• Title/Summary/Keyword: genetic screening

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Screening of conjugated linoleic acid (CLA) producing Lactobacillus plantarum and production of CLA on soy-powder milk by these stains (공액리놀레산 생성 Lactobacillus plantarum 선발 및 이를 이용한 콩-분말 두유에서 공액리놀레산 생산)

  • Kim, Baolo;Lee, Byong Won;Hwang, Chung Eun;Lee, Yu-Young;Lee, Choonwo;Kim, Byung Joo;Park, Ji-Yong;Sim, Eun-Yeong;Haque, Md. Azizul;Lee, Dong Hoon;Lee, Jin Hwan;Ahn, Min Ju;Lee, Hee Yul;Ko, Jong Min;Kim, Hyun Tae;Cho, Kye Man
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.231-240
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    • 2015
  • In this study, a total of 16 conjugated linoleic acid (CLA) producing lactic acid bacteria (LAB) were isolated from fermented foods. Among those strains, the S48 and P1201 strains were capable of producing higher CLA contents than other LABs. The two strains were classified as Lactobacillus plantarum based on morphological, physiological, chemotaxonomic, and molecular-genetic properties. The survival rates of these strain appeared to be 59.57% and 62.22% under artificial gastric conditions after 4 h at pH 2.5, respectively. These strains produced the cis-9, trans-11, and trans-10, cis-12 CLA isomers from 8% skim milk medium supplemented with the different free LA concentration at $37^{\circ}C$ for 48 h and the production of two CLA isomers constantly increased in the growth until 48 h of incubation. After 48 h of fermentation, the levels of CLA appeared highest in steamed soy-powder milk than fresh and roasted soy-powder milks. In particular, the CLA contents were produced $183.57{\mu}g/ml$ and $198.72{\mu}g/ml$ from steamed soy-powder milk after fermentation (48 h) with S48 and P1201 strains, respectively.

Validity Test for Molecular Markers Associated with Resistance to Phytophthora Root Rot in Chili Pepper (Capsicum annuum L.) (고추의 역병 저항성과 연관된 분자표지의 효용성 검정)

  • Lee, Won-Phil;Lee, Jun-Dae;Han, Jung-Heon;Kang, Byoung-Cheorl;Yoon, Jae-Bok
    • Horticultural Science & Technology
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    • v.30 no.1
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    • pp.64-72
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    • 2012
  • Phytophthora root rot has been causing a serious yield loss in pepper production. Since 2004, the year in which commercial cultivars resistant to the disease were firstly commercialized, it has been necessary to introduce the resistance into domestic pepper cultivars for dried red pepper. Therefore, developing molecular markers linked to the resistance is required for an accurate selection of resistant plants and increasing breeding efficiency. Until now, several markers associated with the major dominant gene resistant to Phytophthora root rot have been reported but they have some serious limitations for their usage. In this study, we aimed to develop molecular markers linked to the major dominant gene that can be used for almost of all genetic resources resistant to Phytophthora root rot. Two segregating $F_2$ populations derived from a 'Subicho' ${\times}$ 'CM334' combination and a commercial cultivar 'Dokyacheongcheong' were used to develop molecular markers associated with the resistance. After screening 1,024 AFLP primer combinations with bulked segregant analysis, three AFLP (AFLP1, AFLP2, and AFLP3) markers were identified and converted into three CAPS markers (M1-CAPS, M2-CAPS, and M3-CAPS), respectively. Among them, M3-CAPS marker was further studied in ten resistants, fourteen susceptibles, five hybrids and 53 commercial cultivars. As a result, M3-CAPS marker was more fitted to identify Phytophthora resistance than previously reported P5-SNAP and Phyto5.2-SCAR markers. The result indicated that the M3-CAPS marker will be useful for resistance breeding to Phytophthora root rot in chili pepper.

Non Destructive Fast Determination of Fatty Acid Composition by Near Infrared Reflectance Spectroscopy in Sesame

  • Kang, Churl-Whan;Kim, Dong-Hwi;Lee, Sung-Woo;Kim, Ki-Jong;Cho, Kyu-Chae;Shim, Kang-Bo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.283-291
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    • 2006
  • To investigate seed non destructive and fast determination technique utilizing near infrared reflectance spectroscopy (NIRs) for screening ultra high oleic (C18:1) and linoleic (C18:2) fatty acid content sesame varieties among genetic resources and lines of pedigree generations of cross and mutation breeding were carried out in National Institute of Crop Science (NICS). 150 among 378 landraces and introduced cultivars were released to analyse fatty acids by NIRs and gas chromatography (GC). Average content of each fatty acid was 9.64% in palmitic acid (C16:0), 4.73% in stearic acid (C18:0), 42.26% in oleic acid and 43.38% in linoleic acid by GC. The content range of each fatty acid was from 7.29 to 12.27% in palmitic, 6.49% from 2.39 to 8.88% in stearic, 12.59% of wider range compared to that of stearic and palmitic from 37.36 to 49.95% in oleic and of the widest from 30.60 to 47.40% in linoleic acid. Spectrums analyzed by NIRs were distributed from 400 to 2,500 nm wavelengths and varietal distribution of fatty acids were appeared as regular distribution. Varietal differences of oleic acid content good for food processing and human health by NIRs was 14.08% of which 1.49% wider range than that of GC from 38.31 to 52.39%. Varietal differences of linoleic acid content by NIRs was 16.41% of which 0.39% narrower range than that of GC from 30.60 to 47.01%. Varietal differences of oleic and linoleic acid content in NIRs analysis were appeared relatively similar inclination compared with those of GC. Partial least square regression (PLSR) among multiple variant regression (MVR) in NIRs calibration statistics was carried out in spectrum characteristics on the wavelength from 700 to 2,500 nm with oleic and linoleic acids. Correlation coefficient of root square (RSQ) in oleic acid content was 0.724 of which 72.4 percent of sample varieties among all distributed in the range of 0.570 percent of standard error when calibrated (SEC) which were considerably acceptable in statistic confidence significantly for analysis between NIRs and GC. Standard error of cross validation (SECV) of oleic acid was 0.725 of which distributed in the range of 0.725 percent standard error among the samples of mother population between analyzed value by NIRs analysis and analyzed value by GC. RSQ of linoleic acid content was 0.735 of which 73.5 percent of sample varieties among all distributed in the range of 0.643 percent of SEC. SECV of linoleic acid was 0.711 of which distributed in the range of 0.711 percent standard error among the samples of mother population between NIRs analysis and GC analysis. Consequently, adoption NIR analysis for fatty acids of oleic and linoleic instead that of GC was recognized statistically significant between NIRs and GC analysis through not only majority of samples distributed in the range of negligible SEC but also SECV. For enlarging and increasing statistic significance of NIRs analysis, wider range of fatty acids contented sesame germplasm should be kept on releasing additionally for increasing correlation coefficient of RSQ and reducing SEC and SECV in the future.

Selection of Salt-Tolerant Silage Rice Through in vitro Screening and Saltol QTL Analysis (기내 선발과 Saltol QTL 분석을 통한 내염성 증진 사료용 벼 선발)

  • Cho, Chuloh;Kim, Kyung Hwa;Ahn, Eok-Keun;Park, Hyangmi;Choi, Man-Soo;Chun, Jaebuhm;Seo, Mi-Suk;Jin, Mina;Kim, Dool-Yi
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.3
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    • pp.214-221
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    • 2020
  • Salinity is one of the major abiotic stressors that inhibits the growth, yield, and productivity of crop plants. Therefore, it is necessary to develop crops with increased salt tolerance for cultivation in saline soils such as is found in reclaimed land. The objective of this study was to develop a salt-tolerant silage rice line that grows on reclaimed land. In order to develop this salt-tolerant silage rice, we transferred Saltol, a major QTL associated with salt tolerance, from IR64-Saltol, a salt-tolerant indica variety, into Mogyang, a susceptible elite japonica variety. To determine the effect of salt stress, Mogyang and IR64-Saltol cultivars were grown on a medium containing various concentrations of NaCl in in vitro conditions. Shoot length was found to decrease with increasing salt concentrations, and root growth was almost arrested at NaCl concentrations over 50 mM in the Mogyang cultivar. Based on these preliminary results, we screened five salt-tolerant lines showing superior growth under salt stress conditions. Polymerase chain reaction and sequencing results showed that the introgression types of Saltol QTL were derived from the IR64-Saltol cultivar in almost all selected lines. Based on the observed growth and physiological characteristics, the new Saltol introgression lines showed higher salt tolerance compared to the Mogyang parental cultivar. The salt-tolerant lines identified in this study could be used as a genetic resource to improve rice salt tolerance.

Process Optimization of Dextran Production by Leuconostoc sp. strain YSK. Isolated from Fermented Kimchi (김치로부터 분리된 Leuconostoc sp. strain YSK 균주에 의한 덱스트란 생산 조건의 최적화)

  • Hwang, Seung-Kyun;Hong, Jun-Taek;Jung, Kyung-Hwan;Chang, Byung-Chul;Hwang, Kyung-Suk;Shin, Jung-Hee; Yim, Sung-Paal;Yoo, Sun-Kyun
    • Journal of Life Science
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    • v.18 no.10
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    • pp.1377-1383
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    • 2008
  • A bacterium producing non- or partially digestible dextran was isolated from kimchi broth by enrichment culture technique. The bacterium was identified tentatively as Leuconostoc sp. strain SKY. We established the response surface methodology (Box-Behnken design) to optimize the principle parameters such as culture pH, temperature, and yeast extract concentration for maximizing production of dextran. The ranges of parameters were determined based on prior screening works done at our laboratory and accordingly chosen as 5.5, 6.5, and 7.5 for pH, 25, 30, and $35^{\circ}C$ for temperature, and 1, 5, and 9 g/l yeast extract. Initial concentration of sucrose was 100 g/l. The mineral medium consisted of 3.0 g $KH_2PO_4$, 0.01 g $FeSO_4{\cdot}H_2O$, 0.01 g $MnSO_4{\cdot}4H_2O$, 0.2 g $MgSO_4{\cdot}7H_2O$, 0.01 g NaCl, and 0.05 g $CaCO_3$ per 1 liter deionized water. The optimum values of pH and temperature, and yeast extract concentration were obtained at pH (around 7.0), temperature (27 to $28^{\circ}C$), and yeast extract (6 to 7 g/l). The best dextran yield was 60% (dextran/g sucrose). The best dextran productivity was 0.8 g/h-l.

Resistance Screening to Pepper mild mottle virus Pathotypes in Paprika Cultivars (고추약한모틀바이러스 병원형에 대한 파프리카 품종의 저항성 스크리닝)

  • Choi, Gug-Seoun;Choi, Seung-Kook;Cho, In-Sook;Kwon, Sun-Jung
    • Research in Plant Disease
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    • v.20 no.4
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    • pp.299-302
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    • 2014
  • The Paprika plant infected with Pepper mild mottle virus (PMMoV) do not produce commercial fruit as causing necrotic spots symptom on the fruit. Ten cultivars of paprika were analyzed to select the resistance cultivars against PMMoV pathotypes, $P_{1.2}$ and $P_{1.2.3}$, using bioassay and genetic markers. $L^1$, $L^3$, and $L^4$ genotypes expressing resistance to the pathotypes existed in those cultivars but $L^2$ genotype did not. $L^4L^4$ in cvs. Easy and Magnifico, $L^4L^3$ in cvs. Scirocco and Orange glory F1, $L^4L^1$ in cv. Special F1, $L^3L^3$ in cvs. Fiesta, Piero and Derby, and $L^3L^1$ in Cupra and Mazzona F1 were identified with SCAR and CAPS markers. The resistant cvs. to the 2 pathotypes were Magnipico, Easy, Scirocco F1, Orange glory and Special F1 and the susceptible cvs. were Fiesta, Piero, Derby, Cupra and Mazzona F1. The susceptible cvs. of the absence of $L^4$ genotype showed systemic infection when inoculated with PMMoV-$P_{1.2.3}$. However, those cvs. despite the presence of $L^3$ genotype showed vein necrosis on the inoculated leaf and hypersensitive necrosis symptom on the upper parts when inoculated with PMMoV-$P_{1.2}$.

Cell Death Study in Embryonic Stem Cell-derived Neurons and Its Applications (배아줄기세포 유래 신경계세포에서의 세포사멸 연구와 그 응용)

  • Lee, Chul-Sang
    • Development and Reproduction
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    • v.12 no.1
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    • pp.1-8
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    • 2008
  • Specific protocols to increase the differentiation of neuronal cells from embryonic stem (ES) cells have been well established, such as retinoic acid induction and lineage selection of neuronal cells. For the neuropathological studies, ES-derived neurons (ES neurons) must show normal physiological characteristics related to cell death and survival and should be maintained in vitro for a sufficient time to show insults-specific cell death without spontaneous death. When mouse ES cells were plated onto astrocytes monolayer after retinoic acid induction, most ES cells differentiated into neuronal cells, which were confirmed by the presence of specific neuronal markers, and the cultures were viable for at least four weeks. When these cultures were examined for vulnerability to glutamate excitotoxicity, ES neurons were vulnerable to excitotoxic insults mediated by agonist-specific receptors. The vulnerability to excitotoxic death increased with developmental age of ES neurons in vitro. Specific receptors for Neurotrophin and GDNF family ligands were present in ES neurons. GDNF and NT-3 could modulate the survival and excitotoxic vulnerability of ES neurons. The vulnerability and resistance to toxic insults, which are essential requirements of model culture systems for neuropathological studies, make ES neurons to a useful model culture system. Especially ES cell are highly amenable to genetic modification unlikely to primary neuronal cells, which will give us a chance to answer more complicated neurophysiological questions. Recently there was an outstanding attempt to explore the cellular toxicity using human ES cells (Schrattenholz & Klemm, 2007) and it suggested that ES cells could be a new model system for neurophysiological studies soon and go further a large-scale screening system for pharmacological compounds in the future.

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A Case of Adult-onset Type II Citrullinemia Confirmed by Mutation of SLC25A13 (SLC25A13 유전자 돌연변이로 확진된 성인형 제 2형 시트룰린혈증 1례)

  • Jeung, Min Sub;Yang, Aram;Kim, Jinsup;Park, Hyung-Doo;Lee, Heon Ju;Jin, Dong-Kyu;Cho, Sung Yoon
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.16 no.1
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    • pp.34-41
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    • 2016
  • Adult-onset type II citrullinemia (CTLN2) is characterized by episodes of neurologic symptoms associated with hyperammonemia leading to disorientation, irritability, seizures, and coma. CTLN2 is distinct from classical citrullinemia, which is caused by a mutation of the argininosuccinic acid synthetase (ASS) gene. The serum citrulline level is elevated, while the activity of ASS in liver tissue is decreased. CTLN2 is known to have a poor prognosis if the proper treatment is not taken. We reported a female aged 37 years who developed recurrent attacks of altered consciousness, aberrant behavior, and vomiting. We initially suspected the patient had CTLN2 because of the signs of hyperammonemic encephalopathy, such as altered mentality, memory disturbance, and aberrant behaviors provoked by exercise-induced stress and excessive intravenous amino acid administration. Through her peculiar diet preferences and laboratory findings that included hyperammonemia and citrullinemia, we diagnosed the patient as CTLN2, and SLC25A13 sequencing revealed known compound heterozygous mutations (IVS11+1G>A, c.674C> A). Her parents were heterozygous carriers, and we identified that her older sister had the same mutations. The older sister had not experienced any episodes of hyperammonemia, but she had peculiar diet preferences. The patient and her sister have been well with conservative management. When considering the clinical course of CTLN2, it was meaningful that the older sister could be diagnosed early in an asymptomatic period and that preemptive treatment was employed. Through this case, CTLN2 should be considered in adults who present symptoms of hyperammonemic encephalopathy without a definite etiology. Because of its rare incidence and similar clinical features, CTLN2 is frequently misdiagnosed as hepatic encephalopathy, and it shows a poor prognosis due to the lack of early diagnosis and proper treatment. A high-carbohydrate diet, which is usually used to treat other urea cycle defects, can also exaggerate the clinical course of CTLN2, so proper metabolic screening tests and genetic studies should be performed.

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The current status and control measures of BSE in the worldwide (국내, 외 광우병의 발생 현황과 대응 방안)

  • Yoo, Han-Sang
    • 한국환경농학회:학술대회논문집
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    • 2009.07a
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    • pp.273-282
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    • 2009
  • The transmissible spongiform encephalopathies (TSEs) disease group are fatal neurodegenerative disorders affecting a wide range of hosts. The group includes kuru and Creutzfeldt-Jakob disease (CJD) in humans, scrapie in sheep and goats and Bovine spongiform encephalopathy (BSE) in cattle. The exact nature of the infectious agent involved in the transmission of these diseases remains controversial. However, a central event in their pathogenesis is the accumulation in infected tissues of an abnormal form of a host-encoded protein, the prion protein (PrP). Whereas the normal cellular protein is fully sensitive to protease ($PrP^{sen}$), the disease-associated prion protein ($PrP^d$) is only partly degraded ($PrP^{res}$), its amino-terminal end being removed. BSE was first reported in the mid-80s in the UK. Ten years later, a new form of human prion disease, variant CJD (vCJD) developed in the wake of the BSE epidemic, and there is now strong scientific evidence that vCJD was initiated by the exposure of humans to BSE-infected tissues, thus indicating a zoonotic disease. However, the ban on the feeding of animal-derived proteins to ruminants, and the apparent lack of vertical transmission of BSE, have led to a decline in the incidence of the disease within cattle herd and therefore, an assumed decreased risk for human contacting vCJD. The origin of the original case(s) of BSE still remains an enigma even though three hypotheses have been raised. Hypotheses are i) sheep- or goat-derived scrapie-infected tissues included in meat and bone meal fed to cattle, ii) a previously undetected sporadic or genetic bovine TSE contaminating cattle feed or iii) originating from a human TSE through animal feed contaminated with human remains. A host cellular membrane protein ($PrP^C$), which is abundant in central nervous system tissue, appear to be conformationally altered in the diseased host into a prion protein ($PrP^{Sc}$). This $PrP^{Sc}$ is detergent insoluble and partially protease-resistant ($PrP^{res}$). The term $PrP^{res}$ is normally used to describe the protein detected after protease treatment, in techniques such as Western immunoblotting, and enzyme-linked immunosorbant assay using fresh/frozen tissue. Immunohistochemistry may performed with formalin-fixed tissues. Also, clinical signs of the BSE are one of the major diagnostic indicators. Recently, atypical forms (known as H- and L-type) of BSE have appeared in several European countries, Japan, Canada and the United States. An unusual case was also reported in a miniature zebu. The atypical BSE fall into two groups based on the relative molecular mass (Mm) of the unglycosylated $PrP^{res}$ band relative to that of classical BSE, one of the higher Mm (H-type) and the other lower (L-type). Both types have been detected worldwide as rare cases in older animals, at a low prevalence consistent with the possibility of sporadic forms of prion diseases in cattle. This raises the unwelcome possibility that vCJD could increase in the human population. Now, active surveillance program against BSE is going on in Korea. In regional veterinary service lab, ELISA is applied to screen the BSE in slaughter and confirmatory tests by Western immunoblotting and immunohistochemisty are carried out if there are positive or suspect in the screening test. Also, the ruminant feed ban is rigorously enforced. Removal of specified risk materials such as brain and spinal cord from cattle is mandatory process at slaughter to prevent the infected material from entering the human food chain.

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A Study on the Screening of the Novel Genes Associated with Lysosomal Trafficking and Mutation Detection in Fibroblasts of the Patients with Mucolipidosis type II and III (리소좀 교통 이상을 초래하는 뮤코지방증 2형과 3형 환자의 섬유아세포를 이용한 신규 유전자 탐색 및 돌연변이에 대한 연구)

  • Song, Seng Mi;Chang, Soo Hee;Paik, Kyung Hoon;Jin, Dong-Kyu
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.5 no.1
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    • pp.65-75
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    • 2005
  • Purpose: To understand genetic differences and similarities between mucolipidosis and control. Methods: Using the fibroblast of the mucolipidosis II and control, forward and reverse subtracted libraries were constructed. Among these clones, we investigated mutations in the GNPTA (MGC4170) gene, which codes for the ${\alpha}/{\beta}$ subunits of phosphotransferase, and in the GNPTAG gene, which codes for the ${\gamma}$ subunits in 5 Korean patients with mucolipidosis type II or IIIA. Result: Several differentially expressed cDNAs were cloned and their sequences were determined. Mutation analysis of the interested gene, GNPTA was performed and we identified 7 mutations in the GNPTA gene, but none in the GNPTAG gene. The mutations in type II patients included p.Q104X(c.310C>T), p.R1189X(c.3565C>T), p.S1058X(c.3173C>G), p.W894X(c.2681G>A) and p.H1158fsX15(c.3474_3475delTA), all of which are non-sense or frame shift mutations. However, a splicing site mutation, IVS13+1G>A (c.2715+1G>A) was detected along with a non-sense or a frame shift mutation (p.R1189X or p.E858fsX3(c.2574_2575delGA)) in two mucolipidosis type IIIA patients. Conclusion: This report shows that mutations in the GNPTA gene coding for the ${\alpha}{\beta}$subunits of phosphotransferase, and not mutations in the GNPTAG gene, account for most of mutations found in Korean patients with mucolipidosis type II or IIIA.

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