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Cloning and Characterization of Phosphomannomutase/Phosphoglucomutase (pmm/pgm) Gene of Vibrio anguillarum Related to Synthesis of LPS (Lipopolysaccharide 생합성에 관여하는 Vibrio anguillarum의 phosphomannomutase/phosphoglucomutase 유전자 cloning과 특성)

  • Oh, Ryunkyoung;Moon, Soo Young;Cho, Hwa Jin;Jang, Won Je;Kim, Jang-Ho;Lee, Jong Min;Kong, In-Soo
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.355-362
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    • 2016
  • The phosphomannomutase/phosphoglucomutase gene (pmm/pgm) of Vibrio anguillarum (the causative agent of fish vibriosis) was cloned, and the open reading frame corresponded to a protein with 446 amino acids. The pmm/pgm gene showed a significant degree of sequence homology with the previously reported genes from V. mimicus, V. vulnificus, V. splendidus, and V. harveyi, with 92.3%, 91.4%, 89.9%, and 89.9% amino acid identity, respectively. By reverse transcriptase-polymerase chain reaction, we found that the pmm/pgm gene was upregulated under cold stress condition. The PMM/PGM protein is known to catalyze the interconversion between mannose-1-phosphate and mannose-6-phosphate or glucose-1-phosphate and glucose-6-phosphate, which are important intermediates for lipopolysaccharide (LPS) biosynthesis. To confirm the role of PMM/PGM in the LPS biosynthetic pathway, we constructed a knock out mutant by homologous recombination. The respective LPSs were isolated from the V. anguillarum wild-type and mutant strains, and changes were compared by subjecting them to sodium dodecyl sulfate polyacrylamide gel electrophoresis. Based on the different patterns of the LPSs, we expect the pmm/pgm gene to have an important role in LPS biosynthesis. The pmm/pgm-deficient mutant of V. anguillarum will contribute to further studies about the role of LPS in V. anguillarum pathogenesis.

Anti-Inflammatory Effect of Chondrus ocellatus Holmes Ethanol Extract on Lipopolysaccharide-induced Inflammatory Responses in RAW 264.7 Cells (Lipopolysaccharide로 유도된 RAW 264.7 세포와 마우스모델에 대한 진두발 에탄올 추출물의 항염증 효과)

  • Bae, Nan-Young;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Park, Ji-Hye;Park, Sun-Hee;Sung, Nak-Yun;Byun, Eui-Hong;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.268-277
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    • 2016
  • This study aimed to investigate the anti-inflammatory effect of the ethanol extract from Chondrus ocellatus Holmes (COHEE) in RAW 264.7 cells and in a mouse ear edema model, by measuring the production of lipopolysaccharide-induced inflammatory response mediators. There were no cytotoxic effects on the proliferation of macrophages treated with COHEE compared with the control. COHEE inhibited the production of nitric oxide and pro-inflammatory cytokines [interleukin (IL)-6, tumor necrosis factor-α, and IL-1β]. The extract also reduced the expression of inducible nitric oxide synthase, cyclooxygenase-2, nuclear factor-κB p65, and phosphorylated mitogen-activated protein kinase in a dose-dependent manner. In the croton-oil-induced ear edema model, COHEE decreased the formation of mouse ear edema at the highest dose compared with the control, and histological analysis revealed that the epidermal/dermal tissue thickness and mast cell numbers were reduced. Therefore, these results suggest that COHEE may be a promising topical anti-inflammatory therapeutic material through its action of modulating NF-κB and the MAPK signaling pathway.

Antiviral Activity of Hot-Water Extract and Its Ethanol Precipitate of Phellinus pini Fruiting Body (낙엽진흙버섯 자실체 유래 열수 추출물과 메탄을 침전물의 항바이러스활성)

  • Lee, Sae-Mi;Kim, Sung-Min;Lee, Yoon-Hee;Kim, Woo-Jung;Na, Ye-Seul;Kim, Hyun-Guell;Nam, Jae-Hwan;Shin, Hyun-Dong;Kwon, Doo-Han;Park, Yong-Il
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.33-41
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    • 2009
  • The crushed fruiting body of Alaskan Porodaedalea pini (Brot.) Murrill (syn. Phellinus pini) was extracted in boiling water for 4 h with the yield of 20.5% in dry mass. This hot-water extract showed significant antiviral activity by inhibiting the plaque formation in HeLa cells by coxackievirus B3 (CVB3) and also showed highest inhibitory effect against neuraminidase activity among water extracts of various mushrooms. From the water extract, the ethanol precipitate (EP) and supernatant fraction (ES) were obtained through 75% ethanol precipitation with the yield of 43.3% and 28.3% in dry mass, respectively. Whereas ES did not show any detectable level of antiviral activity, EP showed significant dose-dependent inhibition of plaque formation by CVB3 in HeLa cells with an $EC_{50}$ (50% effective concentration) of 0.45 mg/mL. The cytotoxicity on HeLa cells by EP was relatively low with the $CC_{50}$ (50% cytotoxic concentration) of 2.25 mg/mL. EP also effectively inhibited neuraminidase activity in a dose-dependent manner showing up to 75% inhibition at 1.7 mg/mL. These results suggest that the hot-water extract and its EP of P. pini fruiting body can be a candidate for the development of a potent broad-range antiviral agent against influenza virus(Flu) as well as CVB3. The major active component of EP was shown to be a heteropolysaccharide-protein complex containing glucose as the main sugar residue with mole percentage of 79.8% and other sugars like galactose (19.2%), xylose (17%), mannose (5.8%), and fucose (4.6%) and a small portion (12.7%, in mass) of protein.

Degradation Mechanisms of TCE in Cement/Fe(II) Systems (시멘트/Fe(II) 시스템에서의 TCE 분해 기작)

  • Lee, Yun-Mo;Kang, Wan-Hyup;Choi, Won-Ho;Hwang, In-Sung;Park, Joo-Yang
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.7
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    • pp.778-782
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    • 2007
  • This study investigated the dechlorination mechanisms of TCE by Fe(II) associated with cement. Batch slurry experiments were peformed to investigate the behaviors of selected ions; Fe(II), Fe(III), $Ca^{2+}$, $SO_4^{2-}$ in cement/Fe(II) system. The kinetic experiments of TCE in cement/Fe(II) systems showed that injected Fe(II) was mostly sorbed on cement within 0.5 day and 90% of injected 200 mM sulfate was sorbed on cement within 0.5 day when $[TCE]_0$ = 0.25 mM and $[Fe(II)]_0$ = 200 mM. The kinetic experiments of TCE in hematite/CaO/Fe((II) systems were conducted for simulation of cement/Fe(II) system. Calcium oxide that is one of the major components in cement hydration reactions or has a reactivity in limited conditions. Hematite assumed the ferric iron oxide component of cement. The reactivities observed in hematite/CaO/Fe(II) system were comparable to those reported for cement/Fe(II) systems containing similar molar amounts of Fe(II). The behavior of Fe(II) and $SO_4^{2-}$ sorbed on solid phase at an early stage of reaction in hematite/CaO/Fe(II) system was similar to that of cement/Fe(II) system. Ferric ion was released from hematite at an early period of reaction at low pH. The experimental evidence of kinetic test using hematite/CaO/Fe(II) system implies that the reactive reductant is a mixed-valent Fe(II)-Fe(III) mineral, which may be similar to green rust. Fe(II) sorbed on cement can be converted to new mineral phase having a reactivity such as Fe(II)-Fe(III) (hydr)oxides in cement/Fe(II) systems.

Physicochemical properties of dacquoise made with sugar or sugar replacements, tagatose, and erythritol (타가토스와 에리스리톨로 제조한 다쿠아즈의 물리화학적 특성 연구)

  • Lee, Na Ra;Kang, Suna;Kim, Ji Hyun;Kim, Ha Hyoung;Lee, Jung Ah;Park, Sunmin
    • Journal of Applied Biological Chemistry
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    • v.60 no.1
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    • pp.87-93
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    • 2017
  • The objective of this study was to investigate the physicochemical characteristics and sensory evaluation of dacquoise made with sugar or sugar replacement (tagatose, erythritol). Sugar (S) was replaced with tagatose (T), and sugar powder was replaced with tagatose (TS), and erythritol (TE). S showed the highest sugar contents compared with TS and TE (p <0.05) but there were no significant differences with T. There were not significant differences in specific gravity and spread factor among the sweeteners. S and T tended to have higher loss factors than TS & TE. The more tagatose was increased, the more the redness value increased compared with S, and sucrose showed similar tendency for lightness (redness; TS > T > TE > S, lightness; S > T > TS > TE). TE showed good results for physical characteristics whereas T was better in the sensory test. The above results bear out that tagatose and erythritol both are suitable as alternative sweeteners for making dacquoise, but tagatose was better. Tagatose appeared to be a better replacement for sucrose in baking dacquoise, and dacquoise with 33% of the sugar replaced tagatose (T) could be suitable for use by diabetic and obese people. 95.1% tagatose (TS) maybe appropriate for diabetic patients. In conclusion, tagatose is a suitable sugar replacement for making dacquoise.

Stable Transmission and Continuous Expression of Human Interleukin-10 Transgene in the Offspring of Transgenic Mice (형질전환 생쥐의 후대에서 인간 Interleukin-10 유전자의 안정적 전이와 지속적인 발현)

  • Zheng Z. Y.;Koo D. B.;Han Y. M.;Lee K. K.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.203-207
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    • 2004
  • The transgenic mice carrying human Interleukin-10 (hIL-10) gene in conjunction with bovine (3 -casein promoter express hIL-10 in milk during lactation. In this study, stability of germ line transmission and expression of hIL-10 transgene integrated into host chromosome were monitored up to generation F8 of transgenic mice. When male mouse of generation F8 was crossbred with normal females, approximately half of offspring (50.9±5.8%) were identified as transgenic mice. Generation F9 to F15 mice also showed similar transmission rates (66.0±20.1%, 61.5±16.7%, 41.1±8.4%, 40.7±20.3%, 61.3±10.8%, 49.2±18.8% and 43.8±25.9%, respectively), implying that hIL-10 transgene can be transmitted stably up to long term generation in the transgenic mice. Expression levels of human IL-10 from milk of generation F9 to F14 mice were 3.6± 1.2 mg/ml, 4.2±0.9 mg/ml, 5.7±1.5 mg/ml, 6.3±3.5 mg/ml, 6.8±4.5 mg/ml and 6.8±3.1 mg/ml, respectively, which was showed high-level expression compared with that of generation F1 (1.6 mg/ml) mice. In conclusion, our results suggest that transgenic mice can be continuously passed their transgenes to the progeny through the breeding program with the same productivity of human IL-10 protein in their milk.

Studies on the Efficiency of Piglet Production by Controlling the Farrowing Time I. Factors on the Normal Farrowing Time in Swine (돼지에 있어서 분만시각의 인위적 조절에 의한 자돈생산 효율에 관한 연구 I. 돼지에 있어서 자돈분만 시각에 미치는 요인에 관한 연구)

  • 정영채;김창근;윤종택;이종완;전광주;이석우;이학철;이관순;나광빈
    • Korean Journal of Animal Reproduction
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    • v.24 no.2
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    • pp.171-178
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    • 2000
  • To improve the pig farm management and efficiency of swine industry by inducing the farrowing to day-time from night, In the first experiment, reproduction records of purebred and crossbred pigs were collected and analyzed to estimate the $\varepsilon$ ffec 잉 of factors affecting day and night farrowing. The general linear model was used to estimate the least square means of the factors affecting various reproductive characteristics. And also, chi-square tests were used to examine the independence of the reproductive traits and environmental factors using the SAS (1992). The comparisons between pure and crossbred pigs for total number born, percentage of number born alive, gestation length, time length for farrowing were determined. The results indicated that the gestation length of crossbred (115.11 d) was slightly longer than that of purebred (114.89 d, p<0.05). For the seasonal effects on total number born, the largest was found in spring and no differences were found among summer, fall and winter. The average gestation length was 1 day longer in spring and winter than in summer and fall. The total number born and number born alive were smaller in first, second, and greater or equal to sixth parity than other parities. And also, the percentage of no. born alive was least in greater or equal to sixth parity. For the effect of mating methods, natural and artificial insemination, on total number born and number born alive, no differences between the two methods were found. However, the percentage of number born alive for natural mating was 98.06% and was higher than artificial insemination(93.75%). The time length for farrowing was I hour were found for the 6 hrs of farrowing time. In general situation of pig farms, day-time farrowing was 34.8% and night farrowing was 65.2%, indicating that night farrowing was almost double of the night farrowing.

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Association and Polymorphism of Porcine Candidate Genes with Breeding Values in Litter Size of Large Yorkshire and Landrace Inbred Lines (대요크샤 및 랜드레이스종 근교계통돈의 총산자수와 후보유전자에 대한 다형성과 육종가 간의 연관성 분석)

  • Kim, Myung-Jick;Cho, Kyu-Ho;Kim, Doo-Wan;So, Kyung-Min;Choi, Bong-Hwan;Kim, In-Cheul
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.247-250
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    • 2011
  • The objective of this study was to find out candidate genes associated with litter size trait in pigs of inbred Large Yorkshire and Landrace populations. 86 sows were screened for candidate genotypes along with litter size data recordings. Association of litter size with genotypes of candidate genes were investigated to verify the usefulness of each gene's genotypes as markers for the trait. For the lines of Large Yorkshire, PRLR3 and RBP4 genes were genotyped. Frequency distribution of PRLR3 with genotypes AA, AB and BB were each 0.14, 0.44 and 0.42. And the average litter size by PRLR3 genotypes were 8.83, 10.81 and 10.70 piglets per litter, the average estimated breeding values of which were 0.243, 0.332, 0.365, respectively for AA, AB and BB genotypes. Genotypic frequencies of RBP4 by AA, AB and BB genotypes were 0.10, 0.44 and 0.46. The average litter size by genotypes of RBP4 were 10.40, 10.57 and 10.35 piglets per litter and their corresponding average estimated breeding values were 0.451, 0.353 and 0.261, respectively for genotypes AA, AB and BB. Significance in differences among genotypes were not observed, but B allele of RBP4 seems to be associated with litter size. In Landrace lines, frequencies of RBP4 genotypes, AA, AB and BB were 0.29, 0.55 and 0.16. And the average litter size of these genotypes were 10.50, 11.08 and 11.00 piglets per litter. The corresponding averages of estimated breeding values of each genotypes were 0.172, 0.135 and 0.104. In Landrace lines, allele A was more likely to be associated with litter size, even if differences among average litter size were not significant. We conclude that genotyping of two candidate genes is a helpful tool to identify genetic potentials of litter size in pigs.

Effect of Sucrose Concentration on Survival After Frozen-thawed of Bovine IVF Blastocysts in Ethylene Glycol Based Freezing Medium for Slow-Cooling (소 체외수정란의 Slow Freezing을 위해서 Ethylene Glycol 동결보호제에 Sucrose 첨가 농도에 의한 동결효율)

  • 조상래;김현종;최창용;진현주;손동수;최선호
    • Journal of Animal Science and Technology
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    • v.48 no.6
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    • pp.797-804
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    • 2006
  • The present study was undertaken to investigate the post-thawed survivability of bovine embryo depending on different dose of ethylene glycol and sucrose. Ovaries were collected at local slaughterhouse and the cumulus-oocyte-complexes aspirated from ovaries were in vitro matured, fertilized and cultured at 39°C in an atmosphere of 5% CO2 incubator. For conventional slow-freezing, d 7 or 8 expanded blastocysts were collected. Embryos were equilibrated in 1.5 M and 1.8 M ethylene glycol(EG) with 0.1 M and 0.3 M sucrose in Dulbecco's phosphate-buffered saline(D-PBS) supplemented with 0.5% bovine serum albumin. Embryos were then loaded individually into 0.25ml-straw and placed directly into cooling chamber of programmable freezer precooled to 󰠏7°C, after 2 min, the straw was seeded, maintained at 󰠏7°C for 8 min, and then cooled to 󰠏35°C at 0.3°C/min, plunged and stored in liquid nitrogen for at least 3 days. For thawing, the straw containing embryos were warmed in air for 10 sec and exposed to 37°C water for 20 sec. Straws were then removed from 37°C water. Rates of blastocyst survive and hatching were evaluated at 24 to 72 h post-warming. No difference of the survivability was shown between 1.5 M and 1.8 M EG (71 and 70%, respectively). Addition of 0.1 M sucrose to 1.5 M and 1.8 M ethylene glycol in the freezing solution did not differ significantly embryo survival (74 and 77%, respectively), whereas survival rates was higher(89%) in freezing solution contained 0.3M sucrose to 1.8M EG compared with 0.3M sucrose to 1.5M EG group(71%). However, there was no difference in the overall total cell number between the two groups (122±1.8 vs 131±1.4, respectively). In conclusion, the results suggest that 0.3 M sucrose in 1.8 M EG may be optimal condition for freezing and thawing methods with in vitro produced embryos and may be applied to on-farm conditions for embryo transfer.

Estimation of Environmental Effect and Genetic Parameter on Reproduction Traits for On-farm Test Records (농장검정돈의 번식형질에 미치는 환경효과 및 유전모수의 추정)

  • Jung, D.J.;Kim, B.W.;Roh, S.H.;Kim, H.S.;Moon, W.K.;Kim, H.Y.;Jang, H.G.;Choi, L.S.;Jeon, J.T.;Lee, J.G.
    • Journal of Animal Science and Technology
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    • v.50 no.1
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    • pp.33-44
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    • 2008
  • The purpose of this study was to estimate the genetic parameters and trend of Landrace and Yorkshire pigs, which were raised on private farms from 1999 to 2005 and tested for their reproductive performance by the Korea Animal Improvement Association. Prior to analysis, records without pedigree or having value with larger than±3×standard deviation for the Total number of born were excluded. The effects of breed and environmental factors were estimated with least square method(Harvey, 1979), and estimation of breeding values and genetic parameters were performed on the data of 1’st litter only with GIBBSF90(Misztal, 2001) which was programmed according to Gibbs Sampling method based on Bayesian Inference by Gianola and Fernando(1986), Jensen(1994) and others. Gibbs sampling was performed 50,000 times for each parameter, and the first 5000 samples were regarded as those in burn-in period and thus, excluded for post hoc analysis. Total number of born and total number of accident were statistically significant(p<0.01) for the breed, farrowing year, farrowing season and parity effects, and the number born alive at birth was statistically significantp<(0.01) for the breed, farrowing year, farrowing season and parity effects. No particular trend was observed in the genetic and phenotypic improvement of the total number of born and number born alive at birth before 2001, when the piglet registration system started, but the tendencies of increasing for the total number of born and number born alive and decreasing for the total number of accident were observed since 2001. Somewhat higher heritability estimates of our study seems to be attributed to the situations that first parity records with poor farrowing performances were used in the analyses and it was impossible to obtain accurate reproductive performance due to the absence of criteria for record keeping at the level of individual farms.