The present study was undertaken to evaluate the effect of trisaccharides supplementation in glycerol-free tris (GFT) for the cryopreservation of dog spermatozoa. In the first experiment (E1), dog spermatozoa were resuspended with 50, 75, 100 or 125 mM of raffinose, melezitose or maltotriose and cooled at 4 ℃ for 10 min. To determine the effect of different cooling time, the spermatozoa resuspended with 100 mM of raffinose, melezitose or maltotriose were cooled during 10, 20, 30 or 40 min at 4 ℃ (second experiment; E2). The straws were then aligned horizontally for 10 min on the rack and then plunged into LN2. In the third experiment (E3), to determine the effect of different vapor freezing time, the spermatozoa resuspended with 100 mM raffinose were cooled at 4 ℃ for 20 min and frozen in LN2 for 5, 10, 15 or 20 min and then plunged into LN2. In the fourth experiment (E4), to compare different freezing methods [cooling plus vapor freezing (CV), cooling plus step-down freezing (CS) and direct step-down freezing (SD)], the spermatozoa resuspended with 100 mM raffinose were cooled for 20 min and frozen in LN2 vapor for 5 min in case of CV method. In case of CS method, spermatozoa were cooled for 20 min at 4℃ and then frozen by the step-down freezing method. The straws were then aligned horizontally at 18, 15, 5, and 2 cm respectively from the surface of LN2 for 1, 1, 1.4, and 5 min, respectively in an L shaped straw holder and then plunged into LN2. For SD method, the straws were directly aligned horizontally at the same levels as CS from the surface of LN2 for 1, 1, 1.9, and 5 min, respectively and then plunged into LN2. After thawing at 37℃ for 25 sec, the spermatozoa were then incubated for 30 min in the freezing extender (E1) or in the 50 mM sucrose supplemented GFT (E2, E3, and E4) at 24℃. Following post-thaw incubation, sperm progressive motility and viability were assessed in E1, E2, E3, and E4. In addition, acrosome integrity, and gene expression related to apoptosis (BAX, BCL2, and Caspase10) and sperm motility (SMCP) were evaluated in E4. The results demonstrated that, in E1, using 75 mM trisaccharides resulted in significantly (p<0.05) higher sperm motility in all sugar groups. Using 100 mM melezitose significantly (p<0.05) improved the post-thaw viability than the 100 mM raffinose. The viability in 100 mM maltotriose was similar with 100 mM raffinose and melezitose group. In E2, the different cooling time has no significant effect on post-thaw sperm progressive motility in all the sugar types. In addition, the viability was variable among the different groups. In E3, liquid nitrogen vapor freezing for 5 min resulted in improved motility and viability. The sperm progressive motility was significantly (p<0.05) higher in CV and SD group compared to CS group and the sperm viability was significantly (p<0.05) higher in CV group compared to the other groups in E4. However, the acrosomal integrity of spermatozoa in the group CV was significantly (p<0.05) higher than the group CS and SD. In addition, the expression of SMCP gene was significantly (p<0.05) higher in the CV group than the CS group. In contrast, the expression of Caspase10 significantly (p<0.05) lower in the group CV and SD than the group CS. Furthermore, the ratio of gene expression of BAX and BCL2 was significantly (p<0.05) lower in the group CV than the group CS. Therefore, cryopreservation of dog spermatozoa in 100 mM of raffinose supplemented GFT cooled for 20 min and vapor freezing for 5 min provides better progressive sperm motility, viability, and acrosome integrity with higher expression of SMCP gene and lower expression of caspase10 and BAX/BCL2 ratio following post-thaw incubation in 50 mM sucrose supplemented GFT for 30 min at 24℃.

To preserve the superior genetic resources and restore the endangered species, Somatic cell nuclear transfer (SCNT) has been used widely. In Korea, the research of dog cloning has made outstanding achievements including the production of the world`s first cloned dog. Sapsaree (Sapsalgae), the representative dog of Gyeongsan-si was designated as a Korea natural monument (No. 368). This male dog used in this study has azoospermia due to unknown cause. In this study, the aim was to confirm the cause of infertility in the cell donor dog and to evaluate the reproduction potential of dog cloning using infertile male dog by SCNT. First, to confirm the infertility of the cell donor dog, the reproductive history and the testis were evaluated. The breeding histology was not recorded in individual document. In histopathology, the Sertoli cell tumor was confirmed in biopsy of the cell donor dog after death. But, these tumors are predominantly in older dogs. Second, we produced the cloned dogs with the somatic cells of the infertile dog and the appearance was similar with the cell donor dog. Also, microsatellite analysis confirmed the genetic relationship between the cell donor and clone dogs. Third, the potential breeding capacity of the cloned dog was confirmed. In T4 assay, the normal dog (same age with cloned dogs), cell donor dog, and cloned dogs was investigated. The cell donor dog with azoospermia had very low T4 level, and cloned dogs showed higher level of T4 than normal dogs. In CASA, There was no significant difference in sperm motor ability between normal dogs and cloned dogs. As a result, cloned dogs produced by SCNT had no problem regarding the reproductive function of the testis. In AI experiment, the semen of clone dogs was used to fertilize a natural female bitch and was diagnosed pregnancy by ultrasonography. In total, 7 puppies were born by normal delivery (male: 3, female: 4). In conclusion, this study confirmed that the reproduction problem of non-genetic infertility can generate a normal descendant by SCNT. Also, the first successful research to restore infertile dogs was completed. Furthermore, SCNT would be useful for the restoration of endangered species and application of superior traits.

The recent updates of the North Ecliptic Pole deep (0.5 deg2, NEP-Deep) multi-wavelength survey covering from X-ray to radio-wave is presented. The NEP-Deep provides us with several thousands of 15 μm or 18 μm selected galaxies, which is the largest sample ever made at these wavelengths. A continuous filter coverage in the mid-infrared wavelength (7, 9, 11, 15, 18, and 24 μm) is unique and vital to diagnose the contributions from starbursts and AGNs in the galaxies out to z=2. The new goal of the project is to resolve the nature of the cosmic star formation history at the violent epoch (e.g. z=1{2), and to find a clue to understand its decline from z=1 to present universe by utilizing the unique power of the multiwavelength survey. The progress in this context is brie y mentioned.

We utilize AKARI's slitless spectroscopic capability to detect the 3.3 μm polycyclic aromatic hydrocarbons (PAHs) emission and measure star formation (SF) activity for various AKARI programs. First, we obtain 2∼5μm spectra of 20 flux-limited galaxies with mixed SED classes in order to calibrate the 3.3 μm PAH luminosity (LPAH 3.3) as a star formation rate (SFR) indicator. We find that LPAH3.3 correlates with LIR as well as with the 6.2 μm PAH luminosity ( LPAH 6.2). The correlations does not depend on SED classes. We find that ULIRGs deviate from the correlation between PAH luminosities and LIR, while they do not for the correlation between PAH luminosities. We suggest possible effects to cause this deviation. On the other hand, how AGN activity is linked to SB activity is one of the most intriguing questions. While it is suggested that AGN luminosity of quasars correlates with starburst (SB) luminosity, it is still unclear how AGN activity is connected to SF activity based on host galaxy properties. We are measuring SFRs for the LQSONG sample consisting of reverberation mapped AGNs and PG-QSOs. This is an extension of the ASCSG program by which we investigated the connection between SB and AGN activities for Seyferts type 1s at z ~ 0.36. While we found no strong correlation between LPAH3.3 and AGN luminosity for these Seyferts 1s, LPAH3.3 measured from the central part of galaxies correlates with AGN luminosity, implying that SB and AGN activities are directly connected in the nuclear region.

An overview of the North Ecliptic Pole (NEP) deep multi-wavelength survey covering from X-ray to radio wavelengths is presented. The main science objective of this multi-wavelength project is to unveil the star-formation and AGN activities obscured by dust in the violent epoch of the Universe (z=0.5-2), when the star formation and black-hole evolution activities were much stronger than the present. The NEP deep survey with AKARI/IRC consists of two survey projects: shallow wide (8.2 sq. deg, NEP-Wide) and the deep one (0.6 sq. deg, NEP-Deep). The NEP-Deep provides us with a 15 μm or 18 μm selected sample of several thousands of galaxies, the largest sample ever made at these wavelengths. A continuous filter coverage at mid-IR wavelengths (7, 9, 11, 15, 18, and 24 μ m ) is unique and vital to diagnose the contribution from starbursts and AGNs in the galaxies at the violent epoch. The recent updates of the ancillary data are also provided: optical/near-IR magnitudes (Subaru, CFHT), X-ray (Chandra), FUV/NUV (GALEX), radio (WSRT, GMRT), optical spectra (Keck/DEIMOS etc.), Subaru/FMOS, Herschel/SPIRE, and JCMT/SCUBA-2.

This study was performed to investigate the relationship between PSS-HSP70 gene polymorphism and artificial insemination (AI) reproductivity in the pigs. The RFLP polymorphism of PSS and the SSCP polymorphisms of HSP70 K1, K3 and K4 PCR product were detected different patterns. In the experiment for AI of fresh semen, spring and fall season showed higher litter size born of 10.89 head than 10.47 head of summer season. Landrace was showed higher litter size of 9.96 head than that of Duroc and Yorkshire (p<0.05). Stress relating PSS and HSP70 polymorphism of PSS-Normal, HSP70 K1-BB, K3-AB, K4-AA showd a highest litter size born of 10.97 head and litter size born alive of 10.69 head than that of the other polymorphisms(p<0.05). In the experiment for AI of frozen semen, effects of season and pig breeds were not showed for litter size born. The stress relating polymorphism of PSS-Carrier, HSP70 K1-BB, K3-BB, K4-AB showed highest litter size born of 11.29 head and litter size born alive of 10.82 head and PSS-Normal, HSP70 K1-BB, K3-AB, K4-AA showed the lowest litter size born of 8.48 head and litter size born alive of 7.33 head than that of the other polymorphisms(p<0.05). These results suggest that AI litter size born for the stress of forzen thawed semen may be affected by PSS and HSP70 polymorphism in pigs.

We have made a comprehensive statistical study on the coronal mass ejections(CMEs) associated with helmet streamers. A total number of 3810 CMEs observed by SOHO/LASCO coronagraph from 1996 to 2000 have been visually inspected. By comparing their LASCO images and running difference images, we picked out streamer-associated CMEs, which are classified into two sub-groups: Class-A events whose morphological shape seen in the LASCO running difference image is quite similar to that of the pre-existing streamer, and Class-B events whose ejections occurred in a part of the streamer. The former type of CME may be caused by the destabilization of the helmet streamer and the latter type of CME may be related to the eruption of a filament underlying the helmet streamer or narrow CMEs such as streamer puffs. We have examined the distributions of CME speed and acceleration for both classes as well as the correlation between their speed and acceleration. The major results from these investigations are as follows. First, about a quarter of all CMEs are streamer-associated CMEs. Second, their mean speed is 413 km s-1 for Class-A events and 371 km s-1 for Class-B events. And the fraction of the streamer-associated CMEs decreases with speed. Third, the speed-acceleration diagrams show that there are no correlations between two quantities for both classes and the accelerations are nearly symmetric with respect to zero acceleration line. Fourth, their mean angular width are about 60°, which is similar to that of normal CMEs. Fifth, the fraction of streamer-associated CMEs during the solar minimum is a little larger than that during the solar maximum. Our results show that the kinematic characteristics of streamer-associated CMEs, especially Class-A events, are quite similar to those of quiescent filament-associated CMEs.

This study was carried out to evaluate the effects of washing medium, breed and washing temperature of fresh and frozen-thawed boar sperm on mitochondrial activity and membrane integrity by flow cytometry. More than 80% of fresh sperm washed with mTLP-PVA medium at 20℃ exhibited an intact membrane and a functional mitochondrion. With frozen-thawed samples, a large number of sperm showed both damaged membrane (36.4~46.9%) and nonfunctional mitochondrion (55.1~71.1%) in the mTLP-PVA and BTS washing media at 20℃. There were no breed effects of fresh and frozen-thawed sperm on mitochondrial activity and membrane integrity. The percentages of damaged membrane of fresh and frozen sperm, respectively, were higher at 4℃ washing temperature than at 20℃ washing temperature in the mTLP-PVA medium. We found that washing medium and washing temperature of fresh and frozen-thawed boar sperm were important for the analyses of mitochondrial activity and membrane integrity by flow cytometry.