Co-Fe-Ni-B-Si-Cr based amorphous strips containing nitrogen were manufactured via melt spinning, and then devitrified by crystallization treatment at the various annealing temperatures of for up to 30 minutes in an inert gas atmosphere. The microstructures were examined by using XRD and TEM and the magnetic properties were measured by using VSM and B-H meter. Among the alloys, the amorphous ribbons of containing 121 ppm of nitrogen showed relatively high saturation magnetization. The alloy ribbons crystallized at showed that the grain size of alloy containing 121 ppm of nitrogen was about f nm, which exhibited paramagnetic behavior. The formation of nano-grain structure was attributed to the finely dispersed Fe4N particles and the solid-solutionized nitrogen atoms in the matrix. Accordingly, it can be concluded that the nano-grain structure of 5nm in size could reduce the core loss within the normally applied magnetic field of 300A/m at 10kHz.

This study was conducted to detect the apoptosis incidence in blastocysts and to compare the abundance of Bax, Bcl2L1, VEGF and FGFR2 in in vitro fertilized (IVF), parthenogenetic (PAT) and nuclear transfer (NT) embryos. Oocytes matured for 40 hr were enucleated and reconstructed with confluenced fetal fibroblasts (FFs) derived from a ～45 day fetus. Reconstructed eggs were then fused with 2 DC pulses (2.0 kV/cm, 30 μsec) and cultured with 7.5 μg/ml cytochalasin B for 3 hr. Parthenotes (PAT) were produced with the same electric strength and culture for NT eggs. The embryos were cultured in NCSU-23 medium at 39℃, 5% CO2, 5% O2 in air. In 3 runs, set of 10 embryos at the 4-cell to blastocyst stages were used to extract total RNA for analyzing the gene expression patterns of pro-apoptotic (Bax), anti-apoptotic (Bcl2L1), vasculogenesis (VEGF), implantation (FGFR2III) using real-time quantitative PCR. Cleavage and blastocyst rates were significantly higher (P<0.05) in IVF and PAT (79.3±8.5 and 25.5±6.1, and 85.0±6.4 and 38.6±5.5, respectively) than NT counterparts (65.1±5.2 and 15.6±3.0, respectively). Significantly higher (P<0.05) total cells were observed in IVF controls and PAT (34.7±5.8 and 38.1±4.1) than NT embryos (24.8±3.2). Apoptosis index was significantly lower (P<0.05) in IVF than NT embryos. The Relative abundances (RA) of Bax and VEGF were significantly higher (P<0.05) at blastocyst stage in NT than IVF control. The RA of Bcl2L1 and FGFR2III were significantly higher (P<0.05) at blastocyst stage in IVF than NT. The present study observed the abnormal gene expressions in NT embryos at various developmental stages, suggesting certain clues to find out the cause of the low efficiency of NT to term.

Cyclin B1 is known to reflect the M-phase promoting factor (MPF), a universal regulator of G2/M-phase transition, activity during the process of oocytes maturation. To investigate whether culture condition affects the maturation rate and the expression of cyclin B1 protein, bovine immature oocytes are stimulated and cultured according to the following protocols: Experiment 1: denuded oocytes (denude) only, COC only, denuded oocytes + granulosa cells (denude + GCs) and COC + GCs; Experiment 2: no-activation (control), 7% ethanol for 5 min and 10 l/ml ionomycin for 5 min at immediately before maturation. The maturation rates of denude and no-activation group were significantly lower in both experiments (P<0.05), respectively. Co-culture or stimulation method in bovine immature oocytes culture increases the cyclin B1 expression significantly in both experiments (P<0.05). Based on these results, culture condition affects the maturation rate and the expression of cyclin B1 protein during the first meiotic maturation in bovine immature oocytes.

본 논문의 연구목적은 Choroidal neovascularization (맥락막 신생혈관) 모텔에서 HO-1 발현제와 억제재의 영향을 알아보고자 한다. 30마리의 Brown Norway rat을 각각 10마리씩 세 그룹， Hemin treated group, SnPP treated group, Contorl group으 로 나누어서 실험을 진행하였다. Hemin treat group은 10μmol/kg hemin(Frontier Scientific Inc. USA) 을 SnPP treat group은 10 μmol/kg SnPP(Frontier Scientific Inc. USA)을 laser 시술 2 일에서 14 일까지 복강내 주사하였고 Control군은 0.5 mß씩 식 염 수를 주사하였다. 14 일 후 안저사진촬영과 형광안저촬영을 실시하였다. SnPP treated group에서 Hemin treated group보다 더 많은 선생혈관이 생성되었다. Hemin treated group에서는 맥락막 신생혈관 형성의 정도가 정상대조군에 비교하여 저하됨을 알 수 있었다. 본 연구의 결과 HO-1 의 inducer 인 Hemin 이 맥락막의 신생혈관 억제에 영향을 미치는 것으로 보여진다.

The electro-chemical removal (ECR) of water pollutants by metal-ACF electrodes from wastewater was investigated over wide range of ECR time. The ECR capacities of metallic ACF electrodes were related to physical properties such as adsorption isotherm, surface area and pore size and to reaction time. Surface morphologies and elemental analysis for the metal supported ACFs after electro-catalytic reaction were investigated by scanning electron microscopy (SEM) and energy disperse X-ray (EDX) to explain the changes in adsorption properties. The IR spectra of metallic ACFs for the investigation of functional groups show that the electro-catalytic treatment is consequently associated with the removal of pollutants with the increasing surface reactivity of the activated carbon fibers. The metal-ACFs were electro-catalytically reacted to waste water to investigate the removal efficiency for the COD, T-N, NH4-N, NO3-N and NO2-N. From these removal results of the piggery waste using metallic ACFs substrate, satisfactory removal performance was achieved. The removal efficiency of the metallic ACFs substrate was mainly determined by the properties of the material for adsorption and trapping of organics, and catalytic effects.

The growing oocytes become progressively capable of resuming meiosis, and full meiotic competence appear when they are about 80% of the size of fully grown oocytes. As hormonal influences vary at different stages of reproductive cycle, the size of oocytes may vary according to the reproductive stages. The present study was designed to compare the diameter between the ovulated and freshly collected immature canine oocytes. The ovulated oocytes were collected 72 hr after ovulation by oviductal tube flushing by laparotomy under general anesthesia. Immature oocytes were collected by ovarian slicing method. Diameter of all oocytes was measured directly using epiflurescence microscope with a calibrated micro-eyepiece micrometer at ×200 magnification. The thickness of zona pellucida and diameter of cytoplasm were measured separately and recorded. A total of 2209 zona intact oocytes were collected, among them 628 from anestrus, 675 from follicular, 838 from luteal and 68 by fallopian tubes flushing methods. The average number of oocytes was 104.7, 168.8, 119.7 and 11.3 for anestrus, follicular, luteal and fallopian tubes flushing methods, respectively. The average diameters of the ooplasm and oocyte were significantly varied in different reproductive stages as well as with ovulated oocytes (P<0.05). The average diameter of ooplasm and oocyte was 115.6 and 127.7, 143.0 and 162.0, 134.6 and 150.6, 159.6 and 185.6 for anestrus, follicular, luteal and ovulated oocytes, respectively. Highest number of oocytes with larger diameter could be collected from the follicular and luteal stages. In conclusion, the follicular and luteal ovaries are the best sources of oocytes for canine IVM.

The possibility of producing transgenic embryos expressing the green fluorescence protein (GFP) gene have been evaluated after transfer of exogenous gene into the porcine zygote cytoplasm using the intracytoplasm sperm injection (ICSI) as gene delivery method. For DNA binding to sperm heads, 0.05% Triton X-100 or Lipofectin was used. After injection of the sperm bound to DNA by means of Lipofectin or Triton X-100 triturate, the blastocyst formation rates on day 6 were not significantly different from that of ICSI only group (18.8, 19.2 and 25.3%). In terms of GFP expression, more embryos were in GFP form in Triton X-100 group than in Lipofectin group (40.6 vs 36.4%), while percentage of non-mosaic embryos expressing the GFP gene in all blastomere was higher (P<0.05) in Lipofectin group than in Triton X-100 group (4.2 vs 0.9%). ICSI embryos derived from sperm treated with Lipofectin/DNA complex was transferred into 3 recipients and were collected by uterine flushing on days 5, 7 and 15 after embryo transfer, and then GFP expression was observed by a fluorescence microscopy. Over 26% of the collected embryos were normally expressed GFP gene. These results suggest that foreign gene transfer method with DNA bound sperm caused minimal damage to structure of oocytes that can result to full development of porcine embryos. This was confirmed in this study when the embryos that were transferred after ISCI of DNA bound sperm had a normal development and gene expression until preimplantation.

This study was conducted to examine the effect of IRES controlled reporter gene on screening and production of recombinant human erythropoietin (EPO) proteins from cultured CHO cells. The cDNA was cloned for EPO from human liver cDNA. Using site-directed mutagenesis, we generated recombinant human EPO (rhEPO) with two additional N-glycosylations (Novel erythropoiesis-stimulating protein: NESP). Wild type hEPO and NESP were cloned into expression vectors with GFP reporter gene under regulatory control of CMV promoter and IRES so that the vectors could express both rhEPO and GFP. The expression vectors were transfected to cultured CHO-K1 cells. Under microscopy, expression of GFP was visible. Using supernatant of the culture, ELISA assay, immunocytochemistry and in vitro assay using EPO dependant cell line were performed to estimate biological activity to compare the production characteristics (secretion levels, etc.) between rhEPO and NESP. The activity of NESP protein, obtained by mutagenesis, was described and compared with its rhEPO counterpart produced under same conditions. Although NESP had less secretion level in CHO cell line, the biological activity of NESP was greater than that of rhEPO. These results are consistent with previous researches. We also demonstrated that rhEPO and GFP proteins expressed simultaneously from transfected CHO cell line. Therefore we conclude that use of GFP reporter gene under IRES control could be used to screen and produce rhEPO in cultured CHO cells.

Agaricus blazeiMurill is an edible mushroom distributed in Brazil and presently cultivated in other areas, including Korea, Japan, and China. Its chemical components, including steroids and lectin and various polysaccharides have been widely studied. For this, we used U937/vector and U937/Bcl-2 cells, which were generated by transfection of the cDNA of the Bcl-2 gene. As compared with U937/vector, U937/Bcl-2 cells exhibited a 4-fold greater expression of Bcl-2. Treatment with 0.5 or 4 mg/ml A. blazei Murill for 24 h produced morphological features of apoptosis in U937/vector cells, respectively. This was associated with caspase-3 activation and PARP degradation. In contrast, A. blazei Murill-induced caspase-3 activation and PARP degradation and apoptosis were significantly inhibited by z-DEVD-fmk in U937 cells. Bcl-2 overexpressing cells exhibited sustained caspase-3 activation and expression levels of the Bcl-2 proteins during A. blazei Murill-induced apoptosis. In addition, these findings indicate that Bcl-2 inhibits A. blazei Murill-induced apoptosis by a mechanism that interferes with Bcl-2 degradation and activity of caspase-3 that is involved in the execution of apoptosi.

R-type(Cav2.3) calcium channel contributes to pain sensation in peripheral sensory neurons. Six isoforms of Cav2.3 that result from combinations of presence or deletion of three inserts(insert I and insert in the II-III loop, and insert III in N-terminal regions) have been demonstrated to be present in different mammalian tissues. However, the molecular basis of Cav2.3 in trigeminal ganglion(TG) neurons is not known. In the present study, we determined which isoforms of Cav2.3 are expressed in rat TG neurons using the RT-PCR analysis. Whole tissue RT-PCR analyses revealed that only two isoforms, Cav2.3a and Cav2.3e, were present in TG neurons. From single-cell RT-PCR, we found that Cav2.3e rather than Cav2.3a was the major isoform expressed in TG neurons, and Cav2.3e was preferentially detected in small-sized neurons that express nociceptive marker, transient receptor potential vanilloid 1(TRPV1). Our results suggest that Cav2.3e in trigeminal neurons may be a potential target for the pain treatment.

Malaysian secondary school teachers' readiness to use Information Communication Technology (ICT) is considered a critical skill in terms of national goals for schools that will prepare students to compete in the knowledge economy of the 21st century. The study investigated 303 teachers' ICT readiness in terms of their basic ICT knowledge, skills, and attitudes towards ICT. Data were collected via the Teachers' ICT Readiness instrument which consisted of an ICT knowledge test, an ICT skills test, and attitude towards ICT questionnaire. The results indicated that the majority of teachers had a moderate level of basic ICT knowledge and skills. A majority of the teachers too had a positive attitude towards ICT. Discussion and recommendations focus on the need to capitalize on the positive attitudes to turn these into action, increasing readiness to use ICT.