Yamadazyma kitorensis f.a., sp. nov. and Zygoascus biomembranicola f.a., sp. nov., novel yeasts from the stone chamber interior of the Kitora tumulus, and five novel combinations in Yamadazyma and Zygoascus for species of Candida.

Analysis of D1/D2 large-subunit (LSU) rRNA gene sequences predicted that 17 yeast isolates, mainly from viscous gels (biofilms) taken from the stone chamber interior of the Kitora tumulus in Nara, Japan, were placed in the Yamadazyma and Zygoascus clades. Polyphasic characterization, including morphological, physiological and chemotaxonomic characteristics, multigene sequence divergence and DNA-DNA hybridization, strongly suggested the assignment of one novel species to each of the clades; these are Yamadazyma kitorensis f.a., sp. nov., with the type strain JCM 31005T (ex-type CBS 14158T=isolate K8617-6-8T), and Zygoascus biomembranicola f.a., sp. nov., with the type strain JCM 31007T (ex-type CBS 14157T=isolate K61208-2-11T). Furthermore, the transfer of five known species of the genus Candida as novel combinations to the genera Yamadazyma and Zygoascus is proposed; these are Yamadazyma olivae f.a., comb. nov. (type strain CBS 11171T=ATCC MYA-4568T), Yamadazyma tumulicola f.a., comb. nov. (type strain JCM 15403T=ex-type CBS 10917T=isolate T6517-9-5T), Yamadazyma takamatsuzukensis f.a., comb. nov. (type strain JCM 15410T=CBS 10916T = isolate T4922-1-1T), Zygoascus polysorbophila f.a., comb. nov. (type strain NRRL Y-27161T=CBS 7317T) and Zygoascus bituminiphila f.a., comb. nov. (type strain CBS 8813T=MUCL 41424T).

Early onset and focal spike discharges as indicators of poor prognosis for myoclonic-astatic epilepsy.

BACKGROUND: Myoclonic-astatic epilepsy (MAE) is an epileptic syndrome characterized by unique myoclonus, myoclonic-astatic, or astatic seizures in childhood. MAE prognosis vary from spontaneous remission to intractable seizures with profound mental retardation. AIM: Identifying early risk factors may optimize the treatment of children with MAE. Our hypothesis is early onset age and focal spike discharges on EEG indicate a poor MAE prognosis. METHODS: Using the medical records of 9 children with MAE, we analyzed their clinical histories, EEG findings, and seizure symptoms. All patients were given follow-up observations/treatments by our department for at least 2 years after MAE onset. RESULTS: Five of the patients were given favorable prognoses because their seizures disappeared within 2 years of onset; the other 4 received poor prognoses because their seizures continued more than 2 years. MAE onset in patient with refractory seizures was earlier than that in those with a favorable prognosis (7-24 months vs. 23-38 months). All the patients with refractory seizures showed moderate or severe mental retardation. Among the 5 patients with good prognosis, EEGs showed two with focal spike discharges and three with only generalized spike discharges. In contrast, all cases with a poor prognosis had focal spike discharges. CONCLUSIONS: MAE onset in patients with refractory seizures occurs earlier than in those with favorable prognosis. Prognosis was excellent when EEG findings show no focal spike discharges. Both early seizure onset and the focal spike discharges associated with MAE are indicators of poor prognosis.

A nationwide survey of Aicardi-Goutières syndrome patients identifies a strong association between dominant TREX1 mutations and chilblain lesions: Japanese cohort study.

OBJECTIVES: Aicardi-Goutières syndrome (AGS) is a rare, genetically determined, early onset progressive encephalopathy associated with autoimmune manifestations. AGS is usually inherited in an autosomal recessive manner. The disease is rare, therefore the clinical manifestations and genotype-phenotype correlations, particularly with regard to autoimmune diseases, are still unclear. Here we performed a nationwide survey of AGS patients in Japan and analysed the genetic and clinical data. METHODS: Patients were recruited via questionnaires sent to paediatric or adult neurologists in Japanese hospitals and institutions. Genetic analysis was performed and clinical data were collected. RESULTS: Fourteen AGS patients were identified from 13 families; 10 harboured genetic mutations. Three patients harboured dominant-type TREX1 mutations. These included two de novo cases: one caused by a novel heterozygous p.His195Tyr mutation and the other by a novel somatic mosaicism resulting in a p.Asp200Asn mutation. Chilblain lesions were observed in all patients harbouring dominant-type TREX1 mutations. All three patients harbouring SAMHD1 mutations were diagnosed with autoimmune diseases, two with SLE and one with SS. The latter is the first reported case. CONCLUSION: This study is the first to report a nationwide AGS survey, which identified more patients with sporadic AGS carrying de novo dominant-type TREX1 mutations than expected. There was a strong association between the dominant-type TREX1 mutations and chilblain lesions, and between SAMHD1 mutations and autoimmunity. These findings suggest that rheumatologists should pay attention to possible sporadic AGS cases presenting with neurological disorders and autoimmune manifestations.

Two novel ascomycetous yeast species, Wickerhamomyces scolytoplatypi sp. nov. and Cyberlindnera xylebori sp. nov., isolated from ambrosia beetle galleries.

Thirteen strains of yeasts were isolated from ambrosia beetle galleries at several sites in Japan. Based on the morphological and biochemical characteristics and phylogenetic analysis of the D1/D2 domain of the large subunit (LSU) rRNA gene of the yeasts, 10 strains were shown to represent a novel species of the genus Wickerhamomyces, described as Wickerhamomyces scolytoplatypi sp. nov. (type strain NBRC 11029(T) = CBS 12186(T)), and were closely related to Wickerhamomyces hampshirensis. The three other strains represented a novel species of the genus Cyberlindnera, described as Cyberlindnera xylebori sp. nov. (type strain NBRC 11048(T) = CBS 12187(T)), and were closely related to Cyberlindnera euphorbiiphila. It is suggested that these species are associated with ambrosia beetles and we consider ambrosia beetle galleries as good sources of novel yeasts.

Wickerhamomyces tratensis sp. nov. and Candida namnaoensis sp. nov., two novel ascomycetous yeast species in the Wickerhamomyces clade found in Thailand.

Two closely related yeast strains, ST-382 and ST-392, isolated in Thailand showed intermediate relatedness in the DNA-DNA hybridization experiment suggesting that the two strains represent closely related distinct species. In the tree based on the D1/D2 domain sequences of the large subunit rRNA gene, the two strains are located in a subclade in the Wickerhamomyces clade with high bootstrap support. In the D1/D2 domain, the two strains differed by two nucleotides and are assumed to be very closely related. Strain ST-392(T) (=BCC 15102(T) = NBRC 107799(T) = CBS 12176(T) forming hat-shaped ascospores is described as Wickerhamomyces tratensis sp. nov. and strain ST-382(T) (= BCC 15093(T) = NBRC 107800(T) = CBS 12175(T) is described as Candida namnaoensis sp. nov. because ascospores are not found in this strain. In phenotypic characteristics, W. tratensis and C. namnaoensis are discriminated by the ability of alcoholic fermentation and the assimilation of galactose, D-xylose and D-gluconic acid.

Diagnosing nocturnal frontal lobe epilepsy: a case study of two children.

We describe two children of nocturnal frontal lobe epilepsy (NFLE) diagnosed using carefully observed nocturnal sleep EEGs and detailed patient histories. Case #1, a 14-year-old boy, showed repeated generalized tonic convulsions and frequent eyes opening seizures during sleep. Conventional EEGs - done with the patient awake or in sleep stage I - showed no abnormalities, while a nocturnal sleep EEG - done during in sleep stage II - revealed the repeated, sharp wave bursts predominantly in the right frontal lobe characteristic of NFLE. During these wave bursts, we noticed the boy's eyes opening, although his parents had not been aware this NFLE symptom. Case #2, a 12-year-old boy, showed one daytime generalized convulsion. He had also been suffering from repeated paroxysmal episodes similar to parasomnia - waking up, sitting, walking, screaming, and speaking - which always followed the same patterns lasting several minutes. During the nocturnal sleep EEG, episodes occurred twice, showing abnormal epileptic discharges predominantly in the frontal lobe. His parents did not mention the episodes to us until questioned, as they had recognized them as parasomnia. The previous conventional EEG showed abnormal slow waves in the frontal lobe, which led us to suspect frontal lobe epilepsy and to take a detailed patient history. The frequency and stereotypy of their symptoms during sleep caused us to perform nocturnal sleep EEGs and led us NFLE diagnosis. Detailed patient histories including sleep habits and carefully observed nocturnal sleep EEGs enabled us to recognize these NFLE clinical features.

Pichia porticicola sp. nov., a novel ascomycetous yeast related to Pichia acaciae isolated from galleries of ambrosia beetles in Japan.

Eleven strains of yeasts, isolated from galleries of ambrosia beetles in Japan, formerly identified as Pichia acaciae were found to have different sequences in the D1/D2 domain of the large subunit (LSU) rRNA gene. After detailed taxonomic studies including a DNA-DNA reassociation experiment, 11 strains were found to represent a novel species of the genus Pichia. It is described as Pichia porticicola sp. nov. (type strain NBRC 100302(T) = CBS 11715(T)). The eleven strains were isolated from various samples associated with different kinds of insects and trees which were collected in 11 prefectures, from the north to the south of Japan. This species might be a common species in galleries of ambrosia beetles in Japan.

Candida potacharoeniae sp. nov. and Candida spenceri sp. nov., two novel galactose-containing ascomycetous anamorphic yeast species isolated in Thailand.

Fifteen strains of anamorphic yeasts isolated from various natural substrates collected in various places in Thailand were found to represent two novel species of anamorphic yeast genus Candida based on the sequence analysis of the D1/D2 domain of the large subunit rRNA genes, chemotaxonomic and conventional properties used for the classification of yeasts. These strains are located in the clade including Candida etchellsii and Candida magnoliae. Fourteen strains represented by ST-490(T) (BCC 15176(T)=NBRC 106439(T)= CBS 11674(T)) are closely related to Candida sorbosivorans in the D1/D2 sequences but 11 nucleotides (2.4%) were substituted. The remaining strain, ST-594(T) (=BCC 15278(T)=NBRC 106446(T)=CBS 11673(T)) showed a close relationship to Candida geochares but 21 nucleotides (4.7%) were substituted. Apparently, these strains represent two novel Candida species of the Starmerella clade. The two species are described as Candida potacharoeniae sp. nov. and Candida spenceri sp. nov. in the present paper. Like the most species of this clade, the two species contain galactose in the cells in addition to glucose and mannose and have high mol% G + C of 54.4-55.9 and 54.9, respectively.

Vanderwaltozyma tropicalis sp. nov., a novel ascomycetous yeast species found in Thailand.

Two strains of yeasts isolated in Thailand were found to represent a novel species of the genus Vanderwaltozyma. These strains are close to V. verrucispora but differ by 13 nucleotides (2.3%) in the D1/D2 domain of the LSU rRNA gene. This difference clearly suggests that the two strains represent a distinct species. It is described as Vanderwaltozyma tropicalis sp. nov. Vanderwaltozyma tropicalis is distinguished from V. verrucispora by the assimilation of trehalose, inability to grow in 0.1% cycloheximide and by higher maximum growth temperature.

Millerozyma phetchabunensis sp. nov., a novel ascomycetous yeast species isolated from Nam Nao forest soil in Thailand, and the transfer of Pichia koratensis to the genus Millerozyma.

A total of 4 strains (DR2-4, DW2-1, DS2-1 and DS3-1) of undescribed ascomycetous yeast, isolated from Nam Nao forest soil, were identified as a novel species in the genus Millerozyma. The D1/D2 sequences of the strains differed from the closely related species Millerozyma acaciae and Pichia koratensis by 1.2% (7 nucleotide substitutions) and 1.4% (8 nucleotide substitutions). Phenotypically, all the novel strains were identical, but differed from M. acaciae and P. koratensis by a variety of phenotypic characteristics. Based on phenotypic and phylogenetic data, these four yeast strains were assigned to a single novel species in the genus Millerozyma and the name Millerozyma phetchabunensis sp. nov. is proposed. In addition, we also propose the transfer of P. koratensis, which was placed in the Millerozyma clade based on the analysis of the D1/D2 and ITS sequences, to the genus Millerozyma as M. koratensis comb. nov.

Three new species of bipolar budding yeasts of the genus Hanseniaspora and its anamorph Kloeckera isolated in Thailand.

In the course of a survey of yeast biodiversity in the natural substrates in Thailand, eight strains were found to represent three hitherto undescribed species of Hanseniaspora/Kloeckera. They were isolated from insect frass, flower, lichen, rotted fruit and rotted wood. Based on the morphological and physiological characteristics, and sequences of D1/D2 domain, six strains represent a single species of the genus Hanseniaspora, described as Hanseniaspora thailandica sp. nov. (type BCC 14938(T)=NBRC 104216(T)=CBS 10841(T)), and another strain as Hanseniaspora singularis sp. nov. (type BCC 15001(T)=NBRC 104214(T)=CBS 10840(T)). A further strain, which belongs to Kloeckera and does not produce ascospores, is described as Kloeckera hatyaiensis sp. nov. (type BCC 14939(T)=NBRC 104215(T)=CBS 10842(T)). Strains belonging to H. thailandica sp. nov. differed by 17-19 nucleotide substitutions from Hanseniaspora meyeri, the closest species. DNA reassociation between the two taxa showed 30-48% relatedness. Kloeckera hatyaiensis sp. nov. and H. singularis sp. nov. differed by eight and 16 nucleotide substitutions with one gap from the nearest species, Hanseniaspora clermontiae and Hanseniaspora valbyensis, respectively.

Candida nonsorbophila sp. nov., a new ascomycetous yeast species isolated in Thailand.

Four yeast strains, RS42, SSK10, ST-520 and ST-521, isolated from water in a mangrove forest, bark of a tree and fruit of Ficus sp., respectively, were found to represent a hitherto undescribed anamorphic species. The four strains are related to Candida sinolaborantium in the D1/D2 domain of the large subunit rRNA gene, but differed by eight nucleotide substitutions and two indels, and for this reason are regarded as members of a separate species. Because ascospore formation was not detected, it is described as a new species of Candida, Candida nonsorbophila sp. nov. The type strain is RS42(T) (BCC 25963(T)=NBRC 103860(T)=CBS 10862(T)). This species is distinguished from C. sinolaborantium by the inability to assimilate L-sorbose, L-rhamnose and galactitol, and a higher maximum growth temperature.

Candida wancherniae sp. nov. and Candida morakotiae sp. nov., two novel ascomycetous anamorphic yeast species found in Thailand.

Seven yeast strains isolated from natural substrates of Thailand were found to represent two novel species of Candida, an ascomycetous anamorphic genus. Three strains, ST-233, ST-259 and ST-260, isolated from insect frass and plant leaves were found to represent a single novel species related to Metschnikowia agaves in a tree based on the D1/D2 domain sequences of the 26S rRNA genes. This species is clearly discriminated from M. agaves by the carbon assimilation patterns and required vitamins. The remaining four strains, ST-18, ST-261, ST-606 and ST-658, isolated from the fruit body of a mushroom, insect frass, decayed jack fruit and an unidentified flower, were found to represent a single species which is related to Candida corydali, a recently described insect-associated species, in a neighbor-joining tree based on the D1/D2 sequences. This species is clearly discriminated from C. corydali by the ability to assimilate propane-1,2-diol and the inability to assimilate glucono-delta-lactone. They are described as Candida wancherniae sp. nov. and Candida morakotiae sp. nov., respectively.

Vanderwaltozyma verrucispora sp. nov., a new ascomycetous yeast species.

A new yeast species, Vanderwaltozyma verrucispora, is proposed in this study based on two strains isolated from partially decayed leaves in Japan and one strain from soil in Taiwan. The species is characterized by the fermentation of glucose and galactose, formation of one to four spheroidal to ellipsoidal ascospores with warty surfaces in each ascus, and assimilation of a few carbon and nitrogen compounds. Genus assignment and distinction of the species from the other two recognized species of Vanderwaltozyma is based on the morphological and physiological characteristics, and phylogenetic analysis of nucleotide sequences of the D1/D2 domains of the large subunit (LSU) rRNA gene. From these comparisons, the name V. verrucispora sp. nov. is proposed. Sequence analysis of the D1/D2 domains of the LSU rRNA gene reveals that the phylogenetically closest relative of V. verrucispora is Vanderwaltozyma yarrowii. The type strain of the new species, which was isolated from a partially decayed leaf in Kagoshima Prefecture, Japan, is NBRC 1884(T) (=CBS 10887(T)=BCRC 23141(T)).

Candida kanchanaburiensis sp. nov., a new ascomycetous yeast species related to Pichia nakazawae isolated in Thailand.

Three yeast strains, ST-633, ST-634 and ST-635, isolated from the fruit body of a mushroom, Coprinus sp., and rotted fruit of guava collected in the western region of Thailand, were found to represent a hitherto undescribed species. This yeast is related to Pichia nakazawae var. akitaensis, P. nakazawae var. nakazawae and Pichia philogaea in the D1/D2 domain of 26S rDNA but 12 (2.3%), 13 (2.5%) and 15 (2.8%) nucleotides are different from these taxa, respectively, suggesting the distinctness of the Thai strains at species level. Since ascospore formation was not detected, it is described as a new species of Candida, Candida kanchanaburiensis. This species is distinguished from P. nakazawae by the ability to assimilate 2-ketogluconic acid and L-lysine, and inability to assimilate soluble starch.

Ogataea paradorogensis sp. nov., a novel methylotrophic ascomycetous yeast species isolated from galleries of ambrosia beetles in Japan, with a close relation to Pichia dorogensis.

Two yeast strains isolated from galleries of ambrosia beetles in Japan and maintained in NITE Biological Resource Center (NBRC) as Pichia pini were found to represent a hitherto undescribed species. This species shows close relationship to Pichia dorogensis by the sequence analysis of the D1/D2 domain of 26S rDNA but is clearly differentiated from it by a DNA-DNA reassociation experiment. It is described as Ogataea paradorogensis sp. nov. The vegetative cells and asci of this species are surrounded with distinct capsules like P. dorogensis. One to four hat-shaped ascospores, which tend to be liberated from the asci at maturation, are formed in the ascus.