• Title/Summary/Keyword: PGP 9.5

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Expression of Human Thrombopoietin in Insect Cells by Polyhedrin-gp64 Dual Promoter-Based Baculovirus Vector System

  • Koh, Yeo-Wook;Park, Sang-Kyu;Kim, Kun-Soo;Kim, Seong-Ryong;Lee, Jeong-Kug;Yang, Jai-Myung
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.318-322
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    • 1999
  • A new baculovirus transfer vector (pPGP404) was constructed to increase the expression level of human thrombopoietin (hTPO) in insect cells. In pPGP404, hTPO was cloned next to the AcNPV polyhedrin-gp64 dual promoter and the leader sequence of hTPO was substituted with that of gp64. A recombinant baculovirus, AcPGP404, was constructed by using pPGP404 as a transfer vector. hTPO was expressed in AcPGP404-infected TN5 cells and it was observed that the expression levels of hTPO in TN5 cells increased three-fold ($6.0 {\mu}g/ml^{-1}$) compared to the level expressed under the control of the polyhedrin single promoter. These results indicate that the polyhedrin-gp64 dual promoter system would be useful for expression in large quantities of recombinant proteins in insect cells.

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An Immunohistochemical Study of Pheasant Testis in Active, Inactive and Damaged States (번식기, 비번식기 및 손상상태에 따른 한국꿩 고환의 면역조기화학적 연구)

  • ;Karl-heinz Wrobel
    • Korean Journal of Poultry Science
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    • v.24 no.3
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    • pp.107-116
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    • 1997
  • In order to achieve optimal reproductive performance, reliable morphological and physiological basic data on the reproductive organs are desirable. Adult male Korean ring-necked pheasant in inactive(mid of January) and active state (end of April) were used in this study. In addition, five active state pheasants were received a single dose of 60Co-ray 500 rads each to damage the testes. The objective of this study was to investigate the distribution pattern of protein gene product (PGP) 9.5 and ${\alpha}$-tubulin in the pheasant testes of the active, inactive and ${\gamma}$-ray irradiated active states. The results obtained were summarized as follows 1. The seminiferous tubules collected in inactive states( mid of Jan) showed narrow lumen, and the spermatogonia and the Sertoli cell were well preserved. The PGP 9.5 immunoreactivity of these tubules showed a positive reaction in paranucleus area of the spermatogonia, and a positive reaction in a small number of the Leydig cells in the interstitium of the seminiferous tubules. 2. The seminiferous tubules were dilated in active state(end of April) as compared with the inactive state. The PGP 9.5 reactivity in these tubules showed a positive reaction in many Leydig cells in the interstitium of the seminiferous tubules, and the testes of ${\gamma}$-ray irradiated group showed partially weak reaction in the interstitium of the seminiferous tubules. 3. The ${\alpha}$-tubulin reactivity in the seminiferous tubules of the inactive testes was strongly positive in the cytoplasmic process of the Sertoli cell from the basal stem region to the apical ex-tension. From the broad part of the stem region to the luminal space, the active testes showed a strong positive reaction. The ${\gamma}$-ray irradiated groups showed diminished reaction in the basal region.

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Cytological analysis of pregnancy-associated plasma protein-A expression in porcine neonatal testis (미성숙 돼지 정소에서 pregnancy-associated plasma protein-A의 발현의 세포학적 분석)

  • Kim, Ji-youn;Oh, Keon Bong;Byun, Sung June;Ock, Sun-A;Lee, Hwi-Cheul;Hwang, Seong-Su;Park, SangHyun;Ha, Wootae;Woo, Jae-Seok;Song, Hyuk
    • Journal of Embryo Transfer
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    • v.33 no.3
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    • pp.177-183
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    • 2018
  • The identification of biomarkers of a living tissues is essentially required to understand specific functions of the cells. In previous study, we reported IGFBP 3 as one of the putative biomarkers, by showing specific expression at porcine spermatogonial stem cells (SSCs) of early stage of porcine testis. In this study, we analyzed the expression of seven members of IGFBP family (IGFBPs) in SSCs and histological expression pattern of pregnancy-associated plasma protein-A (PAPP-A), which plays a role on the growth promoting enzyme by cleavage of IGFBPs in testis of 5 days old pig. RT-PCR analysis showed that IGFBP 1, 2, 3, 4, and 6 were expressed at high level specifically in porcine SSCs compared with whole testis. We performed immunohisotochemical staining of testis sections with PAPP-A and protein gene product 9.5 (PGP9.5) which are the known biomarkers for SSCs. We were not able to find co-expression of PAPP-A and PGP9.5; PAPP-A was expressed only in Sertoli cells and PGP9.5 expression was confirmed in spermatogonium. Additionally, we were able to confirm the GATA4 expression in Sertoli and Leydig cells as a regulator of Sertoli cell function was not detected PGP9.5 expressing cells, indicating indirect evidence of that cytolocalization of PAPP-A expression is limited in Sertoli cells. These results suggested that the PAPP-A expressed in Sertoli cells may play role on regulation of development and differentiation of testicular cells through the IGF axis in neonatal porcine testis.

Application of Bacterial Endophytes to Control Bacterial Leaf Blight Disease and Promote Rice Growth

  • Ooi, Ying Shing;Nor, Nik M.I. Mohamed;Furusawa, Go;Tharek, Munirah;Ghazali, Amir H.
    • The Plant Pathology Journal
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    • v.38 no.5
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    • pp.490-502
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    • 2022
  • Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial leaf blight (BLB) disease in rice (Oryza sativa L.) and it is among the most destructive pathogen responsible for severe yield losses. Potential bacterial biocontrol agents (BCAs) with plant growth promotion (PGP) abilities can be applied to better manage the BLB disease and increase crop yield, compared to current conventional practices. Thus, this study aimed to isolate, screen, and identify potential BCAs with PGP abilities. Isolation of the BCAs was performed from internal plant tissues and rhizosphere soil of healthy and Xoo-infected rice. A total of 18 bacterial strains were successfully screened for in vitro antagonistic ability against Xoo, siderophore production and PGP potentials. Among the bacterial strains, 3 endophytes, Bacillus sp. strain USML8, Bacillus sp. strain USML9, and Bacillus sp. strain USMR1 which were isolated from diseased plants harbored the BCA traits and significantly reduced leaf blight severity of rice. Simultaneously, the endophytic BCAs also possessed plant growth promoting traits and were able to enhance rice growth. Application of the selected endophytes (BCAs-PGP) at the early growth stage of rice exhibited potential in suppressing BLB disease and promoting rice growth.

Analysis of Pregnancy-Associated Plasma Protein-A (PAPP-A) in Porcine Neonatal Testis (미성숙 돼지 정소 내에서의 pregnancy-associated plasma protein-A 특성 분석)

  • Lee, W.Y.;Cho, K.H.;Yeo, J.M.;Shin, Y.K.;Park, J.K.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.22 no.1
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    • pp.5-13
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    • 2020
  • The identify of biomarkers in living tissues is useful to understand the characteristics and functions of the cells. Proteins such as protein gene product 9.5, promyelocytic leukemia zinc finger, NANOG, and stage-specific embryonic antigen-1 have been identified as markers for porcine undifferentiated spermatogonia. In this study, the expression of insulin-like growth factor binding proteins (IGFBPs), a newly discovered porcine spermatogonia marker and pregnancy-associated plasma protein-A (PAPP-A), a protein regulator of IGFBPs, were characterized in 5-day-old porcine testis. To analyze the function of IGFBPs, RT-PCR was performed. IGFBP 2, 3, 4, and 6 were detected in porcine spermatogonia and PAPP-A was detected in basement regions in 5day old porcine seminiferous tubules. PAPP-A was not expressed in spermatogonia, but it was expressed in Sertoli cells. These results suggest that the expression of PAPP-A protein in Sertoli cells may regulate the development and differentiation of testicular cells through the IGF axis in porcine neonatal testis.

Comparative Uptake of Tc-99m Sestamibi and Tc-99m Tetrofosmin in Cancer Cells and Tissue Expressing P-Glycoprotein or Multidrug Resistance Associated Protein (P-Glycoprotein과 Multidrug Resistance Associated Protein을 발현하는 암세포와 종양에서 Tc-99m Sestamibi와 Tc-99m Tetrofosmin의 섭취율 비교)

  • Cho, Jung-Ah;Lee, Jae-Tae;Yoo, Jung-Ah;Seo, Ji-Hyoung;Bae, Jin-Ho;Jeong, Shin-Young;Ahn, Byeong-Cheol;Sohn, Sang-Gyun;Ha, Jeoung-Hee;Lee, Kyu-Bo
    • The Korean Journal of Nuclear Medicine
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    • v.39 no.1
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    • pp.34-43
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    • 2005
  • Purpose: $^{99m}Tc$-sestamibi(MIBI) and $^{99m}Tc$-tetrofosmin have been used as substrates for P-glycoprotein (Pgp) and multidrug resistance associated protein (MRP), which are closely associated with multidrug resistance of the tumors. To understand different handling of radiotracers in cancer cell lines expressing Pgp and MRP, we compared cellular uptakes of $^{99m}Tc$-MIBI and $^{99m}Tc$-tetrofosmin. The effects of cyclosporin A (CsA), well-known multidrug resistant reversing agent, on the uptake of both tracers were also compared. Materials and Methods: HCT15/CL02 human colorectal cancer cells for Pgp expressing cells, and human non-small cell lung cancer A549 cells for MRP expressing cells, were used for in vitro and in vivo studies. RT-PCR, western blot analysis and immunohistochemistry were used for detection of Pgp and MRP. MDR-reversal effect with CsA was evaluated at different drug concentrations after incubation with MIBI or tetrofosmin. Radioactivities of supernatant and pellet were measured with gamma well counter. Tumoral uptake of the tracers were measured from tumor bearing nude mice treated with or without CsA. Results: RT-PCR, western blot analysis of the cells and irnrnunochemical staining revealed selective expression of Pgp and MRP for HCY15/CL02 and A549 cells, respectively. There were no significant difference in cellular uptakes of both tracers in HCT15/CL02 cells, but MIBI uptake was slightly higher than that of tetrofosmin in A549 cells. Co-incubation with CsA resulted in a increase in cellular uptakes of MIBI and tetrofosmin. Uptake of MIBI or tetrofosmin in HCT15/CL02 cells was increased by 10- and 2.4-fold, and by 7.5 and 6.3-fold in A549 cells, respectively. Percentage increase of MIBI was higher than that of tetrofosmin with CsA for both cells (p<0.05). In vivo biodistribution study showed that MIBI (114% at 10 min, 257% at 60 min, 396% at 240 min) and tetrofosmin uptake (110% at 10 min, 205% at 60 min, 410% at 240 min) were progressively increased by the time, up to 240 min with CsA. But increases in tumoral uptake were not significantly different between MIBI and tetrofosmin for both tumors. Conclusion: MIBI seems to be a better tracer than tetrofosmin for evaluating MDR reversal effect of the modulators in vitro, but these differences were not evident in vivo tumoral uptake. Both MIBI and tetrofosmin seem to be suitable tracers for imaging Pgp- and MRP-mediated drug resistance in tumors.

Dendritic-like Cells Accumulate in BPD Lung of Baboon (만성 폐질환인 폐이형성증을 보이는 baboon 폐조직에 Dendritic-like cell의 축적)

  • Lee, Jong-Hwan;Kim, Kwang-Hyeon
    • Journal of Life Science
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    • v.18 no.9
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    • pp.1202-1206
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    • 2008
  • The present investigation demonstrates HLA-DR and PGP9.5 double positive cells accumulate thymus cortical region in normal baboon thymus and baboon lung. But, these cells disappeared in thymus and lung of bronchopulmonary dysplasia (BPD) animals. 125d GC animal model is more suitable for BPD than 140d GC animal. Anti-bombesin antibody, 2A11 treated baboon recover normal level of HLA-DR positive cells from BPD animal. In addition, thymocytes show responsiveness for bombesin. These observation suggest that blocking BLPs protects a chronic lung injury by BPD and 2A11 is possible agent for passive therapy of BPD.

A morphological study of vomeronasal organ of Korean black goat (Capra aegagrus hircus) (한국흑염소 보습코기관의 형태학적 관찰)

  • Park, Changnam;Yang, Wonjun;Bae, Yeonji;Lee, Yongduk;Kang, Wanchoul;Ahn, Meejung;Shin, Taekyun
    • Korean Journal of Veterinary Research
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    • v.53 no.1
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    • pp.55-60
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    • 2013
  • The vomeronasal organ (VNO) plays an important role in reproduction and social activities in ruminants including goats. A morphological study on the structure of VNO and its epithelial cells was carried out in Korean black goats. Grossly, the VNO of Korean goats opens into mouth through incisive ducts. Microscopically, the epithelium of VNO consisted of medial sensory epithelium and lateral non-sensory epithelium. Several blood vessels and nerve bundles were observed in the lamina propria encased by vomeronasal cartilage. Immunohistochemical staining showed that protein gene product (PGP) 9.5 was immunostained in the receptor cells of the sensory epithelium and in some cells of the non-sensory epithelium. Galectin-3 was mainly observed in the supporting cells of sensory and non-sensory epithelium. Lectins including wheat germ agglutinin, Ulex europaeus agglutinin, Bandeiraea simplicifolia lectin Isolectin B4, Dolichos biflorus agglutinin and soybean agglutinin used in this study were bound in VNO sensory, non-sensory epithelia as well as in the lamina propria with varying intensity. Collectively, this is a first descriptive morphological study of VNO of Korean black goat with special reference to lectin histochemistry.

Mass Cultivation and Secondary Metabolite Analysis of Rhodobacter capsulatus PS-2 (광합성세균 Rhodobater capsulatus PS-2의 대량배양 최적화 및 대사산물 분석)

  • Bong, Ki Moon;Kim, Jong Min;Yoo, Jae-Hong;Park, In Chul;Lee, Chul Won;Kim, Pyoung Il
    • KSBB Journal
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    • v.31 no.3
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    • pp.158-164
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    • 2016
  • Plant growth promoting (PGP) hormones, which are produced in a small quantity by bacteria, affect in plant growth and development. PGPs play an important role on the crop productivity in agricultural field. In this study, a photosynthetic bacterial strain producing the PGP was isolated from paddy soil. Bacterial isolate was gram negative, rod-shaped and motility positive. From the 16s rRNA gene sequence analysis, the isolate was identified as Rhodobacter capsulatus PS-2. The mass cultivation of R. capsulatus PS-2 was optimized by considering of the carbon, nitrogen and inorganic salt sources. Optimal medium composition was determined as Na-succinate 4.5 g, yeast extract 5 g, $K_2HPO_4$ 1 g, $MgSO_4$ 5 g, per liter. From the result of 500 L fermentation for 2 days using the optimal medium, the viable cells were $8.7{\times}10^9cfu/mL$. R. capsulatus PS-2 strain produced the carotenoid and indole-3-acetic acid (IAA). The carotenoid extraction and quantitative analysis were performed by HCl-assisting method. Total carotenoid contents from R. capsulatus PS-2 culture broth were measured as $7.02{\pm}0.04$ and $6.93{\pm}0.05mg/L$ under photoheterotrophic and chemoheterotrophic conditions, respectively. To measure the productivity of IAA, colorimetric method was employed using Salkowski reagent at optical density 535 nm. The results showed that the highest content of IAA was $197.44{\pm}5.92mg/L$ in the optimal medium supplemented with 0.3% tryptophan.

Skin Biopsy: Emerging Method for Small Nerve Fiber Evaluation (피부조직검사: 소신경섬유 평가의 유용한 방법)

  • Sohn, Eun Hee
    • Annals of Clinical Neurophysiology
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    • v.17 no.2
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    • pp.53-60
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    • 2015
  • Skin biopsy with investigation of small nerve fiber in human epidermis and dermis has been proven to be a useful method for demonstration of small fiber neuropathy. Quantification of intraepidermal nerve fiber density using anti-Protein Gene Product 9.5 (PGP 9.5) antibody is standardized method to diagnose the small fiber neuropathy. Skin biopsy method also makes it possible to differentiate the type of nerve fibers by using different antibodies. Quantification of dermal structures with different type of nerve fibers could be used to invest pathophysiologic mechanism of diseased state.