The aim of this study was to determine the beneficial effect of propofol on human keratinocytes that have undergone hypoxia reoxygenation (H/R) injury and to investigate whether autophagy is associated with the protective mechanism. Thus, we evaluated how propofol influences the intracellular autophagy and apoptosis during the H/R process in the HaCaT cells. The cultured human keratinocyte cells were exposed to 24 h of hypoxia (5% CO2, 1% O2, 94% N2) followed by 12 h of reoxygenation (5% CO2, 21% O2, 74% N2). The experiment was divided into 4 groups: (1) Control=Normoxia ; (2) H/R=Hypoxia Reoxygenation ; (3) PPC+H/R=Propofol Preconditioning+Hypoxia Reoxygenation; (4) 3-MA+PPC+ H/R=3-MA-Methyladenine+Propofol Preconditioning+ Hypoxia Reoxygenation. In addition, Western blot analysis was performed to identify the expression of apoptotic pathway parameters, including Bcl-2, Bax, and caspase 3 involved in mitochondrial-dependent pathway. Autophagy was determined by fluorescence microscopy, MDC staining, AO staining, and western blot. The H/R produced dramatic injuries in keratinocyte cells. In our study, the viability of Propofol in H/R induced HaCaT cells was first studied by MTT assay. The treatment with 25, 50, and 100 μM Propofol in H/R induced HaCaT cells enhanced cell viability in a dose-dependent manner and 100 μM was the most effective dose. The Atg5, Becline-1, LC3-II, and p62 were elevated in PPC group cells, but H/R-induced group showed significant reduction in HaCaT cells. The Atg5 were increased when autophagy was induced by Propofol, and they were decreased when autophagy was suppressed by 3-MA. These data provided evidence that propofol preconditioning induced autophagy and reduced apoptotic cell death in an H/R model of HaCaT cells, which was in agreement with autophagy playing a very important role in cell protection.

Opal glass samples having different chemical compositions were synthesized and transparent glass was obtained after melting. The effects of TiO2, BaF2, and CeO2 content on the color of the opal glass were studied by observing images of the opal samples and analyzing the results via ultraviolet visible spectroscopy and color spectrometry. The aesthetic properties of the opal glass were determined by studying the transmittance of visible light in the 400 nm to 700 nm range. The basic chemical composition of opal glass was SiO2 52.9 wt%, Al2O3 12.35 wt%, Na2CO3 15.08 wt%, K2CO3 10.35 wt%, Ca3(PO)4 4.41 wt%, MgCO3 1.844 wt%, LiCO3 2.184 wt%, and TiO2 0.882 wt%. The glass samples were prepared by varying the weight percentage of TiO2, BaF2, and CeO2. The transmittance of visible light was decreased from 95 % to 75 % in the glass samples in which TiO2 content was increased from 0 to 3.882 wt%. In the blue spectrum region, as the content of TiO2 increased, the reflectance value was observed to become higher. This implies that TiO2 content induces more crystal formation and has an important effect on the optical properties of the glass. The opalescence of opal samples that contained CeO2 or BaF2 is stronger than that in the samples containing TiO2. Opal glass samples comprising TiO2 had tetragonal lattice structures; samples including CeO2 as an additive had cubic lattice structures (FCC, CeO2).

In this study, we examined the effectiveness of in vitro fertilization of porcine immature oocytes on the embryo development of blastocysts or hatched blastocysts and the number of cells according to the in vitro fertilization conditions. In the in vitro fertilization of in vitro matured porcine oocytes, there were no significant differences between treatment groups regarding fertilization rate, blastocyst rate, and embryo development of hatched blastocysts according to the storage periods of liquid sperm of 24, 48, and 72 hours. The embryo development rate of hatched blastocysts after the fertilization according to different spermatozoa concentrations (, , and cells/ml) showed the highest rate in the group with a spermatozoa concentration of cells/ml; in particular, this rate was significantly higher than that in the cells/ml group (p<0.05). The total number of blastocysts cells as well as trophectoderms (TE) that developed in each treatment group were also significantly higher in the cells/ml group than in any other groups (p<0.05). In contrast, the embryo development rate of blastocysts according to different co-incubation periods of sperm and oocyte (1, 3, and 6 hr) was high in the 6-hour group; in particular, the rate was significantly higher than that of the I-hour group (p<0.05). Furthermore, the total number of oocytes cells and TEs that developed was significantly higher in the 6-hour group than any other group (p<0.05). In this study, the most effective treatment conditions for porcine embryo development and high cell number were found to be as follows: a sperm storage period of less than 72 hours, a spermatozoa concentration of cells/ml, and a 6-hour co-incubation period for sperm and ooocyte.

Intracytoplasmic sperm injection (ICSI) is one of the artificial fertilization methods when only a few sperm are available for insemination, and an important tool for the preservation of genetic materials of endangered animal species, especially the male is infertile. Different from other species such as mice and pigs, the conventional ICSI method which uses spiked pipette for injection (Spike-ICSI) is exhibited low success rates in cattle because the bovinesperm head membrane is hard to break during injection procedure. We chose piezo-assisted ICSI (Piezo-ICSI) for the improvement of the injection procedure including sperm head membrane rupture and efficient puncture of the plasma membrane of the oocytes. In this experiment, we compared the efficacy of the bovine ICSI embryo production between the Piezo-ICSI and Spike-ICSI. The second polar body extrusion, pronuclear formation, cleavage and blastocyst formation were evaluated after implementation of two different ICSI techniques. The Piezo-ICSI tended to show comparably higher rates of the second polar body extrusion (41.7%), the pronuclei formation (42.9%) and the two-cell cleavage (41.4%) than Spike-ICSI does (33.3%, 28.6% and 23.5%, respectively) although there is no statistic significance between two groups. In addition, the blastocysts were only obtained from the Piezo-ICSI group (10.3%). Our finding shows that the Piezo-ICSI may be used as an artificial fertilization method in cattle when in vitro fertilization is not applicable.

The early Ordovician graptolites from the upper part (Tumok Member) of the Mungok Formation of the Namaeri and Baeiljae sections at Yeongwol consist of five species belonging to five genera. They are Adelograptus cf. tenellus, Callograptus sp., Dendrograptus suni, Dictyonema sp., and Psigraptus jacksoni. Of these 5 species, Psigraptus is the most excellent index fossil for international correlation with the early Ordovician strata. Adelograptus cf. tenellus from the Mungok Formation of the Namaeri and Baeiljae sections is also recognized with especially short stipe relative to Adelograptus tenellus. Three graptolite zones (such as Adelograptus, Psigraptus, and Dendroid type zones) are correlated with those of Canada, Australia, and China, respectively. The upper part of the Mungok Formation is assigned to the late Tremadocian.

Early Ordovician graptolites from the Mungok Formation of Yeongwol area, Korea comprise seven species belonging to six genera: Callograptus curvithecalis, Callograptus sinicus, Aspidograptus lotolatzensis, Dendrograptus suni, Dictyonema uniforme, Adelograptus cf. tenellus, and Psigraptus jacksoni. Two graptolite zones with three subzones are recognized in the eight sections of the Mungok Formation. The Adelograptus Zone is correlated with (1) La 1b of the Lancefield Formation of Victoria, Australia, (2) the Adelograptus-Clonograptus Zone of the Yehli Formation of Jilin, China, and (3) the Adelograptus Zone of the Road River Formation of Yukon, Canada. It suggests that the upper part of the Mungok Formation corresponds to early Late Tremadocian of Early Ordovician.

Five species of graptolites were discovered from the Jigunsan Shale of Taebaeg area, Korea. They are herein described as Archiclimacograptus riddellensis (Harris, 1924), Pseudamplexograptus distichus(Eichwald, 1840), Hustedograptus teretiusculus(Hisinger) sensu Jaanusson, 1960, Hustedograptus vikarbyensis (Jaanusson, 1960), and Hustedograptus sp. The graptolite assemblage from the Jigunsan Shale corresponds to those of the Pseudamplexograptus distichus zone in the Baltic and German areas and the geologic age of graptolites bearing beds of the Jigunsan Shale might be late Middle Llanvirn.

In recent reports, the multiple reproductive defects such as cryptorchidism, hypospadias, epididymal cysts, low sperm counts, and testicular cancers are increased in humans, and these changes were doubted by the chemicals with estrogenic or antiandrogenic activities in our environment. To compare the effects of neonatal exposure of di (n-butyl) phthalate and flutamide on the development of reproductive organs and to identify the specific mechanisms of these abnormalities related to the male reproducton, Sprague-Dawley neonate male rats were injected subcutaneously during 5-14 days after birth with corn oil (control), flutamide (0.05, 0.1, and 0.5 mg/animal) and DBP (5, 10, and 20 mg/animal). Animals were killed at 31 (immature) and 42 (pubertal) days of age respectively and blood was collected from abdominal aorta for serum testosterone analysis. Testes, epididymides, seminal vesicles, ventral prostate, levator ani plus bulbocavernosus muscle (LABC), cowpers glands and glans penis were weighed. Expression of steroid hormone receptors (AR and ER) was examined in the testes and ventral prostate. At 31 days of age, ventral prostate, seminal vesicles, LABC, and cowpers glands significantly decreased in the flutamide (0.5 mg/animal) and DBP (20 mg/animal), but serum testosterone levels were not changed. Flutamide slightly delayed the testes descent at the high dose (0.5 mg/animal), but DBP did not show any significant effect on the testes descent at all doses. DBP and flutamide decreased the expression of AR protein in the testes but did not affect the expression of ERa and ER protein in the testes. At 42 days of age, ventral prostate, seminal vesicles, and cowpers glands weights were still significantly decreased at the high dose of flutamide (0.5 mg/animal) and DBP (20 mg/animal), but the weights of testes and epididymides were not different. Serum testosterone decreased significantly in DBP treated animals and slightly, not significantly, in flutamide group. While DBP still significantly decreased the expression of AR protein in testis, flutamide recovered from downregulation of AR protein and did not affect the expression of ERa and ER protein in the testes. Based on these results, flutamide and DBP have shown several similar patterns in reproductive abnormalitis, but some marked differences which may be caused by different acting mechanism.

Identification of spermatogonial stem cell-specific surface molecules is important in understanding the molecular mechanisms underlying the maintenance and differentiation of these cells. We have found that spermatogonia from busulfan treated mice expressed an autoantigen that distinguishes between undifferentiated and differentiated spermatogonia. Four to six weeks after busulfan treatment, germ cells located in the basal compartment of seminiferous epithelium show isotype-specific IgG deposits that form due to autoimmunity. Before busulfan treatment, the level of testicular IgG was very low but IgG levels began to increase after week 4 and peaked at week 6. When cells from the busulfan treated testis were analyzed using laser scanning cytomeoy (LSC), the frequency of cells positive for IgG deposits, 6-integrin, and 1-integrin were 16.53.8%, 11.82.6%, and 9.0 1.4%, respectively. Immunofluorescent staining suggested that most, if not all of the cells with IgG-deposits isolated from a laminin-coated dish, were also positive for a spermatogonial stem cell marker \ulcorner6-integrins as well as for a germ cell-specific marker TRA 98. We determined serum and intratesticular IgG levels and the soundness of seminiferous tubule basement membrane from busulfan treated mice using electron microscopy, in order to study the mechanism responsible for IgG deposits in spermatogonia. We found that the basement membranes of seminiferous tubules from busulfan treated mice were severely impaired when compared to those of normal adult, neonates and w/wv mice. Furthermore, new blood cells were observed in the surface of the damaged basement membrane along the seminiferous tubules. These results suggest that the IgG in spermatogonial stem cells accumulates from circulating blood through the impaired basement membranes induced by busulfan treatment. Taken together, our study suggests that IgG can be used as a new marker for undifferentiated spermatogonia cells.