Electron gun using carbon-nanofiber field emitter.

An electron gun constructed using carbon-nanofiber (CNF) emitters and an electrostatic Einzel lens system has been characterized for the development of a high-resolution x-ray source. The CNFs used were grown on tungsten and palladium tips by plasma-enhanced chemical-vapor deposition. Electron beams with the energies of 10

Daily nasal inoculation with the insulin gene ameliorates diabetes in mice.

The present study examined the feasibility of liposome-mediated gene transfer via nasal administration, for treating insulin-dependent diabetes mellitus. The rat insulin gene was packed under control of the CMV promoter, complexed with DC-chol/DOPE-based liposomes and administered daily via the nasal route in mice made severely diabetic by streptozocin. Sustained expression of the insulin gene was achieved and insulinopenia, ketonuria and death were prevented. Hyperglycemia and body weight reduction were significantly suppressed without evidence of hypoglycemia throughout the experimental period. RT-PCR and FISH analysis indicated that insulin was produced in the alveolar epithelial cells of the lung. Liposome-mediated in vivo gene transfer via nasal administration may provide an efficacious route for delivery of hormonal and other gene products into the blood stream.

Summary of the JCO criticality accident in Tokai-mura and a dose assessment.

A criticality accident occurred on September 30, 1999, in a conversion test facility at the JCO Tokai site. The accident was triggered by pouring an 18.8% enriched uranyl nitrate solution into a precipitation vessel beyond the critical mass. The accident continued for about 19 hours before the criticality could be stopped. during which time neutrons and gamma-rays were emitted continuously due to fission reactions. The total number of fission reactions was 2.5 x 10(18), which was estimated by an activity analysis of the fission products in the solution of the precipitation vessel. The accident gave serious radiation dose to 3 employees and fatal dose to 2 of them. Neutrons and gamma-rays emitted by the accident caused meaningful doses to the residents of the surrounding area of JCO. The dominant dose to the residents and JCO employees was brought by neutrons and gamma-rays from the precipitation vessel, while the contribution of radioactive plume was negligible. The individual dose was estimated for 234 resident, 169 JCO employees and 260 emergency personnel. The maximum doses were 21 mSv for the residents, 48mSv for the JCO employees, and 9.4mSv for the emergency personnel, respectively. No deterministic effect, however, has been observed, except for the 3 workers.

Soluble intercellular adhesion molecule-1 (ICAM-1), CD4, CD8 and interleukin-2 receptor in patients with Behçet's disease and Vogt-Koyanagi-Harada's disease.

OBJECTIVES: In a search for serum markers of disease activity in uveitis, we measured the levels of the soluble form of ICAM-1 (sICAM-1), CD4 (sCD4), CD8 (sCD8) and interleukin-2 receptor (sIL-2R) in the serum of patients with Behçet's disease (BD) and Vogt-Koyanagi-Harada's disease (VKH). METHODS: The study population consisted of 20 patients with active BD (treated with tacrolimus), 15 patients with inactive BD, 24 patients with VKH [20 of them successfully treated with systemic corticosteroids (cured group) and 4 of them with two or more episodes of uveitis after withdrawal of systemic steroid (recurrence group)], and 20 normal individuals. The levels of serum sICAM-1, sCD4, sCD8 and sIL-2R were measured by sandwich ELISA. RESULTS: Sera from patients with BD in the convalescent stage showed significantly higher levels of sICAM-1 than those in the acute stage. Patients with active BD in both stages or VKH in the acute stage had significantly higher levels of serum sCD4 and sIL-2R than the controls. The levels of sCD8 in patients with both diseases in both stages differed significantly compared to the controls. No difference was noted in the pattern of decline of these soluble markers after treatment in the cured and recurrence groups of VKH patients. A positive correlation was found between the serum levels of sCD4 and sIL-2R in patients with both diseases in the acute stage. CONCLUSIONS: The results suggest that these soluble markers may represent potentially useful parameters to monitor disease activity or chronic inflammation in certain types of autoimmune uveitis.

Immunolocalization of prolyl 4-hydroxylase subunits, alpha-smooth muscle actin, and extracellular matrix components in human lens capsules with lens implants.

Lens capsules become fibrotic after the extraction of a cataract. To understand this phenomenon, we evaluated the immunolocalization of prolyl 4-hydroxylase (an enzyme involved in procollagen hydroxylation), and extracellular matrix components and cytoskeletal components in a normal human lens capsule and in others with intraocular lenses. Lens capsules containing intraocular lenses were removed from a patient with proliferative vitreoretinopathy and three with proliferative diabetic retinopathy during vitreous surgery. Two circular sections of the anterior capsules with lens epithelial cells were obtained by anterior capsulotomy during cataract surgery. In addition, a lens capsular bag was obtained immediately after phacoemulsification. The lens capsules were processed for light microscopic immunohistochemical detection of the alpha and beta subunits of prolyl 4-hydroxylase, extracellular matrix components (including collagen types, laminin and cellular fibronectin) or cytoskeletal components (such as cytokeratin, vimentin and alpha-smooth muscle actin). Monolayer lens epithelial cells were seen on the inner surface of the normal anterior capsules. Each intraocular lens was found to be fixed in the capsular bag. Light microscopic immunohistochemistry showed that these proliferating cells expressed vimentin and alpha-smooth muscle actin; in contrast, quiescent lens epithelial cells did not stain for alpha-smooth muscle actin. Marked immunostaining for subunits of prolyl 4-hydroxylase was detected in lens epithelial cells proliferating on the capsules, while no or only faint prolyl 4-hydroxylase immunoreactivity was detected in quiescent lens epithelial cells immediately after phacoemulsification. Collagen types I, III and VI and cellular fibronectin were observed diffusely in accumulated connective tissue on a capsule with an intraocular lens. Type IV collagen immunoreactivity was seen both in the capsules and in the connective tissue accumulation on the capsules. Collagen V and laminin were detected in association with cellular proliferation. Collagen VII and VIII and laminin 5 were not seen. We concluded that during wound healing of the lens capsule after cataract extraction, the lens epithelial cells that proliferate on the inner surface of the capsule transform it into a myofibroblastic phenotype, expressing prolyl 4-hydroxylase and alpha-smooth muscle actin. These proliferating cells are involved in the production of collagen on the lens capsule. This results in a postoperative fibrotic process and contraction of the lens capsule.

T lymphopenia in genetically obese rats.

Although obese animals are more susceptible to infection, the underlying causes are not fully known. In this study, long-term measurements were made of lymphocyte subsets in peripheral blood, spleen, and thymus in genetically obese Zucker (fa/fa) rats. Blastogenic response of splenocytes to mitogens was also examined. fa/fa rats developed obesity, hyperlipidemia, and hyperinsulinemia after 5 weeks of age. Flow cytometric analysis revealed that T cells in peripheral blood, spleen, and thymus were all reduced significantly in obese rats after 8 weeks of age compared to nonobese (Fa/-) littermates. All T-cell subsets examined, including CD4+ and CD8+ T cells, were similarly reduced in spleen and thymus as well as in peripheral blood with advance in age. In addition, proliferative responses of splenocytes to mitogens were significantly low in obese rats. These results indicate that long-term obesity may reduce the size of the T-cell pool and impair the responsiveness of splenocytes in rats.

Recombinant TIMP-1 and -2 enhance the proliferation of rabbit corneal epithelial cells in vitro and the spreading of rabbit corneal epithelium in situ.

PURPOSE: The repair of the corneal epithelium is modulated by matrix metalloproteinases. The present study examined the effects of recombinant (r-) tissue inhibitors of metalloproteinases-1 and -2 (TIMP-1 and -2) on the proliferation of cultured epithelial cells from rabbit cornea, and on the spreading of a sheet of squamous epithelium of rabbit cornea placed in an organ culture system. METHODS: DNA synthesis of the cells, with or without r-TIMPs, was determined by an immunoassay for BrdU incorporation. Cell proliferation was assayed by measuring MTT mitochondrial activity. Epithelial spreading was evaluated by culturing small corneal blocks for 24 h in the presence or absence of each agent. Cryosections were prepared and the epithelial growth on the cut stromal surface was measured. RESULTS: Each agent, r-TIMP-1 (at 50 and 100 ng/ml) and r-TIMP-2 (at 50 ng/ml) significantly enhanced the DNA synthesis and MTT activity of the corneal epithelial cells in vitro. Relative to the untreated control cells, DNA synthesis was increased 2.4-fold by r-TIMP-1 and 2.3-fold by r-TIMP-2. r-TIMP-1 (at 100 and 200 ng/ml) and r-TIMP-2 (at 10 and 50 ng/ml) each significantly enhanced the spreading of the corneal epithelium. Relative to the untreated control tissue, spreading of the epithelial sheet was increased 1.7-fold by r-TIMP-1 and 1.4-fold by r-TIMP-2. Higher concentrations of r-TIMP-1 and r-TIMP-2 did not further enhance either the DNA synthesis of the cultured cells or the spreading of the epithelium. CONCLUSIONS: Exogenous TIMPs enhanced the spreading of the corneal epithelium and proliferation of cultured corneal epithelial cells. Findings suggest that endogenous TIMPs may influence the healing of corneal epithelium in vivo.

Construction and performance test of SGM-TRAIN at UVSOR.

A new spherical-grating monochromator with translational and rotational assembly including a normal-incidence mount (SGM-TRAIN) has been constructed at BL5A of the UVSOR facility. The SGM-TRAIN is an advanced version of a constant-length SGM with the following improvements: (i) a wide energy range of 5-250 eV; (ii) a high resolving power; (iii) use of linear and circular polarization; (iv) reduction of second-order light; (v) two computer-controlled driving modes. Part of the performance tests are reported along with a detailed description of the design.

Cationic liposomes are a strong adjuvant for a DNA vaccine of human immunodeficiency virus type 1.

Liposomes have been widely used to enhance the immune response. In the present investigation, we studied their in vivo immunomodulation of an HIV-1-specific DNA vaccine candidate (pCMV160/REV) constructed with the cytomegalovirus (CMV) promoter-conjugated HIV-1 env and rev DNA plasmids. By immunizing with pCMV160/REV and cationic liposomes through various routes (intramuscular, intraperitoneal, subcutaneous, intradermal, and intranasal), we induced higher levels of both antibody production and delayed-type hypersensitivity (DTH) than by using DNA vaccine alone. The HIV-1-specific cytotoxic T lymphocyte (CTL) activity was observed to be stronger on immunization with the DNA vaccine and cationic liposome combination. The intramuscular, intraperitoneal, and intranasal inoculation routes were more effective in inducing strong DTH and antibody responses than the subcutaneous and intradermal routes. Taken together, these results suggest that cationic liposomes can be highly effective when used with DNA vaccines and administered by various routes.

Changes in DNA copy number in primary gastric carcinomas by comparative genomic hybridization.

We examined 33 primary gastric carcinomas using comparative genomic hybridization to detect changes in the DNA copy number and the chromosomal location of these changes. Ninety-four percent (31 of 33) showed 1 or more DNA copy number changes, such as increases at 2p23-p25 (observed in 21% of the total cases), 3q26.3-q27 (24%), 7p15 (24%), 9p22-pter (18%), and 13q22-q34 (21%) and decreases at 1p34.2-p36.2 (18%) and Y (52%). Histological examination indicated that increases at 3q26.1-q26.3 and 7p15 and decreases at 1p36.1-p36. 2 and Y were commonly observed in both differentiated and undifferentiated types. Increases at 3q27, 6q23-q25, and 7cen-p14 and decreases at 1p34.2-p35 and 17p12 were predominantly observed in the differentiated type, and increases at 2p23-pter, 9p22-pter, and 13q31-qter and a decrease at 6p21.3 were predominantly observed in the undifferentiated type. In addition, clinical staging of tumors showed that increases at 2p23-p25, 7p14-p21, 7q31-q32, and 9p22-pter and a decrease at Y were observed in early-stage tumors, whereas increases at 9q32-q33 and 15q26 were observed only in late-stage tumors. Many of the abnormalities detected in this study were not previously reported in gastric carcinomas. Our comparative genomic hybridization results indicate the presence of genetic alterations that may play some important role in the development and progression of gastric carcinomas.

Genetic control of in vivo tumor necrosis factor production in mice.

We report on the genetic effect on in vivo production of tumor necrosis factor (TNF)-alpha induced by lipopolysaccharides (LPS) using various congenic mouse strains. B10.A, Bl0.A(3R), B10.AQR, B10.A(5R), and B10.S(7R) produced significantly high TNF-alpha compared with B10.BR, B10.S, C57BL/10, B10.A(2R), B10.A(4R), B10.G, B10.DA(80NS), and B10.RIII(71NS). This suggests that LPS-induced TNF-alpha production is genetically controlled by H-2. Mice with the same alleles on K, A, E, or S loci produced various (high or low) levels of TNF-alpha, thus indicating that regulatory genes are located outside these loci. All strains with H-2Dd produced significantly high levels of TNF-alpha, but strains with other alleles in the H-2D locus produced low levels. Thus, TNF-alpha production appears to be genetically linked to H-2D itself or H-2D linked genes and the allele d is linked to a high responder gene. This was the case with the A background. C3H/HeN (H-2k), however, showed a high TNF-alpha production, suggesting the presence of another controlling gene outside H-2. In addition, high TNF-alpha productivity was transmitted into F1 mice (B10.A X B10.BR) in a dominant fashion. Both LPS-stimulated and unstimulated TNF-alpha mRNA expression in splenic macrophages were enhanced in high responder strains. Thus, we conclude that TNF-alpha production is closely related to genes within or linked to the H-2D locus as well as others outside H-2.

Use of sodium dodecyl sulphate to clarify the end-point of anodic-stripping complexometric titrations.

It has been found that addition of sodium dodecyl sulphate will completely suppress the dissociation of the copper-EDTA complex at the electrode surface in the anodic-stripping complexometric titration of copper and make the end-point of the titration very clear. The addition of SDS also makes it possible to titrate traces of nitrilotriacetic acid, which forms a copper complex that is less stable than the EDTA complex. The effect of SDS is presumed to be due to electrostatic repulsion between the negative charges of adsorbed SDS and the metal complex at the electrode surface.