Previously, we demonstrated that the shift and/or restriction of feeding time during relatively short-term period (4 weeks) could alter the pituitary gonadotropin expression and the weights of seminal vesicle and prostate in rats. We also found that the reverse feeding (RF) schedule (up to 8 weeks) might induce an adaptable metabolic stress and cause impairment of androgen-dependent reproductive tissues. In the present study, we extended the RF time regimen up to 12 weeks, and measured the reproductive tissue weights. After 4 and 8 weeks of RF, the weights of epididymis were not significantly different. After 12 weeks, however, epididymis weights of RF animals were significantly different (CON 12W : RF 12W = 48.26±0.62mg : 44.05±1.57mg, p＜0.05). After 4 and 12 weeks of feeding, seminal vesicle weights of RF animals were significantly decreased (CON 4W : RF 4W = 79.36±8.34mg : 46.28±2.43mg, p＜0.001; CON 12W : RF 12W = 72.04±3.76mg : 46.71±2.27mg, p＜0.001, respectively). Prostate weights were not changed by RF. Kidney and spleen weights of RF animals were significantly different on weeks 4 and 12 (Kidney, CON 4W : RF 4W = 249.72±4.20mg : 228.41±3.03mg, p＜0.001; CON 12W : RF 12W = 309.15±7.49mg : 250.72±6.13mg, p＜0.001, respectively, Spleen, CON 4W : RF 4W = 111.26±3.76mg : 96.88±4.69mg, p＜0.05; CON 12W : RF 12W = 123.93±10.72mg : 94.68±5.65mg, p＜0.05, respectively). Histology analysis of seminal vesicle revealed that the thinner epithelial cell layers, reduced complexities of swollen papilla folding in the exocrine glands on weeks 4 and 12 of RF. There was no histological difference between control and RF group on week 8. The present study indicates that up to 12 weeks RF induced differential changes in tissue weights of male mice. In particular, seminal vesicle, kidney and spleen seemed to temporarily adapted to the RF-induced metabolic stress on week 8 of feeding schedule. These results confirmed the our previous study that the RF might induce an adaptable metabolic stress and cause impairment of androgen-dependent reproductive tissues such as epididymis and seminal vesicle as well as non-reproductive tissues such as kidney and spleen. Further studies will be needed to achieve a better understanding of the how does mealtime shift affect the reproductive function and exact nature of adaptation.

Ovarian tissue cryopreservation and transplantation causes follicle depletion. To overcome this problem, we investigate the effect of Anti-Müllerian hormone (AMH), a follicle recruitment control hormone, supplementation before and/or after mouse ovarian transplantation. A total of 120 5-week-aged BD F-1 female mice were used. The mice were randomly divided into four groups according to AMH doses (0, 5, 25, 125 μg/mL, respectively). AMH was injected intraperitoneally on every other day for a week before, after, or before and after transplantation of ovaries under kidney capsules was performed. One week after transplantation, follicular normality was evaluated by histological analysis and TUNEL assay. In Group A and C, morphologically intact follicle (G1) ratios of AMH treated groups showed no statistically significant difference. In Group B, G1 ratios of 25 and 125 μg/mL of AMH treated groups were higher than those of 5 μg/mL treated group, but there was no improvement in G1 ratio after AMH treatment. In every group, apoptotic follicle ratios did not show any trend according to AMH treatment. Proportions of primordial follicle were not significantly different according to AMH treatment in all groups. The result of the present study demonstrated that AMH treatment during on transplantation of cryopreserved ovaries has no significant effect on follicle survival and prevention of follicle depletion.

The influence of triploidization on histological characteristics of retina, trunk kidney, liver and midgut tissue, and comparative analysis of cell cycle and expression of cell cycle protein from tail fin and gill tissue in Far Eastern catfish, Silurus asotus were analyzed. In the infertile triploid fish, the nucleus and/or cell size of secondary proximal tubule cells of trunk kidney, hepatocyte and midgut epithelium are much larger than those of the corresponding cells in the diploid fish (P<0.05). However, triploid tissue showed fewer number of outer nuclear layer in retina and nuclei in secondary proximal tubule of trunk kidney than those for diploid tissue. The mean percentages of the G1-, the S- and the G2+M-phase fractions were 92.5%, 3.2% and 4.3% in tail fin tissue of diploid, and 93.4%, 2.6% and 4.0% in those of triploid, respectively. There were no significant differences in the percentages of each cell cycle fraction between diploid and triploid. The differences of cell cycle between tail fin tissue and gill tissue were statistically significant in diploid and triploid (P<0.05). Also, the differences between diploid and triploid were statistically significant in tail fin tissue and gill tissue (P<0.05). Results from this study suggest that some characteristics in the triploid exhibiting larger cell and nucleus size with fewer number of cell than diploid can be used as an indicator in the identification of triploidization and ploidy level in Far Eastern catfish.

This study was performed to investigate the expression of cathepsin B mRNA and protein in eutopic and ectopic endometrial tissues of patients with endometriosis and in normal endometrial tissues and to clarify the association between the cathepsin B expression and endometriosis. A total of 40 women with histologically confirmed endometriosis were recruited for study group. For controls, 20 women undergoing operative treatment for uterine myoma, cervical intraepithelial neoplasia (CIN) or benign gynecologic conditions other than endometriosis were recruited. Eutopic endometrial tissues of both groups and ectopic endometrial tissue of study group were collected during the operations. We employed real time reverse transcriptase - polymerase chain reaction (RT-PCR) to quantify mRNA levels of cathepsin B in these tissues. Then, we performed western blot analysis to measure the protein levels of cathepsin B. The expressions of cathepsin B mRNA and protein were significantly higher in both eutopic and ectopic endometrial tissues of women with endometriosis than in endometrial tissues of controls. These data suggest that the higher expression of cathepsin B in the endometrial tissues might be associated with the development of endometriosis. In addition, eutopic endometrium itself with higher expression cathepsin B may play a pivotal role in the histogenesis of endometriosis.

Notch signaling plays fundamental roles in various animal development. It has been suggested that Hr-Notch, a Notch homologue in the ascidian Halocynthia roretzi, is involved in the formation of peripheral neurons by suppressing the neural fates and promoting the epidermal differentiation. However, roles of Notch signaling remain controversial in the formation of nervous system in ascidian embryos. To precisely investigate functions of Notch signaling, we have isolated and characterized Hr-Numb, a Numb homologue which is a negative regulator of Notch signaling, in H. roretzi. Maternal expression of Hr-Numb mRNAs was detected in egg cytoplasm and the transcripts were inherited by the animal blastomeres. Its zygotic expression became evident by the early neurula stage and the transcripts were detected in dorsal neural precursor cells. Suppression of Hr-Numb function by an antisense morpholino oligonucleotide resulted in larvae with defect in brain vesicle and palps formation. Similar results have been obtained by overexpression of the constitutively activated Hr-Notch forms. Therefore, these results suggest that Hr-Numb is involved in Notch signaling during ascidian embryogenesis.

The management and the use of major mineral nutriments such as nitrogen, phosphorous, and potassium, etc have been practiced and improved in various cultivating methods of Panax ginseng C. A. Meyer. The purposes of this study were to examine the content of major mineral nutrients on different ginseng aging from 1 to 6 years old, to analyze their uptake and utilization in tissues of ginseng, and to find out their proper managing techniques throughout the cultivation of ginseng. In case of the leaves, the N content was not clearly different from 1 to 6 years old, while the content of P and K was generally decreased throughout the cultivating years. In case of the roots, the content of N and K was gradually decreased from 1 to 6 years old, while the P content was increased until 3 years old, decreased at 4 years old, increased again at 5 years old, and decreased again at 6 years old. The uptake amount of N was increased in root of ginseng from 1 to 6 years old, 0.02 to 2.79kg/10a based on dry weight, respectively. Other minerals of P, K, Ca, and Mg were increased for the cultivating year. Comparing the uptake amounts of N, P, K with different cultivating year, they were the highest uptake amount at 4 years old and then were decreased. The management techniques of major mineral in cultivation of ginseng would be studied and evaluated more in order to have better ginseng production.

The objective of this study was to determine the fatty acid composition of newly developed tissues of germinated soybean seeds. Five soybean accessions with varied fatty acid composition were allowed to germinate in sand under greenhouse conditions. Seedlings were picked up after 4, 6, 8, 10 and 12 days of germination and freeze dried. The fatty acid composition of the newly developed tissues was analyzed by gas chromatography. Significant variation in fatty acid composition was observed between accessions, days of germination, and variety × day of germination in whole and the cotyledons. In the case of newly developed five tissues, significant variation in fatty acid composition were observed between days of germination except oleic acid for root, hypocotyl and epicotyl stem and except stearic acid for hypocotyl and unifoliate leaves while all the parameters were significantly different for accession. Significant interactions of accession and days of germination were observed for palmitic, linoleic and linolenic acid in all tissues; only for oleic acid in hypocotyl, epicotyl and unifoliate leaves; and only for stearic acid in root, hypocotyl, epicotyl and unifoliate leaves. During germination, the fatty acid composition of newly developed tissues changed dramatically but whole seedlings and cotyledons changed slightly. These tissues contained five major fatty acids as found in original seeds, but compositions were totally different from that of the seed: higher in palmitic, stearic and linolenic acid and lower in oleic and linoleic acid. New tissues conserved their fatty acid compositions regardless of genotypic variation in the original seeds.

As in the O. mykiss electrophoretic profiles of RNA, the signals of each RNA sample from 9 individual tissues such as liver, muscle, brain, heart, pituitary gland, kidney, intestine, spleen and gill similar to positive control were obtained. The tissue distributions of the complimentary DNA (cDNA) of O. mykiss four genes were analyzed using quantitative real-time PCR with primer sets for tissue expression analysis. In this rainbow trout species, author obtained bands of various sizes, ranged from 700 bp to 1,400 bp. A dissociation curve was made at the end of each run to make sure that there was no non-specific amplification. Supplementarily, the Ct of each DNA was compared. The Ct values of vinculin with rainbow trout tissues were determined in a manner similar to those for agouti-related protein (AgRP) and melanocortin receptors (MC4R I and MC4R II). Further, obtained Cts for standard curve of each DNA were affected by specific product (vinculin, AgRP and MC4R II genes). After several experiments with four individual genes of rainbow trout, author estimated a variation ratio of the mean Ct value of the DNA extracted using the comparative CTt method was 37.27, and the standard deviation was 5.33. The correlation coefficient between the Ct values and the concentration of cDNA was -0.98, -0.99, -0.91 and -0.86, respectively (vinculin, AgRP, MC4R I and MC4R II genes). Since this correlation showed high linearity, the straight line obtained was used as a standard for the O. mykiss tissues reared in aquarium. A PCR efficiency of 100% is ideally achieved when the slopes are close to the theoretical value of -3.31. According to quantification method, the results of quantification are strongly affected by the DNA fragmentation. The size of most DNA fragments obtained from various tissues of rainbow trout used in the experiment was approximately 100 bp. According to the four slopes, an efficiency of nearly 100% was estimated for four genes detection methods. Additionally, further analysis with more individuals and primers will be required to fully establish optimization in rainbow trout.

손바닥선인장 약주의 pH 변화는 발효가 진행됨에 따라 감소하는 경향을 나타내었으나, 총산 함량은 증가 하는 경향을 보였다. 환원당 함량은 발효 2일에는 모든 시료구가 증가하였으나 발효 4일 경과후 감소하는 경향을 나타냈으며, ethanol은 발효 2일째부터 함량이 급속히 증가하여 발효 6일 부터는 완만한 증가를 보였고 발효 종료 시점인 14일에는 16.23~17.10%의 최대치를 나타냈다. 약주에서 확인된 유리당은 sucrose, maltose,

An efficient transformation method for soybean [Glycine max (L.) Merr.] using meristematic tissues of germinating seeds has been established. The embryonic axes were excised from germinating seeds of Korean soybean cultivar, Iksannamulkong and 0.5-2 cm long segment containing meristematic tissues were prepared by cutting hypocotyl region. The explants were inoculated with Agrobacterium tumefaciens strain LBA4404 harboring a binary vector with the bar gene as a selectable marker gene and a β-glucuronidase (GUSINT) reporter gene, and then co-cultured for 7 days on co-cultivation medium (CCM). The meristematic tissues were cultured on shoot induction medium (SIMP6) supplemented with 0.4 mg/l N6-benzylaminopurine (BAP) and 0.1 mg/l indolebutyric acid (IBA) in the presence of 6 mg/l L-phosphinotricin (PPT) for 2 weeks and the surviving explants were transferred to shoot elongation medium (SEMP6). Transformation was confirmed by Southern blot analysis and the transformation efficiencies ranged from 1.48 to 2.07%. The new modified transformation method was successfully implemented for obtaining several transgenic lines with SMV-CP gene. It is expected that this method could efficiently be used for the transformation of recalcitrant soybean cultivars.

방사광 X-선 현미경은 임상실험에 유용한 도구로 높은 배율과 고 해상도로 동물 장기조직 시료의 세부 구 조를 관찰할 수 있다. X-선 영상은 위상 대조도 메커니즘으로 설명할 수 있다. 우리는 쥐의 꼬리, 신경 및 허파 의 in-vivo 및 in-vitro위상 대조도 영상을 8 KeV mono 빔으로부터 10배 현미경대물렌즈와 CCD 카메라를 이용하여 얻었다. 기존의 흡수 X-선 영상 보다 SR 영상이 세밀한 구조의 높은 분해능 영상을 볼 수 있었다. SR 영상은 생물학, 재료 및 임상 연구에 무한한 가능성을 가지고 있다.

인삼포장에서 염류의 집적은 우량인삼의 생산에 많은 장애요인이 되고 있다. 본 연구는 인삼종자의 NaCl에 대한 반응정도를 조사하고, 또한 체세포배의 발생시 및 인삼모상근에서 사포닌의 생성에 미치는 반응을 조사하였다. 인삼 접합자 배를NaCl이 첨가되지 않은 배지에 접종한 결과 84.5%가 발아되었으며 0.1M에서는 3% 만이 발아되었고, 0.2M이상의 처리구에서는 전혀 발아가 되지 않았다. 또한 체세포배 발생 은 미숙배를 이용할 경우 NaCl 무첨가배지에서 가장 양호하였으며 농도가 증가할수록 감소하는 경향을 보였다. 반면에 성숙배에서 는 NaCl 무첨가배지보다 0.05M첨가배지에서 더 양호한 경향을 보였다. 뿌리 상태인 인삼의 모상근의 경우에는 광상태에서 NaCl를 첨가한 배지에 배양시 0.04M에서부터 안토시안이 형성되기 시작하여 0.08M에서는 눈으로 확인이 가능할 정도의 많은 량이 형성되었다. 안토시안을 함유한 세포주는 다른 세포주에 비해 crude saponin뿐만 아니라 total ginsenoside의 함량도 높은 경향을 보였으며 panaxatriol보다 panaxadiol이 2배 이상 많았다.

The effect of carbenicillin on the dedifferentiation and the regeneration efficiency of plant tissues of horseradish(Armoracia rusticana) was evaluated, Inhibition effect for callus initiation was observed when leaf blade, root and petiole segments were grown on MS medium containing 500 mg/L to 2000 mg/L carbenicillin and 0.5 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). The regeneration of horseradish shoots from leaf blade, root and petiole explants were decreased as the addition of carbenicillin increased from 1000 mg/L to 2000 mg/L in MS medium containing 0.5 mg/L of 6-benzylaminopurine (BAP) or kinetin. Especially, 500 mg/L carbenicillin treatment significantly inhibited shoot induction when leaf blade explants were grown on hormone-free MS medium. It was suggested that the toxic effects of combinations of carbenicillin and 2,4-D may be due to high auxin activity levels.

Two rice cultivars (Oryza sativa L.), Hwa-seongbyeo of Japonica type and Taebackbyeo of Indica/Japonica type, were cultivated with hydroponic culture to examine nitrogen effects on the growth responses, contents and utilizations of carbohydrates, and the ripening velocity of grains with three different N levels. Plant height and tiller number were clearly increased to 80 ppm N level compared to 40 ppm N level and then they were slightly decreased in N level of 120 ppm. Higher dry weights were appeared with 80 ppm N level than did with other N levels, showing statistically differences in both cultivars and N levels, while dry weight of roots was heavier with decreasing the N levels. Therefore, T/R ratios were not significantly different among N levels, although there was statistically differences between rice cultivars. After the flowering stage, higher water-soluble carbohydrate (WSC) and water-insoluble carbohydrate (WISC) were contained in stem compared with other parts, showing that WISC of sheath and stem, unlike WSC, was significantly different among N levels. Starch of grain, WISC, was remarkedly increased from 3.0% at just after the flowering to 52.0% and 75.0% at 15 and 30 day after the flowering, respectively, showing that lower N application had faster accumulation of starch in rice grains. N would affect the contents of carbohydrates of each tissue, and starch accumulation in rice grains.