• KOREAN JOURNAL OF CROP SCIENCE
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• v.17
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• pp.1-44
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• 1974

These studies were made at Suwon in 1972 and at Suwon, Iri, and Kwangju in 1973 to investigate grain filling process and variation of grain quality of NB 68513 and Caprock as hard red winter wheat, Suke ＃169 as soft red winter wheat variety and Yungkwang as semi-hard winter variety, grown under-three different fertilizer levels and seeding dates. Other experiments were conducted to find the effects of temperature, humidity and light intensity on the grain filling process and grain quality of Yungkwang and NB 68513 wheat varieties. These, experiments were conducted at Suwon in 1973 and 1974. 1. Grain filling process of wheat cultivars: 1) The frequency distribution of a grain weight shows that wider distribution of grain weight was associated with large grain groups rather than small grain group. In the large grain groups, the frequency was mostly concentrated near mean value, while the frequency was dispersed over the values in the small grain group. 2) The grain weight was more affected by the grain thickness and width than by grain length. 3) The grain weight during the ripening period was rapidly increased from 14 days after flowering to 35 days in Yungkwang and from 14 days after flowering to 28 days in NB 68513. The large grain group, Yungkwang was rather slowly increased and took a longer period in increase of endosperm ratio of grain than the small grain group, NB 68513. 4) In general, the 1, 000 grain weight was reduced under high temperature, low humidity, while it was increased under low temperature and high humidity condition, and under high temperature and humidity condition. The effect of shading on grain weight was greater in high temperature than in low temperature condition and no definite tendency was found in high humidity condition. 5) The effects of temperature, humidity and shading on 1, 000 grain weight were greater in large-grain group, Yungkwang than in small grain group, NB 68513. Highly significant positive correlation was found between 1, 000 grain weight and days to ripening. 6) The 1, 000 grain weight and test weight were increased more or less as the fertilizer levels applied were increased. However, the rate of increasing 1, 000 grain weight was low when fertilizer levels were increased from standard to double. The 1, 000 grain weight was high when planted early. Such tendency was greater in Suwon than in Kwangju or Iri area. 2. Milling quality: 7) The milling rate in a same group of varieties was higher under the condition of low temperature, high humidity and early maturing culture which were responsible for increasing 1, 000 grain weight. No definite relations were found along with locations. 8) In the varieties tested, the higher milling rate was found in large grain variety, Yungkwang, and the lowest milling rate was obtained from Suke ＃ 169, the small grain variety. But the small grained hard wheat variety such as Caprock and NB 68513 showed higher milling rate compared with the soft wheat variety, Suke ＃ 169. 9) There were no great differences of ash content due to location, fertilizer level and seeding date while remarkable differences due to variety were found. The ash content was high in the hard wheat varieties such as NB 68513, Caprock and low in soft wheat varieties such as Yungkwang and Suke ＃ 169. 3. Protein content: 10) The protein content was increased under the condition of high temperature, low humidity and shading, which were responsible for reduction of 1, 000 grain weight. The varietal differences of protein content due to high temperature, low humidity and shading conditions were greater in Yungkwang than in NB 68513. 11) The high content of protein in grain within one to two weeks after flowering might be due to the high ratio of pericarp and embryo to endosperm. As grains ripen, the effects of embryo and pericarp on protein content were decreased, reducing protein content. However, the protein content was getting increased from three or four weeks after flowering, and maximized at seven weeks after flowering. The protein content of grain at three to four weeks after flowering increased as the increase of 1, 000 grain weight. But the protein content of matured grain appeared to be affected by daily temperature on calender rather than by duration of ripening period. 12) Highly significant positive correlation value was found between the grain protein content and flour protein content. 13) The protein content was increased under the high level of fertilizers and late seeding. The local differences of protein content were greater in Suwon than in Kwangju and Iri. 14) Protein content in the varieties tested were high in Yungkwang, NB 68513 and Caprock, and low in Suke ＃ 169. However, variation in protein content due to the cultural methods was low in Suke ＃ 169. 15) Protein yield per unit area was increased in accordance with increase of fertilizer levels and early maturing culture. However, nitrogen fertilizer was utilized rather effectively in early maturing culture and Yungkwang was the highest in protein yield per unit area. 4. Physio-chemical properties of wheat flour: 16) Sedimentation value was higher under the conditions of high temperature, low humidity and high levels of fertilizers than under the conditions of low temperature, high moisture and low levels of fertilizers. Such differences of sedimentation values were more apparent in NB 68513 and Caprock than Yungkwang and Suke ＃ 169. The local difference of sedimentation value was greater in Suwon than in Kwangju and Iri. Even though the sedimentation value was highly correlated with protein content of grain, the high humidity was considered one of the factors affecting sedimentation value. 17) Changes of Pelshenke values due to the differences of cultural practices and locations were generally coincident with sedimentation values. 18) The mixing time required for mixogram was four to six minutes in NB 68513, five to seven minutes in Cap rock. The great variation of mixing time for Yungkwang and Suke ＃ 169 due to location and planting conditions was found. The mixing height and area were high in hard wheat than in soft wheat. Variation of protein content due to cultural methods were inconsistent. However, the pattern of mixogram were very much same regardless the treatments applied. With this regard, it could be concluded that the mixogram is a kind of method expressing the specific character of the variety. 19) Even though the milling property of NB 68513 and Caprock was deteriorated under either high temperature and low humidity of high fertilizer levels and late seeding conditions, baking quality was better due to improved physio-chemical properties of flour. In contrast, early maturing culture deteriorated physio-chemical properties, milling property of grain and grain protein yield per unit area was increased. However, it might be concluded that the hard wheat production of NB 68513 and Caprock for baking purpose could be done better in Suwon than in Iri or Kwangju area. 5. Interrelationships between the physio-chemical characters of wheat flour: 20) Physio-chemical properties of flour didn't have direct relationship with milling rate and ash content. Low grain weight produced high protein content and better physio-chemical flour properties. 21) In hard wheat varieties like NB 68513 and Caprock, protein content was significantly correlated with sedimentation value, Pelshenke value and mixing height. However, gluten strength and baking quality were improved by the increased protein content. In Yungkwang and Suk ＃ 169, protein content was correlated with sedimentation value, but no correlations were found with Pelshenke value and mixing height. Consequently, increase of protein content didn't improve the gluten strength in soft wheat. 22) The highly significant relationships between protein content and gluten strength and sedimentation . value, and between Pelshenke value, mixogram and gluten strength indicated that the determination of mixogram and Pelshenke value are useful for de terming soft and hard type of varieties. Determination of sedimentation value is considered useful method for quality evaluation of wheat grain under different cultural practices.

• Korean Journal of Animal Reproduction
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• v.19 no.3
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• pp.227-234
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• 1995

This study was carried out to investigate the effects according to the time course of glucose exposure on the development of one-cell embryos beyond morula in CR$_laa$ medium. One-cell zygotes from B6CBA F$_1$ mice were recovered at 24 ~ 25h after hCG and cumulus cells were removed with 0.1% hyaluronidase. The embryos were pooled and subsequently divided into each groups and cultured in CR$_laa$ at 37$^{\circ}C$ in 5% CO$_2$ in aIr. The embryos were either, placed in CR$_laa$ containing various concentration (5.5, 16.5, 27.5 and 38.5 mM) of glucose for 1 min. and subsequently returned to the fresh culture medium (without glucose), or were transferred to the same media containing glucose at 72 h post hCG. The results obtained in these experiments were summarized as follows: 1. The development rates of zygotes, recovered from the oviducts in M2 and cultured in CR$_laa$ with 3mg/Im FAF-BSA, to expanded blastocysts (25.7%) and hatching bIastocysts (17.6%) were significantly higher than those of zygotes recovered in TL Hepes (0% and 0%, respectively). 2. The development rates of one-cell embryos exposed to 27.5 mM glucose at 72 h post hCG for 1 min, were 68.8% (CR$_1$+BSA) a and 77.1% (CR$_1$+FBS) of expanded blastocyst stage, but there were no significant differences between the embryos exposed for 1 min. or transferred at 72 h. 3. Regardless of glucose concentration (5.5, 16.5, 27.5 & 38.5mM), 45.7~61.5% of embryos developed to the blastocyst stage. There were no significant differences between any of the treatments on the devel-opment of one-cell embryos. Therefore, the detrimental effect of highly concentration was not appeared.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.4
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• pp.237-243
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• 2006

Objective: The aim of this study was to evaluate the efficacy of frozen-thawed ET in poor prognosis patients such as the old age (38~44 years; OA group) and the patients who did not achieve clinical pregnancy with the first fresh ET cycle (non-pregnant patients; NP group). Methods: Laboratory and clinical data were collected from fresh and frozen-thawed ET cycles of OA and NP group. Controlled ovarian hyperstimulation (COH) and conventional insemination or ICSI, in vitro culture and ET were performed by routine procedures. Supernumerary embryos were frozen by the slow freezing method, and frozen embryos were thawed by the rapid thawing method. Embryo development, pregnancy and implantation rates were statistically analyzed by Student t-test and chi square test Results: Mean ages were similar between fresh ET ($40.0{\pm}1.8$ years, n=206) and frozen-thawed ET ($39.9{\pm}1.9$ years, n=69) cycles in OA group. However, the clinical pregnancy and implantation rate of subsequent frozen-thawed ET significantly higher than those of fresh ET cycles (29.0% and 11.2% vs. 16.5% and 7.0%, p<0.05). In NP group, there was no difference in the mean age between fresh ET ($31.2{\pm}2.3$ years, n=40) and frozen-thawed ET ($31.9{\pm}3.1$ years, n=119) in subsequent cycles. The clinical pregnancy and implantation rates were similar between the subsequent fresh ET (42.5% and 22.6%) and the frozen-thawed ET (40.3% and 18.8%). Conclusion: In old age patients, higher pregnancy rate of frozen-thawed ET compared to fresh ET cycles in this study. It may be related that better uterine environments for implantation in frozen-thawed ET cycles than that of non-physiological hormonal condition in uterus of fresh COH cycles.

• Journal of Aquaculture
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• v.20 no.2
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• pp.96-105
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• 2007

A pair of maroon clownfishes with an indonesian native, reared in recirculation culture system to develope its aquaculture techniques. Courtship, spawning behavior, egg developments and rearing of the maroon clownfish larvae were documented. The larval development were described with illustrative figures. The spawning was occurred 8 times between Feburary and August 2004. The gravid female spawned during 15:00-20:00. The male mainly took care of the eggs supplying oxygen by water currents using their pectoral fins, anal fin and mouth. The fertilized eggs were separative-adhesive and oval in shape, and $1.99{\pm}0.03\;mm$ in longer diameter and $0.88{\pm}0.03\;mm$ in shorter diameter. The fertilized eggs were in deep-orange color. Cleavage occurred in 30 minutes after fertilization, and the egg reached 2 cells stage in 1 hour 10 minutes after fertilization at $27.0^{\circ}{\pm}0.5^{\circ}C$. The embryo was formed in 23 hours 40 minutes after fertilization. Hatching began in between $120{\pm}2$ hours and $150{\pm}12$ hours after fertilization at $27.0^{\circ}C$ in the incubator. Total length (TL) of the newly hatched larvae was 3.22 mm with mouth and anus opened. Ten days after hatching, mean TL of the larvae were 6.21 mm with 28 dorsal fin rays, 17 anal fin rays and 28 caudal fin rays. Nineteen days after hatching, mean TL of the larvae were 9.34 mm. At this stage the larva had three white bands on the body, and they began to feed on commercial diet.

• Journal of Plant Biotechnology
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• v.34 no.1
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• pp.61-68
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• 2007

To develop rice (Oryza sativa L.) cultivars to be planted on salt-affected sites, cell lines with enhanced proline content and resistance to growth inhibition by Azetidine-2-carboxylic acid (AZCA), a proline analogue, were screened out among calli irradiated with gamma ray of 50, 70, 90, and 120 Gy. The calli had been derived from embryo culture of the cultivar Donganbyeo. Selected AZCA resistant lines that had high proline accumulation were used as sources for selection of NaCl resistant lines. To determine an optimum concentration for selection of NaCl resistant lines, Donganbyeo seeds were initially cultured on the media containing various NaCl concentrations (0 to 2.5%) for 40 days, and 1.5% NaCl concentration was determined as the optimum concentration. One hundred sixteen salt-tolerant (ST) lines were selected from bulked 20,000 seeds of the AZCA resistant $M_{3}$ seeds in the medium containing 1.5% NaCl. The putative 33 lines ($M_{4}$ generation) considered with salt-tolerance were further analyzed for salt tolerance, amino acid and ion contents, and expression patterns of the salt tolerance-related genes. Out of the 33 lines, 7 lines were confirmed to have superior salt tolerance. Based on growth comparison of the entries, the selected mutant lines exhibited greater shoot length with average 1.5 times, root length with 1.3 times, root numbers with 1.1 times, and fresh weight with 1.5 times than control. Proline contents were increased maximum 20%, 100% and 20% in the leaf, seed and callus, respectively, of the selected lines. Compared to control, amino acid contents of the mutants were 24 to 29%, 49 to 143%, 32 to 60% higher in the leaf, seed and callus, respectively. The ratio of $Na^{+}/K^{+}$ for most of the ST-lines were lower than that of control, ranging from 1.0 to 3.8 for the leaf and 11.5 to 28.5 for the root, while the control had 3.5 and 32.9 in the leaf and root, respectively. The transcription patterns for the P5CS and NHXI genes observed by RT-PCR analysis indicated that these genes were actively expressed under salt stress. The selected mutants will be useful for the development of rice cultivar resistant to salt stress.

• Journal of Yeungnam Medical Science
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• v.15 no.1
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• pp.55-66
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• 1998

Pathophysiological mechanism of hemorrhagic fever with renal syndrome (HFRS) is not fully understood. Major clinical findings of HFRS patients are widespread hemorrhage, acute renal failure and shock. Basic lesion is vascular injury with microvascular hemorrhage and relatively little inflammation. According to autopsy findings, renal medulla shows focal hemorrhage, tubular necrosis and interstitial mononuclear infiltrates. The predominant cell type in the renal and pulmonary interstitium is a fibroblast and it participates in the healing process at the injury site by secreting a large amount of extracellular matrix proteins. Cultured human lung fibroblasts and Mongolian gerbil fibroblasts were known to be good host cells for the hantaan virus. It is possible that not only the endothelial cell but also the fibroblast is a target of Hantaan virus and the fibroblast might be involved in the pathogenesis and the healing process in HFRS. Integrins are adhesion molecules, and act as receptors for many extracellular matrix proteins. Recently, there are many reports that cell surface integrins influence on some viral infections or reversely viruses influence on the expression of integrins. The ${\alpha}_5{\beta}_1$ integrin is a major receptor for the fibronectin which is an important extracellular matrix protein secreted by fibroblasts. In this study, the role of ${\alpha}_5{\beta}_1$ integrin in the infection of Hantaan virus was examined by using anti-${\alpha}_5{\beta}_1$, integrin, anti-${\alpha}_5$ integrin and anti-${\beta}_1$, integrin antibodies in chicken embryo fibroblasts (CEF) and Mongolian gerbil fibroblasts(MGF). The treatment of anti-${\alpha}_5{\beta}_1$, integrin antibody in CEF reduced the virion titers 26.8% and the amount of nucleocapsid N protein 32.6% when compared with control CEF. When MGF were treated with anti-${\alpha}_5$, anti-${\beta}_1$ and anti-${\alpha}_5{\beta}_1$ integrin antibodies, virion titers were reduced by 26.5%, 29.4% and 28.7% and the amount of nucleocapsid N protein were reduced by 65.2%, 59.7% and 72.6%. These results suggested that ${\alpha}_5{\beta}_1$ integrin might act as a receptor for the Hantaan virus or blocking of ${\alpha}_5{\beta}_1$ integrin influences on the viral replication in CEF and MGF. It is also possible that the blocking of only one subunit of integrin represents similar results in that of whole molecule.

• Korean Journal of Animal Reproduction
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• v.26 no.1
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• pp.41-51
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• 2002

The present study was carried out to examine the effect of cysteamine in vitro maturation (IVM) of porcine oocytes and development of porcine IVM/IVF Embryos. The results were summarized as follows : 1. The rates of nuclear maturation, penetrated oocytes, pronuclear formation, polyspermic oocytes and mean numbers of the penetrated sperm were not different in NCSU23 maturation medium with 0, 25, 50 and 100 $\mu$M cysteamine (P〉0.05). 2. The rates of blastocyst formation at day 7 after in vitro fertilization in 0, 25, 50 and 100 $\mu$M cysteamine were 17.9$\pm$6.1, 17.4$\pm$6.3, 24.2$\pm$1.9 and 16.9$\pm$2.0%, respectively. And the total cells were 30.7$\pm$2.4, 34.9$\pm$2.8, 39.6$\pm$2.3 and 36.8$\pm$3.6, respectively. Fifty $\mu$M cystealnine group was significantly higher than those of any other treatment groups (P＜0.05). 3. The ratios of ICM/total cells in 20~40% category were 20.5, 41.6, 19.5 and 31.5%, respectively. Twenty five $\mu$M cysteamine group was higher than those of other groups. 4. The rates of blastocyst formation at day 7 in the NCSU-23 culture medium of porcine IVF-produced embryos with 0, 25, 50, and 100 $\mu$M cysteamine were 16.0$\pm$0.2, 13.6$\pm$1.7, 25.0$\pm$0.8 and 15.7$\pm$4.5%, respectively. And the total cells were 27.0$\pm$3.7, 36.1$\pm$4.8, 34.0$\pm$3.8 and 25.2$\pm$4.4, respectively. Fifty $\mu$M cysteamine group was significantly higher than those of any other treatment groups (P＜0.05). 5. The ratios of ICM/total cells in 20~40% category were 53.8, 30.0, 16.6 and 11.1%, respectively. The addition groups of cysteamine were lower than those of control group. In conclusion, these results suggested that the addition of 50 $\mu$M cysteamine in the IVM medium and 25~50 $\mu$M cysteamine in IVC medium were effective on the blastocyst formation and total cells of blastocysts.

• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.293-302
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• 1997

Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

• Development and Reproduction
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• v.6 no.1
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• pp.55-66
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• 2002

Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.319-326
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• 1996

The objective of this study was to investigate correlation between the morphology by microscopic assessments of surplus blastocysts produced in human IVF program and their cell number obtained by differential labelling method. For these experiments, 76 surplus human blastocysts were obtained from 36 patients on day 5 after IVF, the embryos were classified to early (ErB), early expanding (EEB), middle expanding (MEB), expanded blastocyst (EdB) according to their blastocoel expansion and zona thickness. When the ovum size and zona thickness of the classified blastocysts were measured using micrometer, although the embryos were produced in the same culture condition, there were significant variances in ovum size ($148.8 217.6{\mu}m$) and zona thickness ($1.2-14.4{\mu}m$). Total blastomere cell number counted after hoechst staining was increased by two to three fold during the transition period from ErB ($39.1{\pm}3.6$) to EdB ($(89.6{\pm}3.3)$) stage on day 5 after IVF. ICM ($11.9{\pm}1.8-22.2{\pm}4.3$) and TE ($24.5{\pm}3.6-70.0{\pm}7.7$) cell numbers using differential labelling were also showed the increased pattern according to the developmental level. Especially, EdB which showed poor ICM morphologically also indicated the low ICM cell number after differential labelling. This demonstrated that there is good correlation between the morphological assessment and the cell number. The count of ICM and TE nuclei using differential labelling can be used as an important criterion, if it is accompanied with morphological assessments, in selecting the better embryos for improving the pregnancy rates in human blastocyst transfer program.