• Asian Pacific Journal of Cancer Prevention
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• 제13권4호
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• pp.1115-1118
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• 2012

Diallyl sulfide (DAS), a flavoring compound derived from garlic, is considered to have cancer chemopreventive potential in experimental animals and humans. This study was designated to examine possible chemopreventive effects of DAS on colon carcinogenesis using genetically engineered transgenic $Apc^{Min/+}$ mice, a well-established animal model for familial adenomatous polyposis (FAP) and sporadic colorectal cancer. Male C57BL/6J-$Apc^{Min/+}$ mice were divided into three groups. Animals of group 1 were placed on the basal diet (AIN-76A) as non-treated controls. Animals of groups 2 and 3 were given DAS-containing diets (in doses of 100 and 300 ppm, respectively). All mice were sacrificed at the end of week 10 of the experiment. Histopathological investigation revealed that the incidence of colonic polyps was decreased dose-dependently by 19% (13/16) in group 2 and by 32% (13/20) in group 3 compared to the 100% incidence (10/10) in group 1. The multiplicity of colonic polyps per mouse was also slightly decreased by DAS treatment ($1.88{\pm}0.35$ in group 2 and $1.63{\pm}0.36$ in group 3) compared to $2.00{\pm}0.39$ in group 1. On the other hand, there were no significant differences in the numbers of total polyps per mouse in the small intestine between the groups. Taken together, we suggest that DAS may exert promising inhibitory effects on colon carcinogenesis in the transgenic $Apc^{Min/+}$ mice.

• 한국수정란이식학회지
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• 제21권2호
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• pp.85-93
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• 2006

본 연구는 소 형질 전환 체세포 핵이식에서 용이한 탈핵을 위해 demecolcine을 이용할 시 탈핵율과 핵이식란의 발육능을 높이기 위한 최적의 조건을 알아보고자 실시되었다. 도축장 유래 미성숙 난자를 18시간 체외성숙 후 제1극체가 확인된 성숙 난자를 0.1, 0.2, 0.4 및 0.8 ug/ml의 demecolcine이 첨가된 배지에서 1시간 더 처리한 다음 세포막이 돌출되어 있는 난자를 체세포 핵이식에 공여하여 각 군간 배반포로의 발육능을 비교하였다. Demecolcine 처리 후 핵이 포함된 셰포막의 protrusion rates를 각 군간 비교한 결과 0.2, 0.4 및 0.8 ug/ml 군에서 0.1 ug/ml 군보다 유의적으로 높았으며 (82.8, 86.2, 90.4 vs. 70.1%), conventional blind 방법과 비교한 결과 demecolcine를 이용한 군에서 유의적으로 높은 탈핵율을 보였다(75.3 vs. 96.2%; p<0.05). 체세포 핵이식란의 발육능 비교에서는 0.1 및 0.2ug/ml 군에서 대조군과 함께 유의적으로 높은 분할율 및 배반포로의 발육능을 보였다(p<0.05). 결론적으로 소 형질전환 체세포 핵이식을 위한 탈핵시 높은 탈핵율과 배반포로의 발육율을 얻을 수 있는 demecolcine의 적정농도는 0.2ug/ml이라고 사료된다.

• Molecules and Cells
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• 제27권2호
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• pp.183-190
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• 2009

The plasma membrane-localized BRASSINOSTEROID-INSENSITIVE1 (BRI1) and BRI1-ASSOCIATED KINASE1 (BAK1) are a well-known receptor pair involved in brassinosteroids (BR) signaling in Arabidposis. The formation of a receptor complex in response to BRs and the subsequent activation of cytoplasmic domain kinase activity share mechanistic characteristics with animal receptor kinases. Here, we demonstrate that BRI1 and BAK1 are BR-dependently phosphorylated, and that phosphorylated forms of the two proteins persist for different lengths of time. Mutations of either protein abolished phosphorylation of the counterpart protein, implying transphosphorylation of the receptor kinases. To investigate the specific amino acids critical for formation of the receptor complex and activation of BAK1 kinase activity, we expressed several versions of BAK1 in yeast and plants. L32E and L46E substitutions resulted in a loss of binding of BAK1 to BRI1, and threonine T455 was essential for the kinase activity of BAK1 in yeast. Transgenic bri1 mutant plants overexpressing BAK1(L46E) displayed reduced apical dominance and seed development. In addition, transgenic wild type plants overexpressing BAK1(T455A) lost the phosphorylation activity normally exhibited in response to BL, leading to semi-dwarfism. These results suggest that BAK1 is a critical component regulating the duration of BR efficacy, even though it cannot directly bind BRs in plants.

• Molecules and Cells
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• 제42권4호
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• pp.301-312
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• 2019

Post-transcriptional regulation underlies the circadian control of gene expression and animal behaviors. However, the role of mRNA surveillance via the nonsense-mediated mRNA decay (NMD) pathway in circadian rhythms remains elusive. Here, we report that Drosophila NMD pathway acts in a subset of circadian pacemaker neurons to maintain robust 24 h rhythms of free-running locomotor activity. RNA interference-mediated depletion of key NMD factors in timeless-expressing clock cells decreased the amplitude of circadian locomotor behaviors. Transgenic manipulation of the NMD pathway in clock neurons expressing a neuropeptide PIGMENT-DISPERSING FACTOR (PDF) was sufficient to dampen or lengthen free-running locomotor rhythms. Confocal imaging of a transgenic NMD reporter revealed that arrhythmic Clock mutants exhibited stronger NMD activity in PDF-expressing neurons than wild-type. We further found that hypomorphic mutations in Suppressor with morphogenetic effect on genitalia 5 (Smg5) or Smg6 impaired circadian behaviors. These NMD mutants normally developed PDF-expressing clock neurons and displayed daily oscillations in the transcript levels of core clock genes. By contrast, the loss of Smg5 or Smg6 function affected the relative transcript levels of cAMP response element-binding protein B (CrebB) in an isoform-specific manner. Moreover, the overexpression of a transcriptional repressor form of CrebB rescued free-running locomotor rhythms in Smg5-depleted flies. These data demonstrate that CrebB is a rate-limiting substrate of the genetic NMD pathway important for the behavioral output of circadian clocks in Drosophila.

• 한국초지조사료학회지
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• 제28권3호
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• pp.165-170
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• 2008

티모시의 colt 품종을 이용하여 최적 조직배양조건을 확립하기 위하여 성숙종자를 배양함에 있어 첨가되어지는 식물 생장조절 물질의종류와 농도가 배발생 캘러스 유도 및 식물체재분화에 미치는 영향을 조사하였다. 배발생 캘러스 유도시 첨가되는 2,4-D는 3 mg/L 첨가 구에서 78%로 캘러스 유도율이 가장 높았으며, 3 mg/L 2,4-D와 0.1 mg/L의 BA를 첨가하여 배양했을 때 형성된 캘러스는 조직적으로 치밀하며 유백색의 배발생 캘러스가 가장 많이 형성되었다. Type B 캘러스만을 이용하여 식물체재분화에는 1 mg/L 2,4-D와 3 mg/L BA가 첨가된 배지에 캘러스를 배양했을 때 25.3%의 재분화율을 나타내었다. 본 연구를 통하여 확립된 효율적인 재분화 시스템은 분자육종을 통한 신품종 티모시의 개발에 유용하게 이용될 수 있을 것이다.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.97-97
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• 2002

Human eryhropoietin (EPO) is acidic glycoprotein hormone that plays key role in hematopoiesis by facilitating differentiation of erythrocyte and formation of hemoglobin (Hb) and is used for the treatment of anemia. Human EPO is consist of 166 amino acids which is modified by three N-glycosylations (24, 38, 83) and single O-glycosylation (126). N-glycosylation is reported to be related to the cellular secretion and activity of EPO. In this study, we examined effects of mutagenesis in glycosylation site of recombinat hEPO for the cellular secretion during production from cultured CHO cell. We produced rhEpo which was cloned by PCR from human liver cDNA (TaKaRa) in cultured CHO cell. Using supernatant of the culture, ELISA assay and western analysis were performed. To estimate biological activity, 20IU of rhuEpo was subcutaneously injected into four ICR mice. After 8 days, HCT level was increased average 13 per cent, RBC was increased ca. 2${\times}$10$\^$6//${\mu}\ell$. In disease model Rat (anemia c-kit, WSRC-WS/WS), HCT was increased ca. 12%, RBC was increased ca. 1.6${\times}$10$\^$6//${\mu}\ell$. These results suggests that rhEpo we produced has biological activity. To remove glycosylation site by substituting 24, 38, 83, and 126th asparagine (or serine) with glutamic acid, overlapping -extension site-directed mutagenesis was performed. To add novel glycosylation sites, 69, 105th leucine was mutated to asparagine. Mutant EPO construct was transfected into CHO cell. Supernatant of the cell culture was analyzed using ELISA assay with monoclonal anti-EPO antibody (Medac, Germany). Since, several reports for mutagenesis of glycosylation sites showed case-by-case results, we examined both transient expression and stable expression. Addition of novel glycosylation sites resulted no secretion while deletion mutants had little effect except some double deletion mutants (24/83 and 38/83) and triple mutant. We suggest that not single but combination of glycosyl group affect secretion of EPO.

• Asian-Australasian Journal of Animal Sciences
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• 제21권9호
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• pp.1318-1323
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• 2008

Corn distiller's dried grains with solubles (DDGS) is a completely new feed ingredient in the Korean feed market. There is an ever increasing need for the Korean feed industry to import and make the best of it as a high protein and high energy feed ingredient. A layer feeding trial was conducted for 10 weeks to investigate the effects of addition of light-colored DDGS to layer diets on laying performance, egg qualities and yolk fatty acid composition. Also, the economics of using DDGS in the Korean situation was analyzed. Nine hundred Hy-line Brown layers, 24 weeks of age, were employed in a feeding trial consisting of four dietary treatments (0, 10, 15, and 20% DDGS), and five replicates per treatment. All experimental diets were prepared as iso-protein (17%) and iso-calorie (TMEn 2,780 kcal/kg). The use of DDGS up to 20% in layer diets did not exert any influence on feed intake, laying rate, total egg mass, mean egg weight and feed conversion ratio (p>0.05). The color and breaking strength of eggshell, as well as the albumin height and Haugh unit were not affected by the addition of DDGS up to 20% in the diet. The yolk color was significantly increased by DDGS supplementation (p<0.05). As the DDGS level increased, the oleic acid content decreased, and the linoleic acid increased (p<0.05) in egg yolk. The degree of saturation of yolk fatty acids was not affected by DDGS supplementation. The inclusion of light-colored DDGS up to 20% in layer diets resulted in a decrease of feed cost per kg without any undesirable effect on laying performance. In conclusion, the light-colored DDGS (L* 56.65) could be used up to 20% in layer diets without any harmful effect on laying performance, and possibly provide economic benefits to the Korean poultry industry.

• 한국임상수의학회지
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• 제27권6호
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• pp.647-651
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• 2010

연속형 $CO_2$ 레이저와 펄스형 $CO_2$ 레이저를 이용한 돼지 피부절개 시 창상 치유에 미치는 영향을 평가하고자 본 실험을 실시하였다. 다섯 마리의 돼지(Landrace x Yorkshire) (45-51 kg, 4-6 개월령, 수컷 3마리, 암컷 2마리)를 이용 하였고, 각각의 돼지에서 우측 및 좌측의 등쪽 피부에 대칭적으로 연속형 $CO_2$ 레이저와 펄스형 $CO_2$ 레이저를 이용하여 절개($2{\times}2{\times}2cm^2$) 하였다. 양측 피부 절개는 Maxon 3-0 를 이용하여 봉합하였다. 수술 후 3, 7, 14, 21일에 병리조직학적 검사를 실시하였다. 창상 부위의 재상피화는 연속형 $CO_2$ 레이저 군에 비해 펄스형 $CO_2$ 레이저군에서 더 많이 이루어졌다. 육아조직 형성은 창상후 경과일 3일에 펄스 $CO_2$ 레이저군에서 유의적으로 높게 나타났다(P < 0.05). 섬유아세포는 창상후 경과일 7일에 펄스형 $CO_2$ 레이져군에서 유의적으로 많게 형성되었다(P < 0.05). 결론적으로 피부절개 시에 있어서 펄스형 $CO_2$ 레이저는 연속형 $CO_2$ 레이저에 비하여 재상피화, 육아조직형성 및 섬유아세포가 더 높게 나타났으며, 레이저 시술에 따른 조직손상을 적게 나타내었다. 따라서 피부절개 시에 있어서 펄스형$CO_2$ 레이저가 연속형 $CO_2$ 레이저 보다 더 적합할 것으로 판단된다.

• 한국가축번식학회지
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• 제27권1호
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• pp.53-59
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• 2003

본 연구에서는 한우 태아의 시기별로 35일령, 50일령, 70일령 및 90일령의 fetal fibroblast cell line을 생산하였고, bovine-specific primer와 Y chromosome-specific primer를 이용하여 PCR에 의해 성을 판별하여 각각 암수 2 line의 한우 fetal fibroblast cell line을 확립하였다. 이들 cell line을 계대배양하여 passage number가 10 이상에서 염색체 분석을 실시하였는데 모두에서 80%이상의 세포가 60개의 정상 염색체수의 나타내어 계대배양이 karyotype에 영향을 미치지 않는 것으로 나타났다. Serum starvation과 confluent 배양 방법을 이용하여 Go 상태로 유도되었는지 확인하기 위해 PCNA antibody를 이용하여 Western blotting 분석을 실시하였는데 PCNA 발현이 현저히 감소되는 것을 확인할 수 있었고, 다시 정상 medium으로 환원시켰을 때 세포분열이 재개되어 Go상태로 유도되었음을 확인할 수 있었다. 또한 serum stravation 방법이 conflent한 배양방법보다 PCNA 발현양이 적은 것으로 나타나 좀더 효율적인 Go 상태 세포 주기 조절방법으로 판명되었다.

• Reproductive and Developmental Biology
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• 제38권3호
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• pp.99-105
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• 2014

In this study, to further understand the mechanism of animal growth and to develop a miniature transgenic animal model, we constructed and tested tetracycline-inducible RNAi system using shRNA targeting the mRNA of mouse insulin-like growth factor (mIGF-1) or mouse growth hormone receptor (mGHR) gene. Quantitative real-time PCR analysis of mouse liver cell (Hepa1c1c7) cells transfected with these vectors showed 85% or 90% of expression inhibition effect of IGF-1 or GHR, respectively. In ELISA analysis, the protein level of IGF-1 in the cells expressing the shRNA targeting IGF-1 mRNA was reduced to 26% of non-transformed control cells. Unexpectedly, in case of using shRNA targeting GHR, the IGF-1 protein level was decreased to 75% of control cells. Further experiments are needed to explain the lower interference effect of GHR shRNA in IGF-1 protein. Accumulated knowledge of this approach could be applicable to a variety of related biological area including gene function study, gene therapy, development of miniature animals, etc.