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Mucosal tissue forms the first line of defense against pathogenic microorganisms. Cellular damage in the mucosal epithelium may induce the interleukin (IL)-22-related activation of many immune cells, which are essential for maintaining the mucosal epithelial barrier. A previous study on mucosal immunity elucidated that mammalian IL-22 contributes to mucus and antimicrobial peptides (AMPs) production and anti-apoptotic function. IL-22 has been identified in several teleost species and is also induced in response to bacterial infections. However, the roles of IL-22 in teleost immunity and mucus homeostasis are poorly understood. In this study, Japanese medaka (Oryzias latipes) was used as a model fish. The medaka il22, il22 receptor A1 (il22ra1), and il22 binding protein (il22bp) were cloned and characterized. The expression of medaka il22, il22ra1, and il22bp in various tissues was measured using qPCR. These genes were expressed at high levels in the mucosal tissues of the intestines, gills, and skin. The localization of il22 and il22bp mRNA in the gills and intestines was confirmed by in situ hybridizations. Herein, we established IL-22-knockout (KO) medaka using the CRISPR/Cas9 system. In the IL-22-KO medaka, a 4-bp deletion caused a frameshift in il22. To investigate the genes subject to IL-22-dependent regulation, we compared the transcripts of larval medaka between wild-type (WT) and IL-22-KO medaka using RNA-seq and qPCR analyses. The comparison was performed not only in the naïve state but also in the dextran sulfate sodium (DSS)-exposed state. At the transcriptional level, 368 genes, including immune genes, such as those encoding AMPs and cytokines, were significantly downregulated in IL-22-KO medaka compared that in WT medaka in naïve states. Gene ontology analysis revealed that upon DSS stimulation, genes associated with cell death, acute inflammatory response, cell proliferation, and others were upregulated in WT medaka. Furthermore, in DSS-stimulated IL-22-KO medaka, wound healing was delayed, the number of apoptotic cells increased, and the number of goblet cells in the intestinal epithelium decreased. These results suggested that in medaka, IL-22 is important for maintaining intestinal homeostasis, and the disruption of the IL-22 pathway is associated with the exacerbation of inflammatory pathology, as observed for mammalian IL-22.

In mammals, interleukin (IL)-17A and IL-17F, mainly produced by Th17 cells, are hallmark inflammatory cytokines that play important roles in the intestinal mucosal immune response. In contrast, three mammalian IL-17A and IL-17F counterparts (IL-17A/F1-3) have been identified in teleosts, and most of their functions have been described in the lymphoid organs. However, their function in the intestinal mucosal immune response is poorly understood. In this study, a recombinant (r) tiger puffer fish fugu (Takifugu rubripes) IL-17A/F1 was produced and purified using a mammalian expression system, and was used to stimulate cells isolated from fugu head kidney and intestines. The gene expression levels of TNF-α, IL-1β, IL-6, and β-defensin-like protein-1 (BD-1) genes were evaluated at 0, 3, 6 and 12 h post-stimulation (hps). Phagocytic activity and superoxide anion production were evaluated at the same time points using an NBT assay. The rIL-17A/F1 protein was shown to induce the expression of pro-inflammatory cytokines and antimicrobial peptides in both head kidney and intestinal cells. Expression levels for IL-1β, TNF-α, and IL-6 were all up-regulated between 3 and 12 hps. In addition, stimulation with rIL-17A/F1 enhanced phagocytic activity at 24 hps. Superoxide anion production was increased at 48 hps in the head kidney cells and moderately increased at 48 hps in intestinal cells. This study suggests that fugu IL-17A/F1 plays an important role in promoting the innate immune response and may act as a bridge between innate and adaptive immunity in the head kidney and intestine.

Anti-mucin1 (MUC1) antibodies have long been used clinically in cancer diagnosis and therapy and specific bindings of some of them are known to be dependent on the differential glycosylation of MUC1. However, a systematic comparison of the binding specificities of anti-MUC1 antibodies was not previously conducted. Here, a total of 20 glycopeptides including the tandem repeat unit of MUC1, APPAHGVTSAPDTRPAPGSTAPPAHGV with GalNAc (Tn-antigen), Galβ1-3GalNAc (T-antigen), NeuAcα2-3Galβ1-3GalNAc (sialyl-T-antigen), or NeuAcα2-6GalNAc (sialyl-Tn-antigen) at each threonine or serine residue were prepared by a combination of chemical glycopeptide synthesis and enzymatic extension of carbohydrate chains. These glycopeptides were tested by the enzyme-linked immunosorbent assay (ELISA) for their capacity to bind 13 monoclonal antibodies (mAbs) known to be specific for MUC1. The results indicated that anti-MUC1 mAbs have diverse specificities but can be classified into a few characteristic groups based on their binding pattern toward glycopeptides in some cases having a specific glycan at unique glycosylation sites. Because the clinical significance of some of these antibodies was already established, the structural features identified by these antibodies as revealed in the present study should provide useful information relevant to their further clinical use and the biological understanding of MUC1.

A mesoporous Pt/SO–ZrO 2 catalyst was prepared by combining sol–gel and surfactant-assisted templating methods. Its pore structure was analyzed by an adsorption technique, and its catalytic behavior in the CH 4 /CO 2 reforming reaction was investigated using a high-pressure flow reactor at 0.1–1.0 MPa and 803 K. The Pt/SO–ZrO 2 catalyst’s surface consists of many mesopores; these yielded a high specific surface area (ca. 130 m 2 /g) and a high Pt dispersion, which was three times that of Pt/ZrO 2 prepared by a conventional method. H 2 formation activity over the Pt/SO–ZrO 2 catalyst at 0.1–1.0 MPa was higher than that over Pt/ZrO 2 , and the H 2 /CO ratio also increased. At a gas hourly space velocity of 60,000 h −1 at 1.0 MPa, the catalytic stability was improved owing to high resistance to coking, and the H 2 space–time yield reached 93 mmol/h g cat .

This paper describes the results of an experimental investigation on the drop off impact test on a range of sandwich panels. The magnesium alloy sandwich panels were fabricated with rolled sheets at different thickness by pressing and bonding method. Out-plane compression test was employed to obtain its basic deformation-force behavior. The impact experiments were carried out in which a steel cylinder was dropped off at various height levels, ranging from 0.5m to 1.5 cm to impact the panel. A high speed camera was employed to take pictures at 20 thousand frames per second and the low-velocity impact response on the sandwich panels is recorded with a dynamic force senor under the panel simultaneously. The shock response with time and the impact absorption energy were analyzed and compared. The results of this study proved that the magnesium alloy honeycomb sandwich panels have good impact energy absorption performance.

About 160 species of pycnogonids have been reported from Japan. Among these, only 18 species have been reported from the extensive archipelago of the Nansei (southwest) Islands, even though this warm‐temperate to subtropical region should have a pycnogonid fauna as rich as or richer than that surrounding the Japanese main islands. We provide descriptions for five species of pycnogonids collected from depths of 52–334 m along the continental shelf adjacent to the Nansei Islands. Rhopalorhynchus tenuissimum (Haswell, 1884) is the first record of this genus and species for Japan, and Pycnogonum tenue Slater, 1879 is a new record for the Nansei Island region; we report some previously unrecognized taxonomic characters for these species. We describe as new species Endeis leviseminentia, Hedgpethia elongata, and Pycnogonum spatium.

脳血管障害例の座位姿勢制御における視覚的垂直定位の影響を身体軸傾斜との関連から検討した。右麻痺7例，左麻痺8例，健常若年者9名及び健常高齢者4名に，視覚的垂直定位と，傾斜した視覚指標への身体軸傾斜を測定した。片麻痺例は身体軸傾斜が健常者より有意に小さく，麻痺側傾斜時にその傾向が著明であったが，運動麻痺の重症度と身体軸傾斜との間に有意な相関を認めなかった。視覚的垂直定位の標準偏差は左麻痺例で他の3群より有意に大きく，身体軸傾斜の標準偏差は左麻痺例の左傾斜時に大きい傾向を認めた。視覚的垂直定位の標準偏差は身体軸傾斜時の動揺に影響し，左麻痺例で顕著であると示唆された。

Summary Background: Epidermolysis bullosa simplex associated with muscular dystrophy is caused by plectin deficiency. Objective To report clinical, immunohistochemical, ultrastructural and molecular features of a 52-year-old Japanese patient affected with this disease, whose muscular disease had been followed-up for 27 years. Methods: We performed histopathological study, immunofluorescence, electron microscopic study and mutation detection analysis for plectin. Results: The patient developed blisters and erosions followed by nail deformity on the traumatized regions from birth. The skin lesions were continuously developed to date. The histopathological study showed subepidermal blister. Electron microscopic study showed blister formation inside the basal cells at the level just above the attachment plaque of hemidesmosome. Immunofluorescence showed complete loss of staining to plectin. The mutation analysis using protein truncation test and DNA sequencing revealed a C-to-T transition at nucleotide position 7006 of the plectin cDNA sequence, which lead a novel homozygous nonsense mutation (R2319X). Conclusion: From the above results, the diagnosis of epidermolysis bullosa simplex associated with muscular dystrophy was made. Slight muscular dystrophy was noticed at the age of 25 years. The muscular dystrophy gradually progressed and she could not walk at the age of 46 years. However, she can still breathe and swallow by herself. This is the patient of this disease with the longest follow-up, and may indicate the slow progress of muscular condition of this disease.

(-)-Epigallocatechin gallate (EGCG), a major component of green tea catechins, is known to inhibit cell growth and to induce apoptosis in a variety of cultured cells. We examined effects of green tea catechins in cultured cells derived from human gastric carcinoma. The proliferation of four cell lines (MKN-1, MKN-45, MKN-74 and KATO-III) was inhibited with EGCG in a dose-dependent manner. The growth of MKN-45 cells was most efficiently inhibited by the treatment (IC(50): 40 muM EGCG) among the four cell lines, while KATO-III cells were most insensitive (IC(50): 80-150 muM) to the EGCG treatment. In addition, (-)-epicatechin (EC) had a major synergistic effect on the induction of apoptosis in MKN-45 cells treated with EGCG; however it had little effect on the inhibition of cell growth induced by EGCG. To study the molecular mechanisms behind the induction of apoptosis by EGCG, the activity of caspases in MKN-45 cells treated with EGCG was examined. Activity levels of caspases-3, -8 and -9 were elevated in EGCG-treated cells, suggesting that these caspases are involved in the apoptosis induced by EGCG. Furthermore, the synergistic effect of EC with EGCG on the induction of apoptosis was specifically canceled by catalase treatment, suggesting that the synergism involves the extracellular production of reactive oxygen species.

In 1997, the members of Japan Stroke Society presented the Japan Stroke Scale (JSS) for the acute stage, and in 2000 the Japan Stroke Scale of Motor (JSS-M) with the items of motor system required for the independence of activities of daily living (ADL). JSS-M consists of eight major items. We classified subjects with nine points or less into group A, with 10-19 points into group B and with 20 points or more into group C according to JSS scores for the acute stage at the baseline. In JSS scores, higher scores means more intense severity of disability. Subjects were 46 cases with cerebral hemorrhage and 63 cases with cerebral infarction, 109 cases in total. Age: cases with cerebral hemorrhage had a mean age of 58.3 ± 14.7, while those with cerebral infarction had a mean age of 69.2 ± 11.0: males 67, and females 42. The rate of improvement of cerebral hemorrhage was higher than that of cerebral infarction in groups A and B. The rate of improvement was the highest in Gait in groups A and B, and Dysphagia and Complex Movement followed in order after that; in group B the rate of improvement of Hand and Arm followed after Complex Movement. The mean rate of improvement of JSS was 98% in group A, 66.7% in group B, and 57.8% in group C. JSS-M is a very useful evaluation, but the study of evaluation methods of trunk and upper limb function required.

The proto-oncogene c-myc is a multifunctional gene that regulates cell division, cell growth, and apoptosis. Here we report a new function of c-myc: induction of autophagy. Autophagy is a bulk degradation system for intracellular proteins. Autophagy proceeds with characteristic morphologies, which begins with the formation of a double-membrane structure called the autophagosome surrounding a portion of the cytoplasm, after which its outer membrane then fuses with the lysosomal membrane to become an autolysosome. Autophagosomes and autolysosomes are generally called autophagic vacuoles. When c-Myc protein was overexpressed in rat 3Y1 fibroblasts or when the chimeric protein c-MycER was activated by estrogen, the number of autophagic vacuoles in cells increased significantly. The formation of autophagic vacuoles induced by c-Myc was completely blocked by a specific inhibitor of autophagosome formation, 3-methyladenine. A c-Myc mutant lacking Myc Box II induced neither apoptosis nor oncogenic transformation, but still stimulated autophagy. An inhibitor of caspases suppressed apoptosis but not autophagy. These results suggest that the autophagy caused by c-myc is not due to the apoptosis or tumorigenesis induced by c-myc. Taken together, our results suggest that the induction of autophagy is a novel function of c-myc.

Evaluation of disorders constitutes an important part of work in rehabilitation medicine. But all of the objective evaluation methods for quantifying ADL have both advantages and disadvantages, and they are often difficult to use. Evaluation and judgment of ADL should be carried out in terms of movements simplified as much as possible. An ADL Score Study Group, consisting of medical co-workers, has been organized, and they have proposed a tentative plan for patients with paraplegia due to spinal cord injury. Method; ADL items were limited to 10, with sub-items limited to two to three per item, to simplify the ADL score as much as possible. The evaluation was carried out in accordance with the followings; 0 point, requiring full assistance, 1 point, partial assistance, 2 points, possible to do activities with insufficient perfectness, 3 points, possible to do activities independently, with a maximum of 99 points. Subjects were 35 patients with complete paralysis due to spinal cord injury. Thirty four persons were males and one was female. Results; Evaluation scores were 81 points for Group A (Spinal cord injury T1-7), 85 points for Group B (Spinal cord injury T8-L1), 95 points for Group C (Spinal cord injury L2 or lower). It was demonstrated that the evaluation of two items, reach and walking, was sufficient for independent patients with paraplegia due to spinal cord injury.

Making a functional prognosis for patients with traumatic incomplete spinal cord injuries is extremely important not only for the patients themselves, in attempting to achieve independence in activities of daily living and social rehabilitation, but also for medical co-workers, the family and almost everyone in the society. However, only very few reports deal with this subject, and rehabilitation is often conducted aimlessly. In the present study we tried at our department to summarize the functional prognosis of patients with spinal injuries and set up a preliminary standard for rehabilitation. The present study included 75 patients with Frankel class C, D, E, incomplete spinal injury who underwent rehabilitation training in the past ten years at our department. The level of the injury was the cervical cord in 53 patients and the thoracic or lumbar cord in 22 patients. The average age of the patients was 36.2 years. Seventy of the patients were male and 5 female. The time of the final evaluation after the injury was on the average 11.8 months (SD=5.9 months). When muscle power of the lower extremities three months after injury reached [4], patients accomplished independent walking 6.8 months after the injury. When muscle power of the lower extremities 6.5 months after injury reached [3-], patients accomplished semi-independent walking with the help of a wheelchair 8.7 months after the injury. Patients with a muscle power of more than [3-] in the upper extremities could walk independently. Among patients with incomplete injury of the cervical cord 64% gained independence in ADL 8.6 months after the injury, but patients with central paralysis or strong spasticity required assistance.

The in vitro activity of the new fluoroquinolone HSR-903 was compared with those of ciprofloxacin, lomefloxacin, sparfloxacin, and levofloxacin. HSR-903 inhibited 90% of methicillin-susceptible and -resistant Staphylococcus aureus (MRSA) clinical isolates at 0.78 and 1.56 microg/ml, respectively, and its activity against MRSA was 16-fold higher than those of sparfloxacin and levofloxacin and 64-fold higher than that of ciprofloxacin. The MICs at which 90% of the isolates are inhibited (MIC90s) of HSR-903 for Streptococcus pyogenes and penicillin G-susceptible and -resistant Streptococcus pneumoniae (PRSP) were 0.10, 0.05, and 0.05 microg/ml, respectively. Against PRSP, the activity of HSR-903 was 4-fold higher than that of sparfloxacin and 32- to 256-fold higher than those of the other quinolones. The MIC90 of HSR-903 for Enterococcus faecalis was 0.20 microg/ml, and HSR-903 was more active than the other quinolones against enterococci. The activity of HSR-903 against members of the family Enterobacteriaceae and Pseudomonas aeruginosa was roughly similar to that of ciprofloxacin and greater than those of the other quinolones. Against Haemophilus influenzae, Moraxella catarrhalis, and Helicobacter pylori, HSR-903 was the most potent of the quinolones tested. The activity of HSR-903 was not affected by the medium, the inoculum size, or the addition of serum, but decreased under acidic conditions, as did those of the other quinolones tested. HSR-903 exhibited rapid bactericidal action and had a good postantibiotic effect on S. aureus and P. aeruginosa. HSR-903 inhibited supercoiling by DNA gyrase from Escherichia coli, but it was much less active against human topoisomerase II.

北海道を中心に大量に廃棄されるホタテ貝殻は, 投棄場所の確保と臭気対策の問題が次第に顕在化している。カルシウム資源の一つとしての用途拡大は, これらの問題の解決と資源の有効利用の両面からもっとも望ましい方策である。本研究ではホタテ貝殻を生石灰として, あるいはそれを再炭酸化しての工業的利用の可能性を探るために, 灯油を燃料とした内熱式の流動層による連続焼成実験を行い焼成条件および装置特性について検討した。原料貝殻を3mm以下に予備粉砕することにより良好な流動化状態が維持でき, 流動層底部から連続的に抜き出した焼成品は温度850℃, 平均滞留時間が80分以上で99%以上の高い焼成率が得られた。焼成品 (生石灰) の反応性と亜硫酸ガスの吸収特性を石灰石の焼成品と比較した結果, ホタテ貝殻からの生石灰は比表面積が小さい一方で細孔径が大きいため, 脱硫反応に対して優れた特性を示すことがわかった。

The early stage of colorectal cancer is divided into two morphological types polypoid and superficial. What percentage of advanced cancer is derived from the polypoid or superficial type is a matter of concern. The ratio of superficial variety among of ulcerated cancers was studied using those 2 cm or less in diameter, which often retain mucosal cancer. The existence of horizontal growing mucosal cancer at the marginal area of ulcerated cancer is a hallmark showing the lesion being of superficial origin. Eight (57 %) out of the 14 cases of ulcerated cancer, 2 cm or less, had this finding. This suggests that more than half of smaller colorectal cancers are of superficial origin. A zig-zag pattern (ZZP) was evident in 3 cases of superficial origin. No cases in the nonsuperficial group had the ZZP. The existence of circumferential ZZP during colonoscopic observation indicates that the cancer is of superficial origin.

In the present study, we examined the production method of the carbonaceous adsorbent, in which acrylonitrile and methyl acrylate copolymers were used as raw materials, and phosphate and zinc chloride were added as carbonization and activation accelerating agents. The raw materials were treated in air for 7hr at temperatures between 120-220°C.Three parts of 50% aqueous solution of phosphate or zinc chloride were added to 1 part of each char (PANACHP, PANACHZ), and the solution was then directly placed in a reaction oven at 600-900°C. The weight reduction rate W/Wo (-) decreased linearly with increased activation time. Both the methylene blue adsorbability (MB) and the internal surface area (S) of the activated material showed a maximum value and then decreased with increased activation time. MB and S of PANACHP and PANACHZ showed the maximum value in conditions with activation temperature of 900°C and activation time of 180min. In these conditions, the following results were obtained: MB: 450mg/g and S: 1, 280m2/g for PANACHP and MB: 310mg/g, S: 1, 080m2/g for PANACHZ, indicating that both PANACHP and PANACHZ were carbonaceous adsorbents which possessed more than twice as much mercaptan gas adsorbability as the activated carbon on the market.

An experimental study was carried out on the preparation for activated carbon from bark wastes in wood industry. This study involved an activation test with the phosphoric acid at temperatures in the range of 400 to 900°C, after the bark materials being dried, ground and sorted for sizes smaller than 4 mm.The Parameters, such as internal surface area, pore size distribution and pore volume of the activated product, were measured, and the adsorption capacity with the methylene blue and benzene was tested separately. It was found that for activation longer than 5 minutes, the methylene blue adsorption capacity was increased with an increase in temperature of the above range, while the internal surface area and the benzene adsorption capacity were decreased at the temperatures above 600°C. Besides both trichloroethylene and tetrachloroethylene adsorption tests have shown that activated products of higher adsorption capacity could be obtained with this phosphoric acid activation process as compared with the commercial activated carbon.

Capsule endoscopy (CE) is used widely for determining the cause of obscure gastrointestinal bleeding (OGIB). However, negative findings still arise from CE examination. The aim of this study was to determine the factors associated with negative findings on CE in patients with OGIB.A total of 134 patients who underwent CE for overt (n = 104) or occult (n = 30) OGIB between October 2007 and April 2010 were included. The clinical backgrounds of the patients (age; sex; the use of anti-coagulant, anti-platelet drugs or NSAIDs; comorbidity and the timing of CE examination after bleeding) were noted.The overall diagnostic yield of CE in detecting the relevant findings was 50% (n = 67). Multivariate analysis revealed that the use of anti-platelet drug and the timing of CE (≥ 16 days) were predictive factors for negative findings on CE (odds ratio 2.69 [1.01-7.21], p = 0.048 and odds ratio 2.32 [1.01-5.33], p = 0.047, respectively). Among the patients with the use of low-dose aspirin (LDA, n = 28) as anti-platelet drug, cessation of it before CE was the only predictive factor for negative findings on CE (odds ratio 12.0 [1.72-83.5], p = 0.012).In the patients with OGIB, the use of LDA and the cessation of it before CE made it difficult to detect the cause of bleeding by CE. This might indicate that the source of OGIB related to LDA heals immediately after cessation of the drugs or is a very small lesion that could not be detected by CE.

As volume of wastes increases at an extraordinary rate, it is often difficult today to find the place for dumping and treatment of large volume of the wastes. In Japan it is an urgent problem to decrease volume of wastes and to recycle them.Therefore we try to utilize organic wastes from timbers as activated carbon material for environmental protection. Bark of Ezomatsu. Karamatsu and Todomatsu imported from USSR are selected for material of production test of activated carbon.Bark material is dried at the temperature 50°C in 48 hours, ground and sorted under 4mm. From thermal analysis (TG, DTG, DTA) data optimum temperature of activation is found to be ranged from 400°C to 600°C.The test of chemical treatment of bark sample by ZnCl2 is performed at the temperature of 400, 450, 500 and 600°C. Yield specific surface area and adsorption of methylene blue, benzene, trimethylamine, stylene, dimethyl sulfide and 1-butanethiol are measured for activated samples.Activated carbon samples produced in these experiments have the same or higher adsorption capacity than the commercial activated carbon.

RAA (Robot Assisted Activities) allow people to feel joy and pleasure and recover through contact with a robot. Considerable research has reported that RAA is useful for elderly persons, children hospitalized in pediatric wards and so on. However no specific evaluation method concerning RAA has yet been established. In this paper, we propose a reaction time measurement system for evaluating RAA. The level of caution or concentration of a subject can be estimated in terms of the reaction time based on visual or acoustic stimuli. We try to evaluate RAA using the variance in the reaction time before and after RAA respectively. We have developed a reaction time measurement system with hardware, following which we compared several reaction time measurement systems concerning performance and operability. We reported the results of comparisons featuring such measurement systems and the basic results of the RAA evaluation experiment using reaction time measurement. The results suggested that selection of the optimal enforcement form in RAA has been enabled through application of this reaction time measurement system.

Background: The in vitro antimicrobial activities of new fluoroquinolones were tested against quinolone-resistant Haemophilus influenzae of clinical isolates. Methods: The nucleotide sequences of the gyrA and parC genes from three ciprofloxacin-resistant strains of Haemophilus influenzae (MIC, 1.56–6.25 µg/ml) were determined. The gyrase was purified from the clinical isolates, and the inhibitory activities of quinolones against the enzyme were tested. Results: These strains possessed at least one amino acid substitution in each of the GyrA (asparagine at residue 88 (Asp-88) to Tyr, Ser-84 to Leu or Ser-84 to Leu and Asp-88 to Asn) and ParC (Glu-88 to Lys). The antibacterial activity of olamufloxacin against the resistant strains was most potent compared with other quinolones, and the inhibitory activities correlated with quinolone resistance of these strains. Conclusions: These results warrant the clinical effects of new types of fluoroquinolones, such as olamufloxacin, against respiratory tract and otolaryngology infections caused by ciprofloxacin-resistant H. influenzae.

A series of erythromycin A 9-O-substituted oxime ether derivatives have been synthesized and evaluated for antibacterial activity against Mycobacterium avium complex (MAC) and Staphylococcus aureus. These compounds possessed stronger in vitro activity against MAC including macrolide-resistant strains than clarithromycin (2), although in vitro antibacterial activities of these compounds were less than that of 2 against Staphylococcus aureus. Our studies found that several factors contribute to the antibacterial activity against MAC. The length and spatial orientation of the substituent at 9-position were found to significantly influenced the anti-MAC activity, especially against macrolide-resistant strains. Of all the compounds prepared, erythromycin A 9-[O-(4-phenylbutyl)oxime] (12q) and erythromycin A 9-[O-(3-phenoxypropyl)oxime] (12t) possessed 16 times stronger antibacterial activity than 2 against clarithromycin-resistant strains. Surprisingly, the minimum inhibitory concentrations (MICs) of 12q and 12t against the resistant strains were almost same as those against the susceptible strains. These results suggest that the erythromycin A 9-O-substituted oxime ether derivatives would be promising macrolide antibiotics.

The in vivo activity of olamufloxacin (HSR-903), a new fluoroquinolone, was evaluated and compared with ciprofloxacin, sparfloxacin and levofloxacin. Olamufloxacin was active against systemic infection in mice inoculated with both Gram-positive and -negative bacteria. Olamufloxacin had equal efficacy for experimental urinary tract infections in mice caused by Pseudomonas aeruginosa.

Ectodomain shedding is an important mechanism to regulate the biological activities of membrane proteins. We focus here on the signaling mechanism of the ectodomain shedding of heparin-binding epidermal growth factor (EGF)-like growth factor (pro HB-EGF). Lysophosphatidic acid (LPA), a ligand for seven-transmembrane G protein-coupled receptors, stimulates the shedding of pro HB-EGF, which constitutes a G protein-coupled receptor-mediated transactivation of the EGF receptor. Experiments using a series of inhibitors and overexpression of mutant forms of signaling molecules revealed that the Ras-Raf-MEK signal is essential for the LPA-induced shedding. In addition, the small GTPase Rac is involved in the LPA-induced shedding, possibly to promote MEK activation. 12-O-Tetradecanoylphorbol-13-acetate is another potent inducer of pro HB-EGF shedding. We also demonstrate that the LPA-induced pathway is distinct from the 12-O-tetradecanoylphorbol-13-acetate-induced pathway and that these pathways constitute a dual signaling cascade that regulates the shedding of pro HB-EGF.

CD9 and CD63 belong to a tetramembrane-spanning glycoprotein family called tetraspanin, and are involved in a wide variety of cellular processes, but the structure-function relationship of this family of proteins has yet to be clarified. CD9 associates with diphtheria toxin receptor (DTR), which is identical to the membrane-anchored form of heparin-binding EGF-like growth factor (proHB-EGF). CD9 upregulates the diphtheria toxin (DT) binding activity of DTR/proHB-EGF, while CD63 does not upregulate the DT binding activity in spite of the fact that this protein also associates with DTR/proHB-EGF on the cell surface. CD9 molecules localize on the cell surface, while those of CD63 localize predominantly at lysosomes and intracellular compartments. We made CD9/CD63 chimeric molecules and then studied their intracellular localization and upregulation activities. The C-terminal regions of CD63, which includes the lysosome sorting motif, showed a strong inhibitory effect on the expression of the chimeric proteins at the cell surface, while mutants lacking the lysosome sorting motif delivered more efficiently on the cell surface, indicating that the lysosome sorting motif contributes to the inhibitory effect of the C-terminal region. However, the N-terminal half of this family of proteins containing the 1st to 3rd transmembrane domains also seems to influence the cell surface expression. For the upregulation of DT binding activity the large extracellular loop (EC2) of CD9 was essential, while the remaining regions influenced the upregulation activity by changing the efficiency of cell surface expression. From these results we discussed the structure-function relationship of this family of proteins.

Sindbis virus (SV), a single-stranded positive-sense RNA virus, multiplies in a variety of cells and causes various outcomes of infection. As we described acute infection of SV induces stress response of small heat shock protein HSP27 and activation of mitogen-activated protein kinase (MAP) signaling pathway (Nakatsue, T., et al., Biochem. Biophys. Res. Commun. 253, 59–64, 1998). In contrast to lytic infection in Vero cells, MRC-5 cells, a human fetus lung cell line, resulted in persistent infection by SV. Here we investigated a cellular factor involved in persistent infection of MRC-5 cells infected with SV. Partial sequence analysis of a 25 kilodalton (kDa) protein, accumulated in large amounts in the cells, showed that manganese-superoxide dismutase (Mn-SOD) was induced during the infections. When Mn-SOD was overexpressed in Vero cells, 20% of the cells survived more than one month, in contrast with the death of 99% of the vehicle-transfected Vero cells at 48 h after infection with SV. These data strongly suggest that a cellular factor which regulates the oxidative pathway modulates the outcome of SV infection.

In general, viral infection is supposed to induce stress responses in the host cell. However, very few detailed observations about virus-induced stress responses have been reported. Here we investigated specific stress responses in Vero cells infected with Sindbis virus (SV), a single-stranded RNA virus, acute infection with which is known to cause apoptotic cell death in the host cells. Prior to the onset of apoptosis, p38 mitogen-activated protein kinase (MAPK) and c-Jun NH2-terminal kinases (JNKs) were activated. Subsequently, a 27-kDa heat shock protein (HSP27) became phosphorylated, and intracellular distribution of HSP27 was changed from the cytoplasm to the perinuclear region. These results indicate that the cellular signaling cascades activated by pro-inflammatory cytokines and environmental stresses are also activated as a result of lytic infection with SV. These responses may contribute to the delayed onset of apoptosis in the host cells and the facilitation of viral replication.

The in-vitro antimicrobial activity of HSR-903, a new fluoroquinolone, was tested against 51 clinical Neisseria gonorrhoeae isolates in comparison with ciprofloxacin, levofloxacin and sparfloxacin. The MICs of HSR-903 for 11 isolates with alterations in both GyrA and ParC, for 19 isolates with alterations only in GyrA and for 21 isolates without alterations in either GyrA or ParC ranged from 0.03 mg/L to 1.0 mg/L (MIC 90 = 0.25 mg/L), from 0.03 mg/L to 0.5 mg/L (MIC 90 = 0.125 mg/L) and from

42歳の女性。6ヵ月前より臍部に結節を生じ,月経時の出血と疼痛を認めるようになった。初診時,径20mm×高さ5mmの表面平滑の暗赤色のドーム状の結節を認めた。組織学的には真皮下層に子宮内膜円柱上皮細胞よりなる腺腔構造が認められ,臍部子宮内膜症と診断した。子宮,骨盤内に他病変は認めなかった。GnRHアゴニスト製剤による保存的治療を行い出血,疼痛の消失と結節の若干の縮小をみている。

Five species of pycnogonid from shallow-water sampling of the hyperbenthos on some sandy beaches in Ecuador in 2001 and 2002 are discussed. Three of the species, Tanystylum isthmiacum Stock, 1955, Anoropallene palpida (Hilton, 1939) and Anoplodactylus californicus Hall, 1912, have been recorded previously from Ecuador waters; one species, Ammothella symbia Child, 1979, is newly recorded for Ecuador; one species of Anoplodactylus is described herein as new. The currently known pycnogonid fauna of Ecuador is listed.

Lunatoic acid A and (–)-mitorubrinic acid have been isolated as chlamydospore-like cell-inducing substances from Cochliobolus lunatus. (+)-Sclerotiorin, (+)-rotiorin, citrinin and sclerin were shown to have the same activity. Although they share no common structural feature, all these active substances react with methylamine. The reaction products do not have this activity. The reaction curves showed a positive correlation between the chemical reactivity of chlamydospore-like cell-inducing substances with methylamine and their biological activities.

Desmoyokin was first isolated from bovine muzzle epidermis and thought to be an epidermal desmosome-related protein. We previously demonstrated that the Desmoyokin gene is identical to the Ahnak gene, which is expressed ubiquitously and downregulated in neuroblastomas. It was assumed Ahnak/Desmoyokin was associated with epidermal cell adhesion, tumorigenesis, cell proliferation and differentiation, and embryonic development. To determine the precise biological function of Ahnak/Desmoyokin, we generated a null mutation in ES cells and mice. The resultant Ahnak/Desmoyokin-deficient ES cells normally differentiated into embryoid bodies and neural cells. The mutant mice were viable and fertile and showed no gross developmental defects. Electron microscopic examination of skin sections demonstrated that the ultrastructure of epidermal intercellular junctions, including desmosomes, of the mutant mice was indistinguishable from that of wild-type mice. Two-stage chemical skin carcinogenesis experiments showed no difference in frequency or onset of cutaneous tumor formation between wild-type and mutant mice. Moreover, no tumorigenesis was observed in other tissues and organs of mutant mice up to 2 y of age. These results lead us to conclude that Ahnak/Desmoyokin deficiency has only a minimal effect on epidermal cell adhesion, tumorigenesis, cell proliferation and differentiation, and overall mouse development.