A novel interferon regulatory factor family transcription factor, ICSAT/Pip/LSIRF, that negatively regulates the activity of interferon-regulated genes

We have isolated a novel cDNA clone encoding interferon (IFN) consensus sequence-binding protein in adult T-cell leukemia cell line or activated T cells (ICSAT); this protein is the human homolog of the recently cloned Pip/LSIRF. ICSAT is structurally most closely related to the previously cloned ICSBP, a member of the IFN regulatory factor (IRF) family of proteins that binds to interferon consensus sequences (ICSs) found in many promoters of the IFN-regulated genes. Among T-cell lines investigated, ICSAT was abundantly expressed in human T-cell leukemia virus type 1 (HTLV-1)-infected T cells. When the HTLV-1 tax gene was expressed or phorbol myristake acetate-A23187 stimulation was used, ICSAT expression was induced in Jurkat cells which otherwise do not express ICSAT. When the binding of ICSAT to four different ICSs was tested, the relative differences in binding affinities for those ICSs were determined. To study the functional role of ICSAT, we performed cotransfection experiments with the human embryonal carcinoma cell line N-Tera2. ICSAT was demonstrated to possess repressive function over the gene activation induced by IFN stimulation or by IRF-1 cotransfection. Such repressive function is similar to that seen in IRF-2 or ICSBP. However, we have found that ICSAT has a different repressive effect from that of IRF-2 or ICSBP in some IFN-responsive reporter constructs. These results suggest that a novel mechanism of gene regulation by "differential repression" is used by multiple members of repressor proteins with different repressive effects on the IFN-responsive genes.     

Determination of horseradish peroxidase concentration using the chemiluminescence of Cypridina luciferin analogue, 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazin-3-one

The chemiluminescence of the Cypridina luciferin analogue, 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazin++ +-3-one (MCLA) was observed at 462 nm in the presence of horseradish peroxidase (HRP) and the total spectrum of light emitted was found to depend linearly on HRP concentration. Methods for the determination of HRP concentration using the chemiluminescence was investigated. HRP could be detected in the range from 100 pmol/L to 100 nmol/L under the optimum condition, H2O2 (10 mmol/L) and MCLA (10 mumol/L) at pH 5.8.     

Development of revolving drop surface tensiometer

A revolving drop surface tensiometer, which measures the surface tension of a small amount of liquid, is proposed. A remarkable feature of this device is that while using the pendant drop method, it employs a centrifugal force to deform the liquid droplet. The centrifugal force induces a large distortion of the droplet, which enables an accurate measurement of the surface tension to be made. In our experimental setup, the centrifugal force can be increased so that the apparent acceleration becomes up to 100 times larger than that due to gravity, and the capability of this method to measure surface tensions was demonstrated with ethylene glycol.     

A size effect law for notched tensile strength of woven carbon/epoxy laminates

Specimen-size effect and notch-size effect on the tensile strength of woven fabric carbon/epoxy laminates are evaluated and modeled. For two different layups of [(0/90)₁₂] and [(±45)₂/(0/90)₅]_S, respectively, static tension tests were performed on two-dimensional geometrically similar unnotched and double-edge notched specimens scaled to three different sizes. Experimental results demonstrate that the notched strength of the woven CFRP laminates depend on the size of specimen as well as the size of notch. The ratio of notched strength to unnotched strength decreases as the length of notch increases, regardless of the size of specimen. For a given size of notch, the notch strength ratio becomes larger with decreasing size of specimen. A notch-size effect law is derived by means of the Neuber interpolation method. A specimen-size effect is embedded into the notch sensitivity parameter involved by the notch-size effect law to establish a size effect law that can cope with these two kinds of size effect. The engineering size effect law proposed can adequately describe the specimen-size effect as well as notch-size effect on the tensile strength of the woven CFRP laminates. It is also demonstrated that the size effect law allows determining the size independent fracture toughness on the basis of notched strengths of small specimens that fail in a quasi-brittle manner.     

Microstructure and high-temperature strength of Al<Subscript>2</Subscript>O<Subscript>3</Subscript>/Er<Subscript>3</Subscript>Al<Subscript>5</Subscript>O<Subscript>12</Subscript>/ZrO<Subscript>2</Subscript> ternary melt growth composite

A new Al₂O₃/Er₃Al₅O₁₂(EAG)/ZrO₂ ternary MGC (Melt Growth Composite) with a novel microstructure has been fabricated by unidirectional solidification. This ternary MGC has a microstructure consisting of continuous networks of single-crystal Al₂O₃, single-crystal EAG and fine cubic-ZrO₂ phases without grain boundaries. The ternary MGC has also characteristic dimensions of the microstructure of around 2–4 m for EAG phases, around 2–4 m for Al₂O₃ phases reinforced with around 0.4–0.8 m cubic-ZrO₂ phases. No amorphous phases are formed at interfaces between phases in the ternary MGC. The ternary MGCs flexural strength at 1873 K is approximately 700 MPa, more than twice the 330 MPa of the Al₂O₃/EAG binary MGC. The fracture manner of the Al₂O₃/EAG/ZrO₂ ternary MGC at 1873 K shows the same intergranular fracture as the Al₂O₃/EAG binary MGC, but is significantly different from the transgranular fracture of the sintered ceramic.     

Middle ear mucosa pathophysiology and healing processes in children with otitis media with effusion (OME)

Most otitis media with effusion (OME) in children heals spontaneously, but it is sometimes persistent and intractable. The major factors determining these different courses of OME are unknown. Identifying such factors would be clinically helpful. Factors affecting on the outcome of OME were studied experimentally and clinically. In animal experiment OME was induced in rabbit by occluding the Eustachian tube. Clinical study was conducted by investigating a histology of the middle ear mucosa, sampled from 64 children with OME (87 ears) who underwent middle ear tube ventilation. Furthermore, these cases studied how the degree of mastoid pneumatization before and after treatment, changes in the volume of the middle ear air cavity and the course after removal of the middle ear ventilation tube affected lesions in the middle ear mucosa. In animals, histology of the middle ear mucosa showed that in the early stages, the subepithelial layer of the middle ear mucosa was characterized by edematous hypertrophy and enlarged blood vessels. In later stages, it became fibrous as a result of chronic inflammation. These results suggest that lesions of the subepithelial layer are unlikely to recover. In clinical cases, histological changes in mucosal samples were rated on a two-point scale: changes in the epithelium and changes in the subepithelial layer. Re-pneumatization, a sign of healing of middle ear mucosal inflammation, was seen 12 months to 18 months after surgery. Among patients in whom the tube was left inserted for more than 18 months, only 11% had unfavorable outcomes and the recurrence rate was low. All patients who had recurrence of disease despite more than 18 months of tube insertion had prolonged inflammation in the middle ear mucosa. There results indicate that the air cavity returns to approximately normal degree of pneumatization more than 18 months after surgery, and that the extent of lesions in the middle ear mucosa, especially the subepithelial layer, is closely related to the healing of OME.     

Gene transfer therapy for heritable disease: cell and expression targeting

Gene therapy is defined as the delivery of a functional gene for expression in somatic tissues with the intent to cure a disease. Different gene transfer strategies may be required to target different tissues. Adenosine deaminase (ADA) deficiency is a good gene therapy model for targeting a rare population of pluripotent hematopoietic stem cells capable of self-renewal. We present evidence for the highly efficient gene transfer and sustained expression of human ADA in human primitive hematopoietic progenitors using retroviral supernatant with a supportive stromal layer. A stem cell-enriched (CD34+) fraction was also successfully transduced. Duchenne muscular dystrophy (DMD) is also a good model for somatic gene therapy. Two of the challenges presented by this model are the large size of the gene and the large number of target cells. Germline gene transfer and correction of the phenotype has been demonstrated in transgenic mdx mice using both a full-length and a truncated form of the dystrophin cDNA. We present here a deletion mutagenesis strategy to truncate the dystrophin cDNA such that it can be accommodated by retroviral and adenoviral vectors useful for somatic gene therapy.