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Summary The distribution of genets of a root and butt rot pathogen Heterobasidion parviporum was studied in a 68-year-old Abies sachalinensis plantation in Japan. A total of 33 pure cultures of H. parviporum were isolated from diseased Todo fir stumps after clear-felling. Individual genets of the fungus were identified by three different methods: somatic incompatibility test, random amplified polymorphic DNA, and DNA microsatellite analyses. The combined results of the three analyses identified at least eight genets within a 60 × 100 m plot. One genet, consisting of only one isolate, was genetically differentiated from the other genets by all three methods; the other 32 isolates, however, were grouped with minor discrepancies, into seven genetically close genets. The genetic differentiation was lower than that recorded previously in Europe. A single genet infected 1–15 trees in the plot. The longest distance of the two most isolated trees of a single genet was 51 m. The close genetic relationship between genets and their large sizes suggest that basidiospores from restricted sources (just a few fruiting bodies) infected the site before establishment of the present Todo fir stand and that the disease has spread mainly by vegetative growth of the mycelia through root contact. Absence of fruiting bodies of the fungus in the study plot also suggests the greater importance of vegetative growth than spore dispersal for the spread of this fungus.

Interstitial lung disease (ILD) is a noteworthy condition in the treatment of systemic sclerosis (SSc) because of its associated mortality and morbidity; however, the efficacy of various treatments for ILD has been controversial in previous reports. In this study, we examined the efficacy and safety of intravenous cyclophosphamide (IVCY) pulse therapy with prednisolone (PSL) for the treatment of ILD with SSc. A total of 121 patients with SSc were screened and evaluated for ILD, using high-resolution computed tomography of the chest, pulmonary function testing, and bronchoalveolar lavage. Thirteen patients with active ILD were enrolled in this study. The treatment protocol for ILD was 0.4 g/m(2) of body surface area of IVCY monthly plus 0.8 mg/kg of body weight of PSL daily. Two to six doses of IVCY were administered, depending on the remission of ILD. Initial PSL doses were maintained for a month and then gradually tapered to 10 mg daily. An activity index of ILD showed improvements in all patients in the 12 months after the initial intervention; however, four patients experienced recurrence of ILD after 24 months, and one additional patient had recurrence of ILD after 36 months. Seven patients reached the 48-month point with no recurrence of ILD. This long observational study for 48 months showed the efficacy of IVCY with PSL for active alveolitis in the first year. However, because five patients had recurrence of ILD more than 1 year after the treatment, it would be necessary to consider maintenance therapy for ILD beyond 1 year.

ナラタケモドキはサクラ類の根株腐朽病菌として近年問題となっている。そこで, 被害林から分離されたナラタケモドキとワタゲナラタケを用いてサクラ苗への接種試験を行った。ナラタケモドキ接種苗4本のうち1本が接種4年後に枯死した。それらすべての接種苗の根系に菌糸束は認められなかったが, 根株の外樹皮, 内樹皮および形成層に菌糸膜が形成され, 枯死苗では地際部の形成層を取り巻いていた。一方, ワタゲナラタケ接種苗は4本すべて枯死しなかった。それらすべての接種苗の根系に多数の菌糸束が絡み付いていたが, 菌糸膜の形成は外樹皮の一部に限られ, 内樹皮と形成層での菌糸膜形成は接種苗1本の細い根数本のみに認められた。接種苗からはそれぞれの接種菌が再分離された。対照苗3本はすべて枯死せず, 菌糸膜や菌糸束の形成も認められなかった。これらの結果から, ナラタケモドキはサクラ苗の内樹皮と形成層に侵入し, さらに枯死させることが明らかになった。

An undetermined Armillaria species was collected on Amami-Oshima, a subtropical island of Japan. The phylogenetic position of the Armillaria sp. was determined using sequences of the elongation factor-1α (EF-1α) gene and the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2) of ribosomal DNA (rDNA). The phylogenetic analyses based on EF-1α and ITS sequences showed that this species differs from known Japanese taxa of Armillaria. The sequences of this species and A. novae-zelandiae from Southeast Asia were contained in a strongly supported clade, which was adjacent to a well-supported sister clade containing A. novae-zelandiae from Australia and New Zealand.

Summary Distribution, host preference and pathogenicity of Japanese Armillaria species on conifers were investigated on the basis of field collections of 65 isolates. We identified seven Armillaria species from 19 conifer species including six major Japanese plantation conifers using mating tests and sequences of the translation elongation-1 α gene. Armillaria mellea, Armillaria ostoyae, Armillaria cepistipes and Armillaria sinapina were frequently collected, whereas Armillaria nabsnona, Armillaria tabescens and a biological species Nagasawa’s E were rare. On the basis of host condition when the isolates were collected, A. mellea, A. ostoyae, A. cepistipes and A. tabescens are considered as moderate to aggressive pathogens of conifers in Japan.

Objective. To determine whether IL-18 is involved in the inflammation of DM and PM. Methods. Thirty-three patients with DM were enrolled in this study, including 25 with interstitial lung disease (ILD). In addition, 16 patients with PM were enrolled, including 6 with ILD. All patients were admitted to our hospital as a result of their condition requiring treatment, and clinical laboratory data including serum IL-18 were recorded on admission. Results. Serum IL-18 was significantly (P < 0.0001) higher in both DM and PM patients than in healthy controls (n = 30). Serum ferritin and IL-18 were significantly (P = 0.003 and 0.0044, respectively) higher in DM than in PM patients. Additionally, ferritin and IL-18 were significantly (P = 0.023 and 0.034, respectively) higher in DM patients with ILD than in DM patients without ILD. Significant positive correlations were found between creatine kinase (CK) and ferritin (rs = 0.39, P = 0.024); CK and IL-18 (rs = 0.48, P = 0.005); and IL-18 and ferritin (rs = 0.54, P = 0.0012) in the DM group as a whole. These findings were different for the DM plus ILD subgroup: significant positive correlations were found between CK and ferritin (rs = 0.40, P = 0.047); CK and IL-18 (rs = 0.63, P = 0.0008); and IL-18 and ferritin (rs = 0.41, P = 0.042). Conclusion. Serum IL-18 was strikingly elevated in patients with DM and was associated particularly with disease activity and ILD complication in DM.

In mammals and probably all vertebrates, receptor editing of TCR alpha genes enhances immature T cell positive selection over a three-day interval in the thymic cortex. Surprisingly we found extensive somatic hypermutation (SHM) operating at the TCRα locus in the nurse shark thymus, implying that SHM contributes to receptor modifications that enhance positive selection. We analyzed mutation in TCRα families of clones with the same VJ rearrangement. Additionally, in situ hybridization showed the strongest activation-induced cytidine deaminase (AID) expression in the central thymic cortex and bordering the corticomedullary junction. The frequency of mutation at TCRα was as high as that seen at B cell receptor (BCR) loci in sharks and mammals. Complementarity determining regions (CDRs) accumulated significantly more mutations than framework regions (FWs), and significantly more mutations in CDRs resulted in amino acid replacement than in silent mutation. We saw a preference for transition mutations as well as a strong bias toward G:C substitutions within AID hotspots, especially within CDR regions. We suggest that TCRα utilizes SHM to boost positive selection and perhaps to broaden diversification of the αβ T cell repertoire in sharks, the first reported use of this process in thymic diversification in vertebrates.

Objective. The aim of this study is to evaluate the clinical manifestation and prognostic factors of anti-melanoma differentiation-associated gene 5 (MDA5) antibody-associated interstitial lung disease (ILD) with DM. Methods. Fourteen patients who presented with anti-MDA5 antibody and 10 patients with anti-aminoacyl-tRNA synthetase (ARS) antibody were enrolled. All patients were diagnosed as having DM with ILD. Clinical manifestations in the patients with anti-MDA5 antibody were compared with those in the patients with anti-ARS antibody. Results. The frequencies of acute/subacute interstitial pneumonia (A/SIP) and fatal outcome were significantly higher in the subset with anti-MDA5 antibody. The creatine kinase (CK) value was significantly lower and the γ-glutamyl transpeptidase and ferritin values were significantly higher in the subset with anti-MDA5 antibody. Significant correlations were found between PaO 2 /F i O 2 and ferritin (r s =-0.59, P=0.035), alveolar-arterial oxygen difference (A-aDO 2 ) and KL-6 (r s =0.73, P=0.016) and A-aDO 2 and ferritin (r s =0.66, P=0.013) in the subset with anti-MDA5 antibody. The most significant prognostic factor was ferritin. The cumulative survival rate was significantly lower (P < 0.0001) in the subset with ferritin ≥ 1600 ng/ml than that in the subset with ferritin

The aim of this study was to evaluate neurological manifestations of primary Sjögren’s syndrome (pSS) and investigate the etiology and pathogenesis of peripheral and central nervous complications in pSS. Thirty-two patients with pSS were enrolled in the present study, 20 of whom had neurological involvement plus sicca symptoms. The clinical features were evaluated by neurological examinations including nerve conduction study, magnetic resonance imaging, cerebrospinal fluid, and electroencephalogram. The frequency of fever was significantly higher (P = 0.006) in pSS with neurological involvement than in pSS without neurological involvement. There was no statistical significance in other factors between the two groups. Peripheral nervous system (PNS), central nervous system (CNS), and both PNS and CNS involvements were revealed in 14, 3, and 3 patients, respectively. Optic neuritis and trigeminal neuralgia were revealed frequently in cranial neuropathy. Anti-aquaporin 4 antibody was detected in one patient with optic neuritis. Of the nine patients with polyneuropathy, eight patients presented pure sensory neuropathy including small fiber neuropathy (SFN). pSS with SFN appeared to have no clinically abnormal features, including muscle weakness and decreasing deep tendon reflex. Skin biopsy revealed epidermal nerve fiber degenerated in one pSS patient with pure sensory neuropathy who was diagnosed as having SFN. Our observations suggest that a number of mechanisms can be attributed to neurological involvements in pSS rather than just the mechanisms previously described (i.e., vasculitis and ganglioneuronitis). Presumably, specific autoantibodies may directly induce injury of the nervous system.

The decay risk of airborne wood-decay fungi was investigated by using an air sampler. Japanese cedar disks 7.8 cm in diameter and about 3 mm in thickness with moisture content of about 100% were placed in a “BIOSAMP” air sampler and exposed to 1000 l air. Air sampling was carried out from June to September at the same sampling site in Tsukuba, Japan. The exposed disks were then incubated for 16 weeks in a damp container kept at 26° ± 2°C. During the incubation period, wood mass loss ranged from −15 to 807 mg with a mean mass loss of 244 mg. Factors affecting mass loss were explored. Wood moisture content and ratio of heartwood area proved to be significant factors. In addition, six weather factors were found to influence mass loss. Disks that were sampled on a cloudy day showed significantly higher mean mass loss compared to those sampled on a sunny day. Subculturing of filamentous fungi from 16-week incubated disks suggested one-third of the isolated fungi produced ligninolytic enzymes.

Relationships among East Asian, North American and European Laetiporus sulphureus s. lat., a cosmopolitan brown rot species complex, were assessed with phylogenetic analyses and incompatibility tests. Three East Asian taxa, Laetiporus cremeiporus sp. nov., Laetiporus montanus and Laetiporus versisporus, are described and illustrated as well as compared with related taxa from Southeast Asia, North America and Europe. Phylogenetic analyses showed that L. cremeiporus and L. versisporus are clearly distinct species among Laetiporus taxa. The three conifer inhabiting species, Laetiporus conifericola, Laetiporus huroniensis and L. montanus, are closely related to each other. The European population of L. montanus exhibits two sequence variants of the EF1alpha: one is the same as observed in L. sulphureus in Europe and the other is that observed in East Asian population of L. montanus. A key to the known species of Laetiporus in the northern hemisphere is provided.

Sixty-two isolates from basidiocarps of Armillaria spp. were obtained from Hokkaido Island, northern Japan. Six species (Armillaria cepistipes, A. gallica, A. nabsnona, A. ostoyae, A. sinapina, and an undescribed species, “Nag. E”) were identified by pairing tests with known tester strains and one subspecies (Armillaria mellea subsp. nipponica, a non-heterothallic form of A. mellea) was identified by its macro- and micro-morphological characters of the basidiocarps. This is the first case of “Nag. E” being reported from Hokkaido Island.

Polyporus longiporus, belonging to group Polyporellus, is newly reported from Japan in what also constitutes the second report of the species since its original description. This species is characterized by radially elongated pores, smooth and brown to dark brown pileus, and cylindrical and slightly curved basidiospores. Phylogenetic analysis based on the ITS region revealed that P. longiporus forms a terminal, strongly supported clade that is closely related to P. ciliatus. A description and illustrations of the present species and a key to the species of group Polyporellus are provided.

We describe and illustrate three Heterobasidion species distributed in East Asia, based on both dried specimens and cultures. Heterobasidion parviporum, formerly known as H. annosum from Asia, is characterized by effused-reflexed basidiocarps with cuticulate pilei, small round pores, thin context and short tubes in each layer. Examination of the type specimen of Trametes insularis revealed that this is distinct from the East Asian species widely known as “H. insulare”. No prior correct name for this taxon exists, and thus we propose the name H. orientale sp. nov. Heterobasidion orientale is characterized by sessile to effused-reflexed basidiocarps covered by a thin crust, reddish brown pileus with a marginal white zone and regular to labyrinthiform pores. Heterobasidion ecrustosum sp. nov. is characterized by convex basidiocarps frequently covered by a crust only at the base, large regular pores and long tubes. Morphological characters of these three species are compared with other Heterobasidion species. Cultural characters of these three species are also described. All isolates show oedocephaloid conidiophores bearing subglobose to ovoid conidia. Conidia of H. parviporum are smaller than those of the other two species. Descriptions and a key to the known species of Heterobasidion are provided.

ならたけもどき病は, 樹木園, 緑地等の樹木に樹勢衰退を引き起こす重要病害である。ならたけもどき病が発生しているフユザクラ林において, ナラタケモドキの土壌中の分布把握を意図し, 杭捕捉法によるナラタケモドキの検出を試みた。枯死木を含む試験区と枯死木を含まない任意に設置した試験区の計11試験区を設定し, コナラ枝で作製した杭を計615本設置した結果, 9試験区の計86本の杭にナラタケ属菌の根状菌糸束や菌糸膜が認められた。そのうち58本の杭から分離されたナラタケ属菌の種は, 交配試験およびPCR-RFLP解析によりすべてワタゲナラタケと同定された。ナラタケモドキは杭捕捉法により検出されなかった。ワタゲナラタケは根状菌糸束を豊富に形成し, 土壌中の広い範囲に生息していると考えられた。

To validate the association of a single nucleotide polymorphism (SNP) of the connective tissue growth factor gene (CTGF) with susceptibility to systemic sclerosis (SSc) in the Japanese population.395 Japanese patients with SSc, 115 patients with rheumatoid arthritis and 269 healthy Japanese volunteers were enrolled in the study. An SNP (rs6918698) at -945 bp from the start codon in the promoter region of the CTGF gene was determined by allelic discrimination with the use of a specific TaqMan probe.The G allele showed a significantly higher frequency in patients with SSc than in controls (p0.001; odds ratio 1.5; 95% confidence interval 1.2 to 1.9). In particular, the clinical subsets of SSc showed a more significant association between the G allele and diffuse cutaneous SSc (p0.001) and the presence of interstitial lung disease (p0.001), the presence of anti-topoisomerase I antibody (p0.001) and anti-U1RNP antibody (p = 0.010). Association analyses using the genotype of the SNP yielded results similar to those of analyses using the allele.This study confirms the association between an SNP in the CTGF gene and susceptibility to SSc, especially in the presence of diffuse cutaneous SSc, interstitial lung disease and anti-topoisomerase I antibody. The results strongly suggest that this SNP may be a powerful indicator of severe skin and lung involvement in patients with SSc.

Summary The occurrence and symptoms of root and butt rot were examined in a 35 × 30 m plot of 68-year-old Todo fir plantation in Hokkaido, Japan. Forty-seven percent of the cut stumps were decayed and 52% of the decayed stumps showed similar decay characteristics with yellowish orange to light brown colouration and expanded pockets in the heartwood. Morphological characteristics of the pure cultures isolated from the decay were similar to the cultures isolated from basidiocarps of Heterobasidion annosum sensu lato, found on fallen logs outside of the research site. Also DNA analysis based on the combined data set of three gene loci (glyceraldehyde 3-phosphate dehydrogenase, heat shock protein 80–1 and elongation factor 1-alpha genes) showed that the isolates from the decay are included in the same clade with the Japanese H. annosum s.l. isolates. They form a subclade to H. parviporum (the European S group of H. annosum s.l.). This is the first report of molecular determination of H. annosum s.l. isolated from root and butt rot in a plantation in Japan.

An unusual mortality of several woody plant species was recently found in the Ogasawara Islands. Here, we show that brown root rot caused by Phellinus noxius was the primary cause of this unusual tree death. Typical symptoms of the disease were confirmed on 41 plant species in 29 plant families. Among them, 23 were novel hosts of P. noxius, including 15 species endemic to the Ogasawara Islands. Six species were new host records in Japan.

Utilizing invisible 2-dimensional codes and digital audio players with a 2-dimensional code scanner, we developed paper-based textbooks with audio support for students with print disabilities, called "multimodal textbooks." Multimodal textbooks can be read with the combination of the two modes: "reading printed text" and "listening to the speech of the text from a digital audio player with a 2-dimensional code scanner." Since multimodal textbooks look the same as regular textbooks and the price of a digital audio player is reasonable (about 30 euro), we think multimodal textbooks are suitable for students with print disabilities in ordinary classrooms.

Phellinus noxius is a pathogenic fungus that causes brown root rot disease in a variety of tree species. This fungus is distributed in tropical and sub-tropical regions of Southeast and East Asia, Oceania, Australia, Central America and Africa. In Japan, it was first discovered on Ishigaki Island in Okinawa Prefecture in 1988; since then, it has been found on several of the Ryukyu Islands. Recently, this fungus was identified from the Ogasawara (Bonin) Islands, where it has killed trees, including rare endemic tree species. For effective control or quarantine methods, it is important to clarify whether the Japanese populations of P. noxius are indigenous to the area or if they have been introduced from other areas. We developed 20 microsatellite markers from genome assembly of P. noxius and genotyped 128 isolates from 12 of the Ryukyu Islands and 3 of the Ogasawara Islands. All isolates had unique genotypes, indicating that basidiospore infection is a primary dissemination method for the formation of new disease foci. Genetic structure analyses strongly supported genetic differentiation between the Ryukyu populations and the Ogasawara populations of P. noxius. High polymorphism of microsatellite loci suggests that Japanese populations are indigenous or were introduced a very long time ago. We discuss differences in invasion patterns between the Ryukyu Islands and the Ogasawara Islands.

Tricholoma matsutake is the most commercially important edible mushroom in pine forests in Japan. Tricholoma bakamatsutake and T. fulvocastaneum, species closely related to T. matsutake, occur in Fagaceae forests. We examined ectomycorrhizal (EM) formation by these Tricholoma species by in vitro synthesis among seven strains (two of T. matsutake, four of T. bakamatsutake, one of T. fulvocastaneum) and axenic plants of pine (Pinus densiflora) and oak (Quercus serrata, Q. phillyraeoides). All strains, except for one of T. matsutake, formed EM associations with both pine and oak. Plant growth and mycelial development were differently affected by EM formation depending on the plant-fungus combination.

Trunk rot poses a substantial threat to Sanbu-sugi, one of the most economically important cultivars of Japanese cedar (Cryptomeria japonica). The etiology of this disease, including its main agents, is incompletely known. This trunk rot was attributed to Fomitiporia (Phellinus) hartigii or F. (Phellinus) punctata. Here we phylogenetically analyzed DNA sequences of four markers from a set of strains isolated from trunk-rot symptoms and recovered a single, monophyletic clade, indicating that a single taxon is involved. This clade was identified as Fomitiporia torreyae, a species described from eastern China. This analysis also proved that trunk rot and/or dieback of other conifers and broadleaf trees, including Sawara cypress (Chamaecyparis pisifera), Japanese umbrella pine (Sciadopitys verticillata) and Japanese pear (Pyrus pyrifolia var. culta), were caused by the same species. The phylogenetic approach to Fomitiporia revealed that the F. torreyae clade was closely related to F. bannaensis but clearly distinct from F. punctata, which originally was thought to be the cause of trunk rot in Sanbu-sugi. Fomitiporia torreyae is redescribed on the basis of more than 40 specimens from multiple hosts from Japan and China. Fomitiporia juniperina comb. nov. is proposed.

Utilizing invisible 2-dimensional codes and digital audio players with a 2-dimensional code scanner, we developed a new type of textbooks for students with print disabilities, called “multimodal textbooks.” Multimodal textbooks can be read with the combination of the two modes: “reading printed text” and “listening to the speech of the text from a digital audio player with a 2-dimensional code scanner.” Since a multimodal textbook looks the same as a regular textbook and the price of a digital audio player is reasonable (about 30 euro), we think multimodal textbooks are suitable for students with print disabilities in ordinary classrooms.

Objective: Adult-onset Still's disease (AOSD) is a clinical entity with heterogeneous etiology. We have encountered patients with AOSD who had severe polyarthritis and who fulfilled the classification criteria for rheumatoid arthritis (RA); however, most patients with AOSD typically exhibit mild arthritis. In this study, we proposed two clinical subsets of AOSD and investigated the clinically significant characteristics of the two subtypes. Methods: We retrospectively analyzed 71 consecutive patients with AOSD. We reviewed the medical records of all patients who were followed up for more than 2 years. We classified all the patients with AOSD into the following 2 subsets: an RA subtype for patients who met the criteria for RA according to the American College of Rheumatology and a non-RA subtype for patients who did not meet the criteria for RA. Results: Our results indicated that the non-RA subtype was accompanied by severe inflammatory complications, including pleuritis and hemophagocytic syndrome. In addition, the serum ferritin and serum IL-18 levels were significantly higher in patients with the non-RA subtype than in those with the RA subtype. Interestingly, only 1 patient with the RA subtype had anti-CCP antibodies, and 1 non-RA subtype patient had rheumatoid factor. These findings distinguish these patients from patients with true RA. Conclusions: There were two subsets of patients with AOSD in the examined population. Patients with high levels of IL-18 or ferritin presented with severe systemic inflammatory disorders (the non-RA subtype), and patients with low levels of IL-18 or ferritin developed severe arthritis (RA subtype).

Changes in two of the elements of myocardial subcellular organelles relating to cardiac energetics, ventricular myosin isozymes and mitochondrial DNA mutations, were examined using left ventricular tissue samples obtained at autopsy from patients with ischemic heart disease. Myosin isozymes were examined in tissues from nine patients with ischemic heart disease and 12 control patients with cancer but no heart disease. Extracted myosin was separated by pyrophosphate gel electrophoresis. The relative concentration of each component was determined by densitometry. Mitochondrial DNA mutations were evaluated in tissues from ten patients with myocardial infarction and 11 control patients with cancer but no heart disease. DNA was extracted and mitochondrial DNA mutations were detected by the polymerase chain reaction. Two bands were revealed by pyrophosphate gel electrophoresis. These contained VM-A, which exhibited faster electrophoretic mobility and was present in lower concentrations, and VM-B, which had a lower mobility and a higher concentration, respectively. SDS polyacrylamide gel electrophoresis showed that these two components contained the heavy chain and light chains 1 and 2 of myosin. VM-A concentrations tended to be higher in patients with ischemic heart disease than in controls. A 7.4-kb deletion was detected between the D-loop and the ATPase 6 genes of mitochondrial DNA from the myocardium of 6 out of 10 patients with myocardial infarction. The relative amounts of the two myosin isozymes could be altered by ischemic heart disease, although the functional significance of these components is unclear. The changes in the two myosin isozymes might be an adaptive change to disordered energy metabolism, but this change was small. The myocardial mitochondrial DNA deletions in patients with myocardial infarction were thought to result from ischemic damage.

The effects of water on viscoelastic properties of human hair were investigated by dynamic mechanical analysis (DMA) and differential scanning calorimetry (DSC). Dynamic modulus (E') of dry human hair decreased linearly with increasing temperature. On the other hand, E' of wet human hair showed an inflection point near 0°C. Moreover, a new dispersion peak was observed in tan δ curves of human hair containing water molecules. The DMA curves of thioglicolic acid-treated human hair were similar to those of untreated human hair. Therefore, it was found that hydrogen bonds between protein chains in human hair acted more prominently than disulfide linkage (S-S linkage). The glass transition temperature (Tg) of human hair decreased with increasing absorbed water content (Wc) and then saturated at about 30% of Wc. The non-freezing water content (Wnf) of human hair measured by DSC was about 30%. This means that physical properties of human hair such as Tg and strength are controlled by bound water which is strongly restricted by hydrophilic groups of proteins.

Objective The complication of interstitial lung disease (ILD) in polymyositis/dermatomyositis (PM/DM) is associated with anti–aminoacyl–transfer RNA synthetase (anti-aaRS) antibody or anti–melanoma differentiation–associated gene 5 (anti–MDA-5) antibody positivity. Anti–MDA-5 antibody is associated with clinically amyopathic DM and fatal outcome due to rapidly progressive ILD in Asian populations. The association between genetic factors and anti–MDA-5 antibody–positive DM is unclear. This study was undertaken to investigate the HLA–DRB1 genotype in patients with anti–MDA-5 antibody–positive DM. Methods We examined genetic differences among 17 patients with anti–MDA-5 antibody–positive DM, 33 patients with anti-aaRS antibody–positive PM/DM, 33 patients with PM/DM without anti-aaRS antibody or ILD, and 265 healthy controls. Results The frequencies of HLA–DRB1*0101 and DRB1*0405 were 29% and 71%, respectively, in patients with anti–MDA-5 antibody–positive DM, which were higher than the frequencies in healthy controls (10% and 25%, respectively). Among the 17 patients with anti–MDA-5 antibody–positive DM, 16 (94%) harbored either the DRB1*0101 or DRB1*0405 allele. The combined frequency of the DRB1*0101 allele and the DRB1*0405 allele was significantly higher in patients with anti–MDA-5 antibody–positive DM than in patients with PM/DM without anti-aaRS antibody or ILD, with an odds ratio (OR) of 42.7 (95% confidence interval [95% CI] 4.9–370.2) (P = 1.1 × 10−5), or in patients with anti-aaRS antibody–positive PM/DM (OR 13.3 [95% CI 1.6–112.6], P = 4.5 × 10−3). Conclusion Our findings indicate that HLA–DRB1*0101/*0405 is associated with susceptibility to anti–MDA-5 antibody–positive DM in the Japanese population.

“Matsutake” mushrooms are formed by several species of Tricholoma sect. Caligata distributed across the northern hemisphere. A phylogenetic analysis of matsutake based on virtually neutral mutations in DNA sequences resolved robust relationships among Tricholoma anatolicum, Tricholoma bakamatsutake, Tricholoma magnivelare, Tricholoma matsutake, and Tricholoma sp. from Mexico (=Tricholoma sp. Mex). However, relationships among these matsutake and other species, such as Tricholoma caligatum and Tricholoma fulvocastaneum, were ambiguous. We, therefore, analyzed genomic copy numbers of σ marY1 , marY1, and marY2N retrotransposons by comparing them with the single-copy mobile DNA megB1 using real-time polymerase chain reaction (PCR) to clarify matsutake phylogeny. We also examined types of megB1-associated domains, composed of a number of poly (A) and poly (T) reminiscent of RNA-derived DNA elements among these species. Both datasets resolved two distinct groups, one composed of T. bakamatsutake, T. fulvocastaneum, and T. caligatum that could have diverged earlier and the other comprising T. magnivelare, Tricholoma sp. Mex, T. anatolicum, and T. matsutake that could have evolved later. In the first group, T. caligatum was the closest to the second group, followed by T. fulvocastaneum and T. bakamatsutake. Within the second group, T. magnivelare was clearly differentiated from the other species. The data suggest that matsutake underwent substantial evolution between the first group, mostly composed of Fagaceae symbionts, and the second group, comprised only of Pinaceae symbionts, but diverged little within each groups. Mobile DNA markers could be useful in resolving difficult phylogenies due to, for example, closely spaced speciation events.

Objective. Interstitial lung disease (ILD) is a serious complication of SSc. We aimed to identify markers associated with SSc-related ILD. Methods. RNA was prepared from the peripheral blood mononuclear cells of 14 SSc patients, divided into four different RNA pools according to the presence or absence of ILD and to the treatment, and subjected to microarray analysis. Real-time quantitative PCR was used to confirm the microarray results in 43 SSc patients, 42 autoimmune controls and 10 healthy controls. Genomic DNA samples were collected from 149 patients with SSc (70 in Hokkaido and 79 in Tokyo) who underwent a high-resolution CT for the evaluation of ILD and from 230 healthy controls. Genotyping was performed using sequence-specific primers. Results. The microarray analysis revealed HLA-DRB5 to be the only gene commonly up-regulated in patients with ILD compared with those without ILD in both comparison groups. High expression levels of HLA-DRB5 in SSc patients with ILD were confirmed by real-time quantitative PCR. The prevalence of HLA-DRB5 gene carriers increased in the SSc patients with ILD relative to those without ILD or to healthy controls in both cohorts. Among the four detected alleles, the HLA-DRB5*01:05 allele was significantly more frequent in SSc patients with ILD than in SSc patients without ILD or in healthy controls. These associations were confirmed in the second cohort. Conclusion. HLA-DRB5 was highly expressed in PBMCs from patients with SSc-related ILD. The HLA-DRB5*01:05 allele is a risk factor for ILD in patients with SSc.

Systemic sclerosis (SSc) is a chronic disease of unknown etiology that is characterized by multiple tissue fibrosis. Transforming Growth Factor-beta (TGF-β) is thought to be the most important mediator that induces fibrosis. However, the molecular mechanisms by which fibrosis is induced have not been fully elucidated. In this study, the role of activin, a member of the TGF-β superfamily, was investigated in the pathogenesis of fibrosis in SSc. Serum activin A levels in patients with SSc were measured by ELISA, and the expression of the activin receptor type IB (ACVRIB/ALK4) and the activity of the signaling pathway via ACVRIB/ALK4 were investigated using western blotting. To evaluate a potential therapeutic strategy for SSc, we also attenuated the ACVRIB/ALK4 pathway using an inhibitor. Serum activin A levels were significantly higher in SSc patients than in normal controls. Activin A and ACVRIB/ALK4 expression were also higher in cultured SSc fibroblasts. Activin A stimulation induced phosphorylation of Smad2/3 and CTGF expression in SSc fibroblasts. Procollagen production and Col1α mRNA also increased upon stimulation by activin A. The basal level of Smad2/3 phosphorylation was higher in cultured SSc fibroblasts than in control cells, and treatment with the ALK4/5 inhibitor SB431542 prevented phosphorylation of Smad2/3 and CTGF expression. Furthermore, production of collagen was also induced by activin A. Activin A-ACVRIB/ALK4-Smad-dependent collagen production was augmented in SSc fibroblasts, suggesting the involvement of this signaling mechanism in SSc. Inhibition of the activin A-ACVRIB/ALK4-Smad pathway would be a new approach for the treatment of SSc.

We analyzed the sequences of three DNA regions-the translation elongation factor-1 alpha (EF-1 alpha) gene and the internal transcribed spacer (ITS) and intergenic spacer (IGS) regions of ribosomal DNA-to compare their accuracy in identifying species of Japanese Armillaria. We studied 49 isolates of eight Armillaria species, A. mellea, A. ostoyae, A. nabsnona, A. cepistipes, A. gallica, A. sinapina, A. tabescens and the biological species Nagasawa E (Nag. E). Phylogenetic analyses of the ITS and IGS data helped in identifying A. mellea, A. ostoyae, A. nabsnona, A. tabescens and Nag. E but could not be used to identify A. gallica, A. cepistipes and A. sinapina. Nevertheless our analysis showed that the EF-1 alpha gene was clearly different in the eight examined species. Restriction fragment length polymorphisms (RFLP) of the IGS-1 region could be used to distinguish most species, but the RFLP profiles of some isolates of A. cepistipes and A. sinapina were the same even with four different restriction enzymes. In conclusion, among the techniques examined in this study, analyzing the EF-1 alpha sequence was found to be the most suitable method for identifying different species of Japanese Armillaria.

Relationships were investigated among North American and Japanese isolates of Laetiporus using phylogenetic analysis of ITS sequences and single-spore isolate incompatibility. Single-spore iso- late pairings revealed no significant compatibility between North American and Japanese isolates. ITS analysis revealed 12 clades within the core Laetiporus clade, seven of which are known to occur in North America (including Hawaii and the Caribbean), three in Japan, two in South America, three in Europe and one in South Africa. The identity of L. sulphureus s.s. has yet to be determined and could be either L. ''sulphureus'' (clade C), which appears to be re- stricted to Europe and occurs on angiosperms and gymnospersm, or L. ''sulphureus'' (clade E), which is found in Europe, North America and South America exclusively on angiosperms. Three clades, one from the Caribbean, one from Hawaii and one from South Africa, have yet to be named formally. Of the three Laetiporus species found in Japan two have been named recently (L. cremeiporus and L. montanus) and one has been epitypified (L. versisporus). The single- spore incompatibility and ITS data support recogni- tion of the three Japanese taxa as distinct biological and evolutionary species.

Acute/subacute interstitial pneumonia (A/SIP) is an intractable and fatal complication of DM. Since a useful indicator predicting the complication of A/SIP has not been found, the aim of this study was to determine whether serum ferritin is a potential predictive indicator of the occurrence of A/SIP in 64 patients with DM.Of the total patients enrolled, 19 had A/SIP, 24 had chronic interstitial pneumonia and 21 were without interstitial lung disease (ILD). Clinical manifestations and laboratory data were obtained from medical records on admission.Serum ferritin levels were extremely high in patients with DM with A/SIP. It was significantly higher in DM with A/SIP than that in DM without A/SIP (median 790 vs 186 ng/ml; P0.0001). The cumulative survival rate for 6 months was 62.7% in patients with DM with A/SIP. Moreover, the cumulative survival rate was significantly (P = 0.016) lower in the group with ferritin levelsor =1500 ng/ml than the rate in the group with ferritin levels1500 ng/ml.Serum ferritin can be useful as a predictor of the occurrence of A/SIP and correlates with the prognosis of A/SIP in DM. The intensive treatment using combination therapy with various immunosuppressant agents should be chosen for patients with ILD with DM showing hyperferritinaemia, especially levels1500 ng/ml.

application/pdf, Journal Article, Regarding lunches served at day nurseries, elementary and junior high schools as well as meals served at dormitories of a high school and a university, a series of survey on actual supplies of fat soluble components, dietary fibers and minerals that are considered important to prevent atherosclerosis was performed. Foodstuffs actyally served from January to March, 1990 for 30 days or 25 days continuously were assayed based on the food tables and the results were summarized : 1)The ratio of fat energy in each meal was from 21.5% to 29.4%. 2) The balance of fatty acid was as follows : P/S 1.57, n-6/n-3 4.4 at the nurseries, P/S 0.64 and 0.71, n-6/n-3 6.8 and 7.1 at elementary and junior high schools, and P/S 1.06 and 1.79,n-6/n-3 6.0 and 5.8 at the dormitories ; high school (females) and university (males). E/PUFA was 0.45～0.71 and supplies of cholesterol 81mg at the nurseries, 91mg and 107mg at the elementary and junior high schools, and 343mg and 240mg at the high school and university dormitories. 3) Dietary fivers were noted at 5.0g at the nurseries, 6.1g and 7.3g at the elementary and junior high schools, and 16.3g and 15.1g at the high school and university dormitories. 4) As to the balance of supplied minerals, a gradual decrease was noted with Ca/Mg from 6.87 to 1.40 while a gradual increase was noted with Na/K from 0.90 to 1.80 with advancing ages of the subjects. Sodium chloride per 1,000 Kcal of supplied energy was within the targeted intake level for adults in either case except that being a little much at the university dormitory.

The objective of this study was to explore the association of single nucleotide polymorphisms (SNPs) of the CD244 gene with several clinical features of systemic lupus erythematosus (SLE). Two hundred and forty-three patients with SLE and 369 healthy controls were enrolled. Two SNPs (rs6682654 and rs3766379) in the CD244 gene were determined by allelic discrimination using a specific TaqMan probe. Only SNP rs3766379 was significantly associated with susceptibility to SLE [P = 0.009; odds ratio (OR) 1.28; 95% confidence interval (CI) 1.04-1.57]. The association was preferentially observed in subsets of SLE patients with nephritis and neuropsychiatric lupus. The frequency of the rs6682654 C allele was strongly associated with nephritis and neuropsychiatric lupus (P = 0.00065; OR 1.99; 95% CI 1.34-2.95, and P = 1.6 × 10(-7); OR 3.47; 95% CI 2.12-5.70, respectively), as was the frequency of the rs3766379 T allele (P = 0.0014; OR 1.86; 95% CI 1.27-2.71, and P = 2.6 × 10(-7); OR 3.15; 95% CI 2.00-4.96, respectively). In this study, an SNP of the CD244 gene was associated with susceptibility to SLE. There was a strikingly strong association in SLE patients with nephritis and neuropsychiatric lupus, suggesting that this genetic marker could predict involvement of those severe complications.