The NSs protein of severe fever with thrombocytopenia syndrome virus differentially inhibits the type 1 interferon response among animal species.

Severe fever with thrombocytopenia syndrome virus (SFTSV), which has been reported in China, Korea, Japan, Vietnam, and Taiwan, is a causative agent of severe fever thrombocytopenia syndrome. This virus has a high mortality and induces thrombocytopenia and leukocytopenia in humans, cats, and aged ferrets, whereas immunocompetent adult mice infected with SFTSV never show symptoms. Anti-SFTSV antibodies have been detected in several animals-including goats, sheep, cattle, and pigs. However, there are no reports of severe fever thrombocytopenia syndrome in these animals. Previous studies have reported that the nonstructural protein NSs of SFTSV inhibits the type I interferon (IFN-I) response through the sequestration of human signal transducer and activator of transcription (STAT) proteins. In this study, comparative analysis of the function of NSs as IFN antagonists in human, cat, dog, ferret, mouse, and pig cells revealed a correlation between pathogenicity of SFTSV and the function of NSs in each animal. Furthermore, we found that the inhibition of IFN-I signaling and phosphorylation of STAT1 and STAT2 by NSs depended on the binding ability of NSs to STAT1 and STAT2. Our results imply that the function of NSs in antagonizing STAT2 determines the species-specific pathogenicity of SFTSV.

Preparation and photo-induced activities of water-soluble amyloid ß-C60 complexes.

We have shown that fullerene (C60) becomes soluble in water by mixing fullerene and amyloid ß peptide (Aß40) whose fibril structures are considered to be associated with Alzheimer's disease. The water-solubility of fullerene arises from the generation of a nanosized complex between fullerene and the monomer species of Aß40 (Aß40-C60). The prepared Aß40-C60 exhibits photo-induced activity with visible light to induce the inhibition of Aß40 fibrillation and the cytotoxicity for cultured HeLa cells. The observed photo-induced phenomena result from the generation of singlet oxygen via photoexcitation, inducing oxidative damage to Aß40 and HeLa cells. The oxidized Aß40 following photoexcitation of Aß40-C60 was confirmed by mass spectrometry.

Early tissue formation on whole-area osteochondral defect of rabbit patella by covering with fibroin sponge.

Large osteochondral defects have been difficult to repair via tissue engineering treatments due to the lack of a sufficient number of source cells for repairing the defect and to the severe mechanical stresses affecting the replacement tissue. In the present study, whole-area osteochondral defects of rabbit patella were covered and wrapped with a fibroin sponge containing chondrocytes, with or without Green Fluorescent Protein (GFP) transgenic marking, on the surface facing the osteochondral defect. Five of eight osteochondral defects that were covered with the chondrocyte-seeded fibroin sponges showed hyaline cartilage-like repair containing no fibroin fragments at 6 weeks after surgery. The repaired tissue showed a layer formation, which showed intensive safranin-O and toluidine blue staining, and which showed positive type II collagen immunostaining. The average surface coverage of the repaired cartilage was 53%. On average, 48% of the cells in the repaired tissue were derived from GFP transgenic chondrocytes, which had been seeded in the fibroin sponge. The fibroin-sponge covering had the potential to allow the early repair of large osteochondral defects. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1474-1482, 2016.

[Validation study of analysis of Malachite green in broiled eels (Kabayaki) by LC-MS/MS].

An improved method for analysis of Malachite green (MG) and its metabolite, Leucomalachite green (LMG), in broiled eels without using dichloromethane was developed. This method was evaluated by means of recovery tests according to the guideline for validation of analytical methods by the Ministry of Health, Labour and Welfare of Japan. MG and LMG were extracted from spiked samples with acetonitrile and citric acid/disodium hydrogen phosphate buffer solution, and were salted-out with sodium chloride. The acetonitrile layer was dried over anhydrous sodium sulfate and purified on C18 and strongly acidic cation exchange columns. MG and LMG in the purified solution were determined quantitatively using LC-MS/MS by the internal standard method with surrogates, MG and LMG labeled with deuterium (MG-d5, LMG-d6). The recovery rates of MG and LMG were 99.2% and 93.6%, respectively. The relative standard deviations of repeatability were 2.2% for MG and 4.4% for LMG, and the relative standard deviations of within-laboratory reproducibility were 3.5% for MG and 5.1% for LMG.

[Simultaneous analysis of residual pesticides in fruit, vegetables, brown rice, and oolong tea by LC-MS/MS].

According to "The validity evaluation guideline of the testing methodology on the residual pesticides in food" of the Ministry of Health, Labour and Welfare of Japan, we carried out validity evaluation of a simultaneous analysis method for residual pesticides in various products with the QuEChERS method using a C18 mini column and/or an EnviCarb/PSA mini column for purification. Multi-ingredient simultaneous analysis with high sensitivity in a short time was achieved with LC-MS/MS. Validation was achieved for 269 pesticides among 302 pesticides in eight kinds of fruit, vegetables, and brown rice and 204 pesticides among 233 pesticides in oolong tea. Finally, 151 pesticides in oolong tea and from 234 to 259 pesticides in fruit, vegetables, and brown rice met the values specified in the guideline.

Monitoring of radioactive substances in foods distributed in Kyoto, Japan (1991-2011). - Comparison of detection rates and concentrations before and after the Fukushima Daiichi Nuclear Power Plant accident - .

Since the Chernobyl Nuclear Power Plant accident, radioactive substances have been continually monitored in foods collected in the city of Kyoto, Japan. The importance of the monitoring was increased by the Fukushima Daiichi Nuclear Power Plant accident in March 2011. Here, the detection rates and concentrations of radioactive substances were compared among food samples collected before and after the accident in Fukushima prefecture. Before the accident, (137)Cs was the only radioactive substance detected in foods. The detection rate was 70% for fish and shellfish samples and the highest concentration was 1.7 Bq/kg. It was also 83% for fresh mushroom samples and the highest concentration was 7.5 Bq/kg. In contrast, the detection rate was low for vegetables and the concentrations were also lower than those of the above samples. On the other hand, after the accident, (131)I was detected in food produced in the Tohoku and Kanto areas. Actually, (131)I (3,400 Bq/kg), (134)Cs (280 Bq/kg), and (137)Cs (280 Bq/kg) were detected in mizuna, a leaf vegetable, on March 23, 2011. These radioactive substances were detected in all leaf vegetable samples examined in March and April 2011, but they were not detected in samples examined in November 2011. (131)I was not detected in any food sample examined after May 2011. However, (137)Cs (average=7.9 Bq/kg) was consistently detected in fish and shellfish samples until November, although the concentrations were less than the regulatory limits. It appears unlikely that foods containing radioactive substances over the regulatory limits are currently being distributed in Kyoto.

Development of a novel emergency hemostatic kit for severe hemorrhage.

Photocrosslinkable chitosan (Az-CH-LA) contains lactose moieties and photoreactive azide groups, and its viscous solution forms an insoluble hydrogel on exposure to UV irradiation. We previously developed an emergency hemostatic kit using the Az-CH-LA solution, calcium alginate, and a UV irradiation apparatus. However, a suitable UV irradiation apparatus is required to effectively convert the Az-CH-LA solution into a hydrogel, and power supply to use the UV irradiation apparatus may not always be available in a disaster area or battlefield. To address this problem, we produced a portable, battery-powered UV irradiation apparatus constituting a novel hemostatic kit for severe hemorrhage. When the hemostatic kit using the UV irradiation apparatus was examined using a rat model of severe hemorrhage, the survival rate increased up to 73%. Hematological values as markers of hemorrhage did not change significantly over the first 3 days. In this study, we describe the characteristics of a portable UV irradiation apparatus and its use in an emergency hemostatic kit prepared using Az-CH-LA and calcium alginate for severe hemorrhage.

Chondrocyte distribution and cartilage regeneration in silk fibroin sponge.

Chondrocytes distribution and cartilage formation in three types of fibroin sponges with different average pore sizes (40-80, 80-120 and 100-140 µm) was measured. The image processing was performed combining two methods to identify cells automatically: extraction of local maximum luminance and multi-threshold analysis. The results showed that initial accumulation of chondrocytes localized at surface area at 3 h in the small and medium-pore groups, however, the difference in the cell distributions become equivalent until 24 h after seeding. Cartilaginous tissue was well formed in each group at 21 days, and that in the smaller pore group tend to distribute at the surface area. Spherical tissues were located at the subsurface (200-600 µm below the surface) of the sponge in the medium- and large-pore groups at 21 days. Local cell aggregation was observed at 24 h at the same depth of the fibroin sponge as the spherical tissues observed at 21 days. These results suggest that the initial cell condensation process till 24 h after seeding play an important role in cartilage tissue formation.

Regulatory impacts on stem cell research in Japan.

Although it is a leader in many fields of stem cell research, Japan's policies on many areas of stem cell research have not been widely reported or analyzed in the international literature. In this report, we provide an overview of Japan's centralized approach to regulation and analyze its policy implementation.

Familiarity and prudence of the Japanese public with research into induced pluripotent stem cells, and their desire for its proper regulation.

The lack of knowledge of current public attitudes towards basic research into induced pluripotent stem cells (iPSCs) is a serious problem when considering appropriate ways of governance regarding research and its clinical applications. We therefore conducted an internet-based survey to determine public opinion regarding the research and development of iPSCs and regenerative medicine (RM). A total of 14,908 valid responses were collected, which revealed that the Japanese public were familiar with the terms iPSCs and RM, and many of them had received information about iPSCs and RM through the television and newspapers. They also generally accepted the need for extra funding for research into iPSCs, but also decided to adopt a "wait and see" approach and thought that research and development of iPSCs and RM should be conducted under proper governance in accordance with an international regulatory framework. It will be necessary to discuss an internationally consistent regulatory system and effective mechanisms for information flow.

Multimedia presentations on the human genome: Implementation and assessment of a teaching program for the introduction to genome science using a poster and animations.

Genome science, including topics such as gene recombination, cloning, genetic tests, and gene therapy, is now an established part of our daily lives; thus we need to learn genome science to better equip ourselves for the present day. Learning from topics directly related to the human has been suggested to be more effective than learning from Mendel's peas not only because many students do not understand that plants are organisms, but also because human biology contains important social and health issues. Therefore, we have developed a teaching program for the introduction to genome science, whose subjects are focused on the human genome. This program comprises mixed multimedia presentations: a large poster with illustrations and text on the human genome (a human genome map for every home), and animations on the basics of genome science. We implemented and assessed this program at four high schools. Our results indicate that students felt that they learned about the human genome from the program and some increases in students' understanding were observed with longer exposure to the mixed multimedia presentations.

Maintenance of meiotic arrest and developmental potential of porcine oocytes after parthenogenetic activation and somatic cell nuclear transfer.

Several studies report that meiotic maturation of porcine oocytes can be reversibly preserved. The present study examined how long meiotic maturation can be suppressed. The first experiment determined the preservation medium suitable for reversibly suppressing meiotic maturation of porcine oocytes. The second experiment examined the in vitro developmental potential of oocytes maintained in meiotic arrest after parthenogenetic activation and nuclear transfer of somatic cells. Preservation of cumulus-oocyte complexes with NCSU-37 medium containing 10% follicular fluid, 1 mM dibutyryl cyclic AMP, and follicular shell pieces for 24-96 h at 39 degrees C did not affect oocyte maturation compared with controls (94-98% vs. 98%). The potential of parthenogenetically activated and nuclear-transferred oocytes maintained in meiotic arrest for 24-48 h to develop into blastocysts was not significantly different from that of controls (20-25% vs. 18% and 8-11% vs. 9%, respectively). The present study demonstrated that meiotic maturation of porcine oocytes can be suppressed after preservation for 48 h at 39 degrees C without decreasing oocyte maturation competence or the ability of oocytes to develop to at least the blastocyst stage.

Effect of demecolcine and nocodazole on the efficiency of chemically assisted removal of chromosomes and the developmental potential of nuclear transferred porcine oocytes.

Brief treatment of metaphase II (MII) stage porcine oocytes with 0.4 microg/mL demecolcine in the presence of 0.05 M sucrose produces a membrane protrusion that contains a condensed chromosome mass. The present study examined the optimal conditions for demecolcine and nocodazole treatment in chemically assisted removal of chromosomes. When matured oocytes were treated with 0.1-0.4 microg/mL demecolcine for 60 min or with 0.4 microg/mL demecolcine for 30 min or 3 microg/mL nocodazole for 30 or 60 min, more than 70% of oocytes had a membrane protrusion containing condensed chromosomes were located. There was no difference in the in vitro developmental potential of enucleated oocytes assisted by 0.1 and 0.4 microg/mL demecolcine or 3 microg/mL nocodazole that received porcine somatic cells. After transfer to 10 recipients, however, two of six recipients that received demecolcine-treated enucleated eggs produced four healthy cloned piglets, but none of the four recipients of nocodazole-treated enucleated eggs produced piglets. Further studies are required to increase the successful development to term because the proportion of live piglets was low (4/2, 672, 0.15%).

Production of cloned pigs from adult somatic cells by chemically assisted removal of maternal chromosomes.

The present study demonstrated that brief treatment of in vitro-matured porcine oocytes with demecolcine results in a membrane protrusion that contains a condensed chromosome mass, which can be easily removed by aspiration. This simple, chemically assisted method for removing maternal chromosomes enabled the production of a large number of nuclear-transferred porcine eggs. The development of eggs whose chromosomes were removed by this procedure following transfer of somatic cell nuclei to the blastocyst stage was not significantly different among groups activated using different procedures (6% to 11%) and was also not different among donor cells of different origins (3% to 9%), except for cumulus cells (0.4%). After transfer of 180 to 341 nuclear-transferred eggs that received somatic cells to 6 recipients, 2 of the recipients produced 8 healthy cloned piglets from the heart cells of a female pig. The chemically assisted method for removing maternal chromosomes was also effective for bovine and rabbit eggs.

A Web-accessible database of characteristics of the 1,945 basic Japanese kanji.

In 1981, the Japanese government published a list of the 1,945 basic Japanese kanji (Jooyoo Kanji-hyo), including specifications of pronunciation. This list was established as the standard for kanji usage in print. The database for 1,945 basic Japanese kanji provides 30 cells that explain in detail the various characteristics of kanji. Means, standard deviations, distributions, and information related to previous research concerning these kanji are provided in this paper. The database is saved as a Microsoft Excel 2000 file for Windows. This kanji database is accessible on the Web site of the Oxford Text Archive, Oxford University (http://ota.ahds.ac.uk). Using this database, researchers and educators will be able to conduct planned experiments and organize classroom instruction on the basis of the known characteristics of selected kanji.

[Effects of kana-level neighbors and phoneme-level neighbors on lexical decision of Japanese katakana words].

This study examined the effect of neighborhood size on the recognition of Japanese katakana words. Neighbors of a given katakana word were defined in two ways; kana-level neighbor is a katakana word that can be constructed by changing one letter of the target word preserving the other letter positions, and phoneme-level neighbor is a katakana word that can be created by changing one phoneme (consonant or vowel) preserving the other phoneme positions of the target. In Experiment 1, 30 university students participated in a lexical decision task and numbers of kana-level neighbor were manipulated. The result showed that the number of kana-level neighbor had an inhibitory effect on lexical decision of low familiarity katakana words. In Experiment 2, 30 university students participated in a lexical decision task and numbers of phoneme-level neighbor were manipulated. The result showed that the number of phoneme-level neighbor had no effect on lexical decision of katakana words. These results suggest that neighbors affect each other on their orthographic representation in the recognition of low familiarity Japanese katakana words.

[Effects of neighborhood size and kanji character frequency on lexical decision of Japanese kanji compound words].

The purpose of this study was to make a clear distinction between the effects of neighborhood size (Coltheart, Davelaar, Jonasson, & Besner, 1977) and that of kanji character frequency on the processing of two kanji compound words (jukugo). Kanji characters found in many compound words tend to be frequent, so neighborhood size of jukugo and the character frequency of the component kanji of these jukugo tend to be interrelated. In this study, the effect of neighborhood size on jukugo with medium subjective frequency was investigated controlling kanji character frequency. In an experiment using a lexical decision task with 30 participants, neighborhood size, front kanji character frequency, and rear kanji character frequency of jukugo were manipulated. The results showed that both the neighborhood size and the kanji character frequency affected the lexical decision of jukugo. The fact that there was no interaction between front and rear kanji character frequencies suggests that front and rear characters of jukugo with medium frequency were processed in a serial fashion.