• Title/Summary/Keyword: Re-RT

Search Result 86, Processing Time 0.03 seconds

Direct Stem Blot Immunoassay (DSBIA): A Rapid, Reliable and Economical Detection Technique Suitable for Testing Large Number of Barley Materials for Field Monitoring and Resistance Screening to Barley mild mosaic virus and Barley yellow mosaic virus

  • Jonson, Gilda;Park, Jong-Chul;Kim, Yang-Kil;Kim, Mi-Jung;Lee, Mi-Ja;Hyun, Jong-Nae;Kim, Jung-Gon
    • The Plant Pathology Journal
    • /
    • v.23 no.4
    • /
    • pp.260-265
    • /
    • 2007
  • Testing a large number of samples from field monitoring and routine indexing is cumbersome and the available virus detection tools were labor intensive and expensive. To circumvent these problems we established tissue blot immunoassay (TBIA) method an alternative detection tool to detect Barley mild mosaic virus (BaMMV) and Barley yellow mosaic virus (BaYMV) infection in the field and greenhouse inoculated plants for monitoring and routine indexing applications, respectively. Initially, leaf and stem were tested to determine suitable plant tissue for direct blotting on nitrocellulose membrane. The dilutions of antibodies were optimized for more efficient and economical purposes. Results showed that stem tissue was more suitable for direct blotting for it had no background that interferes in the reaction. Therefore, this technique was referred as direct stem blot immunoassay or DSBIA, in this study. Re-used diluted (1:1000) antiserum and conjugate up to 3 times with the addition of half strength amount of concentrated antibodies was more effective in detecting the virus. The virus blotted on the nitrocellulose membrane from stem tissues kept at room temperature for 3 days were still detectable. The efficiency of DSBIA and RT-PCR in detecting BaMMV and BaYMV were relatively comparable. Results further proved that DSBIA is a rapid, reliable and economical detection method suitable for monitoring BaMMV and BaYMV infection in the field and practical method in indexing large scale of barley materials for virus resistance screening.

Effect of Bee Venom Acupuncture on the Recovery of Chondrocyte Phenotype in Rabbit Cartilage (봉독약침이 연골세포 활성에 미치는 영향)

  • Zhao Mei-Ai;Lee Seung-Ki;Kim Gun-Ho;Choi Sun-Mi;Shim In-Sop;Kang Sung-Keel;Lee Hye-Jung;Hahm Dae-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
    • /
    • v.19 no.5
    • /
    • pp.1251-1255
    • /
    • 2005
  • Articular cartilage is an important factor for studying the arthritic diseases. The chondrocyte isolated from cartilage has the characieristic of differentiation and rf-differentiation when cultured in monolayer The bee venom (BV) acupuncture was investigated to examine their abilities on chondrocyte re-differentiation via rabbit chondrocyte primary culture. And the expression profiles of type II collagen (COL2) was analyzed using RT-PCR and western hybridization at third passage chondrocyte. In general, the mRNA expression of type II collagen was reduced step by step with the subculture of the chondrocyte. However, after the BV treatment for 48hr at third passage, the expressions of type II collagen were found to be significantly up-regulated, the same result was confirmed by western blotting. These results suggested that the diluted solution of BV for herb-acupuncture was very effective on the recovery of articular chondrocyte phenotype.

Differentially Expressed Genes of Potentially Allelopathic Rice in Response against Barnyardgrass

  • Junaedi, Ahmad;Jung, Woo-Suk;Chung, Ill-Min;Kim, Kwang-Ho
    • Journal of Crop Science and Biotechnology
    • /
    • v.10 no.4
    • /
    • pp.231-236
    • /
    • 2007
  • Differentially expressed genes(DEG) were identified in a rice variety, Sathi, an indica type showing high allelopathic potential against barnyardgrass(Echinochloa crus-galli(L.) Beauv. var. frumentaceae). Rice plants were grown with and without barnyardgrass and total RNA was extracted from rice leaves at 45 days after seeding. DEG full-screening was performed by $GeneFishing^{TM}$ method. The differentially expressed bands were re-amplified and sequenced, then analyzed by Basic Local Alignment Search Tool(BLAST) searching for homology sequence identification. Gel electrophoresis showed nine possible genes associated with allelopathic potential in Sathi, six genes(namely DEG-1, 4, 5, 7, 8, and 9) showed higher expression, and three genes(DEG-2, 3 and 6) showed lower expression as compared to the control. cDNA sequence analysis showed that DEG-7 and DEG-9 had the same sequence. From RT PCR results, DEG-6 and DEG-7 were considered as true DEG, whereas DEG-1, 2, 3, 4, 5, and 8 were considered as putative DEG. Results from blast-n and blast-x search suggested that DEG-1 is homologous to a gene for S-adenosylmethionine synthetase, DEG-2 is homologous to a chloroplast gene for ribulose 1,5-bisphosphate carboxylase large subunit, DEG-8 is homologous to oxysterol-binding protein with an 85.7% sequence similarity, DEG-5 is homologous to histone 2B protein with a 47.9% sequence similarity, DEG-6 is homologous to nicotineamine aminotransferase with a 33.1% sequence similarity, DEG-3 has 98.8% similarity with nucleotides sequence that has 33.1% similarity with oxygen evolving complex protein in photosystem II, DEG-7 is homologous to nucleotides sequence that may relate with putative serin/threonine protein kinase and putative transposable element, and DEG-4 has 98.8% similarity with nucleotides sequence for an unknown protein.

  • PDF

Combined Chemoradiotherapy vs Radiotherapy Alone for Locally Advanced Squamous Cell Carcinoma of the Head and Neck (국소적으로 진행된 두경부 편평상피세포종양의 방사선- 항암화학 병용요법과 방사선단독치료의 비교)

  • Jeong, Hyeon-Ju;Suh, Hyun-Suk;Kim, Chul-Soo;Kim, Re-Hwe;Kim, Sung-Rok
    • Radiation Oncology Journal
    • /
    • v.14 no.1
    • /
    • pp.9-15
    • /
    • 1996
  • Purpose: The traditional approach with surgery and/or radiotherapy(RT) for advanced head and neck cancer Provides anticipated cure rates of $10-65\%$ depending on stages and sites. Recently, combined modality with chemotherapy have been extensively investigated in attempts to improve survival and local control. We retrospectively analysed our experience of 31 patients with advanced head and neck cancer. Materials and Methods : November 1983 to October 1994. 31 Patients with Stage III and IV squamous cell head and neck cancer were treated with RT. Sixteen patients were treated with RT alone, and IS patients were treated with combined RT plus chemotherapy. All patients were treated with 4-MV LINAC and radiation dose ranged from 5000 cGy to 7760 cGy (median 7010 cGy). In combined group, 7 patients were treated with cisplatin plus 5-FU 2 patients were treated with methotrexate plus leucovorin plus 5-FU plus cisplatin or carboplatin, and 6 patients were treated with cisplatin as a radiosensitizer. Results : Median follow up period was 16 months (range 4-134 months). The major responses (CR+PR) were noted in 10 patient ($66.6\%$) of the RT alone group and 14 patient ($93.3\%$) of the chemoradiation group. There was no statistical difference in CR rate between the two groups The overall survival rates at 5 years were $23.4\%$ in the radiation alone group, $23.5\%$ in the chemoradiation group Disease-free survival rates at 3 years were $44.5\%$ in the radiation alone group, $40\%$ in the chemoradiation group. There was no statistical differences in overall survival rates and disease-free survival rates between the two groups. Local recurrences occurred in $71.5\%$ of the radiation alone group, $72.7\%$ of the chemoradiation group and distant metastasis occurred in $14.4\%$ of radiation alone group, $9.1\%$ of the chemoradiation group. The frequencies of complications were comparable in both groups except hematologic toxicity Conclusion : Total response rates in the combined chemotherapy and radiotherapy was relatively higher than radiotherapy alone. But our result failed to show any survival benefit of the combined chemotherapy and radiotherapy. The accrual of large number of patients and long term follow-un may be necessary to confirm the present result of combined chemotherapy and radiotherapy.

  • PDF

Gene Expression Analysis of Inducible cAMP Early Repressor (ICER) Gene in Longissimus dorsi of High- and Low Marbled Hanwoo Steers (한우 등심부위 근육 내 조지방함량에 따른 inducible cAMP early repressor (ICER) 유전자발현 분석)

  • Lee, Seung-Hwan;Kim, Nam-Kuk;Kim, Sung-Kon;Cho, Yong-Min;Yoon, Du-hak;Oh, Sung-Jong;Im, Seok-Ki;Park, Eung-Woo
    • Journal of Life Science
    • /
    • v.18 no.8
    • /
    • pp.1090-1095
    • /
    • 2008
  • Marbling (intramuscular fat) is an important factor in determining meat quality in Korean beef market. A grain based finishing system for improving marbling leads to inefficient meat production due to an excessive fat production. Identification of intramuscular fat-specific gene might be achieved more targeted meat production through alternative genetic improvement program such as marker assisted selection (MAS). We carried out ddRT-PCR in 12 and 27 month old Hanwoo steers and detected 300 bp PCR product of the inducible cAMP early repressor (ICER) gene, showing highly gene expression in 27 months old. A 1.5 kb sequence was re-sequenced using primer designed base on the Hanwoo EST sequence. We then predicted the open reading frame (ORF) of ICER gene in ORF finder web program. Tissue distribution of ICER gene expression was analysed in eight Hanwoo tissue using realtime PCR analysis. The highest ICER gene expression showed in Small intestine followed by Longissimus dorsi. Interestingly, the ICER gene expressed 2.5 time higher in longissimus dorsi than in same muscle type, Rump. For gene expression analysis in high- and low marbled individuals, we selected 4 and 3 animal based on the muscle crude fat contents (high is 17-32%, low is 6-7% of crude fat contents). The ICER gene expression was analysed using ANOVA model. Marbling (muscle crude fat contents) was affected by ICER gene (P=0.012). Particularly, the ICER gene expression was 4 times higher in high group (n=4) than low group (n=3). Therefore, ICER gene might be a functional candidate gene related to marbling in Hanwoo.

Distribution of Human Rotavirus Genotypes in a Tertiary Hospital, Seoul, Korea During 2009-2013 (2009년부터 2013년까지 서울의 일개 대학병원에서 동정된 로타바이러스 유전형의 분포)

  • Han, Tae Hee;Park, Sang-Hun;Chung, Ju-Young;Hwang, Eung-Soo
    • Pediatric Infection and Vaccine
    • /
    • v.22 no.2
    • /
    • pp.81-90
    • /
    • 2015
  • Purpose: Group A rotavirus (RV) is most common etiologic agent of acute gastroenteritis (AGE) in children worldwide. Recently, vaccination has been introduced in several countries to reduce the disease burden caused by RV infections, but continuous surveillance of RV strains is necessary to detect the emergence of potential variants induced by vaccine-immune pressure. This study aimed to investigate the changing pattern of RV genotypes in children with AGE, following the introduction of vaccination in Korea. Methods: Genotyping of RVs by RT-PCR on the basis of VP7 and VP4 gene segment sequence was carried out on 201 rotavirus-positive stool samples, from children hospitalized with AGE between August 2009 and June 2013. We have directly sequenced PCR products and analyzed the phylogenetic tree. Results: The most prevalent G genotype was G9 (33.3%), followed by G1 (22.4%), G3 (15.9%), G2 (6.0%), G4 (3.0%), G10 (1.5%), and mixed G-type (15.4%), with some nontypeable cases (2.5%). The detected P genotypes were P[4] (45.3%), P[8] (43.8%), mixed P-type (10.4 %), and P[2] (0.5%). The G9P[4] genotype was predominantly observed in hospitalized cases in Seoul in 2010/2011, however G1P[8] has been re-emerged as the predominant genotype in the following season (P =0.004). Conclusions: It seems that the periodic fluctuation in predominance of the G1, G3, and G9 strains occurred in Korea during 2009-2013, following the introduction of RV vaccination.

Cornu cervi pantotrichum Pharmacopuncture Solution Facilitate Hair Growth in C57BL/6 Mice

  • Lee, Seon-Yong;Lee, Dong-Jin;Kwon, Kang;Lee, Chang-Hyun;Shin, Hyun Jong;Kim, Jai Eun;Ha, Ki-Tae;Jeong, Han-Sol;Seo, Hyung-Sik
    • Journal of Pharmacopuncture
    • /
    • v.19 no.2
    • /
    • pp.122-128
    • /
    • 2016
  • Objectives: Cornu cervi pantotrichum (CCP) has been widely used in Korean and China, as an anti-fatigue, anti-aging, and tonic agent to enhance the functions of the reproductive and the immune systems. Because CCP has various growth factors that play important roles in the development of hair follicles, we examined whether CCP pharmacopuncture solution (CCPPS) was capable of promoting hair growth in an animal model. Methods: One day after hair depilation, CCPPS were topically applied to the dorsal skin of C57BL/6 mice once a day for 15 days. Hair growth activity was evaluated by using macro- and microscopic observations. Dorsal skin tissues were stained with hematoxylin and eosin. Expressions of bromodeoxyuridine (BrdU), proliferating cell nuclear antigen (PCNA), and fibroblast growth factor (FGF)-7 were examined by using immunohistochemical staining. A reverse transcription polymerase chain reaction (RT-PCR) analysis was also conducted to measure the messenger RNA (mRNA) expression of FGF-7. Results: CCPPS induced more active hair growth than normal saline. Histologic analysis showed enlargement of the dermal papilla, elongation of the hair shaft, and expansion of hair thickness in CCPPS treated mice, indicating that CCPPS effectively induced the development of anagen. CCPPS treatment markedly increased the expressions of BrdU and PCNA in the hair follicles of C57BL/6 mice. In addition, CCPPS up regulated the expression of FGF-7, which plays an important role in the development of hair follicles. Conclusion: These results reveal that CCPPS facilitates hair re-growth by proliferation of hair follicular cells and up-regulation of FGF-7 and suggest that CCPPS can potentially be applied as an alternative treatment for patients with alopecia.

CONTRALATERAL NECK LYMPH NODE METASTASIS OF EARLY TONGUE CANCER : A CASE REPORT (반대측 경부 림프절로 전이된 초기 설암의 치험례)

  • Kwon, Myung-Hee;Kim, Sung-Il;Kim, Pyoung-Soo;Leem, Dae-Ho;Shin, Hyo-Keun;Ko, Seung-O
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
    • /
    • v.35 no.5
    • /
    • pp.376-379
    • /
    • 2009
  • Likely to be the most common oral cancer, squamous cell carcinoma(SCC) of the tongue accounts for about 20% of all oral and pharyngeal cancers. SCC of the tongue frequently arises in the lateral border, and if it metastasize, it occurs on submandibular gland and neck lymph nodes. Location of the primary lesions and neck lymph node metastasis affect the prognosis and decrease survival rate of patients with carcinoma of the tongue. The authors experienced the patient with contralateral neck lymph node metastasis of SCC of the tongue. The patient came to our department with chief complaint of elevated lesion on left lateral border of the tongue. The mass was diagnosed as $T_2N_0M_0$, Stage II invasive SCC of oral tongue. Computed tomography(CT) & magnetic resonance imaging(MRI) which were taken before the operation showed no significant finding of metastasis. Surgical mass removal and preventive neck dissection on the left side were done. While follow up PET/CT, contralateral neck lymph node metastasis(right side, level II) was detected, and re-operation(Rt. side RND) was done. There are few studies concerning the contralateral neck lymph node metastasis related with SCC of the tongue. The purpose of this report is to introduce the uncommon case of contralateral neck lymph node metastasis occurred in the $T_2$-stage of SCC of the tongue treated by surgical resection.

Establishement of Antibody Selection by Ribosome Display (Ribosome Display를 이용한 항체선별 방법의 확립)

  • Lee, Myung-Shin;Kwon, Myung-Hee;Hwang Kim, Kyongmin;Park, Sun;Shin, Ho-Joon;Kim, Hyung-Il
    • IMMUNE NETWORK
    • /
    • v.3 no.3
    • /
    • pp.219-226
    • /
    • 2003
  • Background: Phage display is the most widely used technique among display methods to produce monoclonal antibody fragment with a specific binding activity. Having a large library for efficient antibody display/selection is quite laborious process to have more than $10^9$ members of transformants. To overcome these limitations, several in vitro selection approaches have been reported. Ribosome display that links phenotypes, proteins, directly to genotype, mRNA, is one of the in vitro display methods. Ribosome display can reach the size of scFv library up to $10^{14}$ molecules and it can be further diversified during PCR steps. To select the high affinity scFv from one pot library, we established ribosome display technique by modifying the previously reported eukaryotic translation system. Methods: To establish the antibody selection system by ribosome display, we used 3D8, anti-DNA antibody. A 3D8 scFv was synthesized in vitro by an in vitro transcription-translation system. The translated 3D8 scFv and the encoding 3D8 mRNA are connected to the ribosome. These scFv-ribosome-mRNA complexes were selected by binding to their specific antigens. The eluted mRNAs from the complexes are reverse transcribed and re-amplified by PCR. To apply this system, antibody library from immunized mouse with terminal protein (TP)-peptide of hepatitis B virus DNA polymerase TP domain was also used. This TP-peptide encompasses the 57~80 amino acid residues of TP. These mRNA/ribosome/scFv complexes by our system were panned three times against TP-peptide. The enrichment of antibody from library was determined by radioimmunoassay. Results: We specifically selected 3D8, anti-DNA antibody, against ssDNA as a model system. The selected 3D8 RNAs sequences from translation complexes were recovered by RT-PCR. By applying this model system, we enriched TP-peptide-specific scFv pools through three cycles of panning from immunized library. Conclusion: We show that our translating ribosome complexes are well maintained and we can enrich the TP-specific scFv pools. This system can be applied to select specific antibody from an antibody library.

Image Enhancement of the Weathered Zone and Bedrock Surface with a Radial Transform in Engineering Seismic Data (엔지니어링 탄성파자료에서 방사변환을 통한 풍화대 및 기반암 표면의 영상강화)

  • Kim, Ji-Soo;Jeon, Su-In;Lee, Sun-Joong
    • The Journal of Engineering Geology
    • /
    • v.22 no.4
    • /
    • pp.459-466
    • /
    • 2012
  • A difficulty encountered in engineering seismic mapping is that reflection events from shallow discontinuities are commonly overlapped with coherent noise such as air wave, direct waves, head waves, and high-amplitude surface waves. Here, the radial trace transform, a simple geometric re-mapping of a trace gather (x-t domain) to another trace gather (v-t domain), is applied to investigate the rejection effect of coherent linear noises. Two different types of data sets were selected as a representative database: good-quality data for intermediate sounding (hundreds of meters) in a sedimentary basin and very noisy data for shallow (${\leq}50m$) mapping of the weathered zone and bedrock surface. Results obtained with cascaded application of the radial transform and low-cut filtering proved to be as good as, or better than, those produced using f-k filtering, and were especially effective for air wave and direct wave. This simple transform enables better understanding of the characteristics of various types of noise in the RT domain, and can be generally applied to overcoming diffractions and back-scatterings caused by joints, fractures, and faults commonly that are encountered in geotechnical problems.