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1.  Isolation and characterization of native Cry j 3 from Japanese cedar (Cryptomeria japonica) pollen 
Allergy  2007;62(5):547-553.
Japanese cedar (Cryptomeria japonica) pollinosis is the most prevalent allergy in Japan. Recently, the Japanese cedar pollen allergen Cry j 3 was cloned as a homologue of Jun a 3, which is a major allergen from mountain cedar (Juniperus ashei) pollen. However, native Cry j 3 has not been isolated and there are no reports on its allergenic activity. The aims of this study were to isolate native Cry j 3 and assess its immunoglobulin E (IgE)-binding capacity in patients with Japanese cedar pollinosis.
Native Cry j 3 was purified from Japanese cedar pollen by multidimensional chromatography. We assessed the IgE-binding capacity using sera from patients allergic to Japanese cedar pollen by immunoblot analysis and ELISA. Moreover, we assayed the capacity of Cry j 3 to induce histamine release from the patient’s leukocytes. We cloned cDNA corresponding to purified Cry j 3 from a cDNA library of Japanese cedar pollen.
We isolated native Cry j 3 as a 27-kDa protein. The IgE-binding frequency of Cry j 3 from the sera of patients allergic to Japanese cedar pollen was estimated as 27% (27/100) by ELISA. Cry j 3 induced the release of histamine from leukocytes. We cloned the cDNA and named it Cry j 3.8. Cry j 3.8 cDNA encoded 225 amino acids and had significant homology with thaumatin-like proteins.
Cry j 3 is a causative allergen in Japanese cedar pollinosis and may play crucial roles in the cross-reactivity with oral allergy syndrome.
PMCID: PMC2587458  PMID: 17441795
Japanese cedar; oral allergy syndrome; pathogenesis-related protein; pollinosis; thaumatin-like protein
2.  Malignant peripheral nerve sheath tumour arising within neurofibroma. An immunohistochemical analysis in the comparison between benign and malignant components 
Journal of Clinical Pathology  2001;54(8):631-636.
Aims—To compare the expression of immunohistochemical variables between benign and malignant components of malignant peripheral nerve sheath tumour (MPNST) arising within neurofibroma.
Methods—Eight cases of MPNST arising within a neurofibroma, associated with neurofibromatosis type 1 (NF1), were studied. The areas of MPNST and neurofibroma were compared immunohistochemically with regard to the expression of proliferative activity (MIB-1), growth factors, p53, bcl-2, neural cell adhesion molecule (N-CAM), and CD34.
Results—The expression of transforming growth factor ß1 (TGF-ß1), TGF-ß receptor type II, hepatocyte growth factor α (HGF-α), c-met, p53, and N-CAM was higher in the areas of MPNST than in the neurofibromatous areas in four, five, five, eight, five, and three of the eight cases, respectively. CD34 expression was lower in the areas of MPNST than in the neurofibroma areas in three of the eight cases.
Conclusions—On the basis of these findings, TGF-ß1, HGF-α, and p53 might be involved in the malignant transformation of neurofibroma to MPNST.
Key Words: malignant peripheral nerve sheath tumour • neurofibroma • immunohistochemistry • transforming growth factor ß • hepatocyte growth factor • p53
PMCID: PMC1731495  PMID: 11477120
3.  Effects of topical nipradilol, a β blocking agent with α blocking and nitroglycerin-like activities, on intraocular pressure and aqueous dynamics in humans 
AIMS—To study the effects of topical nipradilol, a non-selective β blocker with α blocking and nitroglycerin-like activities, on intraocular pressure (IOP) and aqueous humour dynamics in normal humans and in patients with ocular hypertension.
METHODS—Nipradilol (0.06%, 0.125%, 0.25%, 0.5%) was applied to normal volunteers (n = 12) to test for IOP lowering effects. In a second group of normal volunteers (n = 11), nipradilol (0.125% and 0.25%) and timolol (0.5%) were compared for IOP lowering effects. After a single administration of 0.25% nipradilol, IOP, flare intensity in the anterior chamber, aqueous flow, uveoscleral outflow, tonographic outflow facility, and episcleral venous pressure were either directly measured or mathematically calculated. Topical nipradilol (0.25%) was administered to 24 patients with ocular hypertension twice daily for 8 weeks.
RESULTS—Administration of 0.25% nipradilol decreased IOP with a maximum reduction of 4.2 mm Hg lasting 12 hours. A single instillation of both 0.25% nipradilol and 0.5% timolol reduced the IOP in normotensive human subjects to the same degree. A single instillation of 0.25% nipradilol decreased the aqueous flow rate in the treated eye by 20%. Nipradilol produced no significant effect in tonographic outflow facility or episcleral venous pressure, but uveoscleral outflow was increased. In patients with ocular hypertension, twice daily instillation of 0.25% nipradilol decreased IOP without tachyphylaxis for the 8 week test period.
CONCLUSION—Topical nipradilol (0.25%) reduced IOP by decreasing the aqueous flow rate and probably also by increasing uveoscleral outflow. Nipradilol should be further investigated as a new antiglaucoma drug.

PMCID: PMC1723399  PMID: 10684841
4.  Tumour-amplified kinase BTAK is amplified and overexpressed in gastric cancers with possible involvement in aneuploid formation 
British Journal of Cancer  2001;84(6):824-831.
Our recent analysis of gastric cancers using comparative genomic hybridization (CGH) revealed a novel high frequent copy number increase in the long arm of chromosome 20. Tumour-amplified kinase BTAK was recently cloned from breast cancers and mapped on 20q13 as a target gene for this amplification in human breast cancers. In the study presented here, we analysed BTAK copy-number and expression, and their relation to the ploidy pattern in 72 primary gastric cancers. Furthermore, wild-type BTAK and its deletion mutants were transfected to gastric cancers to examine changes in cell proliferation and DNA ploidy pattern. Evaluation of 72 unselected primary gastric cancers found BTAK amplification in 5% and overexpression in more than 50%. All four clinical samples with BTAK amplification showed aneuploidy and poor prognosis. Transfection of BTAK in near-diploid gastric cancers induced another aneuploid cell population. In contrast, the c-terminal-deleted mutant of BTAK induced no effect in DNA ploidy pattern and inhibited gastric cancer cell proliferation. These results suggest that BTAK may be involved in gastric cancer cell aneuploid formation, and is a candidate gene for the increase in the number of copies of the 20q, and thus may contribute to an increase in the malignant phenotype of gastric cancer. © 2001 Cancer Research Campaign
PMCID: PMC2363814  PMID: 11259099
ploidy pattern; amplification; overexpression; BTAK; gastric cancer
5.  Factory outbreak of Escherichia coli O157:H7 infection in Japan. 
Emerging Infectious Diseases  1999;5(3):424-428.
To determine the cause of a July 1996 outbreak of Escherichia coli O157:H7 among factory workers in Kyoto, Japan, we conducted cohort and case-control studies. Eating radish sprout salad during lunch at the factory cafeteria had been linked to illness. The sprouts were traced to four growers in Japan; one had been associated with an outbreak of E. coli O157:H7 among 6,000 schoolchildren in Sakai earlier in July.
PMCID: PMC2640759  PMID: 10341179
6.  Cellular autoimmunity to retinal specific antigens in patients with Behçet's disease. 
The notion that autoimmune mechanisms play a role in the pathogenesis of certain uveitic conditions in humans is supported by the observation that lymphocytes from such patients respond in culture against retinal specific antigens which are uveitogenic in animals. A large proportion of uveitis patients with Behçet's disease are reported to respond well to S antigen, to interphotoreceptor retinoid binding protein (IRBP) and to several of their uveitogenic peptides, in particular, the S antigen derived peptide M. Patients with Behçet's disease without ocular involvement were reported not to differ in their responses to S antigen from the responses in the control group, yet 35% of them responded to IRBP and approximately two thirds of them responded to the peptides (peptide M, peptide N, R-4, or R-14). The responses were inhibited by monoclonal antibodies to CD4 and to class II MHC HLA-DR molecules. The presence of lymphocyte responses to retinal antigens in patients with Behçet's disease without uveitis might indicate a preclinical stage of ocular involvement. Thus, these data support the idea that autoimmunity to retinal specific antigens may play a role in the ocular inflammation in Behçet's disease.
PMCID: PMC513957  PMID: 8218058
7.  S-antigen specific T cell clones from a patient with Behçet's disease. 
The isolation and characterisation of T cell clones or lines specific to retinal antigens are valuable tools to clarify the underlying mechanisms of autoimmunity to retinal antigens as a contributing factor in ocular inflammation. Patients with Behçet's disease have been reported to be sensitised to S-antigen (S-Ag). In the present study, four T cell clones established from the peripheral blood of a patient with Behçet's disease were analysed. A CD4+ T cell clone (clone 2) and a CD8+ T cell clone (clone 10) proliferated specifically to bovine S-Ag. Although these S-Ag specific T cell clones proliferated vigorously to the intact antigen, their responses to S-Ag derived synthetic peptides M and G were weak, suggesting that the sites of human T cell recognition of S-Ag may be different from those established in the experimental model. The proliferative responses of both clones (2 and 10) were inhibited by anti-HLA-DR monoclonal antibody but not by anti-HLA-class 1 monoclonal antibody. The other two clones studied, clones 6 and 30, were CD3+, CD4-, CD8-, and they did not proliferate specifically to S-Ag. Clone 6 expressed gamma delta T cell receptors (TCR) and showed non-specific cytotoxic activity toward K562 and Daudi cell lines. Clone 30 expressed alpha beta TCR, and was devoid of cytotoxic activity. Human T cell lines and clones specific to retinal antigens will provide the framework necessary to examine the events that lead to ocular inflammation.
PMCID: PMC504995  PMID: 7529558
8.  Mechanisms of antibacterial action of tachyplesins and polyphemusins, a group of antimicrobial peptides isolated from horseshoe crab hemocytes. 
Tachyplesins I and II and polyphemusins I and II, cationic peptides isolated from the hemocytes of horseshoe crabs, show bactericidal activities with similar efficiencies for both gram-negative and gram-positive bacteria. Tachyplesin I inhibited bacterial growth irreversibly within 40 min. A subinhibitory concentration of tachyplesin I sensitized gram-negative bacteria to the bactericidal actions of novobiocin and nalidixic acid, although polymyxin B-resistant strains which have altered lipopolysaccharides were susceptible to tachyplesin I. This implies that tachyplesin permeabilizes the outer membrane and that the likely target of its action is outer membrane constituents other than lipopolysaccharides. On the other hand, a defensin-susceptible phoP strain of Salmonella typhimurium was also susceptible to tachyplesin I. Tachyplesin I rapidly depolarized the inverted inner-membrane vesicles of Escherichia coli. These results suggest that depolarization of the cytoplasmic membrane, preceded by the permeabilization of the outer membrane for gram-negative bacteria, is associated with tachyplesin-mediated bactericidal activity. The similarity between the actions of tachyplesin and those of defensin was discussed.
PMCID: PMC191604  PMID: 1510441
9.  Striatal blood flow, glucose metabolism and 18F-dopa uptake: difference in Parkinson's disease and atypical parkinsonism. 
Striatal blood flow, glucose metabolism and 18F-Dopa uptake were studied with positron emission tomography (PET) in eight non-demented patients with idiopathic Parkinson's disease and eight with atypical Parkinsonism. Patients with atypical Parkinsonism had no specific cause for the Parkinsonian symptoms and were clinically different from Parkinson's disease with lack of resting tremor and a poor response to dopaminergic drugs. Decreased 18F-Dopa uptake in the putamen was observed in patients with Parkinson's disease and atypical Parkinsonism compared with normal controls. 18F-Dopa uptake in the head of the caudate was also significantly reduced in both conditions but relatively less in Parkinson's disease. Decreased blood flow and glucose metabolism in the striatum associated with a global cerebral decrease were also observed in patients with atypical Parkinsonism compared with controls, while they were preserved in patients with Parkinson's disease, indicating affected neurons not only in the striatum but also in the cerebrum in patients with atypical Parkinsonism compared with patients with Parkinson's disease. The differences in the caudate 18F-Dopa uptake, and blood flow and glucose metabolism in the cerebrum including the striatum between Parkinson's disease and atypical Parkinsonism assessed by PET may be due to the differences in the pathophysiological mechanism between Parkinson's disease and atypical Parkinsonism.
PMCID: PMC1014575  PMID: 1744644
10.  In vitro and in vivo activities of QA-241, a new tricyclic quinolone derivative. 
The in vitro susceptibilities of 1,310 clinical isolates to QA-241, a novel tricyclic quinolone, were evaluated in comparison with susceptibilities to norfloxacin, ofloxacin, enoxacin, and ciprofloxacin. The MICs of QA-241 for 90% of staphylococci, Enterococcus faecalis isolates, and streptococcal species ranged from 1.56 to 6.25 micrograms/ml, and the activity of QA-241 was similar to those of norfloxacin and enoxacin but two to four times less potent than those of ofloxacin and ciprofloxacin. At the concentration of less than or equal to 1.56 micrograms/ml, QA-241 inhibited 90% of Haemophilus influenzae, Bordetella pertussis, Neisseria gonorrhoeae, and gram-negative enteric bacteria except for Serratia marcescens and Citrobacter freundii. QA-241 was moderately active (MIC for 90% of strains tested, 6.25 to 12.5 micrograms/ml) against S. marcescens, Pseudomonas aeruginosa, Xanthomonas maltophilia, and Bacteroides fragilis. The antibacterial activity of QA-241 was roughly comparable to that of enoxacin but two to four times less potent than that of ofloxacin. In systemic infections in mice with gram-positive cocci and gram-negative rods, the efficacy of QA-241 was generally greater than that of norfloxacin and similar to those of ofloxacin and ciprofloxacin. In urinary tract infections in mice with Escherichia coli or Pseudomonas aeruginosa, QA-241 was as active as ofloxacin and more active than norfloxacin but less active than ciprofloxacin. In pulmonary infections in mice with Klebsiella pneumoniae, the effectiveness of QA-241 was similar to that of ofloxacin.
PMCID: PMC172615  PMID: 2679369
11.  Isolation, properties, and reassembly of outer sheath carrying a polygonal array from an oral treponeme. 
Journal of Bacteriology  1982;150(3):1405-1413.
The outer sheath carrying a polygonal array was isolated from an oral treponeme, Treponema sp. strain E-21, by disruption of cells by means of repeated freeze-thawing and by removal of flagella under acidic conditions followed by linear sucrose density gradient centrifugation. Electron microscopy revealed that the outer sheath was isolated as a triple-layered vesicle having a polygonal array, free of flagella and wall membrane complex. Using optical diffraction, negatively stained preparations of the outer sheath fragments showed that the polygonal array appeared to be composed of a hexagonal pattern with a predominant spacing of about 16.3 nm. The isolated outer sheath contained 49.7% protein, 30.8% total lipid, and 11.0% carbohydrate. Phospholipid comprised about 95% of the total lipid. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the outer sheath was composed primarily of one major protein with an apparent molecular weight of about 62,000. The material from the isolated outer sheath solubilized with 1% sodium deoxycholate was reassembled into vesicles having a roughly polygonal array upon removal of the detergent by dialysis against 10 mM Tris-hydrochloride buffer with or without Mg2+.
PMCID: PMC216367  PMID: 7076622
12.  An electromyographic study of esotropia. 
Under general anaesthesia the eye position of esotropia generally moved divergently. When retrobulbar anaesthesia was added bilaterally, the eye position moved further in the same direction. In the electromyogram under general anaesthesia the 4 horizontal recti were silent in controls. In many cases of esotropia, however, both medial recti showed a considerable amount of discharge under general anaesthesia. When retrobulbar anaesthesia was superimposed on one eye, the discharge from its medial rectus tapered off and, reciprocally, that of the opposite eye increased. These facts may suggest that proprioception plays in the development of esotropia.
PMCID: PMC1039456  PMID: 7225308
13.  The eighth Frederick H. Verhoeff Lecture. presented by saiichi mishima, MD Behçet's disease in Japan: ophthalmologic aspects. 
The problems of Behçet's disease in Japan have been reviewed with particular emphasis on the ophthalmologic aspects: the historical background for the Japanese works, diagnostic critieria, epidemiology, some statistics, ocular symptomatology, ocular histopathology, etiology, pathophysiology, and treatment. Behçet's disease is the most frequent entity in endogenous uveitis in Japan. Patients are found throughout the country, and the prevalence rate averages seven to eight per 100,000 population: the rate is higher in the northern than in the southern districts. The diagnosis is made on the basis of a combination of clinical symptoms that are divided into the major and minor criteria symptoms. The major criteria comprise the ocular involvement, aphthous ulcers of the oral mucous membrane, genital ulcers, and skin lesions. These symptoms recur often as attacks and the disease follows a chronic course. The ocular involvement is found in 83% to 95% in males and 67% to 73% in females; the male to female ratio in the number of patients is 1.78. Both into the anterior segment type and the fundus and panophthalmic types. The anterior segment type shows serous iridocyclitis with the classic type of hypopyon appearing in about 12% of the attacks. This type is found in about 20%, more often in females than in males, and the visual prognosis is more favorable than in the fundus and panophthalmic types. In the latter two types, attacks of retinal angitis resulting in intensive retinal edema, yellowish-white exudate, and hemorrhages recur particularly in the macular region, and the visual prognosis is poor. More than 50% of male patients lose visual acuity to less than 0.1 in five years, but this is the case in only 10% of female patients. Consequently, Behçet's disease is the cause of blindness in about 12% of acquired blindness in adults. The ocular histopathology during the attack is characterized by severe angitis with intensive infiltration of neutrophil leucocytes largely in the uveal tract and the retina; the latter is severely affected and loss of visual cells and other neural elements results. The etiology of this disease still remains unknown but genetic predisposition is suggested since this disease is strongly linked with HL-A-B5. Environmental factors are also considered. Various abnormalities are found in the blood chemistry, blood cells (particularly in neutrophil leucocytes), immunologic mechanism, fibrinolytic and blood clotting system, and hormonal system. Chemotractic factors are found in the aqueous humor. These changes are particulary enhanced just before and during the ocular attacks. Systemic corticosteroids are deleterious to the visual prognosis, but cyclophosphamide and colchicine appear to suppress attacks and help patients maintain the visual acuity. However, these drugs are toxic, particulary to the reproductive organs, and the patients must be informed of this side effect and be allowed to make a decision before they are used.
PMCID: PMC1311704  PMID: 397657

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