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It was found that lanthanum metal caused the dehalogenation of o -dihalogen substituted arenes and α,α′-dihalogen substituted o -xylenes to generate the corresponding benzynes and o -quinodimethanes. When o -dihalogen substituted arenes were allowed to react with lanthanum metal in the presence of dienes, the Diels–Alder products between benzyne and dienes were formed in moderate to good yields. Similarly, the Diels–Alder adducts of o -quinodimethane with dienophiles were obtained, in the reaction of α,α′-dibromo- o -xylenes with lanthanum metal in the presence of dienophiles.

The structures and electronic states of acetone–metal complexes (Ac–M, where M=Ga, Al, and B) have been calculated by means of ab-initio DFT and configuration interaction (CI) calculations in order to shed light on the mechanism of the electron conductivity and doping effects. It was found that the electronic states of Ac–Ga and Ac–Al at the ground state are composed of ion-pair state expressed approximately by (Ac δ− )(M δ+ ): the electron is transferred from metal to the carbonyl group, suggesting that the carbonyl compound interacting with Ga and Al behaves as an n-type semiconductor. In the case of the Ac–B complex, on the other hand, the electron on Ac is significantly transferred to the boron atom, expecting that hole is transferred in the boron-doped carbonyl compound (p-type semiconductor). In these complexes, the first electronic transition is a charge-transfer band between metal and carbonyl group. The mechanism of the electronic conductivity was discussed on the basis of theoretical results.

In the new World Health Organization (WHO) classification of malignant lymphoma, anaplastic large cell lymphoma of B-cell phenotype is classified either as the anaplastic large cell variant of diffuse large B-cell lymphoma or as Hodgkin's lymphoma. A 71-year-old Japanese man developed fever and generalized lymphadenopathy. Biopsy of the right axillary node revealed morphology of malignant lymphoma in which large cells with abundant cytoplasm and pleomorphic nuclei were scattered among small lymphocytes. Immunostaining with various monoclonal antibodies revealed the large cells to be CD79+, CD20/L26+, CD45RO/UCHL-(1-), CD3-, CD10-, CD30+, NPM/ALK-, EMA-, CD15-, and bcl-(2-). Amplification of the J region of the immunoglobulin heavy chain by polymerase chain reaction revealed a single rearranged band. Therefore the diagnosis of anaplastic large cell variant of diffuse large B-cell lymphoma, stage IIIB, was made from the standpoint of the new WHO classification of malignant lymphoma. Biopsy led to findings of Epstein-Barr virus (EBV)-associated lymphoma with positive in situ hybridization results for EBV small RNAs, positive results of immunostaining with EBV latent membrane 1 antibody, and negative results of immunostaining with Epstein-Barr nuclear antigen 2. Results of immunostaining of the mass with p53 antibody also were positive for lymphoma cells. The findings in this case may suggest a close relationship between p53 expression and latent EBV infection.

Direct ab initio trajectory calculations have been applied to the ionization (i.e., hole-capture) processes of biphenyl (Bp) in order to shed light on hole-capture processes of Bp and poly(4-vinylb...

Ab-initio DFT and configuration interaction (CI) calculations have been carried out for indium–carbonyl compounds, such as acetone–In (Ac–In), 1,8-naphthalic anhydride–In (In2NA) and 3,4,9,10-perylenetetracarboxylic dianhydride–indium (In4PTCDA), in order to shed light on the mechanism of electronic conductivity in the molecular devices. It was found that the electronic state of these complexes at the ground state is composed of ion-pair states expressed approximately by (Acδ−)(Inδ+), (In2)δ+(NA)δ− and (In4)δ+(PTCDA)δ−, respectively. On the other hand, the interaction at the first excited state was changed to a van der Waals interaction. Namely, the electron returned again to the In atom by electronic excitation. The second excited state is attributed to internal excitation within the carbonyl anion, while the charge on the indium was close to neutral. The mechanism of the electronic conductivity was discussed on the basis of theoretical results.

Structures and electronic states of a molecular complex formed between fluorenone and sodium atom (denoted by FL–Na) have been investigated by means of ab initio density functional theory (DFT). The most stable structure of FL–Na has a Cs symmetry where the sodium atom is located on the Cs plane perpendicular to the molecular plane of FL where all atoms are located. The sodium atom binds to the carbonyl oxygen of FL and the angle of Na–O–C is calculated to be 157° at the B3LYP/6-311+G(d) level. The charge on the Na atom shows a positive value (∼+0.80), and unpaired electron is delocalized over the FL molecule, indicating that the FL–Na complex exists as an ion-pair state expressed by (CO)δ−(Na)δ+ at the ground state (δ∼+0.80). The proton hyperfine coupling constants and excitation energies calculated at the B3LYP/6-311+G(d) and B3LYP/6-31++G(d,p) levels are in good agreement with previous experiments. The electronic states at the ground and first excited states were discussed.

Although the α-chymases of primates and dogs are known as chymotrypsin-like proteases, the enzymatic properties of rodent α-chymases (rat mast cell protease 5/rMCP-5 and mouse mast cell protease 5/mMCP-5) have not been fully understood. We report that recombinant rMCP-5 and mMCP-5 are elastase-like proteases, not chymotrypsin-like proteases. An enzyme assay using chromogenic peptidyl substrates showed that mast cell protease-5s (MCP-5s) have a clear preference for small aliphatic amino acids (e.g. alanine, isoleucine, valine) in the P1 site of substrates. We used site-directed mutagenesis and computer modeling approaches to define the determinant residue for the substrate specificity of mMCP-5, and found that the mutant possessing a Gly substitution of the Val at position 216 (V216G) lost elastase-like activity but acquired chymase activity, suggesting that the Val216 dominantly restricts the substrate specificity of mMCP-5. Structural models of mMCP-5 and the V216G mutant based on the crystal structures of serine proteases (rMCP-2, human cathepsin G, and human chymase) revealed the active site differences that can account for the marked differences in substrate specificity of the two enzymes between elastase and chymase. These findings suggest that rodent α-chymases have unique biological activity different from the chymases of other species.

A 64-yr-old Japanese man presented with mild anemia, leukocytosis, and thrombocytosis in November 1999. A diagnosis of chronic myeloid leukemia was made with a positive Ph chromosome, and interferon α treatment was started, 6 million units a day. Two years later, in October 2001, the patient developed leukocytosis, an increased LDH level, and large blasts with basophilic cytoplasm with cytoplasmic projections appeared in the peripheral blood. Bone marrow aspiration revealed increased blasts (59.6%). These blasts were negative on peroxidase stain, positive on acid phosphatase, and weakly positive on α naphthyl butyrate esterase stain and periodic acid-Schiff stain. Immunohistochemical staining with monoclonal antibodies revealed that these blasts were strongly positive with anti-CD41 (glycoprotein IIb/IIIa), weakly positive with CD7, CD33, and CD34, and negative with other monoclonal antibodies. A diagnosis of megakaryoblastic transformation from chronic myeloid leukemia was therefore made. Two-color fluorescence in situ hybridization (FISH) for portions of the major-bcr and abl genes from bone marrow cells revealed two fused signals in 90.6% and one fused signal in 5.8% of 106 cells. A cytogenetic study revealed that bone marrow cells were 69, XYY, + 6, -7, + 8, -9, t(9;22)(q34;q11), + 11, + 13, - 15, - 16, dic(17;18)(p11;p11), - 18, + 19, + 21, der(22)t(9;22) in six of nine examined cells. These findings confirmed that these megakaryoblasts originated from megakaryocytes of the chronic myeloid leukemia clone.

Summary. Expression of the transferrin receptor (TfR) is controlled essentially by post-transcriptional regulation, which is mediated by the interaction between iron regulatory proteins (IRPs) and iron-responsive elements (IREs) located in the 3′-untranslated region (UTR) of TfR mRNA. We examined whether additional controls of expression might occur by investigating the regulation of TfR in the absence of its' IREs by cloning 3′-UTR truncated TfR cDNA (TfRΔIRE) into pcDNA3·1(+) and stably expressing it in CHO-TRVb (Chinese Hamster Ovary, TfR variant cells, clone b) cells which have no endogenous TfR. Both the surface and the total cell TfR protein levels increased significantly after treating the stably transfected cells with the iron chelator desferrioxamine (DFO) compared with the control cells. In contrast, expression of the transferrin receptor 2 (TfR2), which was also cloned in the same vector and stably expressed in the TRVb-CHO cells, was not regulated by iron chelation. In addition, TfR2 fused to the 3′-UTR of TfR (TfR2-IRE) was cloned into the same vector and stably expressed in the CHO-TRVb cells. A comparison showed that expression of full-length TfR and TfR2-IRE increased by fivefold and twofold, respectively, after addition of DFO. Further experiments showed that TRVb-CHO cells, which were stably transfected with TfRΔIRE and exposed to either normal medium or DFO, resulted in similar levels of TfR mRNA, but the TfR protein biosynthetic rate increased with iron chelation. In summary, expression of TfR in cells placed in an iron-deficient milieu can increase by a mechanism which is independent of the IREs of TfR.

Fish have a complex self-defense mechanism against microbial invasion. Recently, l-lysine α-oxidases have been identified from a number of fish species as a novel type of antibacterial protein in the integument. These enzymes exhibit strict substrate specificity for l-lysine, but the underlying mechanisms and details of their catalytic properties remain unknown. In this study, a synthetic gene coding for Scomber japonicus l-lysine α-oxidase, originally termed AIP (for apoptosis-inducing protein), was expressed in Pichia pastoris, and the recombinant enzyme (rAIP) was purified and characterized. rAIP exhibited essentially the same substrate specificity as the native enzyme, catalyzing the oxidative deamination of l-lysine as an exclusive substrate. rAIP was N-glycosylated and remained active over a wide range of pH, with an optimal pH of 7.5. The enzyme was stable in the pH range from 4.5 to 10.0 and was thermally stable up to 60°C. A molecular modelling of rAIP and a comparative structure/sequence analysis with homologous enzymes indicate that Asp(220) and Asp(320) are the substrate-binding residues that are likely to confer exclusive substrate specificity for l-lysine on the fish enzymes.

L-Pipecolic acid is a key component of biologically active molecules and a pharmaceutically important chiral building block. It can be stereoselectively produced from L-lysine by a two-step bioconversion involving L-lysine α-oxidase and ∆(1)-piperideine-2-carboxylae (Pip2C) reductase. In this study, we focused on an L-lysine α-oxidase from Scomber japonicus that was originally identified as an apoptosis-inducing protein (AIP) and applied the enzyme to one-pot fermentation of L-pipecolic acid in Escherichia coli. A synthetic gene coding for an AIP was expressed in E. coli, and the recombinant enzyme was purified and characterized. The purified enzyme was determined to be a homodimer with a molecular mass of 133.9 kDa. The enzyme essentially exhibited the same substrate specificity as the native enzyme. Optimal temperature and pH for the enzymatic reaction were 70 °C and 7.4, respectively. The enzyme was stable below 60 °C and at a pH range of 5.5-7.5 but was markedly inhibited by Co(2+). To establish a one-pot fermentation system for the synthesis of optically pure L-pipecolic acid from DL-lysine, an E. coli strain carrying a plasmid encoding AIP, Pip2C reductase from Pseudomonas putida, lysine racemase from P. putida, and glucose dehydrogenase from Bacillus subtilis was constructed. The one-pot process produced 45.1 g/L of L-pipecolic acid (87.4 % yield from DL-lysine) after a 46-h reaction with high optical purity (>99.9 % enantiomeric excess).

Hepcidin is the central regulator of systemic iron homeostasis; dysregulation of hepcidin expression causes various iron metabolic disorders, including hereditary hemochromatosis and anemia of inflammation. To identify molecules that modulate hepcidin expression, we developed a bioassay system for hepcidin gene (HAMP) promoter activity by stable transfection of Hep3B hepatoma cells with an expression plasmid in which EGFP was linked to a 2.5-kb human HAMP promoter. Interleukin 6, bone morphogenetic protein 6 (BMP-6), and oncostatin M, well-characterized stimulators of the HAMP promoter, strongly enhanced the green fluorescence intensity of these cells. Dorsomorphin, heparin, and cobalt chloride, known inhibitors of hepcidin expression, significantly suppressed green fluorescence intensity, and these inhibitory effects were more prominent when the cells were stimulated with BMP-6. Employing this system, we screened 1,280 biologically active small molecules and found several candidate inhibitors of hepcidin expression. Apomorphine, benzamil, etoposide, CGS-15943, kenpaullone, and rutaecarpine (all at 10 μmol/L) significantly inhibited hepcidin mRNA expression by Hep3B cells without affecting cell viability. CGS-15943 was the strongest suppressor of BMP-6-induced hepcidin-25 secretion in these cells. We conclude that our newly developed hepcidin promoter bioassay system is useful for identifying and evaluating compounds that modulate hepcidin expression.

Objective To compare the effect of tocilizumab (TCZ) with other biologic therapies in improving anemia of rheumatoid arthritis (RA) patients. Methods We compared the change of hemoglobin (Hb) levels in a cohort of 147 consecutive RA patients who were treated with biologics for more than 12 weeks. Twenty eight patients were treated with TCZ, and 119 patients were treated with biologics other than TCZ (87 with TNF inhibitors and 32 with abatacept). The change of Hb levels from baseline to week 12 was compared between the TCZ and the non-TCZ groups. We performed univariate and multivariate analyses with adjustment of potential confounders such as baseline characteristics, concomitant treatment, and the clinical response to treatment. Results Hb levels generally increased after biologic therapies both in the TCZ and the non-TCZ groups. The increase of Hb levels was greater in the TCZ group than in the non-TCZ groups (1.1 g/dL in the TCZ group vs 0.3 g/dL in the non-TCZ group, p = 0.009). Univariate analysis revealed that increase of Hb levels was also significantly associated with lower Hb, higher Low Hemoglobin Density, and higher CRP levels at baseline and greater reduction in the clinical disease activity index. TCZ therapy was significantly associated with the increase of Hb levels even after adjustment for these factors by multivariate analysis (p

Aneuploidy is a characteristic of the majority of human cancers, and recent studies suggest that defects of mitotic checkpoints play a role in carcinogenesis. MAD1L1 is a checkpoint gene, and its dysfunction is associated with chromosomal instability. Rare mutations of this gene have been reported in colon and lung cancers. We examined a total of 44 cell lines (hematopoietic, prostate, osteosarcoma, breast, glioblastoma and lung) and 133 fresh cancer cells (hematopoietic, prostate, breast and glioblastoma) for alterations of MAD1L1 by RT-PCR-SSCP and nucleotide sequencing. Eight mutations consisting of missense, nonsense and frameshift mutations were found, together with a number of nucleotide polymorphisms. All the alterations in cell lines were heterozygous. Frequency of mutations was relatively high in prostate cancer (2/7 cell lines and 2/33 tumor specimens). We placed a mutant truncated MAD1L1, found in a lymphoma sample, into HOS, Ht161 and SJSA cell lines and found that it was less inhibitory than wild type MAD1L1 at decreasing cell proliferation. Co-expression experiments showed that the mutant form had a dominant-negative effect. Furthermore, this mutant impaired the mitotic checkpoint as shown by decreased mitotic indices in HOS cells expressing mutant MAD1L1 after culture with the microtubule-disrupting agent, nocodazole. Our results suggest a pathogenic role of MAD1L1 mutations in various types of human cancer.

Electric field effects on photoinduced electron-transfer processes have been examined in a polymer film for methylene-linked compounds of phenanthrene and N,N-dimethylaniline, based on the measurements of the field effects on fluorescence spectra. Both intra- and intermolecular electron-transfer processes are enhanced by an electric field (F), depending on the methylene chain length, and LE fluorescence emitted from the locally excited state of phenanthrene chromophore is quenched by F. Exciplex fluorescence is also influenced by F. The magnitude of the change in molecular polarizability following fluorescence process has been determined from the Stark shift both for LE fluorescence and for exciplex fluorescence.

Fluorescence emitted from phenanthrene linked to phthalimide by a short methylene chain is unusually enhanced by an electric field in a polymer film. Back-electron transfer which produces the fluorescent state of phenanthrene occurs through a short methylene bond following photoinduced electron transfer from phenanthrene to phthalimide, and its efficiency is enhanced by an electric field.

Myelodysplastic syndrome (MDS) is a heterogeneous group of hematopoietic stem cell disorders characterized by ineffective hematopoiesis and a high propensity to transform into acute myeloid leukemia (AML). Although hematopoietic stem cell transplantation can cure some MDS patients, most patients are not eligible to undergo the treatment because of advanced age or comorbidities. In order to choose the optimal treatments, predicting prognosis is crucial. Chromosomal karyotype, bone marrow (BM) blast percentage, and cytopenia are well-known prognostic factors for MDS patients. These factors are incorporated into the international prognostic scoring system and its revised version (IPSS-R). However, the impact of morphological abnormalities has not been fully determined. Thus, to elucidate this, we examined the influence of the number of dysplastic lineages and the presence of increased ring sideroblasts (RS) on the survival of MDS patients whose BM blast percentages were Since 2006, the National Research Group on Idiopathic Bone Marrow Failure Syndromes, Japan, has been conducting the Prospective Registration, Central Review and Follow-up Study for Aplastic Anemia and Myelodysplastic Syndrome. Patients with BM blasts of By May 2016, 351 cases had been registered to the depository. After central review, 216 cases were diagnosed as MDS according to definition by the French-American-British Cooperative Group. Cases that were not registered within 1 year after the time of institutional diagnosis and cases with follow-up periods of 15% RS in erythroblasts. The mean BM blast percentage was significantly lower in patients with RCUD than in those with RCMD, consistent with the result of our pervious study that used a different cohort where the BM blast percentages in patients with RA according to the 2008 WHO classification were significantly lower than those in patients with RCMD (Matsuda A, et al. Eur J Haematol. 2013;90:79-80). The 2-year OS of patients with RCUD was significantly higher than that of patients with RCMD (84% vs 76%, P = 0.014; Fig. 1a), which is consistent with the results of a German study (Maassen A, et al. Leuk Res. 2013;37:64-70). The cumulative incidence of progression to AML tended to be higher in RCMD than in RCUD, although the difference was not statistically significant (P = 0.0656, Fig. 1b). Among patients with RCMD, those with increased number of RSs showed significantly lower hemoglobin levels and higher cytogenetic scores according to the IPSS-R definition than those without increased RS (Table 2). The 2-year OS of the former was significantly inferior to that of the latter in our cohort (58% vs 87%, P = 0.009; Fig. 1c). Increased number of RSs is known to be associated with SF3B1 mutations, which in turn have been shown to be associated with favorable prognosis. However, the results of the present study indicate that in RCMD, an increase in the number of RSs may be an unfavorable indicator of patient survival. Disclosures Ozawa: Celgene Japan: Consultancy; Sumitomo Dainippon Pharma Co. Ltd: Research Funding; JCR Pharmaceutical Inc: Consultancy; Takara Bio Inc: Research Funding. Takaori-Kondo:Pfizer: Research Funding; Eisai: Research Funding; Takeda Pharmaceutical: Research Funding; Astellas Pharma: Research Funding; Kyowa Kirin: Research Funding; Cognano: Research Funding; Toyama Chemical: Research Funding; Mochida Pharmaceutical: Research Funding; Shionogi: Research Funding; Alexion Pharmaceuticals: Research Funding; Chugai Pharmaceutical: Research Funding; Janssen Pharmaceuticals: Speakers Bureau; Merck Sharp and Dohme: Speakers Bureau; Bristol-Myers Squibb: Speakers Bureau.

In most cells, transferrin receptor (TfR1)-mediated endocytosis is a major pathway for cellular iron uptake. We recently cloned the human transferrin receptor 2 (TfR2) gene, which encodes a second receptor for transferrin (Kawabata, H., Yang, R., Hirama, T., Vuong, P. T., Kawano, S., Gombart, A. F., and Koeffler, H. P. (1999) J. Biol. Chem. 274, 20826-20832). In the present study, the regulation of TfR2 expression and function was investigated. A select Chinese hamster ovary (CHO)-TRVb cell line that does not express either TfR1 or TfR2 was stably transfected with either TfR1 or TfR2-alpha cDNA. TfR2-alpha-expressing cells had considerably lower affinity for holotransferrin when compared with TfR1-expressing CHO cells. Interestingly, in contrast to TfR1, expression of TfR2 mRNA in K562 cells was not up-regulated by desferrioxamine (DFO), a cell membrane-permeable iron chelator. In MG63 cells, expression of TfR2 mRNA was regulated in the cell cycle with the highest expression in late G(1) phase and no expression in G(0)/G(1). DFO reduced cell proliferation and DNA synthesis of CHO-TRVb control cells, whereas it had little effect on TfR2-alpha-expressing CHO cells when measured by clonogenic and cell cycle analysis. In addition, CHO cells that express TfR2-alpha developed into tumors in nude mice whereas CHO control cells did not. In conclusion, TfR2 expression may be regulated by the cell cycle rather than cellular iron status and may support cell growth both in vitro and in vivo.

四国北東部の七宝山体, 弥谷山体, 我拝師山体を構成する瀬戸内火山岩類は, 下位に流紋岩質火砕岩・溶岩, 上位に安山岩質～デイサイト質溶岩・火砕岩が累重する共通の層序を示す.流紋岩質火砕岩の特徴から, 流紋岩質岩の活動時, 本地域には水域が点在し, 火砕岩は起伏のある基盤地形を埋めるように堆積したと推定される.一方, 安山岩質～デイサイト質岩のうち, 普通角閃石や黒雲母などの含水鉱物を斑晶に含む岩石は, これら火山岩類の活動時期のより初期に集中して噴出した.上記の3山体は, 各山体内の火道からもたらされた噴出物が重なることで形成された複合火山といえる.また, 瀬戸内火山岩類中の複合溶岩を構成する岩石は, 玄武岩からデイサイトに限られており, 流紋岩は複合溶岩の構成メンバーになっていない.弥谷山体にみられる複合溶岩は, 岩脈状のマグマ溜まり内で成層していた組成の異なるマグマが混合することで形成された可能性がある.

The interaction of magnesium (Mg) with a graphene surface has been investigated by means of density functional theory (DFT) method. Also, the direct molecular orbital–molecular dynamics (MO-MD) calculation [Tachikawa, J. Phys. Chem. C, 112 (2008) 10193] have been applied to the dynamics of Mg on the graphene surface. The B3LYP/LANL2MB calculations showed that the Mg atom is bound to a hexagonal site and is located in 2.02 A from the graphene surface. The direct MO-MD calculations showed that the Mg atom vibrates in the hexagonal site and diffusion does not occur even at 1000 K. The nature of the interaction between the Mg atom and the graphene sheet was discussed on the basis of theoretical results.

Excimer fluorescence of 1,3-bis-(1-pyrenyl)propane (Py-Py) doped in a PMMA polymer film with a peak at ∼470 nm is quenched by an external electric field extremely well. It is suggested that a field-assisted charge separation of Py-Py followed by efficient charge transports occurs only in the excimer with a sandwich-type configuration. The electroluminescence spectrum which is similar to the excimer fluorescence is generated at high fields without photoirradiation, probably by a radiative recombination of the hole–electron pair through a short methylene chain. A good correlation is found between the efficient fatigue of the excimer fluorescence and the generation of the electroluminescence.

Transferrin receptor (TfR) plays a major role in cellular iron uptake through binding and internalizing a carrier protein transferrin (Tf). We have cloned, sequenced, and mapped a human gene homologous to TfR, termed TfR2. Two transcripts were expressed from this gene: α (∼2.9 kilobase pairs), and β (∼2.5 kilobase pairs). The predicted amino acid sequence revealed that the TfR2-α protein was a type II membrane protein and shared a 45% identity and 66% similarity in its extracellular domain with TfR. The TfR2-β protein lacked the amino-terminal portion of the TfR2-α protein including the putative transmembrane domain. Northern blot analysis showed that the α transcript was predominantly expressed in the liver. In addition, high expression occurred in K562, an erythromegakaryocytic cell line. To analyze the function of TfR2, Chinese hamster ovary TfR-deficient cells (CHO-TRVb cells) were stably transfected with FLAG-tagged TfR2-α. These cells showed an increase in biotinylated Tf binding to the cell surface, which was competed by nonlabeled Tf, but not by lactoferrin. Also, these cells had a marked increase in Tf-bound 55Fe uptake. Taken together, TfR2-α may be a second transferrin receptor that can mediate cellular iron transport.

An NADPH-dependent aldehyde reductase was purified to homogeneity from Candida magnoliae AKU4643 through four steps, including Blue-Sepharose affinity chromatography. The relative molecular mass of the enzyme was estimated to be 33,000 on high performance gel-permeation chromatography and 35,000 on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The substrate specificity of the enzyme was broad and resembled those of other aldo–keto reductases. The partial amino acid sequences of the enzyme showed that it belongs to the aldo–keto reductase superfamily. The enzyme catalyzed the stereoselective reduction of ethyl 4-chloro-3-oxobutanoate to the corresponding ( R )-alcohol, with a 100% enantiomeric excess. The enzyme was inhibited by 1 mM quercetin, CuSO 4 , ZnSO 4 and HgCl 2 . The thermostability of the enzyme was inferior to that of the ( S )-CHBE-producing enzyme from the same strain.

Multiple ethyl 4-chloro-3-oxobutanoate (COBE)-reducing enzymes were isolated from a cell-free extract of Candida magnoliae. A NADPH-dependent COBE-reducing enzyme, distinct from the carbonyl reductase and aldehyde reductase previously reported, was purified to homogeneity using five steps, including polyethylene glycol treatment. The relative molecular mass of the enzyme was estimated to be 86,000 on high performance gel-permeation chromatography and 29,000 on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme catalyzed the reduction of COBE to the corresponding (S)-alcohol with a 51% enantiomeric excess. The substrate specificity of the enzyme was different from those of the other COBE-reducing enzymes of the same strain. The partial amino acid sequences of the enzyme showed that it belongs to the short chain alcohol dehydrogenase/reductase (SDR) family. This is the first report of multiple COBE-reducing enzymes with various stereoselectivities occurring in the same strain but belonging to different (super)families.

The β-hydride elimination of diethylzinc (DEZn) is investigated as a decomposition process of DEZn from the viewpoint of quantum chemistry. There are two steps in the β-hydride elimination of DEZn, as we determined from experimental results. Transition states are fully optimized at each β-hydride elimination step, and the energies of these transition states are about 45 kcal/mol higher than their reactants. The products of the two β-hydride elimination steps have higher energies of about 20 kcal/mol than their reactants. These energy profiles agree well with our experimental results indicating that DEZn is decomposed by the two β-hydride elimination steps at 300 and 650 °C and that such decomposition requires thermal energy for the reactions to proceed. On the other hand, about 55 kcal/mol is required to eliminate an ethyl radical from DEZn. Therefore, the main decomposition process of DEZn is β-hydride elimination.

The electronic states of potassium ion and atom (K+ and K) adsorbed on the amorphous carbon surfaces have been investigated by means of hybrid density functional theory (DFT) calculation to elucidate the nature of interaction between K+/K and the amorphous carbon. Seven graphene sheets (n = 7, 14, 19, 29, 37, 44, and 52, where n means numbers of rings in each carbon cluster) were considered as models of amorphous carbon in the present study. The B3LYP/LANL2MB calculations showed that the potassium ion is located at ca. 2.80 A from the graphene surfaces and the charge of K+ is not dependent on the cluster size of the graphene sheets. On the other hand, the charge of potassium atom (K) increased with increasing cluster size (n), and it reached a constant value (ca. +0.70) at n = 29–52. Also, direct molecular orbital–molecular dynamics (MO–MD) calculation [H. Tachikawa and A. Shimizu: J. Phys. Chem. B 110 (2006) 20445] was applied to diffusion processes of the K+ ion and K atom on the model surfaces of amorphous carbon. The direct MO–MD calculations showed that the K+ ion and K atom diffuse freely on the surface at 800 K. The nature of the interaction between K+/K and the carbon clusters was discussed on the basis of theoretical results.

Hybrid density functional theory (DFT) calculations have been carried out for the lithium adsorbed on a fluorinated graphene surface (F-graphene, C96F24) to elucidate the effect of fluorination of amorphous carbon on the diffusion mechanism of lithium ion. Also, direct molecular orbital–molecular dynamics (MO–MD) calculation [H. Tachikawa and A. Shimizu: J. Phys. Chem. B 109 (2005) 13255] was applied to diffusion processes of the Li+ ion on F-graphene. The B3LYP/LANL2MB calculation showed that the Li+ ion is most stabilized around central position of F-graphene, and the energy was gradually instabilized for the edge region. The direct MO–MD calculations showed that the Li+ ion diffuses on the bulk surface region of F-graphite at 300 K. The nature of the interaction between Li+ and F-graphene was discussed on the basis of theoretical results.

Quantum chemical study on vapor reactions of ZnO growth by metal–organic vapor phase epitaxy (MOVPE) was carried out. Especially, the reaction natures between diethylzinc (DEZn) and different oxygen sources, nitrous oxide (N2O) and water (H2O), have been comparatively discussed, in terms of the adsorption of oxygen sources to DEZn. The adsorption energy and natural population analysis revealed that H2O attacks Zn 4s orbital of DEZn and interaction between H2O and DEZn becomes strong. On the other hand, N2O does not strongly interacts with DEZn. It is confirmed that the reaction of DEZn and H2O is too strong to control by any external energies, while that of DEZn and N2O is well controllable. The result supports the reaction model proposed from the experiments and it is understood that N2O is a desirable source for well-controlled MOVPE.

In recent years, many ceramics have been developed for the uses for mechanical or electronic parts. The high strength ceramics is necessary for the world of engineering materials. The cutting of the alumina green compact was carried out by a lathe with the condition of the cutting speed 100m/min. The purpose of the study is to clarify relation between surface roughness on the surface of green compact and bending strength of the ceramics. The bending experiment on the ceramics is discussed and then bending strength and the fracture energy were investigated. The main results obtained are as follows; (1) The true bending strength of the alumina ceramics is affected by the surface roughness of the alumina green compact. (2) The fracture energy was 30J/m2 in the case of the surface roughness of 5μm.

A NADPH-dependent carbonyl reductase was purified to homogeneity from Candida magnoliae AKU4643 through four steps, including Blue Sepharose affinity chromatography. The enzyme catalyzed the stereoselective reduction of ethyl 4-chloro-3-oxobutanoate to the corresponding (S)-alcohol with a 100% enantiomeric excess, which is a useful chiral building block for the chemical synthesis of pharmaceuticals. The relative molecular mass of the enzyme was estimated to be 76,000 on high performance gel filtration chromatography and 32,000 on SDS polyacrylamide gel electrophoresis. The enzyme reduced alpha-, beta-keto esters and conjugated diketones in addition to ethyl 4-chloro-3-oxobutanoate. The enzyme activity was inhibited by quercetin and HgCl2, but not by EDTA. The N-terminal amino acid sequence of the enzyme showed no apparent similarity with those of other oxidoreductases.

Endonucleases capable of producing internucleosomal DNA cleavage are one of the key enzymes in apoptosis. We examined endonuclease activities contained in nuclei of CD34 + and erythroid cells in the bone marrow (BM) from 12 patients with the myelodysplastic syndromes. The levels of Mg 2+ -dependent and acidic endonucleases showed little changes as compared with those from normal BM. By contrast, the level of Ca 2+ Mg 2+ -dependent endonuclease was appreciably higher in MDS erythroid cells than normal counterparts, although the activity varied markedly in CD34 + and erythroid cells. Our results suggested that Ca 2+ Mg 2+ -dependent endonuclease is related to ineffective erythropoiesis in MDS.

In recent years, ceramics for a few mechanical and electronic parts are performed on the cutting process of the green-compact. The residual crack on the finished surface is influenced the strength of the ceramics after the sintering. The purpose of this paper is to clarify the cutting performance of the ceramics green compact that used the alumina of the grain size 0.2 μ m. The machining performance of alumina green-compact is discussed from view point of the relation between the shape of tool edge and the surface roughness on the workpiece. In the cutting test by turning, the cutting condition is cutting speed 100m/min, feed 0.1mm/rev, depth of cut 0.2mm. The tool shape on the insert tip are made chamfer angle from -25°to 5° The main results obtained are follows:(1) The surface roughness in feed direction and the in circumference direction was existed an correlative. (2)In the chamfer angle -10°on the tool edge, the surface roughness at cutting could give rise to Rmax 4.0 μm. (3)The tool life of this cutting was lager than other cutting.

In recent years, many ceramics have been to developed for the uses. Some ceramics for mechanical or electronic parts are required to be cut as the green compact. The cutting of the green compact is decreased cracks on the machined surface. The purpose of the paper is to clarify the mechanism at cutting of green compact. The cutting mechanism is investigated and discussed from the results of the fracture test by a new type testing machine and of the cutting test by a lathe. In the cutting test, the radius of cutting edge were varied from 0.8mm to 4mm. A fracture model is proposed from AE(Acoustic Emission) signal caused by fracture and SEM image of fracture surface. The main results obtained are as follows; (1) AE signal shows there stages of fracturing the green compact. (2) The fracture energy is from 4 J/m2 to 7 J/m2. (3) A fracture model is available to clarify the cutting mechanism. (4) The tool with large side cutting edge angle is able to cut a work at high efficiency and to elongate the tool life.

A new method for the reductive debromination of 1,2-bis(bromomethyl)arenes has been developed. The treatment of 1,2-bis(bromomethyl)benzene with tetrakis(dimethylamino)ethylene (TDAE) ( 1 ) in the presence of olefins gave 1,2,3,4-tetrahydronaphthalenes in moderate to good yields.

Transplantation-associated thrombotic microangiopathy (TA-TMA) is a devastating complication of hematopoietic stem cell transplantation. TA-TMA likely represents the final stage of vascular endothelial injury; however, its pathophysiology is largely unknown, making clinical management difficult. Recently, the association of neutrophil extracellular traps (NETs) with the development of thrombotic thrombocytopenic purpura and hemolytic uremic syndrome has been reported. Thus, we explored whether NETs are also relevant to the occurrence of TA-TMA. We retrospectively analyzed post-transplant trends of serum NET levels in 90 patients, 11 of whom developed TA-TMA. Relative to baseline (before the conditioning regimen), elevated serum NET levels either at 4 weeks after transplantation or as early as the day of transplantation were associated with significantly increased risk of TA-TMA. In contrast, thrombomodulin, a potential marker for TA-TMA, was not helpful to predict the occurrence of TA-TMA in our study. In addition, we directly detected glomerular deposition of NETs in 2 TA-TMA patients. Increased NET levels are a significant risk factor for TA-TMA, suggesting that NET level is a useful biomarker for TA-TMA.

In recent years, many ceramics with various kinds of properties have appeared on the market. Those ceramics are used for a few mechanical and electronic parts. The strength of alumina-ceramics piece is influenced by the residual crack of the finished surface on the green compact. The purpose of this paper is to clarify the machining performance of alumina-green compact. The machining performance of alumina-green compact is investigated and is discussed from view point of the relation between the flank wear of tool edge and the roughness of machined surface on the work-piece. In the cutting test by turning, the cutting speed was varied from 35 m/min to 150 m/min. The frequency of tool edge was measured by FFT(Fast Fourier Transform) analyzer, and worn parts of the tools were observed with the SEM and the microscope. The main results obtained are follows: (1) In the finish cutting for alumina-green compact, the best cutting condition was obtained as cutting speed of 100 m/min, feed 0.1 mm/rev, depth of cut 0.2 mm by tool of WC sintered carbide, and roughness Rmax of the machined surface was 7.10μm. (2)The tool life of this cutting was lager than other cutting. (3)The size of chip as cutting increased in proportion to the cutting length for 1/4 period of frequency on the tool edge.

Oligosilane-functionalized C60 fullerenes [namely, C60–(SiH2) n –H, n = 1–4] have been investigated by the density functional theory (DFT) method to elucidate the structures and electronic states of oligosilane-radical added fullerene. The DFT calculation showed that oligosilane radicals bind to the carbon atom of C60 in the on-top site, and a strong Si–C heterojunction is formed. The binding energies of oligosilane radicals to C60 were calculated to be 24.6–28.2 kcal/mol at the CAM-B3LYP/6-311G(d,p) level. The electronic states of oligosilane-functionalized fullerenes C60–(SiH2) n –H are discussed on the basis of theoretical results.

Functionalized graphenes are known as a high-performance molecular device. In the present study, the structures and electronic states of the aryl radical functionalized graphene have been investigated by the density functional theory (DFT) method to elucidate the effects of functionalization on the electronic states of graphene (GR). Also, the mechanism of aryl radical reaction with GR was investigated. The benzene, biphenyl, p-terphenyl, and p-quaterphenyl radicals [denoted by (Bz) n (n = 1–4), where n means numbers of benzene rings in aryl radical] were examined as aryl radicals. The DFT calculation of GR–(Bz) n (n = 1–4) showed that the aryl radical binds to the carbon atom of GR, and a C–C single bond was formed. The binding energies of aryl radicals to GR were calculated to be ca. 6.0 kcal mol−1 at the CAM-B3LYP/6-311G(d,p) level. It was found that the activation barrier exists in the aryl radical addition: the barrier heights were calculated to be 10.0 kcal mol−1. The electronic states of GR–(Bz) n were examined on the basis of theoretical results.

The structures and electronic states of alkyl-radical-functionalized C20 fullerenes (denoted by C20–R) have been investigated using density functional theory (DFT). The different alkyl radicals investigated were methyl, ethyl, propyl, and butyl radicals. The DFT calculation indicated that the alkyl radical binds to the carbon atom of C20 in the on-top site, thus forming a strong C–C single bond. The binding energies of the alkyl radicals to C20 were calculated to be 83.9–86.6 kcal/mol at the CAM-B3LYP/6-311G(d,p) level. The electronic states of the C20–R complex are discussed on the basis of the theoretical results.

Links between hepatitis C virus (HCV) infection and several non-Hodgkin lymphomas have been suggested by epidemiological studies. We herein report the first documented case of a patient with HCV-associated Hodgkin lymphoma who showed a marked regression following interferon-based antiviral therapy. This unique case extends the spectrum of HCV-associated malignant lymphomas, confirms the efficacy of antiviral therapy for this rare extrahepatic manifestation and provides valuable clues for achieving a better understanding of lymphomagenesis in HCV.

Using a registration sheet of a prospective registration system for aplastic anemia (AA)/myelodysplastic syndromes (MDS), by the National Research Group on Idiopathic Bone Marrow Failure Syndromes, Japan, we carried out a survey on examinations for diagnosis of bone marrow failure. Bone marrow trephine biopsy was performed in 66 of 105 cases (63%) [Original diagnosis: AA 51 cases (80%), MDS 12 (32%), undiagnosable 3 (75%)]. Bone marrow aspiration was performed in all cases, and aspiration was performed at least twice in 36 cases (34%). The first-line anatomic site for bone marrow aspiration was the posterior iliac crest (62%). Cytogenetic examination was performed in 93%. The concordance rate between the original and the central review diagnosis was 93% among the studied cases: AA, Idiopathic cytopenia of undetermined significance (ICUS) and MDS in total. Flow cytometry analysis to detect paroxysmal nocturnal hemoglobinuria (PNH)-type blood cells was performed in 32%.

Studies have suggested an association between pretransplant serum levels of ferritin and C-reactive protein (CRP) and complications of allogeneic hematopoietic stem cell transplantation (HSCT). To evaluate the prognostic impact of these biomarkers on the development of acute and chronic graft-versus-host disease (GVHD), we retrospectively studied 211 patients who underwent allogeneic HSCT for hematologic diseases at our institution. The cumulative incidence rate of chronic GVHD at 3 years was 40.7 %. In the multivariate analysis, elevated CRP levels (≥2 mg/L) were significantly associated with a high incidence of chronic GVHD, whereas high ferritin levels (≥880 ng/mL) showed a tendency, though not statistically significant, to association with a low incidence of chronic GVHD. No significant association was observed between the pretransplant serum ferritin or CRP levels and the incidence of acute GVHD. Multivariate analysis indicated that high pretransplant serum ferritin levels were significantly associated with increases in treatment-related mortality and relapse rates. Overall, an elevated pretransplant serum ferritin level, but not an elevated serum CRP level, is a strong risk factor for overall mortality (hazard ratio, 2.16; P = 0.002). Our results also indicate that pretransplant serum CRP levels may be a useful biomarker for predicting the risk of chronic GVHD.