The use of epoxy patch grafts for the repair of experimentally-created diaphragmatic defects in dogs

Canine pericardium which had been treated with polyepoxy compounds (Denacol EX-313) was used as a patch graft for the correction of experimentally-created diaphragmatic defects in five dogs belonging to the same litter. Clinical, macroscopic and histological examinations were conducted every month up to five months after suturing of the patch graft. Clinical examination of the patch graft showed no apparent abnormalities. Macroscopic examination conducted during autopsy showed that the patch graft maintained adequate elasticity for five months after suturing, the surface of the patch graft was covered with a thin membrane and neovascularization was observed. Histological examination showed that the surface of the patch graft was covered with a thin membrane. Inflammatory tissue reactions were observed at one month, but gradually decreased from the second month onwards. In addition, the patch graft had excellent tissue affinity.     

Open/folded bit-line arrangement for ultra-high-density DRAM's

An open/folded bit-line (BL) arrangement for scaled DRAM's is proposed. This BL arrangement offers small die size and good array noise immunity. In this arrangement, one BL of an open BL pair is placed in between a folded BL pair, and the sense amplifiers (SA's) for open BL's and those for folded BL's are placed alternately between the memory arrays. This arrangement features a small 6F² memory cell, where F is the device feature size, and a relaxed SA pitch of 6F. The die size of a 64-Mb DRAM can be reduced to 81.6% compared with the one using the conventional folded BL arrangement. The BL-BL coupling noise is reduced to one-half of that of the conventional folded BL arrangement, thanks to the shield effect. Two new circuit techniques, 1) a multiplexer for connecting BL's to SA's, and 2) a binary-to-ternary code converter for the multiplexer have been developed to realize the new BL arrangement     

Pathogenesis and prognosis of neonatal brain damage: introductory remarks

Neonatal brain damage, including the damage in the fetal period, is caused by many factors such as hypoxia, infection, trauma, intoxication and metabolic disorders. According to epidemiological studies, perinatal hypoxic-ischemic brain damage shows the highest incidence, and needs a new strategy for its prevention and treatment. This symposium is focused on the recent advances of basic research and technological science in the pathogenesis, neuroimagings, near-infrared spectroscopy, developmental neurophysiology, and intervention in perinatal hypoxic-ischemic brain damage, taking into consideration on the long term prognosis in each section.     

Mouse dendritic epidermal T cells exhibit chemotactic migration toward PAM 212 keratinocyte culture supernatants

Dendritic epidermal T cells (DETCs) are Thy-1+, CD45+, CD3+, CD4-, CD8-, and T-cell receptor-V gamma 3/V delta 1+ leukocytes that reside normally in adult mouse skin. We have demonstrated previously that keratinocytes serve as adhesion substrates for DETCs, and that interleukin 7 (IL-7), which is produced by keratinocytes, serves as a growth factor for DETCs. The present study was conducted to address the mechanisms by which DETCs migrate into the epidermis, reasoning that keratinocytes may also be a source of chemotactic activity. Short-term DETC lines were 35S-labeled and tested for migration toward Pam 212 keratinocyte culture supernatants using a modified Boyden chamber method; cell movement from upper chambers toward test samples in lower chambers was traced by counting radioactivity. DETC displayed rapid (within 60 min) and marked (> 50%) migration toward keratinocyte supernatants. The majority of cells that had migrated into keratinocyte supernatants expressed the V gamma 3 T-cell receptor, thus verifying that the migrating cells were DETCs. Addition of keratinocyte supernatants to the upper chambers completely blocked migration, suggesting its chemotactic nature. By contrast, no DETC migration was observed toward 3T3 fibroblast supernatants. Chemotactic activities were 1) produced by Pam 212 cells even in the absence of serum; 2) greater than 12 kD in size; 3) heat and pH labile; 4) trypsin sensitive; and 5) precipitated by 60-100% ammonium sulfate. Several cytokines (e.g., IL-1 alpha and IL-8) failed to mediate DETC migration when added to the lower chambers. Likewise, the same cytokines, when added to the upper chambers, failed to inhibit DETC migration toward Pam 212 supernatants. These results support our hypothesis that keratinocytes facilitate the residence of DETC in epidermis by secreting unique chemotactic factors, by providing adhesion substrates, and by elaborating specific growth factors.     

Isotopic evidence in tree rings for historical changes in atmospheric sulfur sources

Little is understood about the usefulness of sulfur isotopic ratios (sigma 34S) in tree rings because the sulfur content in rings is generally insufficient for analysis using conventional methods. We present sigma 34S values of the water-soluble and the organically bound sulfur fractions in rings of coniferous trees grown in Japan, analyzed using a large-volume oxygen bomb. Comparing the sigma 34S values of the organically bound fraction in tree rings with past atmospheric sulfur concentrations and with those of their sources, we find clear evidence that the sigma 34S values of the organically bound fraction in the rings are dependent upon the values of the atmospheric sulfur sources. The evidence suggests that the sigma 34S values in tree rings are a useful chronological proxy for evaluating possible causes of past atmospheric sulfur pollution.     

A Metal-Free,Disulfide Oxidized Form of Superoxide Dismutase 1 as a Primary Misfolded Species with Prion-Like Properties in the Extracellular Environments Surrounding Motor Neuron-Like Cells

Superoxide dismutase 1 (SOD1) is a metalloenzyme with high structural stability, but a lack of Cu and Zn ions decreases its stability and enhances the likelihood of misfolding, which is a pathological hallmark of amyotrophic lateral sclerosis (ALS). A growing body of evidence has demonstrated that misfolded SOD1 has prion-like properties such as transmissibility between cells and intracellular propagation of misfolding of natively folded SOD1. Recently, we found that SOD1 is misfolded in the cerebrospinal fluid of sporadic ALS patients, providing a route by which misfolded SOD1 spreads via the extracellular environment of the central nervous system. Unlike intracellular misfolded SOD1, it is unknown which extracellular misfolded species is most relevant to prion-like properties. Here, we determined a conformational feature of extracellular misfolded SOD1 that is linked to prion-like properties. Using culture media from motor neuron-like cells, NSC-34, extracellular misfolded wild-type, and four ALS-causing SOD1 mutants were characterized as a metal-free, disulfide oxidized form of SOD1 (apo-SOD1^S-S). Extracellular misfolded apo-SOD1^S-S exhibited cell-to-cell transmission from the culture medium to recipient cells as well as intracellular propagation of SOD1 misfolding in recipient cells. Furthermore, culture medium containing misfolded apo-SOD1^S-S exerted cytotoxicity to motor neuron-like cells, which was blocked by removal of misfolded apo-SOD1^S-S from the medium. We conclude that misfolded apo-SOD1^S-S is a primary extracellular species that is linked to prion-like properties.     

Dual roles of proteasome in the metabolism of presenilin 1

Presenilin 1 (PS1) has been identified as a causative gene for most early-onset familial Alzheimer's disease. Biochemical studies revealed that PS1 exists predominantly as two processed fragments in cells and brain tissues. We prepared stably transfected cells expressing the wild-type and familial Alzheimer's disease-associated mutants of PS1 and investigated the enzyme that participates in the metabolism of PS1. After treatment of the cells with proteasome inhibitors, the full-length PS1 was significantly accumulated. The levels of N- and C-terminal fragments were also increased. The accumulation of PS1 with a deletion of exon 10, which is unable to be processed, on treatment of the transfected cells with lactacystin indicated that proteasome can degrade full-length PS1. A synthetic peptide that includes the processing region of PS1 was cleaved by 20S proteasome at the putative processing sites after Met288 and Glu299. Metabolic labeling experiments showed that the appearance of the N-terminal fragment was attenuated by the inhibitor. Finally, 28-kDa N- and 20-kDa C-terminal fragments were generated by purified PS1 in vitro. These data indicated that the proteasome pathway is involved in PS1 processing. These results demonstrate that the proteasome pathway plays dual roles in processing and degradation of PS1.     

Effect of ultrafine grain refinement on hydrogen embrittlement of metastable austenitic stainless steel

Micro-tensile tests were performed on high-pressure-torsion-processed specimens of type 304 steel with grain sizes in the range of 0.1–0.5 μm to clarify the effect of ultrafine grain refinement on the hydrogen embrittlement (HE) of metastable austenitic steel. The ultrafine-grained (UFG) specimens with average grain sizes < ～0.4 μm exhibited a limited uniform elongation followed by a steady-stress regime in the stress–strain curves, which was attributed to a martensitic transformation. A high yield stress and a moderate elongation to failure were attained for the UFG specimens with an average grain size of ～0.5 μm in the uncharged state. Hall–Petch relationships well hold between the yield stress and the average grain size for each uncharged and hydrogen-charged specimen. Hydrogen charging increased the friction stress by 40% but did not change the Hall–Petch coefficient. Hydrogen-induced ductility loss was mitigated by ultrafine grain refinement. Ductility loss due to hydrogen charging manifested in the local deformation after a martensitic transformation. This indicates that hydrogen does not significantly affect the martensitic transformation, but shortens the subsequent local deformation process.