• Parasites, Hosts and Diseases
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• 제33권4호
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• pp.297-304
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• 1995

요코가와흡충증에 대한 관리책 모색의 일환으로 감마선 조사의 효용성을 관찰하고자 이 연구를 시행하였다. 은어 근육 내에 있는 요코가와흡충의 피낭유충(은어 조사군)과 근육으로부터 분리시킨 피낭유충(피낭유충 조사군)에 감마선을 조사하여 각 군별로 500개씩을 흰쥐에 감염시킨 후 7일째에 생존과 발육상황을 관찰하였다 피낭유충 조사군의 조사량은 5 Gy, 20 Gy, 50 Gy 및 100 Gy 이었으며, 은어에 조사한 량은 5 Gy, 50 Gy, 200 Gy 및 500 Gy로 하였다. 연구 결과, 조사량 증가에 따라 충체 회수율이 급격히 감소하는 양상을 보였으며. 은어 조사군의 경우 같은 조사량에 서 피낭유충 조사군에 비하여 더 많은 충체가 회수되었다 피낭유충 조사군의 $LD_{50}$은 4.5 Gy, 은어 조사군은 62 Gy로 계산되었다 회수된 충체들의 형태학적 변화가 피낭유충 조사군은 100 Gy 에서. 은어조사군은 200 Gy 이상에서 인정되었으며. 변화의 예로는 고환의 발육 부진 또는 소실, 자궁내 충란수 감소 또는 소실 등을 들 수 있었다. 이상의 결과로 볼 때, 은어에 200 Gy 정도를 조사한다면 요코가와흡충 피낭유층의 감염력을 소실시킬 수 있으므로 이 흡충증의 관리방법의 하나로 이용할 수 있을 것으로 기대된다.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.135-141
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• 2010

Embryoid bodies (EBs) generated from human embryonic stem cells (hESCs) include spontaneously induced endodermal lineage cells (ELCs). Activin-A plays important roles in the endoderm differentiation of hESCs. Despite studies on the generation of ELCs from hESCs with treatment of Actvin-A, it was unclear for localization and pattern of ELCs by Activin-A during differentiation of hESCs. Accordingly in this study, we knew that Actvin-A increased the cystic EBs formation, including the highly enriched AFP (endoderm lineage specific marker)-expressing cells in the surface of cystic EBs. To induce the EBs formation from undifferentiated hESCs, cells were transferred onto petri-dish and cultured in suspension condition without bFGF removed hESC media (EB media) for 3 days. Next to investigate the effect of Activin-A, EBs were subsequently cultured in EB media supplement with 100 ng/ml Activin-A for 3 days. After 5~7 days of Activin-A treatment, cystic EBs began to appear which increased in numbers reaching ~60% of initially formed EBs over 5 days. Endoderm lineage marker, AFP were highly expressed and specifically localized at the surface region of cystic EBs comparison with normal EBs. We next attached the cystic EBs onto gelatin-coated plates and cultured for 5 days. In the results of real-time PCR and immunocytochemistry analysis, AFP-expressing cells migrated and localized at the outgrowth region of attached cystic EBs. To obtain the AFP-expressing cells of the outgrowth region, we manually isolated by using micro-dissection and cultured them. These cells strongly express AFP over 70% of isolated cells post re-plating. Here, we first showed an expression pattern of specifically localized ELCs by Activin-A during differentiation of hESCs. From this observation, we could highly purified ELCs from undifferentiated hESCs. Taken together, our system will provide a novel and efficient option to generate ELCs from hESCs.

• BMB Reports
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• 제39권3호
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• pp.297-303
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• 2006

As lipolytic enzymes, GDSL lipases play an important role in plant growth and development. In order to identify their functions and roles, the full-length cDNA of a GDSL lipase gene, designated BnLIP2, was isolated from Brassica napus L. BnLIP2 was 1,300 bp long, with 1,122 bp open reading frame (ORF) encoding 373 amino acid residues. Sequence analysis indicated that BnLIP2 belonged to GDSL family. Southern blot analysis indicated that BnLIP2 belonged to a small gene family in rapeseed genome. RT-PCR analysis revealed that BnLIP2 was a tissue-specific expressing gene during reproductive growth and strongly expressed during seed germination. BnLIP2 expression could not be detected until three days after germination, and it subsequently became stronger. The transcript of this gene was deficient in root of seedlings growing at different stages. When juvenile seedlings were treated by methyl jasmonate (MeJ), salicylic acid (SA) and naphthalene acetic acid (NAA), BnLIP2 expression could not be induced in root. Our study implicates that BnLIP2 probably plays an important role in rapeseed germination, morphogenesis, flowering, but independent of root growth and development.

• Reproductive and Developmental Biology
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• 제34권2호
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• pp.89-94
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• 2010

Melatonin is induced by light information through the retina and leads to growth factor activation. Thus, we investigated the effects of melatonin by controlling the photoperiod of growing young rats. Male Sprague-Dawley rats (n=6; 4 weeks old) were divided into two experimental groups: the L/D group (normal photoperiod; light/dark: 12/12 h; lights on at 9:00 a.m.) and the L/L group (light/light: 24 h). Rat body weight and food consumption were measured daily for 8 weeks. After 8 weeks, the rats were anesthetized with a mixture of ketamine (50 mg/kg) and xylazine (10 mg/kg) and sacrificed. Tissue was then collected for RNA isolation (from brain, heart, liver, kidney, adrenal gland, testis, tibia, hind limb muscles). Also, serum was isolated from blood using a centrifugal separation. The L/L group had significantly lower body weight than the L/D group from 4 to 6 weeks (p<0.05). The L/D group had increased tissue mass, compared with the L/L group, but the difference was not statistically significant. The L/D group had a significantly higher melatonin concentration than the L/L group between the hours of midnight and 2:00 a.m (p<0.01). These results indicate that photoperiod length may affect the secretion of melatonin from the pineal gland. Also, the reduction of nocturnal melatonin secretion may retard the development of growing young rats. In future studies, we plan to compare exogenous melatonin administration with endogenous melatonin concentration induced by photoperiod control. Moreover, we will confirm whether the effects seen in pathological animal models can be reversed by controlling the photoperiod.

• 유기물자원화
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• 제29권3호
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• pp.27-33
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• 2021

CO I 유전자 염기서열을 분석한 결과 경북농약연구소에서 독성실험에 이용되고 있는 경주집단 Eisenia속 지렁이의 '분자생물학적 종'은 줄지렁이(E. fetida)로 동정되었고 유기성 폐기물 재활용에 활용되고 있는 영동집단 Eisenia속 지렁이의 '분자생물학적 종'은 붉은줄지렁이(E. andrei)로 동정되었다. 이들 두 분자생물학적 종의 유기성 폐기물에 대한 생식반응을 비교한 결과 줄지렁이 집단의 산란률과 차세대 성충으로의 성장률이 붉은줄지렁이보다 다소 높게 나타나거나 또는 두 집단 간 유의미한 차이가 없는 것으로 나타났다. 두 집단 간 교잡 후 생산되는 산란수나 차세대 성충 생산수는 같은 집단내 개체들로부터 생산된 것들보다 현저하게 낮은 것으로 나타났다. 따라서 줄지렁이 집단과 붉은줄지렁이 집단 간에 완전한 생식적 격리는 이루어지지 않았지만 이 두 집단 간에 생물학적 종분화가 진행되고 있는 것으로 판단된다.

• Clinical and Experimental Reproductive Medicine
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• 제27권3호
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• pp.235-243
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• 2000

Objective: Our present studies were conducted to examine more effective isolating method of preantral follicles from mouse ovaries. Methods: ICR mice (3-6 weeks old) were sacrificed through cervical dislocation and their ovaries were removed and put into watch glasses containing Hams F-10 supplemented with 10% fetal bovine serum (FBS). Preantral follicles were isolated by three different methods; 1) enzymatical method and 2) mincing method, and 3) scraping method. Enzymatical method was carried out as following. Ovaries were bisected with a pair of fine 30G needles. Bisected ovaries were incubated at $37^{\circ}C$ and 5% $CO_2$ incubator in 2-well dish containing Hams F-10 supplemented with collagenase 600 lU/ml and DNAse 20 lU/ml. After 20 min., follicles were isolated by repeated pipetting. Isolated preantral follicles were collected, and the remnant of tissues was placed in incubator and previous procedure was repeated. Mincing method was carried out with a pair of fine 30G needles attached to 1 ml syringes and minced ovary. Scraping method was carried out with a pair of fine 30G needles and scratched to surface of ovary. The differences between isolating methods were analyzed using Student's t-test and Chi-square. Results were considered statistically significant when ${\rho}$ value was less than 0.05. Results: In handling time, mincing or scraping method ($28{\pm}3.42$ min or $16{\pm}1.58$ min) were significantly (p<0.00001) shorter than enzymatical method ($72{\pm}1.69$ min), and scraping method was significantly (p<0.01) shorter than mincing method. Total number of isolated follicles was significantly (p<0.0001) higher in enzymatical method ($49.8{\pm}3.91$) than in mincing or scraping method ($25.3{\pm}2.33$ or $20.5{\pm}1.75$). Isolated follicles in ${\leq}$90${\mu}m$ were significantly (p<0.005) higher in enzymatical method ($15{\pm}1.71$) than in mincing or scraping method ($7.8{\pm}0.98$ or $8.1{\pm}1.31$). In 91~130 ${\mu}m$, isolated follicles were significantly (p<0.0005) higher in enzymatical method ($33{\pm}3.27$) than in mincing or scraping method ($16.3{\pm}1.82$ or $10.7{\pm}1.38$). In ${\geq}$ 131 ${\mu}m$, isolated follicles were not significantly differences between all groups. In equal sizes, the rate of isolated follicles in ${\leq}$ 90 ${\mu}m$ was highest in scraping method (39.6% vs. enzymatical method: 30.1%, p<0.05; mincing method: 30.9%, p=0.11719, NS). Rate of follicles in $91{\sim}130$ ${\mu}m$ was significantly (p<0.05) lower in scraping method (52.7%) than in enzymatical or mincing method (66.3% or 64.5%). Rate of follicles in ${\geq}$131 ${\mu}m$ was highest in scraping method (8.3% vs. enzymatical or scraping method: 3.6%, p<0.05 or 4.6%, p=0.19053, NS). Conclusions: This study suggests that scraping method is simple and useful for isolation of preantral follicles, because this method reduced handling time and recovered enough follicles. The recovered rate of isolated follicles in diameter of 91 ~ 130 ${\mu}m$ was highest in all methods.

• Clinical and Experimental Reproductive Medicine
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• 제11권1호
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• pp.17-32
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• 1984

자궁내장치 사용군에 있어서 비사용군에 대한 골반장기염의 상대위험도는 많은 논란의 대상이 되고있으며 정확한 골반장기염의 진단을 위하여는 복강경을 통한 골반장기의 관찰 및 그 원인균을 규명하기 위한 병소인 난관으로부터의 균주 채취가 필요하다고 하였다. 이에 저자는 1982년 11월 1일부터 1985년 5월 31일까지 만 6개월간 연세대학교 의과대학 부속 세브란스병원 수태조절실 및 서울시내 성가병원에 복강경을 통한 영구불임술을 시술받기 위하여 내원한 여성중 내원 1개월이내에 항생제나 질정의 사용이 전혀 없는 자궁내장치 사용군 30명과 자궁내장치 비사용군 35명을 대상으로 복강경을 통한 골반장기의 관찰 및 난관에서의 가검물 채취와 동시에 자궁경관에서도 세균검사를 병행하여 균주배양을 시행하므로써 골반장기염의 상대위험도를 연구하고 그 관련 균주를 규명하여 아래와 같은 결론을 얻었다. 1. 자궁내장치 사용군과 비사용군의 평균 연령은 각각 31.8세, 30.7세이었고 평균 임신의 횟수는 양군이 3.4회이며 경제적 상태도 양군의 모든 예에서 중류이하로 양군의 차이가 없었다. 2. 복강경을 통한 골반장기염의 소견을 보인 예는 자궁내장치 사용군에서 2예로 6.6%였으며 비사용군에서는 골반장기염의 소견을 볼 수 없었으나 이들사이의 통계학적인 차이는 없었으며(p>0.005) 자궁내장치 사용군의 적혈구 침강속 는 30%에서 16mm/hr 이상으로 증가를 보였으나 백혈구수의 증가는 3.3%에서 있었다. 3. 복강경을 통한 난관에서의 가검물채취로 균주를 배양한 결과 자궁내장치 사용군 30예와 비사용군 35예의 전예에서 배양된 균주는 없었다. 4. 자궁경관의 균주 배양결과는 자궁내장치 사용군과 비사용군에 있어 골반장기염의 원인균이ㅍ될 수 있는 Hemophilus sp. 가 각각 3예, 1예, alpha-Streptococcus 2예, 1예, Corynebacteria 1예, 0예, Bacteroides 는 4예, 1예로 사용군에서 더욱 자주 배양되는 통계학적 차이는 없었다.(p>0.005).