• Title/Summary/Keyword: 역전사효소

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Detection of Astrovirus Infection from Hospitalized Young Children Feces by Reverse Transcription-Polymerase Chain Reaction (중증 위장관염 소아의 대변검체에서 역전사-중합효소 연쇄반응을 이용한 Astrovirus 검출)

  • Park, Hae-Kyung;Woo, So-Youn;Seoh, Ju-Young;Chong, Young-Hae;Seo, Jeong-Wan
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.5
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    • pp.453-459
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    • 1999
  • Astrovirus is frequently associated with diarrhea in children. It can not be readily isolated by cell culture, and an electronmicroscope is usually used for detection of this agent. Recently in 1995 a combined method of reverse transcription-polymerase chain reaction (RT-PCR) was designed for easier detection of astrovirus, which is based on the conserved sequence in 3'-end of genomes of the 7 known serotypes of human astrovirus. As of yet there has not been any report of astrovirus data in Korea using the RT-PCR methods. The purpose of this study was to detect astrovirus incidence, severity of symptoms, seasonal variation and co infection rate with rotavirus in Korean children inpatients with diarrhea. Fecal specimens from 61 young children hospitalized with gasteroenteritis Korea from Jan. 1996 through Mar. 1997. They were examined for astroviurs infection by RT-PCR method. Results are as follows:1. Astrovirus was detected at 9.8% (6/61) from fecal specimens of children with severe diarrhea by EIA using monoclonal antibody coated plates. 2. Astorvirus was detected at 29.5% (18/61) from fecal specimens of children with severe diarrhea by RT-PCR. 3. The age of the 18 children affected by astrovirus ranged from 2 monthes to 7 years with mean of 3.0 years. 4. Mean hospital stay of the 18 children was 6.1 days. 5. Five (27.8%) astrovirus RT-PCR positive strains were confirmed in November and in December, respectively out of 18 specimens in total. 6. Astrovirus coinfection with rotavirus type G1 was confirmed in 15/16 specimens (93.8%), and with type G2 was in 1/16 specimens (6.3%).

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Epidemiologic Characteristics of Human Bocavirus-Associated Respiratory Infection in Children (소아 보카바이러스 호흡기 감염증의 역학적 특징)

  • Choi, Jae Hong;Paik, Ji Yeun;Choi, Eun Hwa;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.18 no.1
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    • pp.61-67
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    • 2011
  • Purpose : This study was performed to investigate the epidemiologic characteristics of human bocavirus (HBoV)-associated lower respiratory tract infections (LRTIs) in children. Methods : Nasopharyngeal aspirate samples were obtained from 658 children who had been hospitalized for LRTIs in Seoul National University (SNU) Children's Hospital and SNU Bundang Hospital from March 2000 to September 2005. Multiplex RT-PCR was performed to detect 11 respiratory viruses including respiratory syncytial virus, adenovirus, rhinovirus, parainfluenza viruses 1 and 3, influenza viruses A and B, human metapneumovirus, HBoV, human coronavirus (HCoV) OC43/ 229E, and HCoV-NL63. Clinical data were reviewed retrospectively. Results : Overall, respiratory viruses were detected in 325 (49.4%) among 658 patients. HBoV was detected in 62 cases (9.4%) and was responsible for 19.1% of virus-positive cases. HBoV was prevalent among infants and young children aged from 3 months to 5 years with the mean age of 25.3 months. Co-detection of HBoV and other respiratory viruses was observed in 37.1% which is significantly higher than average co-detection rate (12.3%) among overall virus-positive cases (P=0.000). HBoV was identified mainly in late spring and early summer from May to July. Conclusion : This study describes epidemiologic features of HBoV in Korean children compared with those associated with other respiratory viruses. HBoV was prevalent among LRTIs in childhood, especially in late spring and early summer season in Korea.

Expression of Aquaporin-4 and -8 Genes in Mouse Uterus during the Estrous Cycle (발정주기 동안 생쥐 자궁에서의 Aquaporin-4와 -8 유전자의 발현)

  • Lee Ji Won;Kang Han Seung;Gye Myung Chan;Hong Seok Ho;Shin Hyeonsang;Kang Soo Mnn;Lee Sung Eun;Kim Moon Kyoo
    • Development and Reproduction
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    • v.8 no.1
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    • pp.49-55
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    • 2004
  • Aquaporins(AQPs) are a family of transmembrane water channel proteins that are widely distributed in various tissues throughout the body and play a major role in Oanscellular and Oansepithelial water movement. Uterine endometrium undergoes recurrent uterine stromal edema in response to hormonal stimuli, however, the mechanism regulating the fluid transport during the estrous cycle has not been fully understood. To investigate the possible role of AQPs in water movement in uterus during the estrous cycle, expression patterns of AQP -1, -3, -4, -5, -8, and -9 UMh in mouse uterus were analyzed by using semiquantitative reverse transcription- polymerase chain reaction(RT-nR). We employed a combination of laser capture microdissection(LCM) and RT-PCR to examine the expression patterns in specific uterine cell types luminal epithelial cells(LE) and stromal cells(S). Our results showed that the level of AQP-4 mRNA was significantly increased while the level of AQP-3 mRNA was significantly decreased during the proestous through the estrus stage. In addition LCM revealed that AQP-4 and -8 mRNAs were highly expressed in LE compared with S. Taken together, these results suggest that AQPs may have an important function in physiological changes of mouse uterus during the estrous cycle.

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Expressions of Pituitary Adenylate Cyclase-Activating Polypeptide and Its Receptor Gene in the Rat Uterus (흰쥐 자궁에서 Pituitary Adenylate Cyclase-Activating Polypeptide와 수용체 유전자의 발현)

  • 이성호
    • Development and Reproduction
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    • v.2 no.1
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    • pp.21-27
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    • 1998
  • The present study was performed to analyze the gene expressions of pituitary adenylate cyclase-activating polypeptide(PACAP) and its receptor in the rat uterus, a candidate for novel extrahypothalamic source and target. The PACAP cDNA fragments corresponding to the common exon region which is found in both the rat hypothalamus and testis were produced from all tissue samples including the rat uterus by reverse transcriptionpolymerase chain reaction (RT-PCR). No PCR product was amplified from the rat hypothalamic, pituitary, ovarian and uterine samples when the 5' primer corresponding to the testis-specific exon 1 region was used, while the predicted size of product was detected from the testis sample. RT-PCR using the uterine RNA and specific primers for the PACAP receptor yielded products with predicted sizes. Transcripts for the rat uterine PACAP receptor were identified as type I isoforms with hip-hop and hip- or hop-type inserts. After pregnant mare's serum gonadotropin (15 IU) treatment of immature rats (day 25), the level of PACAP mRNA was increased in 24 h and 48 h group, and was declined to the lowest in 72 h group. The present study shows the presence of transcripts for PACAP and its receptor isoform in the rat uterus. These finding ssuggest that the uterine PACAP ight act as a novel autocrine and/or paracrine factor via its specific receptors on the reglulation of rat uterine function and physiology during the reproductive cycle.

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Effects of Mineral Trioxide Aggregate on the Proliferation and Differentiation of Human Dental Pulp Stromal Cells from Permanent and Deciduous Teeth (Mineral trioxide aggregate가 유치 및 영구치의 치수기질세포 증식 및 분화에 미치는 영향)

  • Kim, Seunghye;Jeon, Mijeong;Shin, Dong Min;Lee, Jae Ho;Song, Je Seon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.40 no.3
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    • pp.185-193
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    • 2013
  • Mineral trioxide aggregate (MTA) has recently been used as a pulpotomy medicament for primary molars. The aim of this study was to evaluate and compare the proliferation and differentiation potential of dental pulp stromal cells of permanent teeth and deciduous teeth cultured on MTA-coated surface. Human dental pulp stromal cells were obtained from human permanent premolars and deciduous teeth and cultured on MTA-coated culture plates. The cells were subjected to proliferation assay and cell cycle analysis. Their differentiation potential was evaluated by analysing changes in the mRNA expressions of runt-related transcriptional factor 2 (Runx2) and alkaline phosphatase (ALP). Morphological changes of cells in direct contact with MTA were observed using scanning electron microscopy (SEM). The proliferation rates, distribution of cell cycles and mRNA expression patterns of Runx2 and ALP were similar in both types of pulpal cells. SEM observations revealed that both types changed into more dendrite-like cells. On the surface of MTA, human dental pulp stromal cells from deciduous and permanent teeth were able to both proliferate and differentiate into cells that induce mineralization. MTA is suitable as a biocompatible pulpotomy medicament for primary teeth.

GENE EXPRESSION PROFILING IN HUMAN DENTAL PULP CELLS TREATED WITH MINERAL TRIOXIDE AGGREGATE (Mineral trioxide aggregate가 인간치수세포에서 유전자 발현에 미치는 영향)

  • Kim, Yong-Beom;Shon, Won-Jun;Lee, Woo-Cheol;Kum, Kee-Yeon;Baek, Seung-Ho;Bae, Kwang-Shik
    • Restorative Dentistry and Endodontics
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    • v.35 no.3
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    • pp.152-163
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    • 2010
  • This study investigated the changes in gene expression when mineral trioxide aggregate (MTA) was applied in vitro to human dental pulp cells (HDPCs). MTA in a teflon tube (diameter 10 mm, height 2 mm) was applied to HDPCs. Empty tube-applied HDPCs were used as negative control. For microarray analysis, total RNA was extracted at 6, 24, and 72 hrs after MTA application. The results were confirmed selectively by performing reverse transcriptase polymerase chain reaction for genes that showed changes of more than two-fold or less than half. Of the 24,546 genes, 109 genes were up-regulated greater than twofold (e.g., FOSB, THBS1, BHLHB2, EDN1, IL11, FN1, COL10A1, and TUFT1) and 69 genes were down-regulated below 50% (e.g., SMAD6 and DCN). These results suggest that MTA, rather than being a bio-inert material, may have potential to affect the proliferation and differentiation of pulp cells in various ways.

A Molecular Study of Rice Black-Streaked Dwarf Virus (벼 흑조위축병 바이러스의 분자생물학적 연구)

  • Park, Jong-Sug;Bae, Shin-Chyul;Kim, Young-Min;Paik, Young-Ki;Kim, Ju-Kon;Hwang, Young-Soo
    • Applied Biological Chemistry
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    • v.37 no.3
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    • pp.148-153
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    • 1994
  • Rice black-streaked dwarf virus (RBSDV), a member of the plant reoviridae fijivirus group, causes a serious damage for rice production in Korea. To characterize the RBSDV genome, virus particles were produced by feeding of planthopper (Laodelphax striatellus F.) carring RBSDV to maize plants for 2 days. In $30{\sim}40$ days after feeding, the viral particles were purified from the infected maize roots by using $10{\sim}40%$ sucrose gradient centrifugation. After treatment of 10% SDS to remove the viral coat proteins, ten viral double-stranded RNAs were resolved in agrose gel electrophoresis. Total dsRNA was then used to synthesize cDNA by reverse transcriptase and a cDNA library was constructed in the ${\lambda}gt11$ vector. The phages that contain RBSDV cDNA fragments were selected by hybridizing with the random-primed probe prepared from RBSDV dsRNAs. After subcloning of several cDNA fragments into the pUC19 plasmid vector, one clone (pRV3) was chosen for sequencing. The pRV3 clone was shown to be located on the RBSDV genome fragment No.3 by RNA gel-blot analysis. Sequence analysis of the clone revealed that the pRV3 contains two partial open reading frames.

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Detection of Cymbidium Mosaic Virus and Odontoglossum Ringspot Virus in Seed-Derived Plantlets of Phalaenopsis Imported by One-Step RT-PCR (One-Step RT-PCR 방법에 의한 수입 호접란묘의 심비디움 모자이크 바이러스와 오돈토글로섬 윤문 바이러스의 검정)

  • Yun, Jong Sun;Hong, Eui Yon;Kim, Ik Hwan;Yun, Tae;Kim, Tae Su;Paek, Kee Yoeup
    • Horticultural Science & Technology
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    • v.18 no.4
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    • pp.513-517
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    • 2000
  • This experiment was carried out to detect the cymbidium mosaic virus (CymMV) and the odontoglossum ringspot virus (ORSV) in the seed-derived plantlets of Phalaenopsis imported from Taiwan by one-step reverse transcription-polymerase chain reaction (RT-PCR). Simple and rapid crude plant extracts for RT-PCR were prepared. The reverse transcription step was performed at $42^{\circ}C$ for 45 min and the following thermal cycling scheme was used for 36 reaction cycles: template predenaturation at $96^{\circ}C$ for 2 min, template denaturation at $96^{\circ}C$ for 30 s, primer annealing at $60^{\circ}C$ for 30 s, and DNA synthesis at $72^{\circ}C$ for 1 min. Of the 40 seed-derived plantlets of Phalaenopsis imported from Taiwan, all of them were infected with CymMV, but ORSV was not detected.

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Expression of Luteinizing Hormone (LH) Subunit Genes in the Rat Ovary (흰쥐 난소에서의 Luteinizing Hormone (LH) Subunit 유전자 발현)

  • Lee, Sung-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.199-205
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    • 1998
  • The present study was performed to analyze the expression of LH genes in the rat ovary. Expression of LH subunit genes in the rat ovary was demonstrated by amplification of ovarian RNA by RT-PCR. The ovarian $LH_\beta$ transcripts contained at least two parts of the published cDNA structure, the pituitary exons 1, 2 and 3 and the part of testicular ex on 1 in the major trancripts form in rat testis. Using RIA, significant amount of LH-like molecules were detected in crude ovarian extracts, and the competition curves with increasing amount of tissue extracts were parallel with those of standard peptide, indicating that the ovarian immunoreactive LH-like material is similar to authentic pituitary LH molecule. The administration of PMSG to immature rats resulted in a sharp decrease of the ovarian LH contents after 24 h post-injection. In conclusion, these findings demonstrate that genes for LH subunits are expressed in the rat ovary, and suggest that LH can playa central role in regulation of female reproduction with both endocrine (by pituitary LH) and auto- and/or para-crine (by ovarian LH) manner.

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Expression of Stromal Derived Factor-1 is Upregulated In Macrophages during Thymic Regeneration in Adult Rat (흰쥐 가슴샘 재생과정 동안 대식세포에서 SDF-1의 발현증가)

  • Park, Hyun-Joo;Kim, Jong-Gab;Yoon, Sik;Bae, Moon-Kyoung;Bae, Soo-Kyung
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1067-1072
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    • 2009
  • Stromal derived factor-1 (SDF-1 or CXCL12), one of the CXC chemokines, is widely expressed in many tissues, including the thymus. The thymus can regenerate to its normal mass within 14 days after acute involution induced by cyclophosphamide (CY) in adult rats. Despite the established role of SDF-1 signaling in the development of T and B lymphocytes in the thymus, it has not yet been associated with the regeneration of the adult thymus. The purpose of this study was to investigate whether SDF-1, which is expressed in thymic stromal cells, is modulated during thymic regeneration in adult rats. Here, we showed that SDF-1 mRNAs were expressed in high levels in the thymocyte and thymic stromal cells at day 7 of the CY model. SDF-1 protein was shown to be present at the cortex-medulla junction, paraseptum and within the thymic medulla. Double-immunofluorescence for SDF-1 and ED-1 showed that strong SDF-1 expression was detected in the macrophages of the medulla region displaying immunoreactivity for ED-1 during thymus regeneration. Taken together, our results demonstrated that SDF-1 expression increased in regenerating thymic macrophages, suggesting the roles of SDF-1 as a chemo-attractant for damaged cells produced in the process of thymic regeneration after acute involution induced by CY.