• Development and Reproduction
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• v.11 no.1
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• pp.31-41
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• 2007

The spermiogenesis of Crocidura shantungensis were studied by electron microscope. All process of spermiogenesis was divided into 11 phases 15 steps, based on the morphological features of the nucleus and cell organelles in cytoplasm of spermatids. The spermatids in Golgi and cap phases were a spherical shape. On the other hand, at the early acrosomal phase they changed into an oval shape, and the tail was created in this phase. In maturation phase, the shapes of spermatid head were thin and longish. Until step 7 the direction of spermatids head turned toward the lumen of the seminiferous tubule. From step 8 to step 15 their heads turned toward the basal lamina. In step 12, the nucleus and acrosome shown maximal elongation. From Step 13 the nucleus of spermatids became flat, simultaneously with flat expansion of the acrosome expanded, and the visible whole lengths of spermatids were tend to be shorten. Spermatid heading which arrived to step 14 was taken the final shape. The nucleus was doing the wedge shape, and the nuclear chromatins condensed completely and homogenized. In the spermiation phase, the spermatids were gradually disconnected from the cytoplasm of the Sertoli cell. In this phase, the acrosome of the spermatids were fully shorten and flat, and the spermatozoa completed the process of heading and the tailing. Considering all the results, the spermiogenesis may be useful information to analyze the differentiation of spermatogenic cells.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.67-67
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• 2003

Early development of porcine oocytes fertilized in vitro was examined in different culture conditions. Porcine ovaries were collected from local slaughter-house. Cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cystein, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for 42~44 hrs. The frozen-thawed spermatozoa were washed by centrifigation 2 times at 1, 500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, 1$\mu\textrm{g}$/ml streptomycin and 1ng/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to 2.5$\times$10$^{6}$ cells/ml motile sperm during fertilization in vitro. At 8hrs h after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine and 4 mg/ml BSA and cultured for 7 days. In first experiment, the mean numbers of oocytes collected from 20 ovaries were 674.4 oocytes, and 4.1(27.6), 12.5(84.0), 25.4(171.6) and 57.9%(390.8) for A, B, C and D grade in morphological classification. In the second experiment, when culture medium was supplemented with various concentrations of EGF, the proportions of oocytes cleaved were 56.9, 55.7, 61.9 and 54.7% for 0, 5, 10 and 20ng/ml EGF. The higher proportions(15.1%) of oocytes developed to morular stage were obtained at concentration of 10ng/ml than 0 and 5ng/ml EGF (P<0.05). However, the proportions of embryos developed to blastocyst stage were not significantly different among concentrations of EGF. In another experiment, when the medium supplemented with catalase was used, the proportions of oocytes cleaved were higher in the concentration of 0 unit (56.5%, 61/108) than 100 and 1, 000 unit/ml of catalase (P<0.05). Although the developmental capacity of embryos was improved by medium with 0 unit/ml compared with 100, 500 and 1, 000 units/ml of catalase in oocytes developed to morula and blastocyst stages, were not significantly different among concentrations of catalase.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.331-331
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• 2017

This study was conducted to know the influence of air temperature and sunshine duration on yield, chemical component, pigment color difference and starch characteristics of two colored rice cultivars in the plain area of Yeongnam province in Korea. The $L^*$, $a^*$, and $b^*$ value of brown rice in Hongjinju and Josaengheugchal rice cultivars was significantly different at continuous cultivated years, 2015 and 2016. The $L^*$, $b^*$ value of two colored rice was significantly increased in 2016 compared to 2015. The $a^*$ value of Josengheugchal rice cultivar was also significantly higher at 2016 than at 2015. It can be noticed the $a^*$, $b^*$, $L^*$ values in Josaengheugchal varied more than those in Hongjinju. Air temperature during ripening period in 2016 was higher than 2015, especially minimum temperature was too high to proper maturation for rice quality. In Josaengheugchal rice cultivar, sunshine duration after heading was longer in 2016 than in 2015. On the contrary, Hongjinju rice cultivar was ripened under condition of insufficient sunshine duration in 2016. The short growing duration by high temperature and long shiny duration made the lack of pigment for Josaengheugchal brown rice. In Hongjinju rice cultivar, shorten sunshine duration and higher night temperature were the source of the pigment deficiency. The grain size of rice which produced in 2016 was bigger than that of 2015 in both rice cultivars. The 1,000 grain weight of rice from 2016 was also bigger than that of 2015. Head rice ratio was high in the rice cultivars produced in 2015. Protein of milled rice in 2016 was more decreased than that of 2015 in Josaengheugchal rice cultivar, it showed reverse result in Hongjinju rice cultivar. Amylose contents of milled rice in 2016 were more decreased than that of 2015 in Hongjinju rice cultivar. Branch chain length distribution of amylopectin was shown a distinct difference between Josaengheugchal and Hongjinju rice flours by each produced year. Josaengheugchal rice cultivar produced in 2015 had a higher amount of short chains than that of 2016 rice starches. In Josaengheugchal rice cultivar, the pasting temperature and peak, trough, breakdown, final viscosity increased in rice flour which produced at 2016, whereas the setback viscosity and peak time showed lower value than those of rice from 2015. The most pasting properties (except of setback viscosity) of rice starch in Hongjinju rice cultivar grown in 2015 were higher than those of rice cultivar produced in 2016.

• Asian-Australasian Journal of Animal Sciences
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• v.9 no.5
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• pp.583-590
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• 1996

The purpose of this study was to evaluate some factors in the bovine embryonic development from one-cell to blastocyst using modified synthetic oviduct fluid medium (mSOFM), after maturation and in vitro fertilization of the oocytes. The embryonic development to the blastocyst stage was assessed at 7-10 days after in vitro fertilization, and the total cells in the blastocysts were counted by staining nuclei with fluorochrome. Some commercial calf sera (CS) and a superovulated cow serum had different effects on the embryonic development to the blastocyst stage (8.6-21.4%), dependent upon their product lots, although the development might not be affected at least by serum progesterone levels. ${\beta}$-Mercaptoethanol (${\beta}$-ME) supplemented into mSOFM was effective to the embryonic development (27.8%), as well as the co-culture system with cumulus cells (19.5%). In a serum- and feeder cell-free culture using mSOFM containing several growth factors and ${\beta}$-ME instead of CS plus co-cultured cumulus cells, bovine serum albumin (BSA, fraction V), but not polyvinyl alcohol (PVA), was highly effective in embryonic development to the blastocyst stage, almost comparable to CS in the serum-contained culture (CS, BSA and PVA; 27.8, 19.5 and 5.7%, respectively). However, fatty acid free BSA rather reduced the number of developed blastocysts, compared with fraction V BSA (7.3 vs 29.4%). In the serum- and feeder cell-free culture, supplement of glucose to the medium (final 2.0 mM) stimulated the cell proliferation of developing embryos 120 hr after in vitro fertilization. These results indicated that a serum-free medium supplemented with ${\beta}$-ME could successfully support the development of bovine one-cell embryos to the blastocyst stage. Moreover, supplement of glucose and fatty acids to the medium might support preferably the development and cell proliferation of embryos.

• Journal of Embryo Transfer
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• v.16 no.2
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• pp.99-106
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• 2001

This study was undertaken to compare the efficiency of recovery rate and development rate of follicular oocytes collected either by aspiration or by slicing method. The follicular oocytes collected by the two methods matured in TCM199 supplemented with 10% steer serum at 39$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. After 22 h of culture, the oocytes were inseminated with frozen-thawed semen (2$\times$10$^{6}$ sperm/ml of final concentration) prepared with Percoll-density gradient in IVF-TALP medium for 16 h. Later, sets of 15 presumptive zygotes were transferred into 50 $\mu$L, droplets of CR1aa medium. On day 4 of the culture, embryos were transferred to TCM199 until day 9. The percentages of nuclear maturation to pre-metaphase II in the oocytes collected by aspiration are significantly (P<0.05) higher than that by slicing (83% vs. 62%, respectively). The mean number of oocytes recovered by slicing per ovary is significantly (P<0.05) higher than that by aspiration (15.1 vs. 6.7, respectively). Although the rates of cleavage and development to blastocyst of oocytes collected b)\\\\`aspiration are significantly (P<0.05) higher than that by slicing, the number of transferable embryos obtained by slicing method is significantly (P<0.05) higher than that by aspiration. From the results. we may conclude that slicing method is better than aspiration method for obtaining large number of transferable embryos per ovary.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.3
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• pp.389-398
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• 1999

Objective: This study was designed to evaluate the effects of endogenous LH surge, GnRH agonist (GnRH-a) or human chorionic gonadotropin (hCG) as ovulation trigger on pregnancy rate by intrauterine insemination (IUI). Method: Patients received daily 100 mg of clomiphene citrate (CC) for 5 days starting on the third day of the menstrual cycle followed by human menopausal gonadotropin (hMG) for ovulation induction. Follicles larger than >16 mm in diameter were present in the ovary, frequent LH tests in urine were introduced to detect an endogenous LH surge. Final follicular maturation and ovulation were induced by GnRH-a 0.1 mg (s.c.) or hCG $5,000{\sim}10,000$ IU (i.m.) administration except natural ovulation. Pregnancy was classified as clinical if a gestational sac or fetal cardiac activity was seen on ultrasound. Results: There were no differences in age, duration of infertility and follicle size, but more ampules of hMG were used in GnRH-a group compared to hCG 10,000 IU treated group (p<0.05). Lower level of estradiol ($E_2$) on the day of hCG or GnRH-a injection was observed in hCG 10,000 IU group than other treatment groups (p<0.01). The overall clinical pregnancy rate was 19.8% per cycle (32/162) and 22.2% per patient (32/144). Pregnancy rate was higher in natural-endogenous LH surge group (37.5%, 9/24) than GnRH-a (18.8%) or hCG treated group (20.9% & 13.9%), but this difference was not statistically significant. No patient developed ovarian hyperstimulation. Abortion rate was 22.2% (2/9) in hCG 5,000 IU group. Delivery or ongoing pregnancy rate was 37.5% (9/24), 18.8% (3/16), 16.3% (7/43) and 13.9% (11/79) in endogenous LH surge, GnRH-a, hCG 5,000 IU and hCG 10,000 IU treatment groups, respectively. Conclusion: These results support the concept that use of natural-endogenous LH surge in stimulated cycles may be more effective to obtain pregnancies by IUI than GnRH-a or hCG administration.

• Korean Journal of Food Science and Technology
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• v.34 no.4
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• pp.570-576
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• 2002

For the utilization of the by-products of whelk processing, whelk internal organ with the addition of commercial proteolytic enzymes - Flavourzyme, Neutrase, Protease NP, Prozyme - were used to make jeotgal, Korean traditional salted and fermented seafood sauce. The products were prepared at salt concentration of 25% with enzyme contents 0.05 and 0.1%. The samples were stored at $10^{\circ}C$ and the chemical properties were evaluated for 6 months. The pH in all samples were decreased from near 6.8 in the beginning stage to 6.1-6.4 in the final stage of incubation. Amino nitrogen of jeotgal increased with enzyme concentration and showed maximum value, 646 mg%, at 0.1% of Flavourzyme. Total nitrogen content was increased till four months, but rapidly decreased after that. Protein degradations of whelk internal organ during maturation of jeotgals were investigated by SDS-PAGE. The patterns of degradation were different with added enzymes.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.6
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• pp.531-548
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• 2006

The purpose of this study was to investigate the biomechanical, histologic findings of distracted regenerate and TMJ response in modified distraction osteogenesis (DO) technique combined with compression force as biomechanical stimulation method which has been suggested in 2002, and developed thereafter by authors. This study was performed with two experiments. First experiment was designed to explore the optimal ratio of compression force versus distraction force for the new DO technique. Second experiment was planned to evaluate the reaction of TMJ tissue, especially condyle, disc after application of the DO technique with compression force. Total 52 New Zealand adult male-rabbits with 3.0kg body weight were used for the study. For the first study, 30 adult male-rabbits underwent osteotomy at one side of mandibular body and a external distraction device was applied on each rabbit with same manner. In the control group of 10 rabbits, final 8 mm of distraction with 1 mm rate per day was done with conventional DO technique after 5 latency days. For the experimental group of 20 rabbits, a compression force with 1 mm rate per day was added to the distracted mandible on 3-latency day after over-distraction (over-lengthening). As the amount of the rate of compression versus distraction, experimental subgroup I (10 rabbits) was set up as 2 mm compression versus 10 mm distraction (1/5) and experimental subgroup II (10 rabbits) was set up as 3 mm compression versus 11 mm distraction (about 1/3). All 30 rabbits were set up to obtain final 8 mm distraction and sacrificed on postoperative 55 day to analysis on biomechanical, and histologic findings of the bone regenerates. For second study, 22 adult male-rabbits were used to evaluate TMJ response after the DO method application with compression force. In the control group, 10 rabbits was used to be performed with conventional DO method, on the other hand, in a experimental group of 10 rabbits, 10 mm distraction with 2 mm compression (1/5 ratio) was done. The remaining 2 rabbits served as the normal control group. Histomorphologic examinations on both condyle, histological studies on condyle, disc were done at 1, 2, 3, 4, 7 weeks after distraction force application. The results were as follows: 1. On the bone density findings, the experimental group II (force ratio - 1/3) showed higher bone density than the other experimental group (force ratio - 1/5) and control group (control group - $0,2906\;g/cm^2$, experimental group I - $0.2961\;g/cm^2$, experimental group II - $0.3328\;g/cm^2$). 2. In the histologic findings, more rapid bone maturation like as wide lamellar bone site, more trabeculae formation was observed in two experimental groups compared to the conventional DO control group. 3. In morphologic findings of condyle, there were no differences of size and architecture in the condyle in the control and experimental groups. 4. In histologic findings of condyles, there were thicker fiberous and proliferative layers in experimental group than those of control group until 2 weeks after distraction with compression force. But, no differences were seen between two groups on 3, 4, 7 weeks after compression. 5. In histologic findings of disc, more collagen contents in extracellular matrix, more regular fiber bundles, and less elastin fibers were seen in experimental group than control group until 2 weeks after distraction with compression. But, no differences were seen between two groups on 3, 4, 7 weeks after distraction with compression. From this study, we could identify that the new distraction osteogenesis technique with compression stimulation might improve the quality of bone regeneration. The no remarkable differences on TMJ response between control and experimental groups were seen and TMJ tissues were recovered similarly to normal TMJ condition after 3 weeks.

• Applied Microscopy
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• v.7 no.1
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• pp.21-43
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• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.

• Interdisciplinary Bio Central
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• v.1 no.1
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• pp.3.1-3.6
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• 2009

Introduction: GTPases known as translation factor play a vital role as ribosomal subunit assembly chaperone. The bacterial Obg proteins ($Spo{\underline{0B}}$-associated ${\underline{G}}TP$-binding protein) belong to the subfamily of P-loop GTPase proteins and now it is considered as one of the new target for antibacterial drug. The majority of bacterial Obgs have been commonly found to be associated with ribosome, implying that these proteins may play a fundamental role in ribosome assembly or maturation. In addition, one of the experimental evidences suggested that Bacillus subtilis Obg (BsObg) protein binds to the L13 ribosomal protein (BsL13) which is known to be one of the early assembly proteins of the 50S ribosomal subunit in Escherichia coli. In order to investigate binding mode between the BsObg and the BsL13, protein-protein docking simulation was carried out after generating 3D structure of the BsL13 structure using homology modeling method. Materials and Methods: Homology model structure of BsL13 was generated using the EcL13 crystal structure as a template. Protein-protein docking of BsObg protein with ribosomal protein BsL13 was performed by DOT, a macro-molecular docking software, in order to predict a reasonable binding mode. The solvated energy minimization calculation of the docked conformation was carried out to refine the structure. Results and Discussion: The possible binding conformation of BsL13 along with activated Obg fold in BsObg was predicted by computational docking study. The final structure is obtained from the solvated energy minimization. From the analysis, three important H-bond interactions between the Obg fold and the L13 were detected: Obg:Tyr27-L13:Glu32, Obg:Asn76-L13:Glu139, and Obg:Ala136-L13:Glu142. The interaction between the BsObg and BsL13 structures were also analyzed by electrostatic potential calculations to examine the interface surfaces. From the results, the key residues for hydrogen bonding and hydrophobic interaction between the two proteins were predicted. Conclusion and Prospects: In this study, we have focused on the binding mode of the BsObg protein with the ribosomal BsL13 protein. The interaction between the activated Obg and target protein was investigated with protein-protein docking calculations. The binding pattern can be further used as a base for structure-based drug design to find a novel antibacterial drug.