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Miho Hamaguchi (1915-1988) was the first woman to be a licensed architect in Japan. A pioneer in domestic design during the postwar period, she built and consulted on thousands of houses throughout her prolific career. However, she is a little-known figure both in Japan and in the international debate. Her representation in architectural historiography is limited to her influence on kitchen design, but her writings and work go far beyond. Hamaguchi's legacy is one of bold residential architecture that embodied democratic ideas in spatial configurations. She promoted the house as a fundamental tool for gender equality, leaving behind a feudal and patriarchal system. At the end of her career, she found in Costa del Sol the perfect place to carry out a residential project as a cultural exchange. "Kaiyo Club" became a set of three houses since the first design in 1974 until the subsequent extensions were completed in 1987. Throughout its different stages, the project shows a striking Spanish-Japanese transfer where different architectural languages coexist. The white-walled exterior dialogues with the vernacular, while its interior unfolds Japanese patterns with tatami-floored rooms or ofuro-style bathrooms. These dwellings present a unique hybrid materialization, displaying Hamaguchi's design from a humanistic stance, blending of locally rooted modernist spatial principles and reinterpreted traditions.

Supplementary Figure 1 from IFNγ Markedly Cooperates with Intratumoral Dendritic Cell Vaccine in Dog Tumor Models

Supplementary Figure 2 from IFNγ Markedly Cooperates with Intratumoral Dendritic Cell Vaccine in Dog Tumor Models

Supplementary Figure 3 from IFNγ Markedly Cooperates with Intratumoral Dendritic Cell Vaccine in Dog Tumor Models

Supplementary Figure Legends 1-3 from IFNγ Markedly Cooperates with Intratumoral Dendritic Cell Vaccine in Dog Tumor Models

Rice blast is a devastating fungal disease resulting in major losses to rice crops. Owing to continuous acquisition of resistance by the causal fungus, several fungicide chemicals are no longer effective. Therefore, there is a need to identify natural components and develop new agents to control fungal pathogens. We previously demonstrated that the culture filtrate of Biscogniauxia sp. O821 inhibited infection behavior of Magnaporthe oryzae and subsequent blast lesion formation. In the present study, we isolated a new compound, (3aS,4aR,8aS,9aR)-3a-hydroxy-8a-methyl-3,5-dimethylenedecahydronaphto[2,3-b]furan-2(3H)-one (HDFO), from the culture filtrate of Biscogniauxia sp. O821 and determined its molecular weight as 248. The HDFO structure was determined by electrospray ionization-mass spectrometry and nuclear magnetic resonance spectroscopy after purification with column chromatography and high-performance liquid chromatography. The structure of this antifungal compound was similar to that of alantolactone and isoalantolactone. The growth inhibition zone against M. oryzae in presence of HDFO was observed at Rf 0.5–0.6 on a thin layer chromatography plate. HDFO inhibited conidial germination of M. oryzae in a dose-dependent manner (1–200 ppm). Furthermore, blast lesion formation was significantly suppressed by HDFO at over 5 ppm. These results suggest that HDFO from the culture filtrate of Biscogniauxia sp. O821 can protect rice from rice blast disease caused by M. oryzae. This is the first report that HDFO produced by Biscogniauxia sp. can serve as an antifungal compound against M. oryzae.

An ethyl acetate extract of a culture filtrate (ECF) from an unidentified fungal isolate O821 was evaluated for antifungal activity against the rice pathogen Magnaporthe oryzae. The O821-ECF significantly inhibited spore germination, appressorium formation, and mycelial growth of M. oryzae, and its antifungal activity was heat-stable. It also significantly suppressed the number and size of blast lesions. In an analysis of the ITS sequence of this isolate, it shared similarities with species of the fungus Biscogniauxia. These results suggest that isolate O821 of the genus Biscogniauxia produces a heat-stable antifungal compound(s) in its culture filtrate.

Nicotinamide mononucleotide (NMN), a precursor of nicotinamide adenine dinucleotide (NAD), induces disease resistance to the Fusarium head blight fungus Fusarium graminearum in Arabidopsis and barley, but it is unknown at which stage of the infection it acts. Since the rate of haustorial formation of an obligate biotrophic barley powdery mildew fungus Blumeria graminis f. sp. hordei (Bgh) was significantly reduced in NMN-treated coleoptile epidermal cells, the possibility that NMN induces resistance to the biotrophic stage of F. graminearum was investigated. The results show that NMN treatment caused the wandering of hyphal growth and suppressed the formation of appressoria-like structures. Furthermore, we developed an experimental system to monitor the early stage of infection in real-time and analyzed the infection behavior. We observed that the hyphae elongated windingly by NMN treatment. These results suggest that NMN potentiates resistance to the biotrophic invasion of F. graminearum as well as Bgh.

The fungal strain O821 of Biscogniauxia sp. was isolated from wild mushrooms collected in Japan. Here, we demonstrate that a 24-h pretreatment of rice leaves with an ethyl acetate extract of a culture filtrate from strain O821 (O821-ECF) reduced the severity of rice blast disease caused by infection with Magnaporthe oryzae. To investigate possible mechanisms of this effect, we examined defence responses in the leaves. O821-ECF treatment elicited the production of hydrogen peroxide (H2O2) and increased transcription of the PBZ1 gene. A time course analysis of H2O2 production indicated an increase with time after O821-ECF treatment. Expression of the PBZ1 gene similarly peaked at 24 h after O821-ECF treatment. However, no accumulation of salicylic acid (SA) was observed at 24 h after O821-ECF treatment. These results suggest that SA-independent resistance to rice blast is induced by O821-ECF. Studies of O821-ECF may contribute to the development of new plant activators for plant diseases such as rice blast disease.

Background Mindin (spondin2), a secretory protein related to neural development and immunity, is a member of thrombospondin type I repeat (TSR) superfamily proteins, and has a unique glycosylation of C-mannosylation in its structure. However, it remains unclear whether C-mannosylation plays a functional role in the biosynthesis of mindin in cells. Methods Protein C-mannosylation was analyzed by mass spectrometry. Mindin expression was examined by immunoblot and immunofluorescence analyses in COS-7 cells transfected with the expression vectors for wild type (mindin-WT) or C-mannosylation-defective mutant of mindin (mindin-mutF). The redox status was examined in mindin by using 4-acetoamide-4′-maleimidylstilbene-2,2′-disulfonate. Results When mindin cDNA was expressed in COS-7 cells, C-mannosylation of mindin was confirmed at Trp257 by mass spectrometry. In cells expressing a mindin-mutF, secretion of the mutant was significantly inhibited compared with mindin-WT. In immunofluorescence analysis, mindin-mutF was accumulated in the endoplasmic reticulum (ER), whereas mindin-WT was detected in the Golgi. In addition, mindin-mutF showed an enhanced interaction with calreticulin, an ER-resident chaperone, in cells. In cells, reduced forms were increased in mindin-mutF, compared with a mostly oxidized form of mindin-WT. In the presence of chemical chaperones such as dimethylsulfoxide or 4-phenylbutyrate, inhibited secretion of mindin-mutF was ameliorated in cells, although redox-dependent folding was not affected. Conclusions C-Mannosylation of mindin facilitates its secretion especially through modulating disulfide bond formation in mindin in cells. General significance These results suggest that C-mannosylation plays a functional role in the redox-dependent folding and transport of TSR superfamily proteins in cells.

Spore germination and appressorium formation of Magnaporthe oryzae spores was completely suppressed by an ethyl acetate extract of the culture filtrate from the H921 isolate (H921-EAE-CF). Production of antifungal substance(s) in the H921-EAE-CF began to increase up to 3 days after isolate H921 incubation. Furthermore, heat treatment (105°C or 121°C) of H921-EAE-CF did not alter its inhibitory effect on M. oryzae spore germination compared to non-heat-treated H921-EAE-CF. Blast lesion formation inhibition by H921-EAE-CF was dose-dependent. Internal transcribed spacer (ITS) region sequence analysis indicated that this isolate shared similarities with species of the genera Trichoderma. This study suggests that H921-EAE-CF contains some antifungal substances that could be promising candidates for control of rice blast disease.

Fungal strains isolated from the fruiting bodies of wild mushrooms were evaluated for fungicidal activity against Magnaporthe oryzae, the causal agent of the rice blast disease. Fungal isolates (n = 105) were obtained from 46 samples of wild mushrooms. Infection behaviors of M. oryzae were assessed in the presence of culture filtrates from 90 fungal isolates, of which 20 inhibited spore germination. Heat-treated culture filtrates of these isolates were classified into 3 groups according to biological activity. Blast lesion formation by M. oryzae was significantly inhibited by pretreatment with culture filtrates from 4 fungal isolates. ITS region sequence analysis indicated that these isolates shared similarities with species of the genera Annulohypoxylon, Nigrospora, and Penicillium. Studies of symbiotic and parasitic fungi from wild mushrooms may yield potential control agents for plant diseases such as the rice blast disease.

The existence of Th1 responses in a tumor microenvironment elicits a better prognosis for the patients. Transfection of Th1 polarizing cytokines, such as IFNγ, into tumor cells is an effective way to set up an appropriate microenvironment. Using a novel type synthetic vector composed of polyamidoamine dendrons, we transfected canine IFNγ gene into canine tumor cell lines, and examined direct and indirect effects of dendritic cells (DCs) against tumor growth in vitro. A cloned canine IFNγ gene expressed functional protein that induces maturation of DCs. When the canine IFNγ gene was transfected into canine tumor cell lines using the synthetic vector, most cells secreted canine IFNγ. Secretion of IFNγ reduced with time, but was maintained for 48h. DCs incubated with the IFNγ-transfected tumor cells exhibited greater suppressive activity and induced significantly higher cytotoxic activity against the tumor cells, relative to those incubated with untransfected tumor cells and comparable dose of IFNγ. Successful transfection of IFNγ by the synthetic vector efficiently enhanced the anti-tumor immune function of DCs, and sets up a suitable microenvironment for improvement in tumor therapy.

According to the AHA guidelines, Extracorporeal Cardiopulmonary Resuscitation (ECPR) is recommended in the management of the patients of refractory cardiac arrest. However, developing a system to initiate ECPR inside the hospital is thought to be extremely challenging, and indication criteria and

Microcell-mediated chromosome transfer (MMCT) technology enables a single and intact mammalian chromosome or megabase-sized chromosome fragments to be transferred from donor to recipient cells. The conventional MMCT method is performed immediately after the purification of microcells. The timing of the isolation of microcells and the preparation of recipient cells is very important. Thus, ready-made microcells can improve and simplify the process of MMCT. Here, we established a cryopreservation method to store microcells at −80 °C, and compared these cells with conventionally- (immediately-) prepared cells with respect to the efficiency of MMCT and the stability of a human artificial chromosome (HAC) transferred to human HT1080 cells. The HAC transfer in microcell hybrids was confirmed by FISH analysis. There was no significant difference between the two methods regarding chromosome transfer efficiency and the retention rate of HAC. Thus, cryopreservation of ready-to-use microcells provides an improved and simplified protocol for MMCT.

We investigated the effects of lasers irradiation on the exposed dentinal tubule. Human tooth specimens with exposed dentinal tubule orifices were used. Three types of lasers (CO(2) laser, Er:YAG laser and Ga-Al-As laser) were employed. The parameters were 1.0 W in continuous-wave mode with an irradiation time of 30 s for the CO(2) laser, 30 mJ in continuous-wave mode with an irradiation time of 60 s for the Er:YAG laser, and 1.0 W in continuous-wave mode with an irradiation time of 60 s for the Ga-Al-As laser. A non-irradiated group was used as a control. After laser irradiation, the dentinal surface of each sample was observed using SEM. Afterwards, all samples were immersed in methylene blue dye solution in order to evaluate the penetration of the dye solution and observe the change in dentinal permeability after laser irradiation. SEM observation showed that the control group had numerous exposed dentinal tubule orifices, whereas these orifices were closed in the laser-irradiated groups. There was consistent dye penetration into the pulp chamber in the control group, whereas no dye penetration was evident in the laser-irradiated groups. Therefore, laser appears to be a promising treatment for reducing permeation through exposed dentinal tubules.

We have been studying pulsed electric field (PEF) inactivation of various bacteria, which is based on the destruction of biological membrane due to applying pulsed electric field. In this study, we report a novel treatment system based on the textile electrode, combination of polyester fiber and 0.2 mmφ metal wire. The efficiency of PEF inactivation of Escherichia coli cells depended on the peak voltage and the conductivity of the target solutions. Higher efficiency of PEF sterilization could be achieved with 7 kV of peak voltage and lower solution conductivity. When the solution conductivity was lower, some materials, which seemed to be bacterial residue, were sedimented on high voltage electrode.

Background Human artificial chromosome (HAC) vectors have some unique characteristics as compared with conventional vectors, carrying large transgenes without size limitation, showing persistent expression of transgenes, and existing independently from host genome in cells. With these features, HACs are expected to be promising vectors for modifications of a variety of cell types. However, the method of introduction of HACs into target cells is confined to microcell-mediated chromosome transfer (MMCT), which is less efficient than other methods of vector introduction. Application of Measles Virus (MV) fusogenic proteins to MMCT instead of polyethylene glycol (PEG) has partly solved this drawback, whereas the tropism of MV fusogenic proteins is restricted to human CD46- or SLAM-positive cells. Results Here, we show that retargeting of microcell fusion by adding anti-Transferrin receptor (TfR) single chain antibodies (scFvs) to the extracellular C-terminus of the MV-H protein improves the efficiency of MV-MMCT to human fibroblasts which originally barely express both native MV receptors, and are therefore resistant to MV-MMCT. Efficacy of chimeric fusogenic proteins was evaluated by the evidence that the HAC, tagged with a drug-resistant gene and an EGFP gene, was transferred from CHO donor cells into human fibroblasts. Furthermore, it was demonstrated that no perturbation of either the HAC status or the functions of transgenes was observed on account of retargeted MV-MMCT when another HAC carrying four reprogramming factors (iHAC) was transferred into human fibroblasts. Conclusions Retargeted MV-MMCT using chimeric H protein with scFvs succeeded in extending the cell spectrum for gene transfer via HAC vectors. Therefore, this technology could facilitate the systematic cell engineering by HACs. Electronic supplementary material The online version of this article (doi:10.1186/s12896-015-0142-z) contains supplementary material, which is available to authorized users.

Enthalpies of mixing of (R)- and (S)-enantomers of liquid chiral compounds such as benzyl-(1-phenyl-ethyl)-amine (1), 1-phenylethylamine (2), 1-phenyl-ethanol (3), butyric acid oxiranylmethyl ester (4), 4-methyl-[1,3]dioxolan-2-one (5), 2-Chloromethyloxirane (6) and 3-hydroxyisobutyric acid methyl ester (7) have been measured over the whole range of mole fractions at 298.15 K, albeit very small values. Mixing of heterochiral liquids of R-1 + S-1, R-5 + S-5, and R-7 + S-7 realized enthalpic stabilization over the whole range of mole fractions, whereas that of R-2 + S-2, R-3 + S-3, R-4 + S-4, and R-6 + S-6 realized enthalpic destabilization over entire compositions. The extreme values of enthalpies of mixing and the intermolecular interaction obtained by the molecular mechanics calculations showed a linear correlation, except few the compounds measured.

Enthalpies of mixing of ( R )- and ( S )-enantiomers of liquid chiral compounds such as heptane-2-ol, octane-2-ol, nonane-2-ol, 3-chloro-propane-1,2-diol and 2-methyl-1,4-butanediol have been measured over a range of mole fractions at 298.15 K, albeit very small values. Mixing of heterochiral liquids of heptane-2-ol, octane-2-ol, nonane-2-ol, 3-chloro-propane-1,2-diol, realized enthalpic destabilization over the whole range of mole fractions, whereas that of 2-methyl-1,4-butanediol realized enthalpic stabilization over entire compositions. The maximum values of enthalpies of mixing and the intermolecular interaction of cohesive energy density and entropy of vaporization showed a linear correlation except for the compounds having two chiral centers and others.

Human artificial chromosomes (HACs) have unique characteristics as gene-delivery vectors, including episomal transmission and transfer of multiple, large transgenes. Here, we demonstrate the advantages of HAC vectors for reprogramming mouse embryonic fibroblasts (MEFs) into induced pluripotent stem (iPS) cells. Two HAC vectors (iHAC1 and iHAC2) were constructed. Both carried four reprogramming factors, and iHAC2 also encoded a p53-knockdown cassette. iHAC1 partially reprogrammed MEFs, and iHAC2 efficiently reprogrammed MEFs. Global gene expression patterns showed that the iHACs, unlike other vectors, generated relatively uniform iPS cells. Under non-selecting conditions, we established iHAC-free iPS cells by isolating cells that spontaneously lost iHAC2. Analyses of pluripotent markers, teratomas and chimeras confirmed that these iHAC-free iPS cells were pluripotent. Moreover, iHAC-free iPS cells with a re-introduced HAC encoding Herpes Simplex virus thymidine kinase were eliminated by ganciclovir treatment, indicating that the HAC safeguard system functioned in iPS cells. Thus, the HAC vector could generate uniform, integration-free iPS cells with a built-in safeguard system.

Dendritic cell (DC)–based immunotherapy can trigger effective immune responses against cancer in human patients. Although accompanied by little toxicity, further improvements are needed to optimize immune responses for fully satisfactory clinical outcomes. IFNγ, a potent inducer of T helper type 1 immune responses, is considered an important tool to realize improvements. In this study, we sought to clarify the effect of IFNγ on the maturation and activation of DCs and the clinical outcome of DC-based cancer therapy in dogs. In vitro experiments indicated that IFNγ significantly enhanced the expression of immune stimulatory molecules and interleukin-12 by DCs derived from canine monocytes. IFNγ also significantly strengthened DC-mediated growth suppression against tumor cell lines. DC inoculation with concomitant delivery of IFNγ into primary or recurrent tumors elicited significant clinical responses, including four complete responses and two partial responses against malignant tumors, also eliciting partial responses against benign but actively growing tumors. Together, our results indicate that combining IFNγ and DCs could induce strong immune responses against tumors, significantly improving clinical outcomes. The present study of dogs bearing common types of cancer in humans offers a unique line of support for the development of human cancer therapies. Cancer Res; 70(18); 7093–101. ©2010 AACR.