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Expression and Purification of Recombinant Human Interferon-gamma Produced by Escherichia coli (대장균이 생산한 재조합 인체 감마인터페론의 발현과 정제)

  • Park, Jung-Ryeol;Kim, Sung-Woo;Kim, Jae-Bum;Jung, Woo-Hyuk;Han, Myung-Wan;Jo, Young-Bae;Jung, Joon-Ki
    • KSBB Journal
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    • v.21 no.3
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    • pp.204-211
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    • 2006
  • For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.

Fine Needle Aspiration Cytology of Hepatocellular Carcinoma - A Study on 247 Cases - (간세포암종의 세침흡인 세포학적 소견 - 247예에 대한 연구 -)

  • Lee, Kwang-Gil;Lee, Jong-Tae;Choi, Soo-Im;Park, Chan-Il
    • The Korean Journal of Cytopathology
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    • v.1 no.1
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    • pp.1-17
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    • 1990
  • Hepatocellular carcinoma (HCC) is malignant tumor frequently occurring in Koreans. There have been few reports regarding the cytologic findings of fine needle aspiration (FNA) of HCC. Most have suggested a diagnostic problem in the cytology distinguishing HCC from some benign hepatic lesions-for example, a regeneration nodule in cirrhosis and liver cell adenoma. In spite of its high frequency in Korea, no cytologic study has been reported, concerning the FNA of HCC. In an attempt to achieve cytologic criteria for the diagnosis of HCC, the authors studied retrospectively cytopathologic findings of 247 cases of HCC. These cases were confirmed either by histoiogic examination including lobectomy, biopsy, or ceil block material, or, when tissue diagnosis was unavailable, by a high serum alpha-fetoprotein level (over 400 I. U.). All aspiration smears were stained by the Papanicolaou method. In each case, the smears were analyzed for cell patterns and various cytomorphology of the tumor cells. The smear background was assessed for the presence of tumor cell necrosis and inflammatory components and compared to that of metastatic carcinomas. The cell patterns were classified as trabecular, acinar, dispersed, and irregular. The cytologic parameters analyzed included the degree of nuclear atypia and the presence of mitoses, intranuclear cytoplasmic inclusions, nucleolar prominency, endothelial lining, multinucleated giant cells, eosinophiic globules, bile, and Mallory body. Most of the FNA of HCC showed markedly cellular smears. The tumor cells were most frequently arranged in a trabecular pattern (80.3%). The irregular (12.6%), the acinar (5.5%), and the dispersed patterns (1.7%) followed in decreasing frequency. Individual hepatoma cells were larger than normal liver cells. However, they had morphologic features characteristic of the hepatic cells the cells were round or polygonal, their cytoplasm was abundant and granular with eosinophilic or amphophilic stainability, and their nuclei were round to oval, located centrally, and tended to have prominent nucleoli. Anaplasia and pleomorphism of tumor cells were generally mild to moderate. These findings existed even in very well differentiated cases. Mitotic figures were present in about 85% of the cases. Prominent nucleoli were observed only in about half the cases. The frequency of other cytologic features was as follows intranuclear cytoplasmic inclusion in 86.8% : endothelial lining in 56.1% : bile in 19.8% : and giant cells in 60.1%. Clear cells were often present in 11.7%, Most aspiration smears of HCC displayed clean background without necrosis or inflammatory material in contrast to the dirty, necrotic background of metastatic cancers and cholangiocarcinomas. Based on the above mentioned features, it is suqqested that the cytologic critieria most important for the diagnosis of HCC include a markedly cellular smear, trabecular pattern, hepatocytoid appearance of tumor cells, endothelial lining, the presence of bile, giant cells, intranuclear cytoplasmic inclusions, and prominent nucleoli, Among these, trabecular pattern, endothelial lining, giant cells and clean smear background are points to be considered in differentiating HCC from metastatic and cholangiocellular carcinoma.

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Genetic and Economic Analysis for the Relationship between Udder Health and Milk Production Traits in Friesian Cows

  • El-Awady, H.G.;Oudah, E.Z.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.11
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    • pp.1514-1524
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    • 2011
  • A total of 4,752 monthly lactation records of Friesian cows during the period from 2000 to 2005 were used to estimate genetic parameters and to determine the effect of udder health on milk production traits. Three milk production traits were studied: 305-day milk yield (305-dMY), 305-day fat yield (305-dFY) and 305-day protein yield (305-dPY). Four udder health traits were studied: somatic cell count (SCC), mastitis (MAST), udder health status (UDHS) with 10 categories and udder quarter infection (UDQI) with 7 categories. Mixed model least square analysis was used to estimate the fixed effects of month and year of calving and parity (P) on different studied traits. Sire and dam within sire were included in the model as random effects. Data were analyzed using Multi-trait Derivative Free Restricted Maximum Likelihood methodology (MTDFREML) to estimate genetic parameters. Unadjusted means of 305-dMY, 305-dFY, 305-dPY and SCC were 3,936, 121, 90 kg and 453,000 cells/ml, respectively. Increasing SCC from 300,000 to 2,000,000 cells/ml increased UDQI from 5.51 to 23.2%. Losses in monthly and lactationally milk yields per cow ranged from 17 to 93 and from 135 to 991 kg, respectively. The corresponding losses in monthly and lactationally milk yields return per cow at the same level of SCC ranged from 29.8 to 163 and from 236 to 1,734 Egyptian pounds, respectively. Heritability estimates of 305-dMY, 305-dFY, 305-dPY, SCC, MAST, UDHS, UDQI were 0.31${\pm}$0.4, 0.33${\pm}$0.03, 0.35${\pm}$0.05, 0.23${\pm}$0.02, 0.14${\pm}$0.02, 0.13${\pm}$0.03, and 0.09${\pm}$0.01, respectively. All milk production traits showed slightly unfavorable negative phenotypic and genetic correlations with SCC, MAST, UDHS and UDQI. There were positive and high genetic correlations between SCC and each of MAST (0.85${\pm}$0.7), UDHS (0.87${\pm}$0.10) and UDQI (0.77${\pm}$0.06) and between MAST and each of UDHS (0.91${\pm}$0.11) and UDQI (0.83${\pm}$0.07). It could be concluded that the economic losses from mastitis and high SCC are considerable. The high genetic correlation between SCC and clinical mastitis (CM) suggest that the selection for lower SCC would help to reduce or eliminate the undesirable correlated responses of clinical mastitis associated with selection for increasing milk yield. Additionally, it is recommended also that if direct information on under health traits is not available, measures of SCC can be inclusion in a selection criteria to improve the income from dairy cows.

Utilization of Processed Pine Needle Powder in Broiler Chicken Diets

  • Kim, Eunjoo;Jung, Ji Young;Cho, Hyun Min;Wickramasuriya, Samiru Sudharaka;Maceline, Shemil Pyrian;Shin, Taeg Kyun;Kim, Beomgyu;Heo, Jung Min
    • Korean Journal of Poultry Science
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    • v.45 no.3
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    • pp.167-174
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    • 2018
  • This study examined the effects of dietary processed pine needle tree powder (PPNP) on growth performance, visceral organ parameters, ileal digestibility, blood metabolites and ileal morphology of broiler chickens for 5-week after hatch. In total, 220 1-day-old male Ross 308 broiler chicks were randomly allocated to 30 pens (seven birds per pen), and each pen was assigned to one of five dietary treatments, a diet without PPNP (control) and diets with 0.5%, 1.0%, 1.5% and 2.0% PPNP. The body weight and feed intake were measured weekly. One bird from each pen was selected randomly and gently euthanized to collect the blood, visceral organs and digesta samples on 2, 3 and 5 week of age. Birds fed 2.0% PPNP showed decreased (P<0.05) overall body weight, average daily gain and average daily feed intake compared to those fed control, 0.5 and 1.0% PPNP. The small intestine length increased (P<0.05) as the level of PPNP in the diet increased. A deeper crypt depth was observed (P<0.05) when birds fed 2.0% PPNP compared to those fed the control diet at week 2. A higher V:C ratio was observed (P<0.05) in birds fed 1.0% PPNP than in those fed other treatments at week 3. Ileal protein digestibility improved (P<0.05) when birds fed 0.5% and 1.0% PPNP compared to that in those fed the control diet at week 3. The feed conversion ratio, ileal dry matter digestibility and blood urea nitrogen and cholesterol, and gizzard and ceca weights were not affected (P>0.05) by supplementation of any levels of PPNP. These results demonstrated that inclusion of PPNP in broiler diets could be a possible factor in improving overall growth performance suggesting early gut development when 1% PPNP is supplemented.

A Survey of Particulate Matters and CO2 Levels in Seoul Subway Carriages (서울시 지하철 객차 내 PM과 CO2의 농도 분포)

  • Lee, Choel-Min;Park, Wha-Me;Roh, Young-Man;Kim, Yoon-Shin;Park, Dong-Sun
    • Journal of Environmental Health Sciences
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    • v.34 no.1
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    • pp.34-41
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    • 2008
  • The objective of this study is to provide the research data on the actual concentrations of $PM_{10},\;PM_{2.5},\;PM_1\;and\;CO_2$ in Seoul subway carriages. Mean concentrations of $PM_{10},\;PM_{2.5}\;and\;PM_1,\;and\;CO_2$ in subway carriages were investigated at levels of $215.1{\pm}101.4{\mu}g/m^3,\;86.9{\pm}38.6{\mu}g/m^3,\;27.0{\pm}11.4{\mu}g/m^3,\;and\;1,588{\pm}714ppm$, respectively. The mean concentrations in subway carriages were higher when the train ran on an underground track rather than on an above ground track. The measured concentration of particulate matter varied with the time of day and was highest in the morning, followed by noon and evening while the $CO_2$ concentration was highest in the morning, followed by evening and noon. In relation to correlation among the pollutants: the correlation between $PM_{10}\;and\;PM_{2.5}$ was 0.92, and that between $PM_{2.5}\;and\;PM_1$ was 0.94. The inclusion rate of $PM_{2.5}\;to\;PM_{10}$ was $41{\pm}7%$ and that of $PM_1\;to\;PM_{2.5}\;was\;32{\pm}4%$. In addition, the $CO_2$ concentration had a positive relation with the number of people in a carriage, whereas the concentration of $PM_{10}$ had negative correlation to the number of people. In relation to these two pollutants we calculated using a regression equation (34.06+0.04$CO_2$(ppm)-0.09 PM10$({\mu}g/m^3)$($R^2$=0.30, p<0.01, n=707), that a maximum number of 61 persons would ensure that each pollutant is maintained below the criteria level, applicable to subway stations.

A Whole Genome Association Study to Detect Single Nucleotide Polymorphisms for Carcass Traits in Hanwoo Populations

  • Lee, Y.-M.;Han, C.-M.;Li, Yi;Lee, J.-J.;Kim, L.H.;Kim, J.-H.;Kim, D.-I.;Lee, S.-S.;Park, B.-L.;Shin, H.-D.;Kim, K.-S.;Kim, N.-S.;Kim, Jong-Joo
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.4
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    • pp.417-424
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    • 2010
  • The purpose of this study was to detect significant SNPs for carcass quality traits using DNA chips of high SNP density in Hanwoo populations. Carcass data of two hundred and eighty nine steers sired by 30 Korean proven sires were collected from two regions; the Hanwoo Improvement Center of National Agricultural Cooperative Federation in Seosan, Chungnam province and the commercial farms in Gyeongbuk province. The steers in Seosan were born between spring and fall of 2006 and those in Gyeonbuk between falls of 2004 and 2005. The former steers were slaughtered at approximately 24 months, while the latter steers were fed six months longer before slaughter. Among the 55,074 SNPs in the Illumina bovine 50K chip, a total of 32,756 available SNPs were selected for whole genome association study. After adjusting for the effects of sire, region and slaughter age, phenotypes were regressed on each SNP using a simple linear regression model. For the significance threshold, 0.1% point-wise p value from F distribution was used for each SNP test. Among the significant SNPs for a trait, the best set of SNP markers were selected using a stepwise regression procedure, and inclusion and exclusion of each SNP out of the model was determined at the p<0.001 level. A total of 118 SNPs were detected; 15, 20, 22, 28, 20, and 13 SNPs for final weight before slaughter, carcass weight, backfat thickness, weight index, longissimus dorsi muscle area, and marbling score, respectively. Among the significant SNPs, the best set of 44 SNPs was determined by stepwise regression procedures with 7, 9, 6, 9, 7, and 6 SNPs for the respective traits. Each set of SNPs per trait explained 20-40% of phenotypic variance. The number of detected SNPs per trait was not great in whole genome association tests, suggesting additional phenotype and genotype data are required to get more power to detect the trait-related SNPs with high accuracy for estimation of the SNP effect. These SNP markers could be applied to commercial Hanwoo populations via marker-assisted selection to verify the SNP effects and to improve genetic potentials in successive generations of the Hanwoo populations.

The Effects of Condensed Molasses Solubles(CMS) / Molasses Mixtures on Ruminal Microbial Protein Synthesis (Condensed Molasses Solubles(CMS) / 당밀 혼합물이 반추위 미생물 단백질 합성량에 미치는 영향)

  • Yeo, J.M.;Jeong, S.G.;Kim, H.S.;Ahn, B.S.;Kim, C.H.;Shin, H.T.
    • Journal of Animal Science and Technology
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    • v.46 no.1
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    • pp.61-68
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    • 2004
  • An experiment was conducted to evaluate condensed molasses solubles(CMS, a by-product from monosodium glutamate production) as a source of nitrogen for ruminant with particular reference to its effects on microbial protein synthesis. Four non-lactating dairy cows fitted with rumen cannulas were used in a 4 ${\times}$ 4 Latin square with 14-day periods. The four treatments were (1) basal diet consisting of barley straw ad libitum and 3 kg/d of rolled barley, (2) basal diet plus 200 gld molasses and 300 g/d water, (3) basal diet plus 200 g/d molasses, 100 g/d CMS and 200 g/d water, (4) basal diet plus 200 g/d molasses, 200 g/d CMS and 100 g/d water. Ruminal pH remained at high levels and showed little variation during the day between treatments. The concentration of total and individual VFA in the rumen was similar between treatments. There was no difference in the concentration of ammonia in the rumen between treatments, although the intake of nitrogen in molassesl CMS mixture treatments was higher than that of control and molasses treatment. But there was a suggestion of an increased synthesis of microbial protein with the higher level of inclusion of CMS when the allantoin/creatinine ratio was used as an index of microbial protein production(P <0.10).

Treatment of Vasculopathy in Diabetic Foot by Percutaneous Transluminal Angioplasty (경피적 동맥확장술을 이용한 당뇨족 허혈의 치료)

  • Kim, Hong-Ryul;Han, Seung-Kyu;Rha, Seung-Woon;Kim, Hyon-Surk;Kim, Woo-Kyung
    • Archives of Plastic Surgery
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    • v.37 no.2
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    • pp.148-152
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    • 2010
  • Purpose: In treating diabetic foot ulcers, satisfactory vascularity is an essential prerequisite. To improve vascularity, a bypass graft has long been carried out. Recently, however, percutaneous transluminal angioplasty (PTA) has also been tried since the PTA is less invasive than the bypass graft. However, publication demonstrating the improvement of vascularity after the PTA are lacking. Therefore, this study was designed to show usefulness of the PTA in treating vasculopathy of diabetic foot. Materials: and Methods This study included 30 feet of 24 ischemic diabetic foot patients. Inclusion criteria were diabetes (duration > 5 years) and a significant lower extremity ischemia, as determined by a transcutaneous oxygen pressure ($TcpO_2$) < 30 mmHg. The PTA was carried out in 61 arteries. PTA procedure was considered successful, when residual stenosis was less than 30%. The procedure was considered failed when residual stenosis was more than 50%. Residual stenosis between 30% and 50% was considered acceptable. For evaluation of PTA effect, foot $TcpO_2$ and infrared thermography were measured before and 7th day after PTA. Results: Immediately after PTA performed in 61 arteries, 58 and 3 arteries were evaluated as being successful and acceptable, respectively. Before PTA, average foot $TcpO_2$ was $12.6{\pm}8.8$ mmHg and its value was increased to $44.2{\pm}23.9$ on 7th day after PTA (p<0.01). Average skin temperature was $31.8{\pm}1.2^{\circ}C$ before PTA and it was increased to $33.5{\pm}1.1^{\circ}C$ on 7th day after PTA (p<0.01). Conclusion: PTA procedure increases tissue oxygenation of ischemic diabetic feet which do not have wound healing potential due to low tissue oxygenation, to the level of possible wound healing. In addition, PTA increases skin temperature of ischemic diabetic feet which can imply an improvement of peripheral circulation.

Evaluation of the Feeding Value of Sesame Oil Meal and Effects of Its Dietary Supplementation on the Performances of Laying Hens (호마박의 영양적 가치 평가 및 산란계 사료 내 첨가각 사양 성적에 미치는 영향)

  • Im H. J.;Ahn S. M.;You S. J.;Kim Y. R.;Ahn B. K.;Kang C. W.
    • Korean Journal of Poultry Science
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    • v.31 no.4
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    • pp.255-263
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    • 2004
  • Two experiments were conducted to evaluate the feeding values of sesame oil meal (SOM) and to investigate the effects of its dietary supplementation on egg production in laying hens. In experiment I, the values of true metabolizable energy (TME), nitrogen corrected true metabolizable energy (TMEn) and true amino acid availability (TAAA) were determined by force-feeding 16 ISA-Brown roosters and collecting the total excreta from the birds, The TME and TMEn of SOM were 2.30 and 1.99 kcal/g, respectively, and the average TAAA of 15 amino acids was $76.93\%$. In experiment 2, a total of ninety, 48 weeks old ISA-Brown layer were randomly divided into 9 groups of 10 birds each and assigned to three experimental diets containing 0, 5 and $10\%$ SOM for 4 weeks (30 birds per treatment). The inclusion of SOM into laying hen diets at the 5 and $10\%$ level did not affect production and quality of egg. The C18:3 $\omega$3 content of egg yolks in the $10\%$ SOM group was higher than the other groups, but not significantly. There were no adverse effects on blood parameters in layers fed treated diets containing $5\%$ or $10\%$ SOM, The results indicate that SOM can be used for layers diet up to $10\%$ without any significant negative effects on egg production and quality.

The Study of Biochemical Changes Induced by Fish Oil Diet in Rat ( II ) - Changes in Lipoprotein Lipase Activity and Mitochondrial Respiration and Structure - (어유(魚油)식이에 의한 흰쥐체내의 생화학적 변화연구(II) - Lipoprotein Lipase 활성과 미토콘드리아 호흡계의 변화 -)

  • Ha, Tae-Youl;Jung, Seung-Eun;Im, Jung-Gyo;Cho, Sung-Hee
    • Journal of Nutrition and Health
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    • v.17 no.4
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    • pp.297-304
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    • 1984
  • The effect of dietary fish oil ( mackerel oil : MO, eel oil : EO) on energy utilization in rats was studied with measurements of various tissue lipoprotein lipase( LPL ) and live and heart mitochondrial respiration. Fatty acid composition of mitrochondrial inner membrane matrix was also investigated. Dietary fat level was 10%( w/w) and reference groups were fed soybean oil (SO), repeseed oil ( RO) and beef tallow( BT ). Activity of LPL was about 60% higher in post-heparin plasma and 2 to 3 times higher in adipose tissue of BT group than fish oil or vegetable oil group. But there was no significant difference between fish oil and vegetable oil groups. Inclusion of EO above 2% (w/w) in dietary fat with fille oil of BT, markedly reduced both post -heparin plasma and adipose tissue LPL. Effects of MO and EO were not different in adipose tissue LPL, but EO was more effective than MO, in reducing post -heparin plasma LPL when mixed fat with varying amount of fish oil was used. Hepatic mitochondria isolated from fish oil-fed group showed the lowest rate of respiration but had P/O ratio comparable to SO and BT groups. On the other hand, cardiac mitochondria of fish oil group showed no difference in all the mitochondrial respiration parameters observed RO group had lowest P/O ratio both in hepatic and cardiac mitochondria. Fatty acid compositions of mitochondrial lipid differ between SO, RO, BT and MO groups, notably in the content of $C_{22:1}$ fatty acid.

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