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Byline: Yukashi. Yamashita, Taro. Ikegami, Mikio. Suzuki, Hitoshi. Hirakawa, Hiroyuki. Maeda, Satoshi. Yamada, Zeyi. Deng, Shunsuke. Kondo, Hidetoshi. Kinjyo, Asanori. Kiyuna, Shinya. Agena, Takayuki. Uehara, Akira. Ganaha Background: Several studies [...]

The OLETF rat strain is a useful obese type 2 diabetes model with accompanying hypertriglycemia, hyperinsulinemia, hyperglycemia, insulin resistance and abundant visceral fat. Genome-wide quantitative trait loci (QTL) analyses have [...]

To characterize pharmacologically a new insulin sensitizer, NIP-223, we investigated the effect of NIP-223 on PPAR activation, adipocyte differentiation of 3T3-L1 cells, and blood glucose levels in diabetic KK-[A.sup.y] mice, [...]

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Background Serum squamous cell carcinoma antigen (SCCA) levels are elevated in sinonasal inverted papilloma (IP). However, the relationship between tumor volume and SCCA level, and the influence of skin or pulmonary diseases in which the SCCA level is high, have not been established.Objective To clarify whether the level of serum SCCA can be used as a diagnostic marker of IP.Methods Serum SCCA level was measured in 30 patients with IP (IP group) and 57 with inflammatory disease (inflammatory group).Results Overall, 83.3% in the IP group showed elevated serum SCCA levels regardless of whether they were new patients or patients with recurrent IP, and SCCA levels rapidly decreased after surgery. Only 5.3% had elevated SCCA levels in the inflammatory group. Before surgery, the IP group had a median preoperative SCCA level of 2.4 ng/mL, whereas the median preoperative SCCA level was 0.9 ng/mL in the inflammatory group. Pre- and postoperative SCCA levels were significantly different in the IP group. With regard to the IP diagnosis in the IP and inflammatory groups based on the SCCA level (ng/mL), sensitivity and specificity were 83.3% and 94.7%, respectively. There was no significant correlation between SCCA elevation and respiratory function, and skin disease in the two groups, except for smoking in the IP group. Preoperative SCCA levels were significantly higher in smokers than in never-smokers in the IP group. Tumor volume was significantly correlated with SCCA level in IP. Multivariable logistic analysis showed that tumor volume was a predictor of preoperative SCCA elevation (p = 0.036; 95% confidence interval, 1.027–2.176).Conclusion Serum SCCA level is a reliable diagnostic marker to distinguish new and recurrent IP from inflammatory disease. Because smokers tended to have higher SCCA levels in IP, a different cutoff level might be needed. Although respiratory dysfunction and skin disease were not related to SCCA level, they should be taken into consideration when evaluating SCCA level.

Background The clinical importance of serum squamous cell carcinoma antigen (SCCA) and SCCA subclasses has not been established for treating inverted papilloma (IP). The aim of this study was to clarify the clinical importance of serum SCCA and its subclasses in IP, compared with maxillary squamous cell carcinoma and inflammatory disease.Methods Serum SCCA was measured in 22 patients with IP (IP group), 11 with maxillary squamous cell carcinoma (carcinoma group), and 22 with inflammatory disease (inflammatory group). mRNA expression of SCCA subclasses was examined using quantitative real-time polymerase chain reaction.Results In the IP group, 81.8% showed elevated serum SCCA, and 90.3% with recurrent IP showed elevated SCCA. The preoperative SCCA value (mean ± SD, 3.99 ± 4.39) in the IP group was significantly higher than in the carcinoma (1.28 ± 0.88; p = 0.012) and inflammatory (0.60 ± 0.31; p < 0.001) groups. mRNA expression of SCCA1 and SCCA2 in the IP group was higher than in the carcinoma and inflammatory groups. The SCCA2/SCCA1 ratio of mRNA expression (0.11 ± 0.06) in the IP group was similar to that (0.11 ± 0.09) in the inflammatory group, although the ratio (0.20 ± 0.12) in the carcinoma group was significantly higher than in the IP and inflammatory groups The receiver operating characteristic curve analysis for the SCCA2/SCCA1 ratio to detect carcinoma yielded an area under the curve of 0.760 (95% confidence interval, 0.626–0.894).Conclusion The serum level of SCCA is effective for detecting IP, including recurrent IP. In contrast, the SCCA2/SCCA1 ratio is useful for detecting squamous cell carcinoma among other sinonasal diseases.

To investigate the incidence of habitual sniffing and the function of eustachian tube in middle ear cholesteatoma.

Abstract In this paper, we describe intelligent beamforming antenna systems that can be used in the millimeter-wave band for High-altitude platform systems. We have developed two antenna systems for the millimeter-wave band and have designed experiments to test the efficiency of the developed systems. One is a multi-beam-horn antenna that enables high-speed transmission, and the other is an array antenna that digitally controls antenna beams. These antenna systems are also designed to work in the stratosphere. We also describe our solutions to the problems of low temperature and low pressure and show that the two antenna systems can function well in the stratosphere through tests conducted on the ground.

The main purpose of this study is to search for a viral etiology in Ménière’s disease by examining the presence or absence of herpes family virus DNA in the endolymphatic sac (ES) using the in situ hybridization method. This was a prospective study with the ES from 10 patients with Ménière’s disease and from 7 control cases without any pre-mortem ear diseases except a case of acoustic tumor. These 10 patients underwent the ES surgery. The presence of herpes family virus DNA, such as herpes simplex virus types 1 and 2 (HSV1&2), varicella-zoster virus (VZV), Epstein-Barr virus (EBV) and human cytomegalovirus (CMV), was examined using the in situ hybridization method. Serum antibody titers against these viruses just before the ES surgery were studied in these patients. Of the 10 specimens from the patients with Ménière’s disease, 7 were positive for VZV, 4 for EBV, 1 for CMV and none for HSV1&2, although the serum antibody titers against these viruses did not show any significant elevation in these patients just before the ES surgery. This result suggests that the viral DNA in the ES is inactive and is present in a latent form. From the statistical analysis, it can be postulated that VZV infection in early childhood may reach the ES and play a role in the pathogenesis of Ménière’s disease (p = 0.0235). The double infection with both VZV and EBV tended to be another candidate for the pathogenesis of Ménière’s disease (p = 0.0557).

Telecommunication and broadcasting systems using radio-relay/base stations on board the high altitude platforms are expected to create the largest business market among its possible applications. They could also be integrated with satellite- and terrestrial-based systems in the future to construct the more powerful mobile and/or broadband networks. Prototype onboard equipment and ground equipment are being developed on schedule in the fixed, mobile, and broadcasting services. The performance of them will be demonstrated and evaluated in the preliminary flight tests planned in 2002 using alternative aerial vehicles: a helicopter, a jet, and a solar-powered unmanned aerial vehicle (UAV), before flight tests using the high altitude airships. This paper presents activity in Japan on the R&D of wireless access systems using high altitude platform stations and the detail of the preliminary flight test program.

In the World Radiocommunication Conference 2000 (WRC-2000), use of the 31 GHzand 28 GHz bands was permitted for the fixed service (FS) by using highaltitude platform stations (HAPS) in some countries. This paper examines thesharing and compatibility of the HAPS-based FS with the other services usingthe same and adjacent frequency bands and establishes the conditions of thecoexistence with those services. Feasibility of dynamic channel assignment(DCA) scheme among the different communication systems to facilitate thecoexistence is also studied.

To characterize brain activity in response to auditory stimuli during recovery from acute hearing loss, fMRI was performed at two time points in 11 patients with sudden deafness in the right ear, and 10 subjects with normal hearing. In the acute phase, right-ear auditory stimulation induced only a small response in the auditory cortex, limited to the left hemisphere. In the recovery phase, the auditory response was more extensive than in the acute phase. Stimulation of the left ear induced a more extensive response in the left than right hemisphere in both acute and recovery phases, which differed from the pattern in normal subjects. The changes in cortical activation patterns were seen within 1 week of sudden deafness. Thus, alteration of cortical response in deafness occurs earlier than suggested by previous reports.

We performed fMRI on the human parieto-occipital cortex in order to identify the neural processing regions of stereopsis. Visual stimulation for stereopsis was performed with a random-dot stereogram displayed in the image guides of a new binocular visual stimulation device that we developed. Interestingly, regions from the dorsal portion of the occipital lobe to the superior parietal lobule were activated by binocular disparity, while the inferior parietal lobule was not activated. Moreover, these regions were shown as dominant in the right hemisphere. Functional brain mapping revealed that the processing regions of stereopsis were dorsally located in parieto-occipital cortex, and that the superior parietal lobule is an important region for neural processing of human stereopsis.

Whole-genome scans have identified Dmo1 as a major quantitative trait locus (QTL) for obesity and dyslipidaemia in the Otsuka Long Evans Tokushima Fatty (OLETF) rat. We have produced congenic rats for the Dmo1 locus, using marker-assisted speed congenic protocols, enforced by selective removal of other QTL regions (QTL-marker-assisted counterselection), to efficiently transfer chromosomal segments from non-diabetic Fischer 344 (F344) rats into the OLETF background. In the third generation of congenic animals, we observed a substantial therapeutic effect of the Dmo1 locus on lipid metabolism, obesity control and plasma glucose homeostasis. We conclude that single-allele correction of an impaired genetic pathway can generate a substantial therapeutic effect, despite the complex polygenic nature of type II diabetic syndromes.

Through a combination of radiation hybrid mapping and studies by FISH and zoo-FISH we have made a comparative investigation of the distal portion of rat chromosome 1 (RNO1) and the entire mouse chromosome 19(MMU19). It was found that homologous segments of RNO1 and MMU19 are similar in banding morphology and in length as determined by several different methods, and that the gene order of the 46 genes studied appears to be conserved across the homologous segments in the two species. High-resolution zoo-FISH techniques showed that MMU19 probes highlight only a continuous segment on RNO1 (Iq43-qter), with no detectable signals on other rat chromosomes. We conclude that these data suggest the evolutionary conservation of a chromosomal segment from a common rodent ancestor. This segment now constitutes the entire MMU19 and a large segment distally on RNO1q in the mouse and rat, respectively.

The endolymphatic-perilymphatic shunt operation between the scala media and scala tympani was performed in 22 guinea pigs with endolymphatic hydrops induced by the silver nitrate injection method 2 months before the operation. Two (n = 10), 7 (n = 6), and 28 (n = 6) days after the operation, the condition of the fistula and the inner ear pathology were studied histopathologically. In half of the animals, the fistula was open, whereas in the other half, the fistula was closed. In the group examined after 2 postoperative days, the animals with open fistulas showed a slight collapse of Reissner’s membrane, which might indicate that excess endolymph escaped through the fistula into the scala tympani. In the groups examined later, especially in the group examined after 28 postoperative days, the animals showed distention of hydrops despite preservation of the fistula. Moderate-to-severe degenerative changes of the organ of Corti, macula sacculi, and stria vascularis were observed in the vicinity of the fistula. We conclude from this study that the endolymphatic-perilymphatic fistula created inside the cochlea apparently was not sufficient to reduce the hydrops. Rather, it caused degenerative changes of the inner ear structure. This type of surgery is not suitable for Meniere’s disease. (Otolaryngol Head Neck Surg 2000;122:119-23.)

Poly(1,2-dihydroxyethylene-co-vinyl alcohol)s (P(DHE-VA)s) as polymer homologues of methanol/ethanol mixtures were prepared by alkaline hydrolysis of poly(vinylene carbonate-co-vinyl acetate)s. The obtained P(DHE-VA)s were soluble in water reflecting their repeating structures. P(DHE-VA)s were found to show higher thermal stability than poly(vinyl alcohol) (PVA). Membranes composed of P(DHE-VA)s and phosphoric acid exhibited higher ionic conductivity than PVA/phosphoric acid under unhumidified condition, reflecting denser OH groups on the main chains of P(DHE-VA)s.

The Ikaros gene (symbol ZNFN1A1) encodes the hematopoietic zinc finger DNA binding protein, which is now recognized as a central regulator of lymphoid differentiation and has been implicated in leukemogenesis. Recently, an Ikaros-related zinc finger protein, called Aiolos (ZNFN1A3), has been identified and characterized, thus establishing the presence of a gene family whose members may be hematopoietic transcription factors. Among Aiolos-mutant mice, development of B-cell lymphoma was frequently seen. As an initial approach to examining the possible involvement of Aiolos in the pathogenesis of human lymphoid proliferative disease, we isolated cDNA clones for human Aiolos from a B-cell cDNA library. The human Aiolos protein predicted from the cDNA sequence consists of 509 amino acid residues and shares 86% sequence identity with its mouse counterpart. As in the case with mouse Aiolos, no isoform for human Aiolos has been found. Northern blot analysis of various human tissues revealed that the Aiolos transcripts are expressed most strongly in peripheral blood leukocytes, the spleen, and the thymus, supporting the notion that Aiolos plays an important role in lymphoid lineages. Fluorescence in situhybridization using a BAC clone established that the Aiolos gene is mapped to human chromosome band 17q11.2.

TheLy6family of genes encodes glycosylphosphatidylinositol-anchored cell surface glycoproteins expressed on various types of cells. Intriguing patterns of expression ofLy6genes on specific subpopulations of lymphoid and myeloid cells suggest that Ly6 molecules may be involved in the development and homeostasis of hematopoietic cells. We have isolated a new member of the humanLy6gene family,LY6H,from a human fetal brain cDNA library. Fluorescencein situhybridization and radiation hybrid analyses assignedLY6Hto chromosome 8, where other members of theLy6gene family are also located. Northern analysis revealed thatLY6His highly expressed in particular subdivisions of human brain and also in MOLT-3 and -4 acute lymphoblastic leukemia cells. These data suggest that LY6H may play a role(s) in both the central nervous system and the immune system.

Experimental allergic encephalomyelitis is an animal model of a T cell—mediated autoimmune disease — for example, multiple sclerosis. We demonstrated that mice with experimental allergic encephalomyelitis developed retrocochlear hearing loss, and that the lesion of the auditory pathway might be related to T cell receptor Vß8–expressing T cells. To investigate whether anti-Vß8 antibody could prevent hearing loss, we carried out brain stem auditory evoked potential testing, histologic examinations, and flow cytometry in antibody-treated and control myelin basic protein—immunized B10.PL mice. The antibody was administered just before immunization of myelin basic protein. The disease incidence and severity were significantly reduced in the mice injected with the antibody. The results of brain stem auditory evoked potential testing, histologic examinations, and flow cytometry indicated that the depletion of Vß8-expressing T cells brings the prevention of hearing loss, as well as prevention of other neurologic deficits. The development of T cell receptor—specific antibody therapy might help treat retrocochlear hearing loss in multiple sclerosis.

AbstractThe long-term results of endolymphatic sac (ES) shunt surgery in Ménière’s disease have been reported to be unsatisfactory compared to the short-term results, probably because of fibrosis and/or reclosure of the incised ES. To solve this problem, we tried to apply mitomycin C (MMC) intraoperatively to the incised ES as used in trabeculectomy for glaucoma. MMC has an antiproliferative effect as well as an antineoplastic effect. Thus, it is expected that MMC would have a benefit to prevent rapid fibrosis and/or reclosure of the ES. MMC did not show any ototoxicity in our animal study and we started a clinical trial after being approved by the Ethics Committee for Human Research at our university. Fourteen patients with Ménière’s disease underwent ES mastoid shunt surgery with intraoperative application of MMC and they were followed up more than 6 months. No patients experienced vertigo, although some complained of slight dizziness. Five patients out of 14 showed remarkable hearing improvement by more than 10 dB, accompanied with decrease of tinnitus.

We have cloned a novel gene encoding a human ubiquitin-specific protease (USP1). The product, which consists of 785 amino acids with a deduced molecular mass of 88.2 kDa, possesses His and Cys domains that are highly conserved in all members of the ubiquitin-specific processing (UBP) family of proteases. Recombinant USP1 protein showed genuine UBP activity, correctly cleaving Ub-β-galactosidase to produce ubiquitin and β-galactosidase. Chromosomal mapping by fluorescencein situhybridization and radiation hybrid analyses localized the USP1 gene to the p31.3–p32.1 band of chromosome 1. As losses of heterozygosity or amplifications have been observed in the distal region of the short arm of chromosome 1 in some neuroblastomas, breast cancers, and pancreatic adenocarcinomas, the USP1 gene may be a candidate for either the tumor-suppressive or the oncogenic activities associated with that chromosomal region.

Patients with Meniere's disease were investigated for possible immunologic abnormalities. A total of 104 cases—55 bilateral and 49 unilateral—were examined. Immune activity was assessed using four types of immune parameters: (1) the erythrocyte sedimentation rate (ESR) and C-reative protein levels (CRP); (2) serum immunoglobulin levels (IgG, IgM, and IgA); (3) complement levels (C3, C4, and CH50); and (4) autoantibody levels (rheumatoid factor, anti-DNA antibody, and antinuclear antibody). Patients with severe immunologic abnormalities (16% of the bilateral cases and 2% of the unilateral cases) typically displayed bilateral severe hearing loss, bilateral decreased vestibular response, and excellent responsiveness to steroid treatment. As patients with bilateral involvement account for approximately 30% of all cases of Meniere's disease in Japan, immunologic involvement may be assumed in about 6% of the total. The severity of these patients' symptoms and their responsiveness to steroid treatment suggest that the possibility of immunologic involvement is well worth investigation in Meniere's disease.

BAI1 (brain-specific angiogenesis inhibitor 1), a p53-target gene specifically expressed in brain, encodes a seven-span transmembrane protein considered to be a member of the secretin receptor family. Using a two-hybrid system, we isolated a cDNA encoding a product that interacts with the cytoplasmic region of BAI1 and designated it BAP3 (BAI1-associated protein 3). The BAP3 product is a novel C2 domain-containing molecule with homology to Munc13 and synaptotagmin. As with Munc13, BAP3 is expressed predominantly in brain. Deletion-mutant analysis revealed that the interaction between BAI1 and BAP3 was not mediated by the C2 domains. Its predominant expression in brain and homology to Munc13 indicate that BAP3, by interacting with BAI1, might be involved in some neuronal function such as regulating release of neurotransmitters.

To obtain DNA markers on human chromosome 1, we first isolated 500 cosmid clones from mouse A9 cells containing a human chromosome 1 tagged with pSV2neo. Of these, 186 were localized on each band of human chromosome 1 by R-banding fluorescence in situ hybridization; 118 and 68 were on the short and long arms, respectively. We performed restriction fragment length polymorphism (RFLP) analysis of these cosmid clones, and polymorphism was recognized with one or more enzyme in 43 of them. Two markers proved to have variable numbers of tandem repeats. Since several tumor suppressor genes, as well as genes responsible for hereditary disorders, may be located on this human chromosome, the DNA markers will be useful for RFLP analysis or the isolation of new genes related to various disorders. Copyright 1995, 1999 Academic Press

The 26S proteasome is a eukaryotic ATP-dependent protease functioning as a protein death machine. It is a large multisubunit complex, consisting of a catalytic 20S proteasome and two regulatory modules, named PA700. The PA700 complex is composed of multiple subunits of 25-110 kDa, which are classified into two subgroups, a subgroup of at least 6 ATPases that constitute a unique multi-gene family encoding homologous polypeptides conserved during evolution and a subgroup of approximately 15 non-ATPase subunits, most of which are structurally unrelated to each other. In the present study, we report the chromosomal localization and immunological properties of six members of the human 26S proteasomal ATPase family. By use of the fluorescencein situhybridization method, the S4 (PSMC1), MSS1 (PSMC2), TBP1 (PSMC3), TBP7 (PSMC4), p45 (PSMC5), and p42 (PSMC6) genes were mapped to human chromosomes 19p13.3, 7q22.1-q22.3, 11p11.2, 19q13.11-q13.13, 17q23.1-q23.3, and 12q15, respectively, indicating that the genes for multiple ATPases of the 26S proteasome are located on different chromosomes. Immunoblot analysis revealed that all these ATPases were associated with the purified 26S proteasome and that some of them showed striking heterogeneity in their electrical charges.

C-TYPE natriuretic peptide (CNP) is the third member of the natriuretic peptide family which plays an important role in body fluid homeostasis. To determine a possible role of CNP in regulation of an inner ear fluid, we investigated the expression of CNP and atrial natriuretic peptide B receptor (ANP-B receptor) mRNAs in rat inner ear using a reverse transcription polymerase chain reaction (RT-PCR) method. Amplification products with sizes expected for CNP and ANP-B were detected in the inner ear. After cloning and analysis, the sequences for PCR products were identical to those of CNP or ANP-B receptor in the brain. These results indicate that both CNP and ANP-B receptor are expressed in the inner ear of the rat and suggest that CNP may play a role in inner ear function (such as regulation of inner ear fluid) in an autocrine and/or paracrine manner.

From a human fetal-brain cDNA library we isolated two novel genes encoding peptides containing six EGF-like repeats. Both showed significant homologies with nel, a gene strongly expressed in neural tissues of chicken. The cDNAs, designated NELL1 (nel-like, type 1) and NELL2 (nel-like, type 2), contained open reading frames encoding 810 and 816 amino acids, respectively. NELL2 is strongly expressed in brain of adult and fetus but only weakly in fetal kidney. NELL1 and NELL2 were mapped by FISH to chromosomal bands 11p15.1–p15.2 and 12q13.11–q13.12, respectively.

The YL-1gene, encoding a novel nuclear protein with transcription factor-like features, has been isolated from the human chromosome 1q21, one of the regions supposedly carrying a transformation suppressor gene(s) for Kirsten sarcoma virus-transformed NIH3T3 (DT) cells. To test the suppressive activity of the YL-1gene product, we forced the expression of human YL-1cDNA in DT cells. The anchorage-independent growth (colony-forming ability in soft agar medium) was markedly suppressed in cells highly expressing the exogenous human YL-1 protein. Moreover, the soft agar clones, which were rarely originated from these cells, expressed reduced levels of exogenous YL-1or none, with or without the loss/rearrangement of the introduced cDNA. In control experiments, cells carrying an introduced vector alone or an antisense-strand expression plasmid grew in soft agar as efficiently as parental DT cells. In contrast to the suppression of anchorage-independent growth, the forced expression of YL-1did not affect the transformed phenotypes in adherent culture and tumorigenicity in nude mice. These findings not only indicated that the YL-1 protein functions as a transformation suppressor, but also suggest that it may be important for elucidating anchorage independence under separate genetic control from other transformed phenotypes.

We have isolated and characterized a cDNA and the gene of the human homologue of CAAF1, a novel member of the S100 calcium-binding protein family. The 276-bp open reading frame encoded a 92-amino acid polypeptide with a predicted molecular mass of 10,575 Da. The deduced amino acid sequence of human CAAF1 showed 66% homology to bovine CAAF1. The human CAAF1 gene consisted of three exons, with the two EF hand motifs of the CAAF1 protein separately encoded by exons 2 and 3. This gene was expressed at a high level in polymorphonuclear leukocytes and at an intermediate level in esophageal mucosa. The tissue distribution of CAAF1 mRNA was different from that of other S100 proteins. Direct R-banding fluorescence in situ hybridization revealed that the human CAAF1 gene was mapped to chromosome 1q21.2-q22, where most of the S100 genes form a cluster.

Some immortal human cell lines lack telomerase activity. These cell lines were found to contain small dispersed DNA hybridizing to TTAGGG repeats. Such DNA was located in their cytoplasm and nuclei. Normal human fibroblasts or telomerase-positive cell lines did not contain such DNA. Upon cloning and sequencing, it was shown to consist of TTAGGG repeats. When electrophoresed on neutral and alkaline agarose gels, it behaved as double-stranded and linear DNA. These results suggest that telomeric DNA is released from chromosomes in association with maintenance of telomeres in telomerase-negative cell lines.

Irradiation of azobis(4-spiro[2.4]heptane) and azobis(4-spiro[2.5]octane) afforded homoallylically rearranged products via the corresponding titled radicals, in which the rigid geometry locks the cyclopropylcarbinyl radical in the stable bisected conformation.

Sensitivity induced by house dust mite (HDM) extract in mice was investigated in this study. Sensitized B10.RIII mice (H-2rbackground) had T-cell proliferative responses to HDM extract in vitro and an HDM-specific IgE response. When mice were immunized by injection and intranasal inhalation with HDM extract, a histologic study showed eosinophils and mononuclear cell infiltration in the lung tissue and bronchial wall. Tcrαβ-positive cells were also found in the cell infiltration area of the lung lesions. In the control mice that were immunized by injection or intranasal inhalation (but not both), we did not observe cell accumulation in the lung tissue or in the bronchial wall. Epitope studies suggest that T cells recognize multiple epitopes. Molecular analysis of these HDM-specific T-cell hybridoma clones suggest that T-cell receptor use is restricted to members of the Vα 8 and Vβ 6 subfamilies. (J Allergy Clin Immunol 1998;101:51-9)

We have employed cDNA cloning to deduce the complete primary structure of a new subunit, designated p27, of the modulator trimer complex that stimulates the association of the PA700 regulator with the catalytic 20S proteasome to form the ATP-dependent active 26S proteasome. We found two distinct cDNAs encoding two highly homologous proteins except in the C-terminal region, which are termed tentatively p27-1 and p27-2. The short p27-2 cDNA has a deletion of 65 bp near the 3′-end region of the long p27-1 cDNA, which encodes a large protein with an extended C-terminal region, designated p27-L, whereas the long p27-1 cDNA encodes a small protein named p27-S. The polypeptides of p27-L and p27-S consist of 223 and 209 amino acid residues with calculated molecular masses of 24,852 and 22,764 and isoelectric points of 6.50 and 5.28, respectively. Immunoblot analysis with anti-p27 antibody revealed that p27, together with two other ATPase components, TBP1 and p42, was associated with not only the modulator complex but also significantly with the 26S proteasome complex, suggesting that the three are common/sharing subunits in these two complexes. By the fluorescencein situhybridization method, the p27 (PSMD9) gene was mapped to the q24.2–q24.3 band of human chromosome 12. Computer-assisted homology analysis revealed the high sequence similarities of p27-L with a possible counterpart inCaenorhabditis elegansandSaccharomyces cerevisiaewhose function is yet unknown, the yeast gene that is here termedNAS2(non-ATPase subunit 2). Disruption ofNAS2had no effect on cell viability, indicating that the subunit is not essential for proliferation of yeast cells.

A spectrophotometric method has been developed for the determination of tetrathionate, thiosulfate and sulfide in mixtures. The conditions for tetrathionate to be stoichiometrically converted to thiosulfate were established by varying reaction time, reaction temperature, pH and amount of sulfide. The present method consists of three procedures: I, II and III. The absorbance obtained by Procedure I corresponds to the sum of the thiosulfate and twice the tetrathionate in the mixture. The absorbance obtained by Procedure II, where tetrathionate was not sulfitolyzed and sulfide was removed, corresponds to the amount of thiosulfate only. The absorbance obtained by Procedure III corresponds to the sum of the amount of thiosulfate and twice that of sulfide. The method is applicable to the determination of tetrathionate, thiosulfate and sulfide in amounts more than 0.1 μmol and gave a relative standard deviation of 1.2% at 1 μmol level of tetrathionate in the presence of 1 μmol of sulfide and 2 μmol of thiosulfate.

By using a transmission electron microscope equipped with an energy dispersive spectrometer, it was possible to detect the morphological, structural, and chemical characteristics of individual asbestos fibers and clay minerals without any realignment of the equipment. A rapid and convenient procedure for semiquantitative analysis is proposed. Analyses are given for 21 hydrous silicates, asbestos and clay minerals, by both ordinary chemical and energy dispersive methods. The energy dispersive results were comparable to those obtained by chemical analysis. Application of this procedure to asbestos fibers proved this method to be practical and valid for characterization of asbestos in environmental and tissue samples. С помощью трансмиссионного электронного микроскопа, оборудованного рассеивающим энергию спектрометром, стало возможным определять морфологические, структурные и химические характеристики отдельных асбестовых волокон и глинистых минералов без повторной центровки оборудования. Приводятся данные анализов 21 водных силикатов, асбестов и глинистых минералов, полученных с помощью обычных химических и рассеивающих энергию методов. Результаты использования рассеивающих энергию методов были сопостовимы с результатами химических анализов. Применение этой процедуры для изучения асбестовых волокон доказало его практичность и пригодность для получения характеристики асбестов в природных и тканевых образцах. Indem ein Transmission-Elektronen-Mikroskop, ausgestattet mit einem Energie-Streuungsspektrometer, benutzt wurde, war es möglich, die mor -phologischen, strukturellen und chemischen Eigenschaften von einzelnen Asbestfasern und Tonmineralien zu ermitteln, ohne die Apparatur wieder zu eichen. Eine schnelle und bequeme Methode für semi-quantitative Analyse wird vogeschlagen. Analysenresultate, erhalten durch allgemeine, chemische und Energie-Streuungsmethoden, für 21 wasserhaltige Silikate, Asbest und Tonmineralien sind angegeben. Die Resultate der Energie-Streuungsmethode waren vergleichbar mit denen, erhalten durch chemische Analyse. Anwendung dieser Prozedur auf Asbestfasern, hat bewiesen, daß diese Methode praktisch und brauchbar für die Charakterisierung von Asbest in Umwelt-und Gewebeproben ist. Par l’emploi d’un microscope électronique à transmission équippé d’ un spectromètre dispersant l’énergie, il a été possible de détecter les caractéristiques morphologiques, structurales et chimiques de fibres d’asbeste individuelles et de minéraux argileux sans réaligner l’équipement. Un procédé rapide et commode pour l’analyse semi-quantitative est proposé. Des analyses sont données pour 21 silicates hydratées, asbestes et mineraux argileux, à la fois par des méthodes chimiques ordinaires et des méthodes dispersant l’énergie. Les résultats des méthodes dispersant l’énergie sont comparables à ceux obtenus par analyse chimique. L’application de ce procédé aux fibres d’asbeste prouvent que cette méthode est pratique et valable pour la caractérisation d’asbeste dans les échantillons de tissu et du milieu ambiant.

2-Aza-3,7,8,12,13,17,18-heptaalkylporphyrins were synthesized by use of improved ‘3+1’ condensation under non-acidic conditions.

The responses of primary vestibular neurons and perilymphatic pressure changes to middle ear pressure were investigated in guinea pigs with obstructed vestibular or cochlear aqueduct (closed VA or closed CA group) in order to clarify the influence of VA and CA on pressure-induced vestibular response. Although the neural response rates and the amount of perilymphatic pressure change in the closed VA group resembled those in the control group, these values in the closed CA group were higher than in the control group. Patency of the CA had a more significant effect on the vestibular response to middle ear pressure change than patency of the VA.

The responses of primary vestibular neurons and perilymphatic pressure changes to middle ear pressure stimuli in guinea pigs were investigated in order to clarify the direct effects of pressure stimulus on the vestibular apparatus. The vestibular response was related to the amount of middle ear pressure change applied at a rate of ± 100 mmH2O/s. The neural response rates of vestibular units to positive pressure in the middle ear were significantly larger than those to negative pressure. The time course pattern of the perilymphatic pressure change resembled that of the response of the vestibular units, indicating that the vestibular response is elicited by middle ear pressure via the pressure transmitted in the inner ear.

MAD (mothers against decapentaplegic)-related proteins (MADRs) are intracellular components that play critical roles in signal-transduction pathways involving the transforming growth factor β (TGFβ) superfamily. Some Mad genes are candidates for tumor-suppressor functions. From a human fetal brain cDNA library we have isolated a novel Mad-related gene. Two alternatively transcribed mRNAs encode deduced 430- and 467-amino-acid peptides that showed high levels of similarity to MADR1/Smad1/hMAD1 (about 80% identity at the amino acid level). This gene, which we designated MADH6, resides on 13q12–q14 between BRCA2 and RB, a region that frequently displays loss of heterozygosity in breast, liver, and prostate cancers.

Irradiation of 3-oxidopyridazinium betaines (4) and (9) in acetonitrile gave fused diaziridines (5) and (10) which are stable enough to isolate. The amido bond of (5) underwent alcoholysis and aminolysis with ease. Thus, apparent phototranspositions of (4) and (9) to pyridazinones (6) and (11) in water were proved facile hydrolysis of the amide bonds of the intermediates (5) and (10).

AbstractLaser Doppler flowmetry was used to assess cochlear blood flow (CoBF) in guinea pigs with experimental endolymphatic hydrops following intravenous infusion of 5 types of drugs: 50% glycerol, 70% isosorbide, 20% mannitol, 7% sodium bicarbonate, and 1% diphenidol. The magnitude of the CoBF changes following infusion tended to be smaller in the hydropic ears than in the normal control ears. A significant reduction in CoBF changes was observed in hydropic ears infused with isosorbide and sodium bicarbonate. These results suggest that the cochlear microvascular sensitivity to various stimuli such as drug infusion is reduced in hydropic ears. This may result from atrophy of the stria vascularis which is often observed in the hydropic ears of guinea pigs. Thus it seems likely that the same reaction occurs in the inner ear of patients with Ménière’s disease in whom atrophy of the stria vascularis is also presumed to exist in conjunction with extensive endolymphatic hydrops. Therefore, it seems probable that the function of the microvasculature of the stria vascularis is impaired in the inner ear of patients with Ménière’s disease, resulting in the slow progressive deterioration of the inner ear with time.

To study the function of the B cell signal transduction molecule α4 (IGBP1), we isolated a human α4 (IGBP1) gene that has sequence similarity to the yeast protein (TAP42) involved in the rapamycin-sensitive signal transduction pathway. The human α4 has sequence identities with murine α4 of 83.4% nucleotide and 82.9% amino acid sequence, and a stretch of consensus motifs in the carboxyl terminal is conserved among the related genes of human, mouse, yeast, and rice. The gene is expressed as a 1.4-kb mRNA in the spleen, lymph node, thymus, appendix, peripheral blood leukocytes, bone marrow, fetal liver, heart, brain, placenta, skeletal muscle, kidney, and pancreas. The anti-human α4 antibody detected a 45-kDa protein in human lymphoid cell lines. Moreover, human α4 (IGBP1) gene is located at q13.1–q13.3 on chromosome X.

Selectivities in halogenation of 2,3-dimethylbutane, 1-chlorobutane and substituted toluenes with N-chloro-or N-bromo derivatives of 4,4-dimethyl-2-oxazolidinone (NXDMO), 2-oxazolidinone (NXO), and succinimide (NXS) were examined. In the presence of olefin, halogenation of the substituted butane by these N-halo reagents were found to proceed involving hydrogen abstraction by the N-centered radicals, since chlorination by the N-chloro reagents and bromination by the corresponding N-bromo reagents showed the same selectivities. The relative reactivity of a t-C-H bond compared with a prim-C-H bond RSptmarkedly depends on the structure of the N-radical, being 200, 70, and 11 at 80 °C toward the radicals derived from NXDMO, NXO, and NXS, respectively. Reaction constant ρ of hydrogen abstraction from the toluenes by the N-radical from NXDMO was −1.0 at 130 °C.

Background Intractable posterior epistaxis sometimes requires intensive treatment, such as surgery or embolization. Maxillary artery ligation has been widely used for the treatment of intractable posterior epistaxis. It is highly effective, but significant complications may occur, including an oroantral fistula and damage to the infraorbital nerve. Embolization is less invasive and can be performed in poor surgical candidates. However, it has more serious complications, such as facial nerve paralysis and hemiplegia. This investigation evaluates the effectiveness and complications of endoscopic ligation of the sphenopalatine or maxillary artery for the treatment of intractable posterior epistaxis.Methods Between April 2003 and March 2007, 46 patients were hospitalized for the treatment of severe posterior epistaxis in our University Hospital. Thirty patients were successfully treated by anterior and/or posterior nasal packing, and five patients were treated by electrocoagulation. Endoscopic ligation was performed under general anesthesia in 11 patients (6 men and 5 women; age range, 50-80 years).Results Eight patients underwent endoscopic ligation of the sphenopalatine artery, and three patients underwent endoscopic ligation of the maxillary artery through the middle meatus and posterior antral wall opening. There were no complications, and the patients’ postoperative courses were uneventful. Recurrent epistaxis occurred in one patient on oral anticoagulants 15 months after ligation of the sphenopalatine artery, and it was successfully treated by anterior nasal packing.Conclusion Endoscopic ligation of the sphenopalatine or maxillary artery is safer than arterial embolization and is less invasive than transantral ligation of the maxillary artery. This technique appears to be a simple and highly effective surgical treatment for patients with intractable posterior epistaxis.

A new cationic monoazaporphyrin, zinc 2-aza-8,12,13,17-tetraethyl-2,3,7,18-tetramethylporphyrinium iodide 3was synthesized. Photodynamic activity of 3in degradation of 2′,3′-isopropylideneguanosine 4was compared with 2-aza-8,12,13,17-tetraethyl-3,7,18-trimethylporphyrin 1, zinc 2-aza-8,12,13,17-tetraethyl-3,7,18-trimethylporphyrin 2, and hematoporphyrin 5. The quarternary ammonium 3showed a remarkable increase of photodynamic activity compared with 5, although no appreciable difference in the activity was observed between 1and 5.

THE anti-diuretic hormone vasopressin has been shown to be important in regulating inner ear fluid. The diuretic hormone, CNP, and its receptor, ANP-B receptor, may also function in the regulation of inner ear fluid. To determine whether vasopressin directly affects the fluid level, we infused this hormone to rat and assay of V2–AVP receptor mRNA by semiquantitative RT-PCR demonstrated a significantly lower level of this transcript in vasopressin-infused animals than in saline-infused animals. The levels of CNP and ANP-B receptors mRNA, however, were the same in both groups of rats. Results suggest that high plasma levels of vasopressin may be a principal causal factor of endolymphatic hydrops in Meniere's disease, perhaps by down-regulating the number of vasopressin receptors.

The 14-3-3 family of proteins exerts diverse influences on the signal transduction pathways of cells. We have newly identified a human cDNA encoding the γ subtype of the 14-3-3family of genes. The deduced amino acid sequence of human 14-3-3γ was identical to that of rat 14-3-3γ. The human 14-3-3γ gene (HGMW-approved symbol YWHAG) is highly expressed in brain, skeletal muscle, and heart. By fluorescence in situhybridization analysis, the human 14-3-3γ gene was mapped to chromosome 7q11.23. Radiation hybrid mapping has shown that this gene is localized 2.33 cR telomeric to D7S1870, a polymorphic marker located at the most telomeric end of the common deletion region of Williams–Beuren syndrome (WBS). This suggests that haploinsufficiency of 14-3-3γ may not contribute to the WBS phenotype. However, information regarding the precise chromosomal location of a member of the 14-3-3family of genes will aid in examining the relationship between this family of proteins and human disorders.

THE cause of endolymphatic hydrops, a characteristic finding in Menière's disease, is not known. To study the possible involvement of the neurohormones vasopressin and oxytocin in this condition, we investigated whether transcripts of the genes encoding the arginine vasopressin (AVP) and oxytocin receptors are expressed in the rat inner ear. Utilizing the reverse transcription-polymerase chain reaction (RT-PCR) method, primers specific for each receptor showed a single message band of the expected size in the rat inner ear. When the PCR products were cloned, the sequences were identical to those of the real-type (V2) AVP receptor and oxytocin receptor transcripts. The finding of vasopressin and oxytocin receptor mRNAs in the inner ear suggests that these neurohypophyseal hormones may have roles in the regulation of inner ear fluid. In particular, the presence of vasopressin receptor mRNA in the inner ear supports the hypothesis of a relationship between high plasma vasopressin levels and endolymphatic hydrops.