of bar gene as measured by tolerance to damage by glufosinate ammonium, the active ingredient in the herbicide BASTA, was investigated. Dose damage curves were generated by leaf paint tests with BASTA on four genotypes of sorghum. Transgenic plants were characterized in terms of sensitivity to the concentration of glufosinate ammonium. In transgenics, symptoms of BASTA swab tests at different growth stages and PCR analysis for cry1B were carried out and correlated. Germination tests could not be employed for large scale evaluation of transgenic progeny because of mortality of tolerant seedlings after transplantation to soil. Based on the above findings, a simple, inexpensive, time-saving, two-step scheme for effective evaluation of transgenics and their progeny containing bar gene as selection marker using BASTA swab tests is described.

The Role of Genetic Diagnosis in Hemophilia A

  • Lee, Ja Young
    • Journal of Interdisciplinary Genomics
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    • 제4권1호
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    • pp.15-18
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    • 2022
  • Hemophilia A is a rare X-linked congenital deficiency of clotting factor VIII (FVIII) that is traditionally diagnosed by measuring FVIII activity. Various mutations of the FVIII gene have been reported and they influence on the FVIII protein structure. A deficiency of or reduction in FVIII protein manifests as spontaneous or induced bleeding depending on the disease severity. Mutations of the FVIII gene provide important information on the severity of disease and inhibitor development. FVIII mutations also affect the discrepant activities found using different FVIII assays. FVIII activity is affected differently depending on the mutation site. Long-range PCR is commonly used to detect intron 22 inversion, the most common mutation in severe hemophilia. However, point mutations are also common in patients with hemophilia, and direct Sanger sequencing and copy number variant analysis are being used to screen for full mutations in the FVIII gene. Advances in molecular genetic methods, such as next-generation sequencing, may enable accurate analysis of mutations in the factor VIII gene, which may be useful in the diagnosis of mild to moderate hemophilia. Genetic analysis is also useful in diagnosing carriers and managing bleeding control. This review discusses the current knowledge about mutations in hemophilia and focuses on the clinical aspects associated with these mutations and the importance of genetic analysis.

Trunk Injection of Citrus Trees with a Polymeric Nanobactericide Reduces Huanglongbing Severity Caused by Candidatus Liberibacter asiaticus

  • Ramiro Guerrero-Santos;Gabriela Cabrales-Orona;John Paul Delano-Frier;Judith Cabello-Romero;Jose Roman Torres-Lubian;Jose Humberto Valenzuela-Soto
    • The Plant Pathology Journal
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    • 제40권2호
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    • pp.139-150
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    • 2024
  • Huanglongbing (HLB) is a disease caused by the phloem-limited Candidatus Liberibacter asiaticus (CLas) that affects the citrus industry worldwide. To date, only indirect strategies have been implemented to eradicate HLB. Included among these is the population control of the psyllid vector (Diaphorina citri), which usually provides inconsistent results. Even though strategies for direct CLas suppression seem a priori more promising, only a handful of reports have been focused on a confrontation of the pathogen. Recent developments in polymer chemistry have allowed the design of polycationic self-assembled block copolymers with outstanding antibacterial capabilities. Here, we report the use of polymeric nano-sized bactericide particles (PNB) to control CLas directly in the phloem vasculature. The field experiments were performed in Rioverde, San Luis Potosí, and is one of the most important citrusproducing regions in Mexico. An average 52% reduction in the bacterial population was produced when PNB was injected directly into the trunk of 20 infected trees, although, in some cases, reduction levels reached 97%. These results position PNB as a novel and promising nanotechnological tool for citrus crop protection against CLas and other related pathogens.

호흡기 감염병 진단 기술 동향 (Trends in Diagnostic Technology for Respiratory Infectious Disease)

  • 박정원;서홍석;허철;박수준
    • 전자통신동향분석
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    • 제39권4호
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    • pp.54-62
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    • 2024
  • The emergence and resurgence of novel respiratory infectious diseases since the turn of the millennium, including SARS, H1N1 flu, MERS, and COVID-19, have posed a significant global health threat. Efforts to combat these threats have involved various approaches, however, continued research and development are crucial to prepare for the possibility of emerging viruses and viral variants. Direct detection methods for viral pathogens include molecular diagnostic techniques and immunodiagnostic methods, while indirect diagnostic methods involve detecting changes in the condition of infected patients through imaging diagnostics, gas analysis, and biosignal measurement. Molecular diagnostic techniques, utilizing advanced technologies such as gene editing, are being developed to enable faster detection than traditional PCR methods, and research is underway to improve the efficiency of diagnostic devices. Diagnostic technologies for infectious diseases continue to evolve, and several key trends are expected to emerge in the future. Automation will facilitate widespread adoption of rapid and accurate diagnostics, portable diagnostic devices will enable immediate on-site diagnosis by healthcare professionals, and advancements in AI-based deep learning diagnostic models will enhance diagnostic accuracy.

한국인의 신생아 황달과 UGT1A1 및 CYP1A2 유전자 다형성과의 연관성 (The Association of Neonatal Hyperbilirubinemia with UGT1A1 and CYP1A2 Gene Polymorphism in Korean Neonates)

  • 강훈;임준호;김지숙;김은령;김성도;이희제;정주호
    • Clinical and Experimental Pediatrics
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    • 제48권4호
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    • pp.380-386
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    • 2005
  • 목 적 : 신생아 황달은 백인에 비해 중국, 일본, 한국 등 동아시아인에서 2배 이상 많이 발생하는 것으로 보아 유전적 연관성이 있을 것으로 생각되어 왔고, 최근 일본인, 타이완 중국인, 한국인에서 UGT1A1 유전자의 Gly71Arg 다형성이 신생아 황달과 연관성이 있다고 보고되었다. 선천적으로 UDP-glucuronosyltransferase(UDPGT)에 결함이 있는 경우에는 부경로(alternative pathway)로 CYP1A2 효소를 자극하여 빌리루빈 대사가 이루어진다. 출생 후 6-14주가 되어야 성인 UDPGT 정상치에 도달하기 때문에 신생아 황달에서 빌리루빈 대사에 CYP1A2 부경로가 중요한 역할을 할 것으로 생각된다. 이에 저자들은 UGT1A1과 CYP1A2 유전자의 다형성이 한국인 신생아 황달의 발생과 어떤 연관성이 있는지 알아보고자 본 연구를 시행하였다. 방 법 : 혈중 빌리루빈 수치가 12 mg/dL 이상의 건강하고, 황달의 다른 위험인자가 없는 만삭아 79명과 대조군 68명으로부터 혈액 0.5 cc를 채취하여 DNA을 분리하였다. UGT1A1 유전자는 Polymerase chain reaction(PCR) 후에 염기서열 분석을 통해서 Gly71Arg 유전자 다형성을 확인하였으며, CYP1A2는 제한효소인 MboII를 이용하여 PCR-restriction fragment length polymorphism 방법과 염기서열 분석을 통해서 T2698G 유전자 다형성을 확인하였다. 결 과 : UGT1A1 유전자의 Gly71Arg 다형성은 변이형 대립 유전자 분포가 환자군에서 32%로 대조군 11%보다 높았다(P<0.0001). CYP1A2 유전자의 다형성은 변이형 유전형 분포가 환자군에서는 41.8%, 대조군에서 32.3%로 환자군이 높았으며 통계학적으로 유의하였다(P=0.015). 변이형 대립유전자의 빈도는 환자군에서 21%로 대조군 19%보다 높았으나 통계학적 유의성은 없었다(P=0.706). Gly71Arg와 T2698G의 변이형 발생의 연관성은 없었다(P=0.635). 결 론 : 한국인의 신생아 황달에서 체내의 빌리루빈 대사의 주경로와 부경로에 작용하는 효소의 유전자인 UGT1A1과 CYP1A2의 다형성이 확인되었고, UGT1A1 유전자의 Gly71Arg 다형성은 신생아 황달과 연관이 있었으나 CYP1A2 유전자의 T2698G 다형성은 신생아의 황달과 연관이 없었다.