• Title/Summary/Keyword: PAP-II

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자리공 항바이러스 단백질 II 유전자의 형질전환에 의한 연초의 바이러스 저항성 품종 개발 (I)

  • 강신웅;이영기;이기원;박성원;이청호
    • Journal of the Korean Society of Tobacco Science
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    • v.21 no.1
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    • pp.57-63
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    • 1999
  • Pokeweed antiviral protein II (PAP-II) encoding cDNA was synthesized by reverse-transcriptase polymerase chain reaction (RT-PCR) from Phytolacca american a leaf. The PAP-II cDNA fragment of 974bp was subcloned to pBluescript II SK- SmaI site and the inserted PAP-II cDNA fragment was sequenced by dideoxy sequencing method. The number of nucleotides of PAP-II cDNA coding region containing start and stop codon was 933bp. To develop a virus-resistant tobacco plant, PAP-II cDNA fragment was inserted to pKGT101B and the insertion of PAP-II cDNA fragment was confirmed by restriction enzyme analysis and colony PCR.

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Deletion Mutation of Pokeweed Antiviral Protein II Gene and Development of PVY-VN Resistant Tobacco Plants (미국자리공 항바이러스 단백질 II 유전자의 돌연변이 및 PVY-VN 저항성 담배식물체 생산)

  • 강신웅;이영기;박성원;한규웅;김선원;이종철;이청호
    • Journal of the Korean Society of Tobacco Science
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    • v.23 no.2
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    • pp.123-132
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    • 2001
  • In order to transform pokeweed antiviral protein cDNA to tobacco plant, total RNA was extracted from Phytolacca americana. PAP-II cDNA was synthesized from purified total RNA via RT-PCR and subcloned to recombinant vector pBluescript II SK-. 10 deletion mutant PAP-II cDNA fragments which were sequentially deleted from N-terminal by 90bp were synthesized from PAP-II cDNA except leading frame by PCR with primers designed in our laboratory. To select non-cytotoxic clone, pAc55M was constructed with yeast expression vector pAc55 and multicloning site(MCS). Sequentially deleted mutant PAP-II cDNAs were cloned on downstream of gall promoter of pAc55M. 6 non-cytotoxic deletion mutant PAP-II cDNA were selected. Selected cDNAs were cloned into plant expression vector pKGT101BH for transformation of these clones to plant through Agrobacterium tumefacience. After cloning, recombinant pKGT101BH carrying deleted mutant PAP-IIcDNA were transformed to Nicotiana tabacum cv. NC567. Transformed tobacco plants cultured on shooting and rooting media were transfered to green-house. About four weeks later, these plants were infected with physically infection using carborandum with PVY-VN strain. After 4 weeks, plants resistant to virus were selected , and seeds of these plants were gathered. Southern blot hybridization showed deleted fragments by 220bp and 420bp, so resistant ability of these plants is due to mutant PAP-II cDNA.

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Cytotoxicity Test of Pokeweed Antiviral Protein Type I Gene by Using Yeast Expression System (효모 발현 시스템을 이용 PAP(Pokeweed Antiviral Protein) 유전자의 세포독성 연구)

  • 김선원;박성원;강신웅;이영기;이종철;최순용;이청호
    • Journal of the Korean Society of Tobacco Science
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    • v.23 no.2
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    • pp.133-140
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    • 2001
  • PAP-I cDNA was synthesized from total RNA of Phytolacca americana leaves by RT-PCR, and then subcloned to recombinant vector pBluescript II SK-. Using PCR with primers designed in our laboratory, we could get the 9 deletion mutant PAP-I cDNA fragments. The first of the fragments was deleted by 66bp from immature N-terminal and then the rest were deleted by 90bp sequentially. Sequentially deletion mutant PAP-I cDNAs were inserted to pAc55M, on down-stream of gall promoter. Recombinant pAc55M was transformed to yeast cells, psy1 and the cells were spreaded on SC_urn-/glucose plate media. Colonies on SC_ura-/glucose plate were streaked on the same position of SC_ura-/glucose and SC_ura-/galactose plate, and we selected colonies growing on both plates, which carry non-cytotoxic deleted mutant PAP-I cDNA. We selected 4 deletion mutant PAP-I cDNAs which have not cytotoxicity.

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First Trial of Cervical Cytology in Healthy Women of Urban Laos using by Self-sampling Instrument

  • Nabandith, Viengvansay;Pholsena, Vatsana;Mounthisone, Phouthasone;Shimoe, Kyoko;Kato, Saiko;Aoki, Kunio;Noda, Sadamu;Takamatsu, Reika;Saio, Masanao;Yoshimi, Naoki
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.9
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    • pp.4665-4667
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    • 2012
  • Cervical cancer is the most common cancer in Laos women and a screening programme, even with the PAP smear test (PAP test), has yet to be established for routine use. The Pap test is accepted as the most appropriate for cervical cancer screening in some settings but it is not commonly available in Laos hospitals, because there are few cytopathologists and gynecologists have little experience. As a pilot program, seminars for the PAP test were given in 2007 and 2008, and then PAP tests were carried out using self-sampling instrument (Kato's device) with 200 healthy volunteers in Setthathirath hospital, Laos, in 2008. The actual examination number was 196, divided into class I 104 (53.1%), class II 85 (43.3%), class IIIa 4 (2.0%), class IIIb 1 (0.5%), and class V 1 (0.5%) by modified Papanicolau classification. Four cases had menstruation. There were 6 cases with epithelial cell abnormalities including malignancy. There were 7 cases with fungus and 2 cases with trichomonas in Class II. More than 70% volunteers felt comfortable with the Kato's device and wanted to use it next time, because of the avoidance of the embarrassment and a low cost as compared with pelvic examination by gynecologists. This first trial for PAP test for healthy Laos women related to a hospital found three percent to have abnormal cervical epithelial cells. Therefore, this appraoch using a self-sampling device suggests that it should be planned for cervical cancer prevention in Laos.

Perception of Cervical Cancer Screening Among Japanese University Students Who Have Never Had a Pap Smear: A Qualitative Study

  • Oshima, Sumiko;Maezawa, Masaji
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4313-4318
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    • 2013
  • This study aimed to explore attitudes towards cervical cancer screening among Japanese university students who had never had a Pap smear. Four focus-group discussions, each with 15 female university students, took place in November and December 2009. Discussions were recorded and transcripts were analyzed to extract attitudes of young women towards cervical cancer screening. The four themes that emerged were: i) a low sense of reality about cervical cancer; ii) a lack of knowledge about both cervical cancer and Pap smears; iii) a lack of motivation to get screened, and iv) a reluctance to visit the gynecologist. Participants who were interested in undergoing screening for cervical cancer cited the influence of conversations with friends and family, a diagnosis of cancer within their family, and relevant information from the media. The results indicate the importance of getting young women more interested in cervical cancer screening and overcoming their tendency to avoid visiting a gynecologist.

Introduction and Expression of PAP gene using Agrobacterium in Scrophularia buergeriana Miquel (Agrobacterium을 이용한 PAP 유전자의 현삼으로 도입 및 형질발현)

  • Yu, Chang-Yeon;Seong, Eun-Soo;Lim, Jung-Dae;Huang, Shan-Ai;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.2
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    • pp.156-165
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    • 2001
  • Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

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Hypoxic pulmonary vasoconstriction and vascular contractility in monocrotaline-induced pulmonary arterial hypertensive rats

  • Kim, Hae Jin;Yoo, Hae Young
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.6
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    • pp.641-647
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    • 2016
  • Pulmonary arterial hypertension (PAH) is a progressive disease characterized by vascular remodeling of pulmonary arteries (PAs) and increased vascular resistance in the lung. Monocrotaline (MCT), a toxic alkaloid, is widely used for developing rat models of PAH caused by injury to pulmonary endothelial cells; however, characteristics of vascular functions in MCT-induced PAH vary and are not fully understood. Here, we investigated hypoxic pulmonary vasoconstriction (HPV) responses and effects of various vasoconstrictors with isolated/perfused lungs of MCT-induced PAH (PAH-MCT) rats. Using hematoxylin and eosin staining, we confirmed vascular remodeling (i.e., medial thickening of PA) and right ventricle hypertrophy in PAH-MCT rats. The basal pulmonary arterial pressure (PAP) and PAP increase by a raised flow rate (40 mL/min) were higher in the PAH-MCT than in the control rats. In addition, both high $K^+$ (40 mM KCl)- and angiotensin II-induced PAP increases were higher in the PAH-MCT than in the control rats. Surprisingly, application of a nitric oxide synthase inhibitor, L-$N^G$-Nitroarginine methyl ester (L-NAME), induced a marked PAP increase in the PAH-MCT rats, suggesting that endothelial functions were recovered in the three-week PAH-MCT rats. In addition, the medial thickening of the PA was similar to that in chronic hypoxia-induced PAH (PAH-CH) rats. However, the HPV response (i.e., PAP increased by acute hypoxia) was not affected in the MCT rats, whereas HPV disappeared in the PAH-CH rats. These results showed that vascular contractility and HPV remain robust in the MCT-induced PAH rat model with vascular remodeling.

Three Cases of Pulmonary Alveolar Proteinosis (폐포 단백증 3예)

  • Kim, Yeon-Jae;Han, Chun-Duk;Cha, Seung-Ick;Kim, Chang-Ho;Lee, Yeung-Suk;Park, Jae-Yong;Jung, Tae-Hoon;Park, Tae-In;Sohn, Yun-Kyung
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.4
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    • pp.416-423
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    • 1993
  • Pulmonary alveolar proteinosis(PAP) is thought to be a rare disease of unknown etiology characterized by the accumulation of strong PAS-positive lipoproteinaceous material in the pulmonary alveolar spaces. The defect in the clearance and degradation of intra-alveolar phospholipoproteinaceous material in PAP likely represents dysfunction of type II pneumocytes. Although the causative treatment of PAP is not well known, yet whole lung bronchopulmonary lavage is a relatively safe and effective treatment. We experienced three cases of PAP, which were confirmed by light and electron microscopic examinations of lung tissues obtained by open lung biopsy, transbronchial lung biopsy and lung lavage, so we present 3 cases of PAP with a review of the literature.

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Interaction of Nitric Oxide and Renin Angiotensin System in Pulmonary Arterial Circulation of RHR

  • Lee, Byung-Ho;Shin, Hwa-Sup
    • Archives of Pharmacal Research
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    • v.20 no.5
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    • pp.389-395
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    • 1997
  • We investigated the interaction between nitric oxide and the renin angiotensin system in regulating isolated pulmonary arterial tension and pulmonary arterial pressure (PAP) in renal hypertensive rats (RHR) made by complete ligation of left renal artery. Losartan induced a depressor response that was smaller in RHR than in normotensive rats (NR) (3.3 and 7.0 mmHg, respectively, at 3.0 mg/kg, p<0.05), and the response was significantly reduced by $N^{G}$-nitro-Larginine methyl ester (L-NAME). Angiotensin II elevated the PAP (7.6 and 10.8 mmHg at $0.1 {\mu}g/kg$; 20.3 and 23.6 mmHg at $1.0{\mu}g/kg$, respectively) and contracted the isolated pulmonary artery ($pD_2$: 8.79 and 8.71, respectively) from both NR and RHR with similar magnitude, and these effects were significantly enhanced by L-NAME in NR, but not in RHR. Acetylcholine lowered the PAP slightly less effectively in RHR than in NR (3.8 and 6.0 mmHg at 10 .mu.g/kg, respectively) and relaxed the pulmonary artery precontracted with norepinephrine in both rats with similar magnitude ($E_max$: 60.8 and 63.6%, respectively), and the effect being completely abolished after pretreatment.with L-NAME or removal of endothelial cells. These results suggest that nitric oxide interacts with renin angiotensin system to control the pulmonary vascular tension and pulmonary arterial circulation of RHR.R.

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Subglottic Air Pressure in Different Phonetic Context (음성학적 문맥에 따른 성문하압의 차이에 관한 연구)

  • 박상희;정옥란;석동일
    • Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
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    • v.13 no.1
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    • pp.23-27
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    • 2002
  • The purpose of the study is to examine differences in subglottic air pressure as a function of phonetic context. The phonetic contexts consisted of $/i:{p^h}i:{p^h}i:/,/{p^h}i:{p^h}i:/, and /{p^h}{p^h}/$. The aerodynamic and phonatory parameters are investigated in 20 female normal adults. All measurements are taken and analysed using Aerophone II voice function analyzer. The aerodynamic parameters are Peak Air Pressure(PAP) and Mean Air Pressure(MAP), and the phonatory parameters are Phonatory Flow Rate(PFR) Maximum SPL(MSPL), Phonatory SPL(PSPL), Phonatory Power (PP), Phonatory Efficiency(PE), and Phonatory $Resistance^*$ 10-5(PR). A one-way ANOVA revealed the following results. First, the aerodynamic parameters are not significantly different. Second, Peak Air Pressure(PAP) and Mean Air Pressure(MAP), as well as the phonatory parameters such as Phonatory Flow Rate(PFR) Maximum SPL(MSPL), Phonatory SPL(PSPL), and Phonatory Efficiency(PE) were significantly different. Therefore, it is advised that clinicians use only aerodynamic parameters but phonatory parameters when using Aerophone II.

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