• 한국수정란이식학회지
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• 제17권1호
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• pp.13-21
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• 2002

Intracytoplasmic Sperm Injection (ICSI) has been widely used fur both human infertility and basic research. However, the high incidence of chromosomal abnormality is severe problem in cattle. Various oocyte activation stimuli, therefore, were compared by assessment of developmental capacity and chromosome analysis. Motile sperm selected by Percoll-density gradient were treated with 5 mM dithiothreitol (DTT) and injected into an oocyte matured fur 24 h. Eggs were then allocated into 5 treatment groups. Group 1 (control), sperm injection was performed without any further activation stimuli to the oocytes. Group 2 (handled control), sham injection was performed without sperm. In Group 3, oocytes exposed to 5 (M ionomycin for 5 min at 39(C. Group 4. ionomycine + 1.9 mM demethylaminopurine (DMAP, 3 h) and Group 5, ionomycine + 3 h culture in Ml99 + DMAP. Cleavage and the later development rate in Groups 1, 2 and 3 were significantly (P<0.05) lower than those in Groups 4 and 5. The incidence of chromosomal abnormality in the embryos treated directly with DMAP after ionomycine was relatively higher than in the embryo of Group 3 h, delayed DMAP treatment. From this results DMAP caused to be arrested the release of the 2nd polar body, resulting in changes of chromosomal pattern. Therefore, the time interval between ionomycin and DMAP is a crucial role in bovine ICSI.

• Journal of Plant Biotechnology
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• 제31권1호
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• pp.73-78
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• 2004

본 연구에서는 백합의 배수성 육종 체계를 확립하기 위한 기초자료를 얻기 위하여 인위적 체세포 염색체 배수화 방법에 대한 실험을 수행하였다. 체세포 배수화를 위한 in vitro처리의 경우 4배체 뿐 아니라 혼수체의 비율이 처리에 따라 높게 나타나는 경우도 있으며, 무엇보다 4배체 획득율을 기준으로 그 효율성 조사해 본 결과 colchicine의 경우 1,000ppm농도에서 3시간 처리 (효율성=11.2％), oryzalin의 경우 30ppm에서 3시간 처리 (효율성=10.0％),caffeine의 경우 9,000ppm에서 9시간 처리 (효율성=11.7％)가 가장 양호하였다. 처리에 따른 4배체 획득 요율성은 다소 차이가 있을 수 있으나 인체에 대한 유해성, 혼수체 빈도 및 식물체에 대한 toxicity등을 고려 할 때 oryzalin 또는 caffeine사용이 바람직할 것으로 판단된다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.6-6
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• 2001

Oocyte freezing has become a prevalent source for related reproductive technologies. This study was carried out to evaluate viability of post-thawed bovine oocyte injected DTT-treated sperm following by two different activation stimuli (Group 1, 5 M ionomycin, 5 min ＋ CR1aa, 3 h . 1.9 mM dimetylaminopurine (DMP), 3 h; group 2, ionomycin ＋ 10 $\mu\textrm{g}$/$m\ell$ cycloheximide(CHX), 5h). The techniques of ultra-rapid freezing used in this study were essentially similar to those of described by Vajta et al (Theriogenology 1999; 52:939-948), Denuded oocytes at 22 h of culture were exposed to cryoprotectant (3.2 M Ethylene glycol, 2.36 M DMSO, 0.6 M sucrose), and followed by freezing in electron microscopic grid. After thawing the oocytes were transferred back into the drop of maturation medium and cultured for additional 2 h before being subjected to ICSI. All eggs were then cultured in CRlaa medium, and transferred into M199+10% FCS on day 4. The culture was maintained until day 9. In Experiment 1, frozen-ICSI eggs were compared on development into blastocyst to those of unfrozen and IVF control. Those eggs were activated with the method of group 2. A higher proportion of unfrozen-ICSI and IVF eggs developed into cleavage and blastocysts than of frozen-ICSI eggs (65% and 13%; 71% and 23% vs. 39% and 8%; P<0.05). In Experiment 2, development and ploidy of embryos made from group 1 were compared to those from group 2. Between groups there did not differ on the rates of development, however, chromosomal abnormality in group 1 was significantly higher than in group 2 (49% vs. 30%; P<0.05). The present result suggests that frozen bovine oocytes can be used for ICSI.

• 식물분류학회지
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• 제41권1호
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• pp.51-57
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• 2011

본 연구에서는 닭의덩굴속 호장근절의 한반도산 분류군을 중심으로 잎 표피세포 및 털의 미세구조를 관찰하여, 이들 형질의 분류군간/분류군내 변이 양상을 구명하고 식별형질로서의 분류학적 유용성을 검증하고자 하였다. 본 절 분류군들의 표피세포의 형태는 향축면 표피세포의 경우 다각형, 배축면 표피세포는 수층벽이 파상인 부정형으로 근본적으로 유사하였으나, 표피상납질의 발달 정도는 분류군간/지역집단 간에 변이가 있는 것으로 밝혀졌다. Fallopia sachalinensis는 본 절 다른 분류군과는 달리 모든 잎 배축면의 모든 표피세포에 표피상납질이 조밀하게 발달하며, 또한 울릉도 및 독도에 분포하는 본 종 개체들은 일본 및 사할린에 분포하는 개체들에 비해 표피상납질이 상대적으로 가늘게 발달하는 특징을 나타내었다. 기공의 크기는 분류군내에서도 변이가 존재하였으며, 일반적으로 다배체(6, 8, 12배체)인 개체들이 4배체인 개체에 비해 기공이 큰 경향을 나타내었다. 한편, 본 연구에서 조사된 본 절 분류군에는 크게, (1) 원추형 단세포 털, (2) 단세포 또는 2-8개의 세포가 한 줄로 연결된 선형의 털, (3) 다세포로 구성된 방패상 선모의 3 종류가 분포하였으며, 털의 종류와 분포 양상은 분류군 및 지역집단을 구분하는데 유용한 형질인 것으로 판단되었다.

• Clinical and Experimental Reproductive Medicine
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• 제44권1호
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• pp.40-46
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• 2017

Objective: To describe in vitro development of human embryos derived from an individual with a homozygous pathogenic variant in NLRP7 (19q13.42) and recurrent hydatidiform mole (HM), an autosomal recessive condition thought to occur secondary to an oocyte defect. Methods: A patient with five consecutive HM pregnancies was genomically evaluated via next generation sequencing followed by controlled ovarian hyperstimulation, in vitro fertilization (IVF) with intracytoplasmic sperm injection, embryo culture, and preimplantation genetic screening. Findings in NLRP7 were recorded and embryo culture and biopsy data were tabulated as a function of parental origin for any identified ploidy error. Results: The patient was found to have a pathogenic variant in NLRP7 (c.2810+2T>G) in a homozygous state. Fifteen oocytes were retrieved and 10 embryos were available after fertilization via intracytoplasmic sperm injection. Developmental arrest was noted for all 10 embryos after 144 hours in culture, thus no transfer was possible. These non-viable embryos were evaluated by karyomapping and all were diploid biparental; two were euploid and eight had various aneuploidies all of maternal origin. Conclusion: This is the first report of early human embryo development from a patient with any NLRP7 mutation. The pathogenic variant identified here resulted in global developmental arrest at or before blastocyst stage. Standard IVF should therefore be discouraged for such patients, who instead need to consider oocyte (or embryo) donation with IVF as preferred clinical methods to treat infertility.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.176-176
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• 2017

Hydropriming had positive effects on the time for germination to reach 50%, the germination index, the time to final germination percentage, and the number of uniform seedlings with enlarged cotyledons in in vitro germination of small watermelon. In addition, the highest shoot initiation was obtained from hydroprimed cotyledonary nodes ($95{\pm}6%$), followed by non-primed cotyledonary nodes ($78{\pm}6%$), hydroprimed cotyledons ($72{\pm}4%$), and non-primed cotyledons ($48{\pm}4%$). Meanwhile, no shoots were initiated from hypocotyls. The total number of shoots that initiated from cotyledonary nodes and cotyledon explants was insignificant, indicating that both cotyledons and cotyledonary node were good sources for the in vitro culture. Choosing explant sources that favor tetraploidy should be the key for producing higher polyploidy plants; a total of 10.5% of tetraploid regenerants were entirely identified from cotyledon explants. Cotyledons with highly differentiated cells might show higher variations than cotyledonary nodes with more preexisting meristematic cells. Cells of cotyledon tissue might undergo changes in ploidy level during differentiation of the culture, or it might be that some of the variations were already present in the tissues of the donor plant. Morphological changes in fruit length of tetraploid regenerants are genotype-dependent.

• Journal of Plant Biotechnology
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• 제47권3호
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• pp.185-193
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• 2020

최근, 유전자교정 작물의 상업화 승인 건수가 급속하게 늘어나고 있으며, 국내에서도 유전자교정 작물의 개발에 대한 집중적인 투자를 통하여 국제경쟁력을 높이기 위하여 노력하고 있다. 그러나 기존의 유전자변형작물의 상업화 과정에서 끊임없이 제기되어온 인체 및 환경에 대한 잠재적인 위해성 논란이 유전자교정 작물에 대하여서도 제기되고 있다. 특히, 비의도적 돌연변이(off-target)가 가장 큰 논란의 중심이 되고 있다. 따라서 본 리뷰는 식물이 내포하고 있는 장점인 배수체, 체세포 돌연변이 그리고 자연 상태에서 아그로박테리아의 T-DNA 단편의 수평 전이로 창출된 자연적인 유전자변형작물과 기존에 상업화가 승인된 유전자변형작물 이벤트들의 게놈 내 비의도적 돌연변이 사례 등을 검토한 결과 유전자교정 작물에서 나타나는 대부분의 비의도적 돌연변이는 인체 및 환경에 미칠 수 있는 위해성을 우려할 만한 수준이 아니라고 할 수 있었다. 이에, 유전자교정 작물의 안전성 평가를 위하여 새로운 규정을 제정할 필요가 없으며 기존의 유전자변형작물의 안전 관리규정을 일부 "용어의 정의" 등만 개정하여 적용하면 충분할 것으로 사료 되었다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권7호
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• pp.3295-3300
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• 2016

Background: Different types of cancer exhibit abnormalities in cell cycle regulators. The murine double minute-2(MDM2) cell cycle regulator is a proto-oncogene that negatively regulates the P53 tumour suppressor gene. Surface epithelial tumours constitute approximately two thirds of ovarian neoplasms. Each histologic type can be classified as benign, borderline and malignant. This study aimed to examine immunohistochemical expression of the MDM2 protein in ovarian serous and mucinous epithelial tumours (benign, borderline and malignant). Materials and Methods: This study included forty five ovarian tumours, subdivided into fifteen cystadenomas (5 serous and 10 mucinous), fifteen borderline tumours (11 serous and 4 mucinous) and fifteen cystadenocarcinomas (9 serous and 6 mucinous). Paraffin sections were stained with haematoxylin and eosin for histopathologic study, and with mouse monoclonal anti-MDM2 antibody for immunohistochemistry. Results: MDM2 positivity was detected in 28.9% of the studied ovarian tumours. All benign tumours were negative and positivity was significantly higher in malignant than borderline tumours (P value of chi-square test =0.000). Significantly, all MDM2 positive mucinous tumours were malignant with no positive mucinous borderline tumours. Malignant tumours showed positive MDM2 expression in 83.3% of mucinous type and in 55.6% of serous type. Borderline serous tumours showed negative MDM2 in 72.7% of cases (P value of Z test =0.04). Conclusions: Alterations in the expression of the cell cycle regulator (MDM2) occur early in the process of tumourigenesis in serous and mucinous ovarian tumours. We suggest that MDM2 may be used in those tumours as a marker for risk stratification and identification of cases with cancer development and progression. We recommend further studies on MDM2 immunohistochemistry, in conjunction with adjuvant methods as DNA ploidy and FISH gene amplification, focusing on the mucinous tumours and differentiating between the three tumour categories, benign, borderline and malignant.

• 한국발생생물학회지:발생과생식
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• 제20권4호
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• pp.305-314
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• 2016

The marine medaka, Oryzias dancena is a suitable sample as a laboratory animal because it has a small size and clearly distinguishes between female and male. Data on the growth and maturity of the diploid and triploid sea cucurbit species suitable for laboratory animals are very useful for studying other species. Triploidy was induced in the marine medaka by cold shock treatment ($0^{\circ}C$) of fertilized eggs for 45 min, applied two minutes after fertilization. The diploid and triploid male fish were larger than their female counterparts (P<0.05), and the concentrations of thyroid stimulating hormone (TSH) and thyroxine (T4) were higher in the induced triploids over 1 year (P<0.05). In both the diploid and tri-ploid groups the concentrations of TSH and T4 were higher in the male fish than in the females (P<0.05), while the testo-sterone and estradiol-$17{\beta}$ concentrations in the induced triploids were lower than in the diploids (P<0.05). The gonadosomatic index (GSI) of the triploid fish was lower than that for the diploids, and the GSI for females in each ploidy group were higher than that for the males. For both groups the GSI was highest at 4 months of age, and decreased thereafter to 12 months. Analysis of the gonads of one-year-old triploid fish suggested that the induction of triploidy probably causes sterility in this species; this effect was more apparent in females than in males.

• Clinical and Experimental Pediatrics
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• 제50권7호
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• pp.601-605
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• 2007

The cure rate of acute lymphoblastic leukemia (ALL) in children dramatically improved over past 5 decades from zero to about 80%. The main cause of improvement is owing to the development of chemotherapy by multicenter clinical trial of large study groups with the understanding of leukemia biology. Recently, pediatric ALL protocols were applied to the treatment of adolescent and even adult ALL patients. For nearly 30 years, clinical factors have been used to risk-stratify therapy for children with ALL, so that the most intensive therapies are reserved for those patients at the highest risk of relapse. The risk groups of ALL are divided as standard- (low- plus intermediate-), high- and very high-risk group according to the prognostic factors, and treatment results improved by this risk based treatment. The factors used to risk-stratify therapy include age, gender, presenting leukocyte count, immunophenotype, cytogenetic aberrations including ploidy and translocations, and initial response after 1 to 2 weeks of therapy. But treatment efficacy is the most important determinant and can abolish the clinical significance of most, if at all, prognostic factors. Today, in the era of intensive, multiagent regimens, there is increasing evidence that we have reached the limits of prognostic significance of currently applied clinical risk factors in childhood ALL. As the cure rate of ALL is about 80%, introducing new prognostic factors such as new molecular prognostic markers, new methods of assessment about minimal residual disease, and pharmacogenetic study, with the development of stem cell transplantation and molecular targeted therapy are needed to cure residual 20% of childhood ALL patients without short and long term complications.