• 한국식품과학회지
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• 제37권5호
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• pp.822-826
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• 2005

본 연구에서는 천년초 줄기 물추출물의 전처치가 사염화탄소로 유발한 흰쥐의 간 손상에 대해 보호효과를 나타내는지 알아보고자 하였다. 천년초 추출액은 건조 고형분량을 측정하고 1g/kg 및 0.5g/kg 체중이 되는 양을 일정시간에 1일 1회 14일동안 경구투여 하였다. $CCl_4$는 올리브유로 희석하여(1:4) 0.5ml/kg체중이 되도록 시료 마지막 투여 종료 3시간 후에 대조군을 제외한 모든 군에 복강주사하였다. 사염화탄소 처치에 의해 혈청 AST, ALT 및 ALP 활성이 대조군과 비교하여 각각 4.3, 4.7 및 1.2배 증가하였다. 반면, 천년초 줄기 추출물을 1g/kg 투여한 군의 AST, ALT 및 ALP의 활성은 $CCl_4$군과 비교하여 각각 약 36%, 41% 및 22% 감소하였다. 사염화탄소에 의해 간의 지질과산화 반응이 증가하였으며 SOD 및 GST활성이 감소하였다. 1g/kg의 천년초 줄기 추출물의 투여는 간의 지질과산화 반응을 감소시키고 SOD 및 GST의 활성을 회복시키는 결과를 보였다. 또한, 천년초 줄기 추출물은 $132.2{\mu}mol$ vit. C eq/g의 OH 라디칼 소거활성을 나타내었으며 페놀화합물의 함량이 6.52mg/g이다. 이러한 결과는 천년초 줄기 추출물의 항산화 활성 및 사염화탄소로 부터 간손상을 예방하는 효과가 있음을 보여주고 있다.

• 생명과학회지
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• 제17권10호
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• pp.1441-1446
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• 2007

본 연구는 서로 다른 상염색체에 위치하고 있는 초위성체유전 표지를 활용하여 제주마 집단의 타 품종과의 차별적 유전특성분석과 제주마 집단에서 활용할 수 있는 효율적인 개체 식별 시스템 설정을 위해 수행되었다. 공시재료로서는 5품종에서 총 191두가 사용되었으며 13종의 좌위에 대한 개체별 유전자형을 분석하였다. 이들 13종에서 출현된 총 대립 유전자수는 제주마의 경우 155종이 나타났다. 한국 제주마 집단에서 나타난 평균 이형접합도는 0.317-0.902였으며 marker 다형성 정보량은 0.498-0.799로 나타났다. 제주마 집단에서 나타난 대립 유전자 발현 특성은 다른 외래 품종 집단과 매우 상이한 결과를 나타냈다. ATH4 좌위의 경우 제주마 집단에서는 5종의 대립 유전자가 고른 분포를 나타낸 반면 QUA종의 경우와 THO종집단에서 특정 좌위의 극단적 발현 빈도를 나타냈다. 품종특이성을 나타내는 분자표지의 대립유전자 발현양상이 확인되었으며 이러한 품종특이성 발현 분자표지는 집단 내 개체에 대한 품종식별 유전자표지로 활용이 가능한 것으로 나타났다. 9종의 초위성체 유전 표지를 활용할 경우 누적 개체 식별력은 99.9%를 나타냈으며 두 마리의 서로 다른 개체가 서로 같은 유전자형을 가질 짝확률은 $0.60\;{\times}\;10^{10}$으로 추정되었다. 따라서 9종의 선정된 유전표지는 제주마 품종 집단에서 적정 신뢰도를 제공할 수 있는 개체 식별 시스템을 설정할 수 있을 것으로 생각된다.

• Journal of Pharmaceutical Investigation
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• 제30권4호
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• pp.241-246
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• 2000

To develop a sustained-release preparation containing ketorolac tromethamine, two sustained-release pellet formulations were evaluated with a pharmacokinetic study as compared with a conventional commercial tablets (10 mg $Tarasyn^{TM}$, Roche Korea Ltd.). Two sustained-release formulations were as follows; formulation A was composed of an inner layer containing 75% of drug coated with $Eudragit^{TM}$ RS 100 membrane and an outer layer containing 25% of drug mixed with $Eudragit^{TM}$ NE30D, and formulation B was composed of only an inner layer containing 100% of drug coated with $Eudragit^{TM}$ RS 100 membrane. The dissolution test was performed for two formulations. In case of conventional tablets, 2.5 mg of drug per a dose was administered orally into male Albino rabbit (2.0-2.3 kg of body weight) 3 times at intervals of 4 hours. In case of two sustained formulations, 7.5 mg of drug was administered once orally. Blood samples were withdrawn periodically after the administration, and the blood concentration was determined by HPLC. The conventional tablets showed very high peak-trough fluctuation between administered doses, but two sustained formulations showed less fluctuation. Formulation A with the loading dose showed the time to reach minimum effective concentration (MEC) i.e. the onset time was less than 20 min, while Formulation B had more than 1 hr of the onset time. Formulation A had the more constant plasma level than formulation B. However, formulation B had a time lag, so the plasma level was less than MEC for an initial period of 1 hr. In formulation A, the plasma level was maintained within the therapeutic window $(0.3-5\;{\mu}g/ml)$ for a long period. Formulation A was thought to be an ideal sustained-release formulation for ketorolac tromethamine oral delivery system.

• 한국수정란이식학회지
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• 제18권1호
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• pp.51-59
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• 2003

본 실험은 개의 인공수정에 사용할 정자의 보존에 있어서, 개 정액 동결시 동결속도와 응해온도에 따른 정자의 생존율, 운동성 그리고 intact acrosome의 비율을 조사하였던 바 결과는 다음과 같다. 1. 본 실험에서 실험견의 사출된 평균 신선정액의 농도는 3.44$\times$$10^{8}$ /ml로 정상범위에 들었으며, 정자의 형태학적 판정에서 정상적인 정자의 농도는 평균 59.45$\pm$3.45%로 상대적인 기형율은 약 30~40% 정도 나타났으며, 이는 정상적인 상태의 개 정액이라 할 수 있다. 2 개 정액의 동결속도는 동결하는 높이가 6, 10 및 17 cm 일 때 각각 최저온도는 -11$0^{\circ}C$, 7$0^{\circ}C$, -35$^{\circ}C$로 감소하는 경향을 보였으며 이때 최저온도로 감소하는데 소요되는 시간은 각각 6분, 8분 20초 그리고 12분 50초로 결과적으로 분당 동결속도는 각각 19$^{\circ}C$/min, 8.9$^{\circ}C$/min 그리고 3$^{\circ}C$/min로 나타났다. 3. 정액을 동결속도와 융해온도에 따른 정자의 생존율, 운동성 그리고 intact acrosome의 비율은 동결속도가 3$^{\circ}C$/min일 때 가장 높았으며, 융해 온도는 37$^{\circ}C$일 때 효율성이 가장 높은 것으로 나타났다.4. 동결의 방법에 있어서는 액체질소의 표면으로부터 17 cm 높이에서 동결하는 분당 -3$^{\circ}C$의 동결속도에서 동결하여 37$^{\circ}C$에서 2분간 융해하는 방법이 가장 좋은 결과를 보였으며, 생존성과 운동성은 문제없이 사용할 수 있을 것으로 판단되나, 첨체의 intact한 비율은 저조한 결과를 나타내었다.

• 한국수정란이식학회지
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• 제24권4호
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• pp.265-269
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• 2009

The purpose of this study was to determine the effect of different feeding ratios of whole crop barley silage on the embryo production in Hanwoo donors. All donors were basically fed 2.5 kg concentrate daily. Donors were divided into three groups according to the different feeding of forage; hay 70% and rice straw 30% (control, n = 21), whole crop barley silage 80% and rice straw 20% (T1, n = 25), and whole crop barley silage 60% and rice straw 40% (T2, n = 23) fed based on TDN 6.70/ BW 500 kg. All Hanwoo donors received a CIDR together with injections of 1 mg estradiol benzoate and 50 mg progesterone ($P_4$, Day 0). Four days later, they were superovulated with 28 mg FSH twice daily IM in decreasing doses over 4 days. Then donors received 2 doses of $PGF_2{\alpha}$ (25 and 15 mg) with the 5th and 6th injections of FSH on Day 6. CIDR were withdrawn at the 6th FSH injection and the donors received $100\;{\mu}g$ GnRH 36 h after the second $PGF_2{\alpha}$ injection. The donors were artificially inseminated twice, at 8 and 24 h after GnRH, and embryos were recovered 7 or 8 days after the 1st insemination. The flush rate of the donors following positive superovulation responses did not differ among groups (76.2~96.0%, p>0.05). The number of corpus luteum (CL) at embryo recovery also did not differ among groups (10.6~14.0, p>0.05). Furthermore, the mean numbers of total ova (9.4, 10.5 and 12.0) and transferable embryos (5.3, 12.0 and 6.5) did not significantly differ among the control, T1 and T2 groups, respectively (p>0.05). However, mean concentrations of serum $P_4$ of the T1 (64.2 ng/ml) and T2 groups (55.7 ng/ml) were higher than that of control group (43.3 ng/ml, p<0.01), while serum cholesterol concentrations in the control (105.8 mg/dl) and T2 groups ($96.9\;{\pm}\;mg/dl$) were significantly lower than in the T1 group (121.1 mg/dl, p<0.05). Conclusively, whole crop barley silage can be fed a good substitute for hay forage for Hanwoo donors. Furthermore the ratios of whole crop barley silage 60% and rice straw 40% might be more worthful for embryo production.

• 한국수정란이식학회지
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• 제30권3호
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• pp.121-128
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• 2015

The objective of this study was to evaluate the efficiency of sperm cryosurvival in boar sperm separated by Percoll containing antioxidant enzymes. The boar semen was collected into a pre-warmed ($37^{\circ}C$) thermos bottle by gloved-hand method and was separated by 65% Percoll with superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) before freezing. The frozen sperm was thawed at $38.5^{\circ}C$ for 45 sec in water-bath for sperm characteristic analysis. The sperm were estimated with SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction, Rhodamine123/PI double staining for mitochondrial integrity and were analyzed using flow cytometry. In results, sperm viability, acrosome reaction and mitochondrial integrity were improved in separated sperm groups compared with unseparated sperm by Percoll (UP) group. Especially, viability was significantly higher in sperm separated by Percoll containing 400 IU CAT group compared with other groups (P<0.05). And acrosome reaction was decreased in sperm separated by Percoll with 300 IU SOD, 400 IU CAT and 0.5 mM GSH groups compared with other groups, however, there were no significantly difference mitochondrial integrity among sperm separated by Percoll with antioxidant enzymes. In conclusion, we suggest that use of Percoll containing antioxidant enzymes for sperm separation will be beneficial for sperm cryopreservation in pigs.

• 한국수정란이식학회지
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• 제29권3호
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• pp.207-219
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• 2014

Three-dimensional (3D) culture system is useful technique for study of in vivo environment and it was used various experiments. This study was investigated to establish of embryo co-culture system and changes of PAs activity in 3D cultured endometrial cells of pigs. In results, growth of stromal cells into gel matrix were detected only with endometrial and myometrial cells. The most rapid growth of stromal cells were confirmed in $2.5{\times}10^5cells/ml$ and gel matrix containing 15% FBS. Expression of urokinase-PA (uPA) after treatment of hCG (0.5, 1.0, 1.5 and 2.0 IU/ml) were higher than without hCG, but, there are not significant difference among the treatment. On the other hand, expression of uPA after treatment of $IL-1{\beta}$ (0.1, 1, 10 and 100 ng/ml) were higher than without $IL-1{\beta}$, but, there are not significant difference. Expression of uPA after treatment of estrogen (0.2, 2, 20 and 200 ng/ml) were not difference, but PA activity was significantly decreased (p<0.05). Blastocyst was producing in PZM-3 medium containing FBS and endometrial cells were grown in PZM-3 medium. When embryos development with cultured endometrial cells, cleavage rates were not significant difference and blastocyst were not produced in co-culture with stromal cells and 3D culture system. 3D culture system had similar activity to in vivo tissue and these features are very useful for study of in vivo physiology. Nevertheless 3D culture system was not proper in embryo co-culture system. Therefore, we suggest that 3D culture system with embryo co-culture need continuous research.

• 한국수정란이식학회지
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• 제29권3호
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• pp.283-287
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• 2014

Sexed semen is commonly used for the production of calves of the desired gender. Gender selection is important in animal production industries. For example, female cattle are required for the dairy industry while males are preferred in the beef cattle industry. The present study was to assess the in vivo embryo production efficiency using the semen separated according to sex during superovulation in Hanwoo. Seventy Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered on 7 days after the third insemination by flushing the uterus with embryo collection medium. KPN semen straws used artificial insemination contained 20 million sperm (total number 60 million per donor). Sex-sorted semen straws contained 4 million sperm (total number 12 million per donor). The results obtained were as follows: No differences were observed in the efficiency of superovulation rates on KPN semen 87%, and sexed semen 100%, respectively. The mean numbers of total embryos are each $12.58{\pm}8.31$ and $13.25{\pm}7.86$. The mean numbers of transferable embryos, sexed semen were significantly lower than KPN semen ($3.75{\pm}1.98$ vs. $8.23{\pm}6.07$, P<0.05). The rates of unfertilized embryos from superovulation using sexed semen were significantly higher than KPN semen (50% vs. 15%, P<0.05). The rate of degenerated 2-cell embryos from sexed and KPN semen was 60.87% and 11.11%, respectively (p<0.05). In conclusion, these results indicate that superovulation using sexed semen was useful, but efficient embryo production was important to reducing the damage caused by the Flowcytometer-based sperm sorting procedure.

• Journal of Microbiology and Biotechnology
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• 제16권10호
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• pp.1529-1536
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• 2006

We cloned a gene of ompH(D:4) from pigs infected with P. multocida D:4 in Korea [16]. The gene is composed of 1,026 nucleotides coding 342 amino acids (aa) with a signal peptide of 20 aa (GenBank accession number AY603962). In this study, we analyzed the ability of the ompH(D:4) to induce protective immunity against a wild-type challenge in mice. To determine appropriate epitope(s) of the gene, one full and three different types of truncated genes of the ompH(D:4) were constructed by PCR using pET32a or pRSET B as vectors. They were named ompH(D:4)-F (1,026 bp [1-1026] encoding 342 aa), ompH(D:4)-t1 (693 bp [55-747] encoding 231 aa), ompH(D:4)-t2 (561 bp [187-747] encoding 187 aa), and ompH(D:4)-t3 (540 bp [487-1026] encoding 180 aa), respectively. The genes were successfully expressed in Escherichia coli BL21(DE3). Their gene products, polypeptides, OmpH(D:4)-F, -t1, -t2, and -t3, were purified individually using nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. Their $M_rs$ were determined to be 54.6, 29, 24, and 23.2 kDa, respectively, using SDS-PAGE. Antisera against the four kinds of polypeptides were generated in mice for protective immunity analyses. Some $50{\mu}g$ of the four kinds of polypeptides were individually provided intraperitoneally with mice (n=20) as immunogens. The titer of post-immunized antiserum revealed that it grew remarkably compared with pre-antiserum. The lethal dose of the wild-type pathogen was determined at $10{\mu}l$ of live P. multocida D:4 through direct intraperitoneal (IP) injection, into post-immune mice (n=5, three times). Some thirty days later, the lethal dose ($10{\mu}l$) of live pathogen was challenged into the immunized mouse groups [OmpH(D:4)-F, -t1, -t2, and -t3; n=20 each, two times] as well as positive and negative control groups. As compared within samples, the OmpH(D:4)-F-immunized groups showed lower immune ability than the OmpH(D:4)-t1, -t2, and -t3. The results show that the truncated-OmpH(D:4)-t1, -t2, and -t3 can be used for an effective vaccine candidate against swine atrophic rhinitis caused by pathogenic P. multocida (D:4) isolated in Korea.

• Journal of Microbiology and Biotechnology
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• 제26권6호
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• pp.1115-1123
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• 2016

_L-Asparaginase (E.C. 3.5.1.1) is an enzyme involved in asparagine hydrolysis and has the potential to effect leukemic cells and various other cancer cells. We identified the _L-asparaginase gene (_L-ASPG86) in the genus Mesoflavibacter, which consists of a 1,035 bp open reading frame encoding 344 amino acids. Following phylogenetic analysis, the deduced amino acid sequence of _L-ASPG86 (_L-ASPG86) was grouped as a type I asparaginase with respective homologs in Escherichia coli and Yersinia pseudotuberculosis. The _L-ASPG86 gene was cloned into the pET-16b vector to express the respective protein in E. coli BL21 (DE3) cells. Recombinant _L-asparaginase (r-_L-ASPG86) showed optimum conditions at 37-40℃, pH 9. Moreover, r-_L-ASPG86 did not exhibit glutaminase activity. In the metal ions test, its enzymatic activity was highly improved upon addition of 5 mM manganese (3.97-fold) and magnesium (3.35-fold) compared with the untreated control. The specific activity of r-_L-ASPG86 was 687.1 units/mg under optimum conditions (37℃, pH 9, and 5 mM MnSO₄).