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In Vitro Wheat Immature Spike Culture Screening Identified Fusarium Head Blight Resistance in Wheat Spike Cultured Derived Variants and in the Progeny of Their Crosses with an Elite Cultivar

  • Huang, Chen;Gangola, Manu P.;Kutcher, H. Randy;Hucl, Pierre;Ganeshan, Seedhabadee;Chibbar, Ravindra N.
    • The Plant Pathology Journal
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    • v.36 no.6
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    • pp.558-569
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    • 2020
  • Fusarium head blight (FHB) is a devastating fungal disease of wheat (Triticum aestivum L.). The lack of genetic resources with stable FHB resistance combined with a reliable and rapid screening method to evaluate FHB resistance is a major limitation to the development of FHB resistant wheat germplasm. The present study utilized an immature wheat spike culture method to screen wheat spike culture derived variants (SCDV) for FHB resistance. Mycotoxin concentrations determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) correlated significantly (P < 0.01) with FHB severity and disease progression during in vitro spike culture. Selected SCDV lines assessed for FHB resistance in a Fusarium field disease nursery in Carman, Manitoba, Canada in 2016 showed significant (P < 0.01) correlation of disease severity to the in vitro spike culture screening method. Selected resistant SCDV lines were also crossed with an elite cv. CDC Hughes and the progeny of F2 and BC1F2 were screened by high resolution melt curve (HRM) analyses for the wheat UDP-glucosyl transferase gene (TaUGT-3B) single nucleotide polymorphism to identify resistant (T-allele) and susceptible (G-allele) markers. The progeny from the crosses were also screened for FHB severity using the immature spike culture method and identified resistant progeny grouped according to the HRM genotyping data. The results demonstrate a reliable approach using the immature spike culture to screen for FHB resistance in progeny of crosses in early stage of breeding programs.

Study on the Shelf Life of Sterilized Products according to Packaging Materials (포장재에 따른 멸균품의 유효기간에 관한 연구)

  • Chang, Song Ja;Jeong, Jeong Hee;Choi, Kyoung Mi;Kim, Mi Young;Park, Joo Hee;Jeong, Na Yeon
    • Journal of Korean Clinical Nursing Research
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    • v.25 no.3
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    • pp.333-341
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    • 2019
  • Purpose: The purpose of this study was to determine the most appropriate shelf life for sterilized products according to their packaging material. Methods: Samples were prepared to target six nursing units in one general hospital in Seoul. After steam and E.O gas sterilization, sterilized product, samples were supplied to wards. Data collection was conducted for 3 months, after the expiration date of 3 months had passed for samples packaged with crepe paper and nonwoven wraps. For samples packaged with paper-plastic pouches, data collection conducted for 3 months when the expiration date of 9 months had passed. The sterilized products were collected and tested for microbial contamination. Identification of the storage environment was done as samples were collected. Results: This study confirmed that the storage environment met international standards such as CDC, except for temperature. For steam sterilized crepe paper packaging samples and steam and E.O gas sterilized for nonwoven packaging samples no contamination in all products was found for 3 months past the expiration date. However, in the E.O gas sterilized paper-plastic pouch packaging sterile samples, Gram-positive bacilli were detected in one sample from a surgical intensive care unit at 45 weeks and another sample from an operating room at 47 weeks. Furthermore, the results did not show any microorganisms for up to 52 weeks in all products. Conclusion: According to the results of this study, sterilized product packaging made with crepe paper and nonwoven wraps is better able an extended shelf life from 3 months to 6 months, reducing unnecessary costs.

Hsp90 Inhibitor Induces Cell Cycle Arrest and Apoptosis of Early Embryos and Primary Cells in Pigs

  • Son, Myeong-Ju;Park, Jin-Mo;Min, Sung-Hun;Hong, Joo-Hee;Park, Hum-Dai;Koo, Deog-Bon
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.33-45
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    • 2011
  • Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of cancer cell. Inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the cancer cell was reported. However, its role during oocyte maturation and early embryo development is very insufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on meiotic maturation and early embryonic development in pigs. We also investigated several indicators of developmental potential, including structural integrity, gene expression (Hsp90-, cell cycle-, and apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Then, we examined the roles of Hsp90 inhibitor on viability of primary cells in pigs. Porcine oocytes were cultured in the NCSU-23 medium with or without 17-AAG for 44 h. The proportion of GV arrested oocytes was significantly different between the 17-AAG treated and untreated group (78.2 vs 34.8%, p<0.05). After completion of meiotic maturation, the proportion of MII oocytes was lower in the 17-AAG treated group than in the control group (27.9 vs 71.0%, p<0.05). After IVF, the percentage of penetrated oocytes was significantly lower in the 17-AAG treated group (25.2%), resulting in lower normal pronucleus formation (2PN of 14.6%). Therefore, the inhibition of meiotic progression by Hsp90 inhibitor played a critical role in fertilization status. Porcine embryo were cultured in the PZM-3 medium with or without 17-AAG for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (7.5 vs 4.4, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. The mRNA expressions of cell cycle-related genes were down-regulated in the 17-AAG treated group compared with control. Also, the expression of the pro-apoptotic gene Bax increased in 17-AAG treated group, whereas expression of the anti-apoptotic gene Bel-XL decreased. However, the expression of ER stress-related genes did not changed by 17-AAG. Cultured pESF cells were treated with or without 17-AAG and used for MIT assay. The results showed that viability of pESF cells were decreased by treatment of 17-AAG ($2{\mu}M$) for 24 hr. These results indicated that 17-AAG decreased cell proliferation and increased cell death. Expression patterns Hsp90 complex genes (Hsp70 and p23), cell cycle-related genes (cdc2 and cdc25c) and apoptosis-related genes (Bax and Bcl-XL) were significantly changed by using RT-PCR analysis. The spliced form of pXbp-1 product (pXbp-1s) was detected in the tunicamycin (TM) treated cells, but it is not detected in 17-AAG treated cells. In conclusion, Hsp90 appears to play a direct role in porcine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with cell cycle- and apoptosis-related genes expression in developing porcine embryos.

A Study on the Correlation between Categorization of the Individual Exposure Levels to Agent Orange and Serum Dioxin Levels Among the Korean Vietnam Veterans (베트남 참전 제대 군인의 범주화된 에이전트 오렌지 개인 폭로량과 혈청 다이옥신 측정치와의 상관성에 관한 연구)

  • Kang, Han-K.;Lim, Hyun-Sul;Cheong, Hae-Kwan;Lim, Min-Kyung;Kim, Joung-Soon
    • Journal of Preventive Medicine and Public Health
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    • v.34 no.1
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    • pp.80-88
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    • 2001
  • Objectives : In an epidemiologic study on the health impact of Agent Orange exposure, the valid estimation of exposure level is the most important step. Based on recent studies, we examined the correlation between exposure levels categorized by personal exposure estimates and serum 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD, Dioxin), exploring the possibility of utilizing the exposure level as a surrogate for the estimate of exposure to agent orange. Methods : During the study period (Jan 1996-Feb 1996), blood specimens of 745 subjects taken randomly among 1,329 persons and kept frozen, were analyzed for 2,3,7,8-TCDO and six other dioxin congeners. The serum dioxin and congeners were measured in 1998 by CDC, adjusted for serum lipids. We categorized the total exposure scores into five groups based on Agent Orange exposure data collected by interview and military records. Pearson and Spearman's correlation coefficients & multiple regression analysis were used to identify the relationship of the exposure level categorized with serum concentration of 2,3,7,8-TCDD, and six other dioxin congeners. Results : Dioxin and the other congeners, except 1,2,4,6,7,8-HpCDD, showed significant correlations to exposure categories (p<0.005): 2,3,7,8-TCDD and OCDD showed positive correlations, whereas the other congeners did negative. The values of 2,3,7,8-TCDD differed according to exposure category and proportionally increased from the low exposure group to the high, a dose-response relationship, even after other possible confounding variables were adjusted for. In multiple regression analysis, age$(\beta=0.033)$, dioxin$(\beta=0.433)$, 1,2,3,7,8-PeCDD$(\beta=-0.998)$, 1,2,3,4,7,8-HxCDD$(\beta=-0.773)$, 1,2,3,6,7,8-HxCDD$(\beta=0.255)$, 1,2,3,7,8,9-HxCDD$(\beta=-3.468)$, 1,2,3,4,6,7,8-HpCDD$(\beta=0.109)$ we re found to be significantly related to the total exposure score(p<0.005). Conclusion : This study demonstrated that the use of such categorizations as a surrogate measure of agent orange exposure in identifying exposure degrees in a health impact study is valid.

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The Study on Carbon Budget Assessment in Pear Orchard (배 재배지의 탄소수지 산정에 관한 연구)

  • Suh, Sanguk;Choi, Eunjung;Jeong, Hyuncheol;Lee, Jongsik;Kim, Gunyeob;Lee, Jaeseok;Sho, Kyuho
    • Korean Journal of Environmental Biology
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    • v.33 no.3
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    • pp.345-351
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    • 2015
  • This study was conducted to find out the methodology of carbon budget assessment among soil, atmosphere and plant. Soil respiration, net ecosystem productivity of herbs and net ecosystem productivity of woody plants have been measured in 30 years old pear orchard at Naju. Closed Dynamic Chamber (CDC) method was used to measure soil respiration and net ecosystem productivity of herbs. Net ecosystem productivity of woody plant (pear) was determined by eddy covariance method using the EddyPro (5.2.1) program. As for soil respiration, $429.1mgCO_2m^{-2}h^{-1}$ was released to atmosphere and sensitivity of soil temperature ($Q_{10}$) was 2.3. In case of herbs, respiration was superior to photosynthesis during measurement period. From 20 to 24 Jun 2015, the sum of absorbed and released $CO_2$ by herb's photosynthesis and respiration was $156.1mgCO_2m^{-2}h^{-1}$. Woody plants showed the $680.1mgCO_2m^{-2}h^{-1}$ of absorption by photosynthesis. In a farm scale, the sum of soil respiration, and net ecosystem productivity of herbs and woody plants was $0.04tonCO_2ha^{-1}$ during the measurement period, and it showed that pear orchard act as a $CO_2$ sink. This study using various approaches is expected to present a methodology for evaluating the carbon budget of perennial woody crop plantations.

Sensitization of the Apoptotic Effect of ${\gamma}$-Irradiation in Genistein-pretreated CaSki Cervical Cancer Cells

  • Shin, Jang-In;Shim, Jung-Hyun;Kim, Ki-Hong;Choi, Hee-Sook;Kim, Jae-Wha;Lee, Hee-Gu;Kim, Bo-Yeon;Park, Sue-Nie;Park, Ok-Jin;Yoon, Do-Young
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.523-531
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    • 2008
  • Radiotherapy is currently applied in the treatment of human cancers. We studied whether genistein would enhance the radiosensitivity and explored its precise molecular mechanism in cervical cancer cells. After co-treatment with genistein and irradiation, the viability, cell cycle analysis, and apoptosis signaling cascades were elucidated in CaSki cells. The viability was decreased by co-treatment with genistein and irradiation compared with irradiation treatment alone. Treatment with only ${\gamma}$-irradiation led to cell cycle arrest at the $G_1$ phase. On the other hand, co-treatment with genistein and ${\gamma}$-irradiation caused a decrease in the $G_1$ phase and a concomitant increase up to 56% in the number of $G_2$ phase. In addition, co-treatment increased the expression of p53 and p21, and Cdc2-tyr-15-p, supporting the occurrence of $G_2/M$ arrest. In general, apoptosis signaling cascades were activated by the following events: release of cytochrome c, upregulation of Bax, down regulation of Bcl-2, and activation of caspase-3 and -8 in the treatment of genistein and irradiation. Apparently, co-treatment downregulated the transcripts of E6*I, E6*II, and E7. Genistein also stimulated irradiation-induced intracellular reactive oxygene, species (ROS) production, and co-treatment-induced apoptosis was inhibited by the antioxidant N-acetylcysteine, suggesting that apoptosis has occurred through the increase in ROS by genistein and ${\gamma}$-irradiation in cervical cancer cells. Gamma-irradiation increased cyclooxygenase-1 (COX-2) expression, whereas the combination with genistein and ${\gamma}$-irradiation almost completely prevented irradiation-induced COX-2 expression and $PGE_2$ production. Co-treatment with genistein and ${\gamma}$-irradiation inhibited proliferation through $G_2/M$ arrest and induced apoptosis via ROS modulation in the CaSki cancer cells.

Association between soluble forms of the receptor for advanced glycation end products and periodontal disease: a retrospective study

  • Kim, Keun-Suh;Lee, Yun Jong;Ahn, Soyeon;Chang, Yoon-Seok;Choi, Yonghoon;Lee, Hyo-Jung
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.47 no.6
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    • pp.445-453
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    • 2021
  • Objectives: Periodontitis is the most common chronic disease that causes tooth loss and is related to systemic diseases such as cardiovascular disease and diabetes. An objective indicator of the current activity of periodontitis is necessary. Soluble forms of the receptor for advanced glycation end products (sRAGE) are markers that reflect the status of inflammatory diseases. In this study, the relationship between sRAGE and periodontitis was analyzed to determine whether it can be used to diagnose the current state of periodontitis. Patients and Methods: Eighty-four patients without any systemic diseases were diagnosed with periodontitis using three classifications of periodontitis. Demographics and oral examination data such as plaque index (PI), bleeding on probing (BOP) index, and probing pocket depth (PPD) were analyzed according to each classification. In addition, correlation and partial correlation between sRAGE and the values indicating periodontitis were analyzed. Results: In each classification, the level of sRAGE tended to decrease if periodontitis was present or severe, but this change was not statistically significant. sRAGE and periodontitis-related variables exhibited a weak correlation, among which the BOP index showed a relatively strong negative correlation (ρ=-0.20). Based on this, on analyzing the correlation between the BOP index and sRAGE in the group with more severe periodontitis (PPD≥5 mm group, severe group of AAP/CDC [American Academy of Periodontology/Centers for Disease Control and Prevention], periodontitis group of López), the correlation further increased (ρ=-0.23, -0.40, -0.50). Partial correlation analysis of the sRAGE and BOP index showed a stronger negative correlation (ρ=-0.36, -0.55, -0.45). Conclusion: sRAGE demonstrated a tendency to decrease upon increased severity of periodontitis according to the classifications used. Above all, the correlation with the BOP index, which reflects the current state of periodontitis, was higher in the group with severe periodontitis. This indicates that the current status of periodontitis can be diagnosed through sRAGE.

Fully Automated Liquid Culture System Compared with Lowenstein-Jensen Solid Medium for Rapid Recovery of Mycobacteria in Sputums (완전 자동화된 액체배양법과 기존의 고체배양법을 이용한 객담 내 mycobacterium의 신속검출에 대한 비교)

  • Park, Seung-Kyu;Kim, Seung-Chul;Kim, Deuk-Mi;Lee, Chang-Woon;Kim, Young;Cho, Sang-Nae
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.6
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    • pp.635-643
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    • 2002
  • Background : The Aim of this study was to compare the recovery of mycobacteria from sputum samples of pulmonary tuberculosis patients using the MB/BacT rapid culture system(Organon Teknika, USA) with that obtained using Lowenstein-Jensen solid medium. Methods : The two culture systems were compared using sputum samples of 99 pulmonary tuberculosis patients. Culture media were incubated at $35-37^{\circ}C$ for six weeks in the MB/BacT system and for 12 weeks in Lowenstein-Jensen solid medium. Solid media were examined macroscopically once a week, and the MB/BacT system positive vials were unloaded from the machine as soon as possible after positive signal from the connected computer was detected Confirmation of growth for mycobacteria was done by Ziehl-Neelson stained smears. Isolates were identified to differentiate Mycobacterium tuberculosis from mycobacterium other than tuberculosis(MOTT) by phenotypic and molecular methods. Results : Of the sputum samples of the 99 patients, 58 samples were smear positive and 41 in negative smear. Mycobacteria were recovered from 67(67.7%) samples by using both culture systems. The yield with MB/BacT was higher than that with Lowenstein-Jensen [67(67.7%) vs. 52(52.5%), p<0.001]. Moreover, 15(15.2%) samples were positive only in the MB/BacT, whereas none of samples was positive only in Lowenstein-Jensen. In smear-positive and smear-negative samples, the recovery rate with MB/BacT was also higher than that with Lowenstein-Jensen [sputum-positive; 56/58(96.6%) vs. 46/58(79.3%), p=0.005, sputum-negative; 6/41(14.6%) vs. 5/41(12.2%), p<0.001]. The mean times to detection of Mycobacteria were 13.3 and 27.2 days with MB/BacT and Lowenstein-Jensen respectively(p<0.001). Conclusion : This results indicate that the the MB/BacT is more efficient and faster than Lowenstein-Jensen for the recovery of mycobacteria.

The Significance of SDF-1α-CXCR4 Axis in in vivo Angiogenic Ability of Human Periodontal Ligament Stem Cells

  • Bae, Yoon-Kyung;Kim, Gee-Hye;Lee, Jae Cheoun;Seo, Byoung-Moo;Joo, Kyeung-Min;Lee, Gene;Nam, Hyun
    • Molecules and Cells
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    • v.40 no.6
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    • pp.386-392
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    • 2017
  • Periodontal ligament stem cells (PDLSCs) are multipotent stem cells derived from periodontium and have mesenchymal stem cell (MSC)-like characteristics. Recently, the perivascular region was recognized as the developmental origin of MSCs, which suggests the in vivo angiogenic potential of PDLSCs. In this study, we investigated whether PDLSCs could be a potential source of perivascular cells, which could contribute to in vivo angiogenesis. PDLSCs exhibited typical MSC-like characteristics such as the expression pattern of surface markers (CD29, CD44, CD73, and CD105) and differentiation potentials (osteogenic and adipogenic differentiation). Moreover, PDLSCs expressed perivascular cell markers such as NG2, ${\alpha}-smooth$ muscle actin, platelet-derived growth factor receptor ${\beta}$, and CD146. We conducted an in vivo Matrigel plug assay to confirm the in vivo angiogenic potential of PDLSCs. We could not observe significant vessel-like structures with PDLSCs alone or human umbilical vein endothelial cells (HUVECs) alone at day 7 after injection. However, when PDLSCs and HUVECs were co-injected, there were vessel-like structures containing red blood cells in the lumens, which suggested that anastomosis occurred between newly formed vessels and host circulatory system. To block the $SDF-1{\alpha}$ and CXCR4 axis between PDLSCs and HUVECs, AMD3100, a CXCR4 antagonist, was added into the Matrigel plug. After day 3 and day 7 after injection, there were no significant vessel-like structures. In conclusion, we demonstrated the perivascular characteristics of PDLSCs and their contribution to in vivo angiogenesis, which might imply potential application of PDLSCs into the neovascularization of tissue engineering and vascular diseases.

Analysis on Geographical Variations of the Prevalence of Hypertension Using Multi-year Data (다년도 자료를 이용한 고혈압 유병률의 지역간 변이 분석)

  • Kim, Yoomi;Cho, Daegon;Hong, Sungok;Kim, Eunju;Kang, Sunghong
    • Journal of the Korean Geographical Society
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    • v.49 no.6
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    • pp.935-948
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    • 2014
  • As chronic diseases have become more prevalent and problematic, effective cares for major chronic diseases have been a locus of the healthcare policy. In this regard, this study examines how region-specific characteristics affect the prevalence of hypertension in South Korea. To analyze, we combined a unique multi-year data set including key indicators of health conditions and health behaviors at the 237 small administrative districts. The data are collected from the Annual Community Health Survey between 2009 and 2011 by Korea Centers for Disease Control and Prevention and other government organizations. For the purpose of investigating regional variations, we estimated using Geographically Weighted Regression (GWR) and decision tree model. Our finding first suggests that using the multi-year data is more legitimate than using the single-year data for the geographical analysis of chronic diseases, because the significant annual differences are observed in most variables. We also find that the prevalence of hypertension is more likely to be positively associated with the prevalence of diabetes and obesity but to be negatively associated with population density. More importantly, noticeable geographical variations in these factors are observed according to the results from the GWR. In line with this result, additional findings from the decision tree model suggest that primary influential factors that affect the hypertension prevalence are indeed heterogeneous across regional groups. Taken as a whole, accounting for geographical variations of health conditions, health behaviors and other socioeconomic factors is very important when the regionally customized healthcare policy is implemented to mitigate the hypertension prevalence. In short, our study sheds light on possible ways to manage the chronic diseases for policy makers in the local government.

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