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Pre-existing antibodies against adeno-associated virus (AAV), caused by natural AAV infections, interfere with recombinant AAV vector-mediated gene transfer. We studied the prevalence of neutralizing antibodies against AAV serotypes 1, 2, 5, 8, and 9 in healthy subjects (n = 85) and hemophilia patients (n = 59) in a Japanese population. For healthy subjects, the prevalence of neutralizing antibodies against AAV serotypes 1, 2, 5, 8, and 9 was 36.5%, 35.3%, 37.6%, 32.9%, and 36.5%, respectively, while that in hemophilia patients was 39.7%, 28.8%, 35.6%, 32.9%, and 27.4%, respectively. There was no difference in the prevalence of neutralizing antibody against each AAV serotype between the healthy subjects and the hemophilia patients. The prevalence of neutralizing antibodies against all AAV serotypes increased with age in both healthy subjects and hemophilia patients. High titers of neutralizing antibodies against AAV2 (≥1:224) and AAV8 (≥1:224) were more evident in older individuals (≥42 years old). Approximately 50% of all screened individuals were seronegative for neutralizing antibodies against each AAV tested, while approximately 25% of individuals were seropositive for each AAV serotype tested. The prevalence of seronegativity for all AAV serotypes was 67.0% (healthy subjects, 68.6%; hemophilia patients, 65.0%) and 18.6% (healthy subjects, 20.5%; hemophilia patients, 15.7%) in young (

Drug-induced hypersensitivity syndrome (DIHS), also known as drug reaction with eosinophilia and systemic symptoms (DRESS), is a severe multiple-organ condition caused by drug treatment. The current report describes a Japanese boy who underwent aspirin treatment for Kawasaki disease, and who subsequently presented with the manifestations of DIHS/DRESS syndrome. He had been treated with a single high dose of intravenous immunoglobulin and aspirin orally for Kawasaki disease. One month after the onset of Kawasaki disease, he developed a generalized maculopapular eruption, high-grade fever, leukocytosis with eosinophilia, and an increased number of atypical lymphocytes, severe liver dysfunction, lymphadenopathy, and prominent increases in antihuman herpesvirus-6 immunoglobulin G titer. The activity of 2',5'-oligoadenylate synthetase was elevated at the onset stage. Hypersensitivity to aspirin was confirmed by skin patch test and by lymphocyte stimulation test. Based on these findings, the patient was diagnosed with DIHS/DRESS caused by aspirin. To our knowledge, there have been no previous reports of aspirin-induced hypersensitivity syndrome subsequent to Kawasaki disease. The activity of 2',5'-oligoadenylate synthetase might be useful as a diagnostic marker of DIHS/DRESS syndrome and for exploring its pathogenesis.

凝固第VIII因子（FVIII）に対するインヒビター測定は，Bethesda（B）法が広く用いられてきたが，低力価域における定量性が乏しいことから，近年，Nijmegen（N）法が推奨されている．インヒビター力価はBethesda単位で表わされるが，現実にはこれら2法以外の測定法も混在し，各測定法間で感度が異なることも考えられる．今回，インヒビター陰性血友病A患者血漿を対象にB法， N法，さらに，B法の変法であるS法およびM法の4法を比較検討した．S法およびM法は，患者血漿中の凝固因子を56℃で不活化処理し，S法は56℃処理FVIII欠乏血漿を，M法は56℃処理正常血漿を対照とする方法である．各反応液のpH変化とFVIII：C残存率について検討した．pHは，S法，M法では加温前から高く，加温により4法全てが上昇した．また，B法の対照反応液はpH変化をほとんど認めなかったが，測定と対照反応液間で最も大きいpH差（>0.5）が観察された．一方，FVIII：Cは各法ともほぼ同様であったが，加温後のB法対照反応液のみFVIII：C残存率が90.4％と顕著に高値を示した．pHとFVIII：Cには，反応液のpHが高いとFVIII：Cが低値となる負の関係性 （r=0.878）が認められた．インヒビター陰性血友病A患者検体で実施した4法の比較では，B法は61.5％が偽陽性となり，他の3法は全て陰性で一致した．また，FVIII：Cが10％以上の4検体では，N法のインヒビター力価はより低値を示した．以上の成績より，インヒビター評価においては各測定方法の特性を認識した上で解釈することが重要と考えられた．

最近，出血性および血栓性疾患両者の凝固能を評価する新たな方法としてトロンビン生成試験（TGT）が注目されている．小児では中心静脈カテーテル（CVC）の利用も多く，通常その閉塞を防止する目的でヘパリンが使用される．それゆえCVC血にはヘパリンが多かれ少なかれ混入し，正確な凝固能の評価が不可能となる．我々はCVC血を使った凝固能測定に際し，ヘパリンの影響と，硫酸プロタミン，ヘパリナーゼのヘパリン中和効果についてTGTを中心に検討した．その結果，TGTはPT，APTTに比べ，ヘパリンに鋭敏であった．硫酸プロタミンおよびヘパリナーゼは共に至適中和量でヘパリンを完全に中和した．過剰量において，前者はその用量依存性にトロンビン生成抑制作用が認められたが，後者ではその作用は認められなかった．以上より，ヘパリナーゼを利用したTGT測定は，ヘパリン混入血の凝固能を評価する上で有用と思われた．

This paper describes geometric change of the substrate surface when Al2O3 particles of 0.5μm diameter, which is larger than ions and smaller than the particles used in blasting and soon, were jetted on the copper, steels and glass. As Al2O3 particles were jetted at a jet angle of 0°, vertically on the surface, Al2O3 particles were deposited on the substrate surface except glass substrate. As jet angle increased, ripple spacing perpendicular to the jet direction with spacing about 2-15 u m was generated on each material and that spacing grows larger with the increase in the jet angle. The spacing varies in the material and harder material shows the tendency to become smaller in the spacing. The ripple spacing grows rapidly larger with the increase in the jet angle. At a jet angle of 80°, the trace which appears in the removed surface along the jet direction becomes remarkable. That was applicable to polishing of the fine pipe.

A 15-yr-old boy presented with an anterior mediastinal mass, multiple lung metastases and obstruction of the left brachiocephalic vein, the superior vena cava and the subclavian vein. Tumor biopsy by CT guidance confirmed a diagnosis of GCT. Five courses of BEP therapy were performed, and CT of the chest revealed reduction in the anterior mediastinal mass and disappearance of the multiple lung metastases. We performed the anterior mediastinal mass extraction followed by adjuvant chemotherapy consisting of ICE and TIP. However, the AFP levels became elevated soon after. Abnormal accumulation was observed in the right upper lung by DW-MRI. After the operation, two courses of TI chemotherapy and two courses of HDCT followed by auto-PBSCT were performed. He was complicated with auditory disorder and renal dysfunction. Although HDCT followed by auto-PBSCT was effective for the relapsed primary mediastinal GCT, a treatment strategy avoiding late complications is warranted.

To investigate the binding specificity of turnip mosaic virus (TuMV) viral protein-genome linked (VPg) with translation initiation factor 4E, we evaluated here the kinetic parameters for the interactions of human eIF4E, Caenorhabditis elegans IFE-3 and IFE-5 and Arabidopsis eIFiso4E, by surface plasmon resonance (SPR). The results indicated that TuMV VPg does not show a binding preference for Arabidopsis eIFiso4E, even though it is from a host species whereas the other eIF4E orthologues are not. Surprisingly, the effect of m 7 GTP on both the rate constants and equilibrium binding constants for the interactions of VPg differed for the four eIF4E orthologues. In the case of eIFiso4E and IFE-3, m 7 GTP increased k on , but for eIF4E and IFE-5, it decreased k on . To provide insight into the structural basis for these differences in VPg binding, tertiary structures of the eIF4E orthologues were predicted on the basis of the previously determined crystal structure of m 7 GpppA-bound human eIF4E. The results suggested that in cap-bound eIF4E orthologues, the VPg binds to the C-terminal region, which constitutes one side of the entrance to the cap-binding pocket, whereas in the cap-free state, VPg binds to the widely opened cap-binding pocket and its surrounding region. The binding of VPg to the C-terminal region was confirmed by the SPR analyses of N- or C-terminal residues-deleted eIF4E orthologues.

The interaction between turnip mosaic virus (TuMV) viral protein linked to the genome (VPg) and Arabidopsis thaliana eukaryotic initiation factor (iso)4E (eIF(iso)4E) was investigated to address the influence of potyviral VPg on host cellular translational initiation. Affinity chromatographic analysis showed that the region comprising amino acids 62-70 of VPg is important for the interaction with eIF(iso)4E. In vitro translation analysis showed that the addition of VPg significantly inhibited translation of capped RNA in eIF(iso)4E-reconstituted wheat germ extract. This result indicates that VPg inhibits cap-dependent translational initiation via binding to eIF(iso)4E. The inhibition by VPg of in vitro translation of RNA with wheat germ extract did not depend on RNase activity. Our present results may indicate that excess VPg produced at the encapsidation stage shuts off cap-dependent translational initiation in host cells by inhibiting complex formation between eIF(iso)4E and cellular mRNAs.

Coxsackievirus B1 (CVB1) 2A proteinase (2A(pro)) is a cysteine proteinase that cleaves the viral monocistronic polyprotein between the C-terminus of the VP1 region and the N-terminus of the 2A region, and also shuts off translational initiation in host cells by cleavage of eukaryotic initiation factor 4G (eIF4G) isoforms. We expressed in Escherichia coli a series of fusions in which various C-terminal fragments of VP1 were linked to the N-terminus of 2A(pro), and we also employed site-directed mutagenesis to introduce mutations of several amino acid residues. Our results showed that the presence of the C-terminal three amino acid residues of VP1 at the N-terminus of 2A(pro) is sufficient for specific self-cleavage between VP1 and 2A(pro) to generate mature 2A(pro), but the P4 amino acid also plays an important role. We further found that 2A(pro) cleaves the amino acid sequence Leu-Val-Pro-Arg-( *)Gly-Ser (LVPRGS motif), which is the target sequence of thrombin.

The turnip mosaic virus (TuMV) genome-linked protein (VPg) and Arabidopsis thaliana translation initiation factors were expressed and purified in order to investigate their binding properties and kinetics. Affinity chromatography on m(7)GTP-sepharose showed that bound A. thaliana eIF(iso)4E was eluted with crude TuMV VPg. Further column studies with purified VPg and other A. thaliana eIF4E isoforms showed that VPg preferentially bound eIF(iso)4E. Structural data implicate Trp-46 and Trp-92 in eIF(iso)4E in cap recognition. When Trp-46 or Trp-92 were changed to Leu, eIF(iso)4E lost the ability to form a complex with both VPg and m(7)GTP-sepharose. This suggests that the VPg-binding site is located in or near the cap-recognition pocket on eIF(iso)4E. Affinity constants for the interactions with eIF(iso)4E of VPg and capped RNA oligomer were determined using surface plasmon resonance (SPR). The K(D) values showed that the binging affinity of VPg for eIF(iso)4E is stronger than that of capped RNA. This suggests that viral VPg can interfere with formation of a translational initiation complex on host plant cellular mRNA by sequestering eIF(iso)4E. Further experiments with affinity chromatography showed that VPg forms a ternary complex with eIF(iso)4E and eIF(iso)4G. Thus, VPg may participate in viral translational initiation by functioning as an alternative cap-like structure.

Twenty-seven tropolone derivatives were investigated for their tumor-specific cytotoxicity, using 3 normal human cells and 3 human oral tumor cell lines. Tropolone derivatives with phenolic OH group, hinokithiol, its tosylate and methyl ethers have relatively higher tumor specificity. 5-Aminotropolone showed the highest specificity, whereas 2-aminotropone and its derivatives showed little or no specificity. 5-Aminotropolone induced apoptotic cell death characterized by internucleosomal DNA fragmentation and caspase 3 activation in the human promyelocytic leukemic HL-60 cell line. ESR spectroscopy showed that 5-aminotropolone produced radical under alkaline condition, and efficiently scavenged O2- and NO produced by HX-XOD reaction and NOC-7, respectively. These data suggest that 5-aminotropolone may induce cytotoxicity by radical-mediated redox reaction.

朝食の欠食は日周リズムを変え, これによって肥満, 高コレステロール血を増加させ, 各種の知的機能テストの成績を低下させると報告されている。そこで寮内学生 (19～21歳) につき分析を行い下記の成績を得た。1978 (人数=102) 1979 (人数=106)朝食 欠食者(85名) 摂取者(17名) P 欠食者(64名) 摂取者(42名) P〔摂取量/日〕エネルギー (kcal) 1,916 2,180