• Journal of Animal Science and Technology
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• 제55권5호
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• pp.417-425
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• 2013

동결 닭 PGCs의 생식계열 키메라를 이용한 생체에의 복원을 실용화 하기 위해서는, 닭 PGCs의 동결보존기술의 향상에 의해 동결 및 융해 후의 많은 생존세포를 확보 하는 것과, 생식계열 키메라의 제작효율을 높이는 것이 반드시 필요하다. 닭 PGCs는 배양 5.5일령의 닭 원시생식선으로부터 채취하고, ACS 방법에 의해서 순수 닭 PGCs를 분리했다. 닭 PGCs의 동결보존실험결과 다음의 4종류의 동결방법을 각각 비교 검토했다. 1. 플라스틱 스트로에 의한 완만동결법 (SF), 동결보호물질은 2M 에틸렌 글리콜 (EG), 2. 스트로에 의한 급속동결법 (RF), 8M EG + 7% PVP, 3. 동결용 Cryotube에 의한 SF, 2M EG, 4. 튜브에 의한 SF, 10% DMSO. 동결 및 융해 후의 PGCs의 생존율은 각각 76.4%, 70.6%, 80.5%, 78.1%로 관찰되었다.

• 한국수정란이식학회지
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• 제28권3호
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• pp.193-198
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• 2013

This study was designed to evaluate the effect of L-cysteine on sperm characteristics and oocyte cleavage in vitro in Korean native cattle. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk and/or 0, 5, 10 and 20 mM L-cysteine before cryopreservation. The viability in frozen-thawed sperm were estimated by SYBR14/PI double stain, acrosome damage with FITC-PNA, mitochondria intact with Rhodamin123 and hydrogen peroxide($H_2O_2$) level with carboxy-DCFDA by flow-cytometry. The developmental capacity was also assessed with cleavage rates in oocytes fertilized in vitro by frozen-thawed sperm. In results, the sperm viability was significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). In addition, acrosome damage was significantly decreased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). The mitochondria intact was also significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). On the other hand, the cleavage rates were significantly increased in 0 mM, 5 mM and 10 mM groups than 20 mM concentration of L-cysteine (p<0.05). The oocyte degeneration of oocytes were significantly decreased in 0 mM, 5 mM and 10 mM groups than in 20 mM L-cysteine group (P<0.05). However, there are no significantly differences among the L-cysteine treatment groups. We suggest that concentration of 10 mM L-cysteine have beneficial impact for sperm cryopreserved in Korean native cattle. This result also could be recommended for artificial insemination program if supported by an improvement in the fertility results and required further study.

• 한국수산과학회지
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• 제11권2호
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• pp.115-121
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• 1978

부산연안의 해수오탁정도를 조사하기 위하여 생물검정법에 의해 지표종으로서 개량조개, Mactra chinensis를 사용하여 1977년 7월 12일부터 1977년 7월 17일 사이에 3회에 걸쳐 총 12개 지점에 조사를 실시하였다. 판정의 단계로는 수정모형성기, 상실기, 담륜자유생기로 나누어 각각 정상발생율가 기형발생율을 조사 하였다. 1. 부산 연안의 해수욕장의 애수오탁은 해운대, 송도, 광안리, 송형, 다대포의 순으로 그 정도가 심하게 나타났는데 이중 해운대의 경우는 조사당시 7월 7일에 인근 해역의 사고선박으로부터 유출된 유류의 경향이 있었던 것으로 간주된다. 2. 횟집 밀집지역인 동삼동의 경우 총 기형발생율이 $47.17\%$이고, 어패류시장이 있는 충무동의 경우는 $61.14\%$ 총 기형발생율을 나타내고 있다. 3. 공장폐수의 직접적인 영향을 받고있는 수영만과 용호만의 경우는 $70\%$ 이상의 총 기형발생율을 나타내고 있어 생물의 발생이 거의 불가능한 지역으로 나타나고 있다.

• Reproductive and Developmental Biology
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• 제39권3호
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• pp.77-81
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• 2015

본 연구에서 배아의 생식세포 동결에 가장 흔히 쓰이고 있는 두 가지 동결 보호제, 즉 DMSO와 EG의 독성을 비교하고자 생쥐 수정란 모델을 이용한 실험을 하였다. 생후 6주령의 암컷 생쥐 $F_1$ hybrid mice에 10 IU의 PMSG를 복강 주사하여 과배란을 유도하고, 2-세포기 배아를 획득하고 DMSO와 EG 각각 노출시킨 후, 배양을 하였다. 배반포의 전체 세포수는 2-세포기 단계에서 DMSO($68.1{\pm}24.1$)로 EG($81.2{\pm}27.0$) 혹은 control($99.0{\pm}18.3$)(p<0.001) 처리구에 비해서 유의적으로 낮았다. DMSO 처리구가 EG 처리구에 비해 세포수가 적었다. DMSO($15.4{\pm}1.5$)와 EG($10.2{\pm}1.4$) 두 처리구는 대조구($6.1{\pm}0.9$, p<0.0001)와 비교해서 배반포에서 세포사 비율이 더 높음을 확인했다. 또한, DMSO 처리구는 EG 처리구(p<0.001)보다 더 많은 세포사멸된 세포가 확인되었다. DMSO 또는 EG 처리군과 대조군 사이에는 배아 부화율에 있어서 차이가 있었으며, 이는 배아에 대한 동결 보호제의 잠재적인 독성을 확인한 결과였다. 이번 연구에서 장기간 처리했을 때 EG 처리군보다 DMSO 처리군에서 배아발달과 세포수가 저하된 것은 DMSO의 독성이 더 높을 것으로 사료된다.

• 한국가축번식학회지
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• 제17권3호
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• pp.233-242
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• 1993

The effects of in vitro maturation and sperm treatment condition on the in vitro fertilization (IVF) and developmental capacity of bovine oocytes were investigated and the development of embryos was compared under the 2 different co-culture system, with GC or BOEC. The cultured embryo to 16 cell or morula wre transferred into recipients or frozen by 2 different freezing method. The results obtained were summarized as follows; 1. In vitro maturation rates of vovine follicular oocytes cultrued in TCM199 with 10% FCS or ECS were 64.0% and 72.7%, but the case of addition of 10% FCS or ECS to TCM199 co-cultured with granulosa cells were 81.3% and 84.0%, respectively. IVM rate of three TCM199 added to granulosa cells was higher than that of media without granulosa cells. 2. When bovine follicular oocytes were matured in TCM199 with 10% FCS and GC and then fertilized in vitro by sperm treated with caffeine, embryo developments of bovine oocytes co-cultured with BOEC were 38.4% and 51.4%, respectively. But those of bovine oocytes co-cultured with GC were 52.2% by sperm treated with caffeine-heparin. 3. Cleavage rates of bovine oocytes cultured with 10% FCS alone and fertilized in vitro by sperm treated with caffeine-heparin was 33.0%. 4. When bovine follicular oocytes were matured in TCM199 with 10% FCS and GC, embryo developments of bovine ooctyes co-cultured with BOEC of GC were 46.0% and 50.2%, respectively. 5. When bovine follicular oocytes were matured in TCM199 with 10% ECS and GC, embryo developments co-cultured with BOEC or GC were 45.2% and 51.4%, respectively. 6. When Korean Native cow's follicular oocytes matured in TCM199 with 10% FCS and GC, embryo developed co-cultured with BOEC or GC were 45.2% and 51.4%, respectively. 6. When Korean Native cow's follicular oocytes matured in TCM199 with 10% FCS and GC, embryo developments of the bovine oocyte co-cultured with BOEC and GC were 41.8% and 60.1%. But with FCS 10% those of the bovine oocytes co-cultured with BOEC and GC were 42.0% and 48.4%, respectively. 7. When Holstein's follicular oocytes were matured in TCM199 with 10% ECS and GC, embryo developments fo the bovine oocytes co-cultured with BOEC and GC were 50.0% and 57.7%, but with ECS 10% those of the bovine oocytes co-cultured with BOEC and GC were 52.2% and 56.5%, respectively. 8. The viability of frozen-thawed embryos ranged from 60~80% and those of frozen-thawed embryos from vitrification was lower than that from conventional metiod. 9. The selected fresh embryos were transferred nonsurgically to 7 recipients but did not result in pregnancy.

• 한국수학교육학회지시리즈C:초등수학교육
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• 제14권3호
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• pp.237-259
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• 2011

수학 학습장애는 정상적인 수학적 역량 획득을 저해하는 학습 장애의 한 유형으로, 아동, 청소년의 5- 10% 정도가 겪는 학습 장애의 한 분야이다. 현재 수학학습장애를 진단하기 위하여 기초학력평가나 표준화된 검사가 사용되고 있다. 검사 결과를 토대로 아동의 사전지식을 파악하고, 취약한 영역을 찾는 것 역시 중요하다. 본 연구는 수학학습장애의 유형을 파악하고, 중재의 출발점을 알려줄 수 있는 포괄적인 진단 검사가 포함해야 하는 구성요소를 찾는 것을 목표로 하고 있다. 이를 위하여 신경심리학적 이론에 근거하여 수학학습에 영향을 주는 기본적인 인지적 요인들을 찾고, 관련되어 활성화 되는 두뇌 영역과 그들의 세부적 기능을 살펴본다. 또 수학학습장애 아동의 신경심리학적 특징을 살펴 본 다음 수학학습장애의 유형을 분류한다. 그 결과를 바탕으로 교육현장에서 사용될 수 있는 심층 진단 검사의 개발을 위한 기초연구를 수행하고자 한다.

• 한국응용곤충학회지
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• 제54권4호
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• pp.419-423
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• 2015

애멸구(Laodelphax striatellus)는 rice stripe virus (RSV)의 매개충으로 벼에 큰 피해를 주는 해충이다. 본 연구에서는 애멸구의 장 단시형, 암 수, 약 성충에 대한 RSV 보독률과 이병률을 비교하였다. 애멸구의 장 단시형의 RSV 보독률은 각각 60.7%, 63.1%로 크게 차이는 없었다. 암 수에 대한 RSV 보독률은 각각 61.9%, 52.2%로 암컷의 보독률이 더 높았으나 유의성은 없었다. 약 성충의 보독률을 비교한 결과 각각 51.2%, 58.7%로 역시 크게 차이가 나지는 않았다. RSV에 감염된 애멸구에 노출된 건전한 벼의 이병률은 장시형은 53.3%, 단시형은 48.2%를 보였으며, 약 성충의 이병률은 각각 38.2%, 42.6%를 보여 유의성은 없었다. 반면 암컷은 50.5%의 이병률을 보이고 수컷은 22.3%의 이병률을 보여 암컷이 수컷에 비해 22.3% 이병률이 높아 유의성이 있었다. 또한 벼와 애멸구의 RSV 감염여부에 의한 애멸구의 발육기간은 건전한 벼에 RSV 감염 애멸구를 접종 했을 때 가장 긴 것으로 나타났으며, 건전한 벼에 건전한 애멸구를 접종 했을 때 발육기간이 가장 짧은 것으로 나타났다.

• Clinical and Experimental Reproductive Medicine
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• 제25권1호
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• pp.35-41
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• 1998

This study was carried out to investigate the development rates of human embryos co-cultured with cumulus cells to each blastocyst stage. Human zygotes were co-cultured on cumulus cell monolayer in YS medium supplemented with 20% hFF. On day 2, if patient had four or more "good" embryos (regular blastomeres without fragmentation), embryos were further cultured for 72hrs. Blastocysts on day 5 were classified into early blastocyst (ErB), early expanding blastocyst (EEB), middle expanding Blastocyst (MEB), and expanded blastocyst (EdB) on the basis of their morphological aspects of trophectoderm cells and blastocoele. Subsequently, maximum 3 of best blastocysts were transferred in 486 cycles. The results in this study were as follows: Patients who had four or more "good" embryos on day 2 were 498 persons, but patients whose embryos could not be transferred due to failure in development to the blastocyst stage on day 5 were 12 persons (2.4%). The development rate of embryos to the blastocyst stage was 58.2% (2,885/4,957) on day 5, and the rates that developed to the ErB, EEB, MEB, and EdB stage were 15.0% (743/4,957), 14.9% (739/4,957), 14.4% (714/ 4,957), and 13.9% (689/4,957), respectively. Total 1366 blastocysts were transferred in 486 cycles (mean number=2.81). The implantation rate and the ongoing implantation rate obtained by observing the number of G-sac and FHB were 29.9% (409/1,366) and 22.5% (308/1,366), respectively. The clinical pregnancy rate was 51.2% (249/486), and the ongoing pregnancy rate' was 39.1% (190/486). Among women showing ongoing pregnancy, women with singleton were 50% (95/190), women with twin were 37.9% (72/190), and women with triplet were 12.1% (23/190). Although triplet pregnancy rate in this study was high such as 12.1%, because many blastocysts with high viability were produced in our co-culture system using cumulus cells on day 5, we really believe that a multiple pregnancy except twin should not occur by selecting good embryos for maximum two blastocyst transfer. These results demonstrate that autologous cumulus cells may be used for the production of blastocysts with high developmental competence, and the use of autologous cumulus cells to be collected easily, and to be treated conveniently at OPU must be an effective means for obtaining high implantation and pregnancy rate.

• Clinical and Experimental Reproductive Medicine
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• 제30권4호
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• pp.333-340
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• 2003

Objective: This study was performed to examine the maturation and the development to the blastocyst stage of immature oocytes collected from patients with high risk of ovarian hyperstimulation syndrome (OHSS). Materials and Methods: Cumulus-oocyte complexes (COCs) were collected following only HCGpriming for non stimulated IVF-ET cycles of the patients. At the time of oocyte collection, COCs were classified into three groups in accordance with their appearance (Group I: oocytes with dispersed cumulus cells; Group II: oocytes with compacted cumulus cells; Group III: oocytes with sparse cumulus cells). The in vitro maturation and blastocyst development rates of the COCs were compared among these groups. From August 2001 to June 2002, 48 IVM/IVF-ET cycles from 42 patients (mean age: $32.4{\pm}3.8$ years) were performed. To prevent the occurrence of OHSS, the patients were primed with 10, 000 IU HCG alone 36 h before oocyte collection without gonadotropin stimulation. Oocytes were aspirated on cycle days from 7 to 13. The normal COCs were classified into three groups according to their appearance. The aspirated immature oocytes were cultured in YS maturation medium containing 30% (v/v) human follicular fluid (HFF), 1 IU/ml FSH, 10 IU/ml HCG and 10 ng/ml rhEGF. Fertilization was induced by intracytoplasmic sperm injection (ICSI). All zygotes were co-cultured with cumulus cells in $10{\mu}l$ YS medium containing 10% HFF until day 7 after oocyte collection. Blastocyst transfer was performed on day 5 after ICSI. Results: Th e mean number of oocytes cultured in the IVM/IVF cycles was $24.7{\pm}10.6$. Of 1185 COCs, those assigned to Group I, II and III were 470 (39.7%), 414 (35.0%) and 301 (25.4%), respectively. The maturation rate (94.5%, 444/470, p<0.05) in Group I was significantly higher than those of Group II (62.8%, 260/414) and Group III (73.1%, 220/301). Especially, 30.9% of COCs in Group I (145/470) was matured on the day of oocyte aspiration. There were no differences in the rates of fertilization and cleavage among the three groups. The development rate to the blastocyst stage in Group I (54.6%, 206/377, p<0.05) was also significantly higher than those in Group II (33.0%, 68/206) and Group III (30.1%, 52/173). Twenty-four clinical pregnancies (50.0%) was obtained and 22 pregnancies (45.8%) are ongoing. Implantation rate in the present study was 24.6%. Conclusion: These results suggest that there is a positive correlation between the appearance of COCs and the developmental competence of the immature oocytes in non stimulated IVM/IVF cycles.

• Ecology and Resilient Infrastructure
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• 제2권2호
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• pp.147-153
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• 2015

독성 소재를 활용하여 하천생태복원 사업을 추진할 경우, 수생 생물이 직접적으로 독성물질에 노출되어 다양한 생물학적 문제를 야기할 수 있다. 따라서, 본 연구에서는 하천사업에 활용되는 콘크리트 소재의 독성을 분석하기 위해 한국산 무당개구리(Bombina orientalis)배아를 이용하여 생존율과 기형발생률 측면에서 발생독성 분석을 수행하였다. 콘크리트 바인더들의 독성을 평가한 결과, 무당개구리 배아의 생존율은 대조군 (control), 식물성 폴리우레탄(P1), 우레탄(P2), 시멘트(C) 실험군에서 각각 100%, 94-95%, 66-89%, 0%로 나타났다. 기형발생률은 대조군, P1, P2, C 실험군에서 각각 10.5%, 5.3-10.5%, 26.3-27.8%, 35.7%로 나타났다. 시멘트의 독성이 가장 강하게 나타났으며, 상대적으로 식물성 폴리우레탄의 독성이 가장 약한 것으로 나타났다. 또한, 충분한 양생시간을 가짐으로써 용출되는 독성물질이 저감되는 것을 확인하였다. 이상의 결과들로서 시멘트와 폴리우레탄은 양서류의 초기발생을 저해하는 것이 확인되었다. 본 연구는 하천복원을 위한 소재의 선택에 있어 생물학적 안전성을 검토하는 것이 매우 중요하며, 친환경 소재 및 친환경 공정 선택의 중요성을 보여준다.