The objectives of this study were to identify QTLs for agronomic traits using introgression lines from a cross between a japonica weedy rice and a Tongil-type rice. A total of 75 introgression lines developed in the Tongil-type rice were characterized. A total of 368 introgressed segments including 285 homozygous and 83 heterozygous loci were detected on 12 chromosomes based on the genotypes of 136 SSR markers. Each of 75 introgression lines contained 0-9 homozygous and 0-8 heterozygous introgressed segments with an average of 5.8 segments per line. A total of 31 quantitative and 2 qualitative loci were identified for 14 agronomic traits and each QTL explained 4.1% to 76.6% of the phenotypic variance. Some QTLs were clustered in a few chromosomal regions. A first cluster was located near RM315 and RM472 on chromosome 1 with QTLs for 1,000 grain weight, culm length, grain width and thickness. Another cluster was detected with four QTLs for 1,000 grain weight, grain length, grain width and grain length/width ratio near the SSR marker RM249 on chromosome 5. Among the 31 QTLs, 9 (28.1%) Hapcheonaengmi3 alleles were beneficial in the Milyang23 background. ILs would be useful to confirm QTLs putatively detected in a primary mapping population for complex traits and serve as a starting point for map-based cloning of the QTLs. Additional backcrosses are being made to purify nearly isogenic lines (NILs) harboring a few favorable Hapcheonaengmi3 alleles in Milyang23 background.
Purple acid phosphatase is important for phosphorus remobilization in plants, but its role in plant adaptation to low phosphorus availability is not known. The cDNA encoding O. sativa purple acid phosphatase (OsPAP1) has 1008 bp with an open reading frame of 335 amino acid residues. The amino acid sequence of OsPAP1 cDNA shows of 50-51% identity with other plant purple acid phosphatases. OsPAP1 was expressed in rice plants and in cell cultures in the absence of phosphate (Pi). The expression was organ-specific with the strongest expression in Pi-deprived roots. Functional expression of the OsPAP1 gene in the transgenic Arabidopsis line was confirmed by northern and western blot analysis. OsPAP1 overexpression lines had higher phosphatase activity than wild-type. Overexpression of OsPAP1 in Arabidopsis plants resulted in increased Pi accumulation under Pi sufficient condition. These results show that the OsPAP1 gene represents more efficient Pi uptake and can be used to develop new transgenic dicotyledonous plants.
Riptortus clavatus, one of the many insects in major crops, damages pods and seeds, which reduces seed vigor and viability in soybeans. This study was conducted to examine the effect of diversely damaged seeds by R. clavatus on seed germination and seedling emergence and to determine the association of damaged seed with quality and yield of soybean sprouts. All seeds damaged by R. clavatus significantly (P<0.05) reduced seed vigor as measured by the rates of seed germination, germination speed, and seedling emergence. Mean seed germination rate of non-damaged seeds in sprout-soybean varieties was 97.8%, whereas the rates of seeds damaged at different levels, 31-50% and 51-80%, were 23.0 and 5.4%, respectively. The rates of seedling rot and abnormal, incomplete germination significantly (P<0.05) increased as the amount of seeds damaged by R. clavatus increased to 5, 10 and 15% against the total seeds for sprout production. Yield of soybean sprouts from seeds damaged at different levels decreased up to 13% as compared to that in normal seeds. In customer preferences on soybean sprout produce, 84% of customers participated in survey preferred to purchase sprouts from seeds with 5% of damaged seeds, but sprouts produced from seeds with 15% of damaged seeds were intended to purchase only by 22% of the customers. Areas of the seed damaged by R. clavatus were readily infected by pathogens as the seed germinated, resulted in deteriorated quality and reduced yield of sprout produce.
English is the accepted common working language of the maritime world and being competent in its use is essential to the safety of ships, their crews and the marine environment. This paper is a response to the urgent need to find a suitable solution to the problem of providing maritime students with quality instruction in Maritime English. This paper will show what type of English instructor is best suited to help cadets have at least a basic grasp of Maritime English communication, with a view to possessing the level required by STCW 95 within the shortest time. It presents ways that maritime institutes can develop their own qualified or 'marinated' English Instructors and what qualifications should be required. It is concluded that by further essential research, interviews and questionnaires etc., the language needs of the university and shipping industry in Korea as a whole can be clearly verified. By examining such data, the present language education systems can be evaluated as to efficacy and relevance, allowing the establishment and implementation of 'best practice' within the training institute. This will result in making excellent informed decisions and choices about how best to improve the language competencies of graduating cadets, thereby creating the catalyst for the success of future seafarers whilst raising the image of the institute and Korean shipping worldwide.
A neurotoxin, 6-hydroxydopamine (6-OHDA) has been widely used to create animal model for Parkinson's disease (PD). The present study was undertaken to examine whether depletion of brain dopamine (DA) stores with 6-OHDA can make alteration in the activities of the testicular steroidogenesis in adult rats. Young adult male rats (3 months old) were received a single dose of 6-OHDA (200 in 10 /animal) by intracerebroventricular (icv) injection, and sacrificed after two weeks. The mRNA levels of steroidogenesis-related enzymes were measured by qRT-PCRs. Serum testosterone levels were measured by radioimmunoassay. Single icv infusion of 6-OHDA significantly decreased the mRNA levels of CYP11A1 (control:6-OHDA group= AU, p<0.05), CYP17 (control:6-OHDA group= AU, p<0.05). There were no changes in the mRNA levels of -HSD (control:6-OHDA group= AU) and -HSD (control: 6-OHDA group= AU), though the levels tended to be decreased in the 6-OHDA treated group. Administration of 6-OHDA decreased significantly the mRNA level of StAR when compared to the level of saline-injected control animals (control:6-OHDA group= AU, p<0.05). Treatment with single dose of 6-OHDA remarkably lowered serum testosterone levels compared to the levels of control group (control:6-OHDA group=, p<0.05). Taken together with our previous study, the present study demonstrated that the activities of hypothalamus-pituitary-testis hormonal axis could be negatively affected by blockade of brain DA biosynthesis, and suggested the reduced reproductive potential might be resulted in the animals. More precise information on the testicular steroidogenic activities in PD patients and PD-like animals should be required prior to the generalization of the sex steroid hormone therapy to meet the highest standards for safety and efficacy.
The canine major histocompatibility complex (MHC) is referred to dog leukocyte antigens (DLA), which is known to be the most polymorphic genetic system in canine species. Many cloned dogs have been produced since Snuppy, first cloned dog, there was no research about genetic identity of MHC among cloned animals. Recently in Lee’s group, two non-transgenic cloned beagles (BG1, 2) were produced by somatic cell nuclear transfer (SCNT) using fetal fibroblast (BF). Also, four transgenic cloned beagles (Ruppy 1-3, 5) were generated using transgenic BF transfected with Red fluorescent protein (RFP) gene. We hypothesize that non-transgenic (BG1, 2) and transgenic (Ruppy 1-3, 5) cloned beagles derived from identical donor cells have the same immunological genetic characteristic except for RFP gene insertion in the genome. Thus, the aim of this study is to confirm the immunological identity of DLA class II in cloned beagles produced using same nuclear donor cell. Genomic DNA was extracted from blood of BG1, BG2, Ruppy 1, 2, 3 and 5. Genomic DNA of normal two control beagle, no correlation with BF was also investigated for rulling out the possibility that beagles were inbred. Forward and reverse primers used for DLA-DQA1 and DQB1 respectively were DQAF: 5’-TAAGGTTCTTTTCTCCCTCT-3’ and DQAR: 5’-GGACAGATTCAGTGAAGAGA-3’ DQBR:5’-CTCACTGGCCCGGCTGTCTC-3’ and DQBR: 5’-CACCTCGC CGCTGCAACGTG-3’. Polymerase Chain Reaction (PCR) products were purified, sequenced directly using the Big Dye Terminator kit. Sequencing analysis was performed on an automated 3730xl DNA analyzer. In experiment 1, sequence of DLA-DQ alpha 1 (DQA1) and DLA-DQ beta 1 (DQB1) exon 2, hypervariabel region, was compared in BG1 and BG2. Experiment 2 also compared the sequence of DQA1 and DQB1 among Ruppy 1, 2, 3 and 5. Experimental 3 compared sequence of DQA1 and DQB1 among all cloned dogs (BG1, BG2 and Ruppy 1-3, 5). As a result, BG1 and BG2 have same allele for DQA1 and DQB1 as we expected. They share DQA1*00101 and DQB1*02901 in experiment 1. In experiment 2, Ruppy 1, 2, 3 and 5 also have identical DQA1*00101 and DQB1*02901 allele. No discrimination between transgenic dogs and cloned dogs was seen in DQA1 and DQB1 Allele in experiment 3. DQA1, DQB1 allele was identified as *00101 and *02901 in all dogs. We provided the allele identity of DQA1and DQB1 in cloned beagles, which can be used as preliminary data for immunological related studies. In conclusion, transgenic cloned dogs despite of red fluorescent protein genes being inserted in their nuclear DNA were immunologically compatible with non-transgenic cloned dogs. We demonstrated that cloned beagles produced using identical nuclear donor were immunologically compatible.
Mechanisms that regulate the number of cells that constitute the body have remained largely elusive. We approached this issue in the ascidian, Halocynthia roretzi, which develops into tadpole larva with small number of cells. Embryonic cells divide 11 times on average from fertilization to hatching. The number of cell division rounds varies between tissue types. For example, notochord cells divide 9 times and give rise to large postmitotic cells in the tadpole. The number of cell division rounds in the partial embryos that were derived from tissue-precursor blastomeres isolated at the 64-cell stage also varied between tissues, and coincided with their counterparts in the intact whole embryos to some extent, suggesting tissueautonomous regulation of cell division. Manipulation of cell fates in notochord, nerve cord, muscle, and mesenchyme lineage cells by inhibition or ectopic activation of the inductive FGF signal changed the number of cell division according to the altered fate. Knockdown and missexpression of Brachyury (Bra), an FGF-induced notochord-specific key transcription factor for notochord differentiation, indicated that Bra is responsible not only for notochord differentiation but also regulates the number of cell division rounds in the notochord lineage cells, suggesting that Bra activates a putative machinery to stop cell division at the specific stage. Results of precocious expression of Bra suggested that the machinery refers the developmental clock that is likely shared in other blastomeres than notochord, and functions to terminate cell division at three rounds after the 64-cell stage. Bra does nothing about the progression of developmental clock itself.
The transcription factors, DMRT1 and FOXL2, play a role in fish sex differentiation of the bipotential precursor into the male and female pathway, respectively. In order to provide the molecular background for understanding hormonal regulation in sexual determination and differentiation in the red-spotted grouper, Epinephelus akaara, one of commercially important epinephelines, and is often used to study protogynous sex change. First, we amplified the partial sequence of two genes (DMRT1 and FOXL2) from the gonad of red-spotted grouper. Also, we surveyed the tissue-specific and sex-specific expression pattern of each genes by RT-PCR. The effects of 17α-methyltestosterone (MT) in the sexually immature gonad of red-spotted grouper were investigated by Real-time quantitative RT-PCR. Fish were treated with MT-dipping method from around 70 days after hatching (DAH) for two month. DMRT1 and FOXL2 cDNA flagments consist of 489 and 836 base pairs (bp) and encodes a protein of 162 and 278 amino acids, respectively. RT-PCR revealed that DMRT1 mRNA was expressed higher level in the testis. Foxl2 was expressed extensively in the neural and peripheral tissues with its highest level in the ovary, indicating a potential role for Foxl2 in the brain-pituitary-gonad axis. Real-time quantitative RT-PCR analyses showed that DMRT1 mRNA expression was upregulated in the MT-treated fish. These results suggest that the sex inversion of red-spotted grouper by MT might be due to the suppression of FOXL2 gene expression, and resulting in the induction of the 11-KT secretion.
Controllable transgenic expression systems in transgenic animal model are valuable to the development of therapeutic approaches in human medical fields. The aim of this study was to 1) produce a transgenic cloned dog using inducible tetracycline vector system, and 2) investigate whether the transgenic cloned dog could be induced the transgene expression using doxycycline (Doxy). Canine fetal fibroblasts were infected with retroviral vectors designed to express the enhanced green fluorescent protein (eGFP) gene under the control of tetracycline-inducible promoter. For somatic cell nuclear transfer (SCNT), nucleus of an in vivo matured oocyte was removed and an eGFP expressed cell cultured with 1 ㎍/㎖ of Doxy was injected. After electrical fusion and chemical activation, the reconstructed embryos were transferred to a recipient and pregnancy diagnosis was performed by ultrasonography. Experiment I evaluated the mean fluorescence intensity (MFI) of infected cells while the cells were cultured in the presence of 1 ㎍/㎖ of Doxy for 5 days, and then in the absence of Doxy for 7 days using fluorescence-activated cell sorter. Experiment II was designed to produce an eGFP controllable transgenic cloned dog via SCNT. For verification of transgenic dog, experiment III was performed Southern Blot analysis and observation in vivo regulation of eGFP expression in the cloned dog treated with 100 ㎎/㎏ of Doxy every 2 days for 2 weeks under ultraviolet light. In experiment IV, western blot was used to detect eGFP increase and decrease in skin tissues of transgenic dog under the presence or absence of Doxy. In the results of experiment I, the MFI for infected cells was rapidly increased to approximately 42.3 times after 3 day-treatment compared to pre-treatment and quickly decreased 3 days after ceasing the treatment. In experiment II, a total of 203 embryos were transferred to nine recipients and three pregnant delivered three pups (Tet-on eGFP 0, Tet-on eGFP 1, and Tet-on eGFP 2) by C-sec and Tet-on eGFP 2 among them is still alive. All cloned pups were genetically identical to the donor cell. Tet-on eGFP 2 showed an apparent in vivo eGFP expression on her body after Doxy administration in experiment III. The result of Sothern blotting showed that the transgene insertion was detected from the three cloned dogs and all organs of Tet-on eGFP 1. Experiment IV indicated that a robust eGFP expression in skin tissue of Tet-on eGFP 2 rapidly increased after Doxy treatment and gradually decreased to basal level on 9 weeks after ceasing the treatment. In conclusion, we report here for the first time an inducible transgenic system in canine species and it can stably induce the transgene expression at intended time. This study has demonstrated the capacity to generate transgenic model dog which could regulate the transgene and it would contribute to human medical research fields.
The objective of this study was to investigate the effects of oxygen tension during in vitro maturation of porcine oocytes on the nuclear maturation and differences in gene expression. Cumulus-oocyte complexes (COCs) were collected from ovaries obtained at a local slaughterhouse, matured for 44 hours in TCM199 supplemented with porcine follicular fluid (pFF) under 5% or 20% oxygen concentration. In results, oxygen tension had no significant effects on nuclear maturation. Relative poly(A) mRNA abundance of MnSOD, CCNB1, LDHA, G6PD, BCL, GPX1, IGFR2, GLUT1, BAX, GREM, PTGS2 was analysed in cumulus cells. GLUT1, G6PD, LDHA were up-regulated in the cumulus cells matured in low oxygen, suggesting a higher glucose uptake and an increase in anaerobic glycolysis, whereas CCNB, MnSOD were up-regulated in the cumulus cells matured in high oxygen, which suggest a higher activity of mitosis-promoting factor and antioxidant response. In conclusion, cumulus cells increase in glucose metabolism via anaerobic glycolysis under low oxygen concentration and show significant change in antioxidant against oxidant damage or apoptotic response under high oxygen concentration. For such an effect of cumulus cells, oocytes could be matured normally regardless of various oxygen concentration.
Obesity is often associated with an impairment of the hypothalamic-pituitary-gonadal axis and also unites with reproductive defects and reduced fertility. The leptin-deficient ob/ob mouse is characterized by a morbid obesity and infertility. In the present study, we examined when ob/ob mouse lost their fertility activity according to aged dependent were investigated. The major organ systems in the body, the ovaries of females are the first to exhibit impaired function with advancing age. Here, ob/ob and ob lean various (4, 6, 9, 12 and 24 wks) aged mice were used. Histology (H & E staining) of ovary, vaginal smear and Reverse transcription polymerase chain reaction (RT-PCR) analysis were carried out. Hematoxylin/eosinstained sections show normal histology in ob lean type mice characterized by the presence of multiple primary and secondary follicles and well developed corpora lutea (CL). Ovaries from ob/ob mice, showed 4 and 6 wks mice similar to ob lean type but according to aged dependent ob/ob mice few primary and secondary follicles was seen. Many large empty follicles were present and no proper developed CL, especially 24 wks mice. Vaginal smear showed abnormal estrous cycle in ob/ob mice. Ovaries of ob/ob mice, mRNA results showed that ovulated gene TIMP1, Pgsh2 and Lhcgr expression levels are decreased according to aged dependent compared to ob lean type mice. These data suggest that aged dependent specifically when 24 wks of ob/ob mice lost their reproductive function.
핀란드 Okiluoto 섬 중앙부에 방사성폐기물 처분장으로 예정된 부지에서 블록 규모 지하수 유동 모의를 수행하였다. 현장에 설치된 심부 관측공에서 관찰된 단열대에 관한 자료를 이용하여 단열망을 구성하였다. 이 단열망을 이용하여 3차원 유한 요소 격자망의 수리전도도장를 생성하고, 이를 지하수 유동 모의에 이용하였다. 현장에서 이루어진 양수시험 전과 후에 심부 관측공에서 측정된 수위와 구간별 유입, 유출량을 이용하여 시추공과 교차하는 단열대의 투수량계수와 부지의 함양률을 조절하며 지하수 유동 모형을 보정하였다. 양수 시험 전과 후를 순차적으로 보정해가며 모의한 결과, 보정된 지하수 유동 모형으로 계산한 지하수위는 관측 자료와 비교적 일치하지만, 관측공에서의 지하수 유입, 유출량은 상당한 차이를 보이는 구간도 있는 것으로 확인되었다. 이런 불일치는 지하수 유동로가 될 수 있는 구조가 지하수 유동 모의를 위한 개념모형에 충분히 반영되지 않아 생기는 것으로 생각되며, 이에 배경단열과 같은 국지적인 유동로 구조가 개념모형에 반영되어야 할 것으로 판단되었다.
In the 1910s and 1920s, T. S. Eliot had frequently made interesting remarks on politics and commented on political issues through literary journalism. In his writings, he expressed a strong dislike for liberal democracy, a main current of British as well as universal political movements then. After taking up the editorship of The Criterion in 1922, Eliot advanced his opinions persistently against liberal democracy on political and literary issues through the quarterly review.This paper examines Eliot’s political ideas reflected in his various writings in The Criterion right after his conversion to the National Church of England in 1927. This examination finds that the political ideas of Eliot and other contributors in the review have close affinities with those expounded by the French political thinker, Charles Maurras, who was a champion of French monarchism, Classicism, Catholicism, and who condemned liberal democracy after the French Revolution. Although Eliot retained some of his family politics, much of his reactionary ideas against liberal democracy germinates from the thoughts of Maurras. For Eliot, liberal democracy in politics was not conducive to the excellence in the of arts as well as modern culture. However, after the condemnation of Maurras by the Vatican, he turns to Christian politics, which must be in service of the Church.
강섬유를 혼입한 콘크리트(Steel Fiber Reinforced Concrete, SFRC) 보는 강섬유의 우수한 인장강도로 인하여 일반 철근콘크리트 보에 비하여 높은 전단강도를 가진다. 이 연구에서는 강섬유 혼입율에 따른 SFRC 보의 전단거동을 규명하기 위하여 실험을 수행하였으며, 특히, 압축영역에서의 비균열 콘크리트 단면의 전단저항 분담율을 분석하였다. 또한, 이 연구의 실험결과 및 기존에 보고된 87개의 실험 데이터를 수집하여 SFRC보의 전단예측식들에 대한 정확도를 평가하였다. 강섬유의 혼입율이 증가할수록 전단강도는 증가하는 경향성을 나타내었다. 그러나, 강섬유 혼입율이 0.5%인 실험체는 사인장 균열 이후 갑작스럽게 파괴되었고, 강섬유의 혼입율이 2.0%인 실험체에서는 전단보강효율이 감소하는 것으로 관찰되어 최대 전단보강효율을 가질 수 있는 혼입율은 1~2% 사이에 있을 것으로 추정된다. 또한, 압축영역에서의 비균열 콘크리트 단면의 전단저항 분담율은 약 21% 이상으로 관찰되었으며, 이 연구에서 평가된 SFRC보의 전단강도에 대한 기존 제안식 중에서 오영훈 등이 제안한 식이 비교적 정확하게 전단강도를 예측하였다.