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Background and Objectives Ceria, one of the rare earth resources, is a potent catalytic agent using in many industrial fields. Interestingly, after converting ceria into biocompatible nanoparticle via chemical engineering, ceria nanoparticle may be used as anti-oxidant agent to treat diseases related to oxidative injuries. However, few studies have elaborated on the association between ceria nanoparticle and ischemia-reperfusion injury in experimental stroke model. Methods We used 3 nm ceria nanoparticle via the combining process using thermal decomposition and reverse micelle method. To make biocompatible, ceria nanoparticle was coated with phospholipids-PEG. We induced ischemia-reperfusion injury in SD rats via modified Longa method, and ceria nanoparticle or PBS was administered intravenously. Infarct volume and the number of apoptotic cells (TUNEL-positive cells) were measured at 24 hours. To identify the anti-oxidant role of ceria nanoparticle, hydroethidine, a marker of superoxide activity, was administered 15 minutes before inducing focal ischemia, and detect ethidine, the product of reaction between superoxide and hydroethidine, using fluorescent microscopy at 3 hours. Results We compared three doses of ceria nanoparticle and the infarct volumes were decreased dose-dependently (ρ = -0.39, p < 0.05). The infarct volumes were 130.8 mm3 in ceria-treated group (0.5 mg/kg) and 236.2 mm3 in control group (p < 0.05). The ceria-treated group exhibited a lower number of TUNEL-positive apoptotic cells (ceria-treated group: 32.5 ± 18.3 cells/mm3 ; control group: 63.4 ± 2.4 cell/mm3; p < 0.05). Ethidine (oxidezed hydroethidine) signals were weakly observed in the ischemic cortices of ceria-treated rats compared to control group. Conclusion Our study documented that ischemia-reperfusion injury was decreased by ceria nanoparticle via superoxide scavenging effects. This result may indicate the potentially protective effect of ceria nanoparticle on ischemic stroke.

Periodic alternating nystagmus (PAN) is characterized by periodic reversal of horizontal jerky nystagmus with a null period of several seconds. Damage to the uvulonodulus or to their connections with the brainstem vestibular nuclei has been suggested as a mechanism of PAN.1,2 However, PAN has rarely been reported in circumscribed cerebellar lesion.3 We report a patient with isolated nodular infarction who developed PAN without fixation in association with perverted head-shaking nystagmus (HSN) and loss of tilt suppression of the postrotatory nystagmus. This is the first report of PAN in circumscribed cerebellar infarction, and provides further evidence that PAN occurs due to dysfunction of cerebellar nodulus. A 69-year-old man with a history of hypertension developed acute vertigo with nausea/vomiting and postural imbalance. On examination, we observed spontaneous horizontal nystagmus in the primary position which reversed its direction with a cycle of approximately 2 minutes and a transition period of several seconds by using video Frenzel goggles (SLMED, Seoul, Korea) (see the video on the Neurology Web site at www.neurology.org). The …

In this study, magnetic and microwave absorbing properties of M-hexaferrites have been investigated in Ka-band frequencies with the substitution of Ti+4 and Co+2 ions into Fe+3 sites. The powders of composition BaFe12-2xTixCoxO19 were prepared by calcining at 1250 ringC in air. Saturation magnetization and coercivity were measured using a vibration sample magnetometer.

Mesenchymal stem cells (MSC) have been known as useful donor cells for somatic cell nuclear transfer (SCNT). It has been suggested that the culture condition of donor cells causes different results on preimplantation development of SCNT embryos. In this study, we investigated the patterns of gene expression of adipose-derived mesenchymal stem cells (ad-MSC) in different culture media (DMEM and RKME), and examined the effect of ad-MSC, with the gene expression changed, used as donor cells on the preimplantation development of cloned embryos. Canine ad-MSC were isolated from fat tissue of 3-year-old female beagle and were cultured in DMEM supplemented with 10% fetal bovine serum (MSC-DMEM) and RKME (MSC-MSC) provided from RNL Bio Corp. (Seoul, Korea). Total RNA was extracted from ad-MSC cultured in each culture medium. After synthesising cDNA of each sample, quantitative RT-PCR was done according to the Takara Bio Inc. guidelines and using the 7300 Real Time PCR Cycler System (Applied Biosystems, Carlsbad, CA, USA). The level of all tested gene transcription was normalized to β-actin expression levels. The relative quantification of gene expression was analysed by the 2–ΔΔCt method. The data from all experiments were analysed by Student’s t-test using a statistical analysis GraphPad Prism 4.02 (GraphPad Software Inc., San Diego, CA, USA). Significance was determined at P 0.05). These results demonstrate that the gene expression of ad-MSC can be modified, by culture media, into a state where reprogramming is easily done. Even so, ad-MSC with gene expression changed by culture medium did not influence the developmental ability of blastocysts. In conclusion, the alteration of gene-related stemness and reprogramming in canine ad-MSC would not be able to effectively control reprogramming in SCNT. This study was supported by RDA (#PJ0089752012), RNL Bio (#550-20120006), IPET (#311062-04-1-SB010), Research Institute for Veterinary Science, and Nestlé Purina Korea.

It is well known that very early development of the mammalian pre-implantation embryo is regulated by gene transcripts and proteins stored in the oocyte and that the embryonic genome gains control of development following 1 to 3 cleavage divisions. An active transcription and translation is required for chromatin condensation and germinal vesicle breakdown in pig oocyte. The transition from maternal to embryonic control of development is a gradual event, and following this transition, the maternally derived transcripts and proteins are gradually degraded. Successful embryonic development is dependent on the temporal and stage-specific expression of proper genes, but information on specific gene expression during early stages before zygotic gene activation (ZGA) is limited. Before activation of the embryonic genome, mRNA and proteins synthesized during oocyte growth and maturation contribute to early development. In this study, we compared the mRNA transcripts level among porcine immature, in vitro-matured and cleaved 2- to 4-cell stage embryos after in vitro fertilization to identify genes that show differential mRNA transcript levels during maturation and very early embryonic development. For the first strand cDNA synthesis, oligo (dT) primers were added to the total RNA isolated from each sample. Using annealing control primer (ACP)-based GeneFishing PCR, we detected tens of different genes showing differential mRNA transcript level (DRTL) and nine DRTL genes were identified to be KCRF, CAMSAP1, SMP1, FLJ20647, LOC132321, NADH1, NADH6, HERC3, and TEGT. Of 9 DRTL genes, TEGT showed higher mRNA transcript level at the immaturation stage, and mRNA transcript levels of the other 8 genes were increased after in vitro maturation. Therefore, we focused on TEGT (testis enhanced gene transcript), which is highly expressed in testis and also in oocytes before in vitro maturation. Differential mRNA transcripts pattern of CAMSAP1 and TEGT were confirmed using RT-PCR and real-time RT-PCR. Porcine TEGT (pTEGT) was cloned and sequenced to have an ORF of 714 bp nucleotides and to encode an integral membrane protein. When overexpressed in HEK293 cells, pTEGT suppressed apoptosis induced by etoposide. We found that pTEGT, but not TEGT-C (C-terminal deletion mutant), inhibited etoposide- and staurosporine-induced cell death. Next, we found that introduction of TEGT siRNA suppressed the anti-apoptotic effect of TEGT. Interestingly, expression of TEGT suppressed etoposide-induced ERK activation, suggesting that ERK phosphorylation is involved in the anti-apoptotic function of the gene. Several reports showed that apoptosis and MAP kinase signaling pathways play important roles in oocyte maturation and early embryo development. Therefore, the anti-apoptotic effect of TEGT was suggested to play a key role in the normal ooctye maturation and early embryo development. This work was supported by the Research Project on the Production of Bio-organs, Ministry of Agriculture and Forestry, Republic of Korea.

The complement system is composed of a complex group of soluble proteins that have important roles in the immune response against foreign cells such as xenografted tissue. Some of the cell surface regulators, also known as membrane complement regulatory proteins (CRPs), are the membrane cofactor protein (CD46), the decay-accelerating factor (CD55), and protectin (CD59). The CD55 is a 70 kDa glycolipid-anchored membrane-bound protein that has regulatory activity by preventing C3 convertase formation and is reported to be expressed in blood and vascular endothelial cells. Its high expression in leukocytes and endothelium is thought to safeguard against locally augmented C3 activation that can occur with inflammation. In this study, we originally cloned the 1.1-kb promoter region of the CD55 gene and constructed a luciferase reporter plasmid, pGL3-1.1CD55. The pGL3-1.1CD55 plasmid and its control plasmid, pGL3-control (SV40 promoter), were transfected into endothelial (MS-1 and BAEC) and epithelial (HaCaT and HEK293) cells, and the soluble fraction of the cell lysates was assayed for luciferase activity. Its promoter activity was compared with that of the promoters of endothelial-specific genes such as MCP, Flk-1, ICAM-2, and throbomodulin. Luciferase activity in each sample was normalized to the �-galactosidase activity of the same sample and the data were analyzed by Sigma Plot program (P < 0.01 versus control). Our results showed that the 1.1 kb CD55 promoter was the strongest among the endothelial cell-specific promoters. To define the important region for the strong expression, the deletion constructs containing 0.96, 0.86, and 0.74, CD55 promoter regions were prepared. Relative to the activity of the control SV40 promoter, decreased luciferase activity was obtained with these deletion constructs, suggesting that the about 0.2 kb 52-flanking region (between -1125 and -967) of the 1.1 kb CD55 promoter region was important for the strong gene expression. Interestingly, on the 52-end of the 0.2 kb region, we could detect two GATA-1 binding sites (from -1122 to -1118 and from -1111 to -1107); the deletion of the sequences between -1125 and -1100 (ΔGATA-1) significantly reduced the promoter activity. Furthermore, ectopic expression of GATA-1 dose-dependently induced the 1.1 kb CD55 promoter activity, but not the ΔGATA-1 activity, strongly implying that the GATA-1 binding site is important for the strong expression. Finally, we confirmed the binding of GATA-1 on the CD55 promoter region with the electrophoretic mobility shift assay (EMSA) using the 28 base pair oligonucleotide probes corresponding to the GATA-1 binding site and chromatin immunoprecipitation (Ch-IP) using anti-GATA-1 antibody. Taken together, these results strongly suggested that GATA-1 plays a critical role in endothelial cell-specific expression of the 1.1 kb CD55 promoter region. This work was supported by the Research Project on the Production of Bio-organs, Ministry of Agriculture and Forestry, Republic of Korea, and by the Korea Research Foundation Grant funded by the Korean Government (MOEHRD) (KRF-2005-070-C00095).