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Background and objectives Meta-iodobenzylguanidine (MIBG) myocardial scintigraphy is an effective tool for distinguishing Parkinson's disease (PD) from other diseases accompanied by parkinsonism. Unlike other Parkinsonian diseases, in PD, MIBG accumulation in the heart tends to decrease. However, previous studies have reported that a decrease in MIBG accumulation also occurs in progressive supranuclear palsy (PSP). Thus, we analyzed the relationship between the degree of MIBG accumulation decrease, clinical symptoms, and brainstem atrophy in PSP. Methods We retrospectively collected data from patients who underwent MIBG myocardial scintigraphy and compared MIBG indices (heart to mediastinum [H/M] ratio, washout rate) between subjects with PSP and other diseases including PD. In addition, we evaluated the relationship between clinical characteristics, MIBG accumulation, and brainstem atrophy in patients with PSP. Results Patients with PSP had a significantly lower early H/M ratio compared with multiple system atrophy with predominant parkinsonism (MSA-P) patients, and a control group. In PSP patients there was a correlation between the decrease in delay H/M ratio, atrophy of the pons, and clinical severity as evaluated by Hoehn and Yahr score. Conclusion Unlike in PD, PSP patients exhibited a mild decrease in MIBG accumulation in MIBG myocardial scintigraphy, which may be related to brainstem atrophy.

There are no specific radiologic features in MOG-Ab (autoantibodies directed against myelin oligodendrocyte glycoprotein)-associated diseases. We present two MOG-Ab-positive patients with symmetrical lesions in the bilateral cingulate cortex of the frontal and parietal lobes. Those lesions showed hyperperfusion in acute phase and hypoperfusion in chronic phase on brain SPECT. In both patients, steroid therapy was effective in acute phase and for prevention of recurrence. High signal in the bilateral cingulate cortex on MR T2-weighted and FLAIR images might to be one of the unique findings considered MOG-Ab associated diseases.

OBJECTIVE To determine the characteristics of olfactory dysfunction in patients with temporal lobe epilepsy (TLE). METHODS Odor identification was assessed using the odor stick identification test for Japanese (OSIT-J, full score 12 points) in 65 patients with TLE and in 74 controls. RESULTS The mean OSIT-J score was significantly lower in patients with TLE (mean ± SD = 8.1 ± 2.8; median = 9) than in the control subjects (mean ± SD = 10.6 ± 1.1; median = 11) (P

The chemical composition of three Laurencia spp., Laurencia sp., L. okamurae and L. saitoi, which were collected from the Seto Inland Sea of Japan, has been examined. Laurencia sp. collected from the coast of Matoba Park, Takehara, Hiroshima Prefecture, contained a brominated chamigrane-type sesquiterpene (1), named matobol, as the main metabolite. The structure of matobol was determined as (+)-(2R,3R,6R,10S)-2,10-dibromochamigr-7(14)-en-3-ol (1). This is the first time that the optically active 1 has been isolated from Laurencia. On the other hand, L. okamurae from the coast of Ikunoshima Island, Hiroshima Prefecture, produced laurinterol (2) that is a known cyclolaurane-type sesquiterpene characteristic to this species in Japan. L. saitoi from the coast of Matoba Park contained a known bromoallenic C15-acetogenin, neolaurallene (3).

Polyamide membranes with a controlled surface morphology were prepared by the interfacial polymerization of 1,3-phenylenediamine (MPD) with 1,3,5-benzenetricarbonyl trichloride (TMC) on polysulfone ultrafiltration supports. In a novel polymerization method, co-solvents, which included acetone, ethyl acetate, diethyl ether, toluene, isopropyl alcohol (IPA) and N,N ′-dimethyl formamide (DMF), were added into the organic phase which made it possible to control the surface morphology and polyamide network structures. As-prepared, the membranes showed multi-layered ridge-and-valley structures, and the types of co-solvent successfully controlled permeate flux and rejection. Polyamide membranes prepared by the addition of 2 wt% ethyl acetate, showed the best performance with NaCl rejection of 99% and permeate flux of more than 1 m 3 /(m 2 d) at 1.5 MPa, which was about twice that of a membrane prepared without a co-solvent.

Polyamide membranes with controlled surface morphology were prepared by the interfacial polymerization of 1,3-phenylenediamine (MPD) with 1,3,5-benzenetricarbonyl trichloride (TMC) on polysulfone ultrafiltration supports. The addition of co-solvents (acetone, ethyl acetate, and diethyl ether) into the organic phase was used to control both the surface morphology and the polyamide network structures. The as-prepared membranes had multi-layered polyamide structures, which consisted of large (1 μm) and ordinal (100–300 nm) ridge-and-valley formations. Permeate flux and rejection reactions were successfully controlled by the types and amounts of co-solvents that were added. The optimal membrane conditions included the addition of 3 wt% ethyl acetate, with a NaCl rejection of more than 99% and a permeate flux of more than 1.8 m3/(m2 d) at 1.5 MPa, which was more than three times higher than the membranes prepared without a co-solvent.

A spray-assisted, 2-step interfacial polymerization (IP) of trimesoyl chloride (TMC)/ m -phenylenediamine (MPD) was proposed for the preparation of polyamide (PA) membranes. In the first step, TMC/hexane solutions were sprayed onto MPD-impregnated polysulfone (PSf) supports for 10–60 s, followed by a second step where the first-step membranes made contact with the TMC/hexane solutions. It is noteworthy that water permeability was increased with spray time in the first step of the 2-step IP, showing approximately doubled values at the spray time of 20–30 s, compared with 1-step PA/PSf composite membranes, while NaCl rejections were practically unchanged. With longer spray time (30–60 s), water permeability decreased while NaCl rejection remained at the same level. A characterization of 1- and 2-step PA membranes, including FE-SEM and ATR-FTIR, revealed the formation of multilayered ridge-and-valley structures of polyamide from the spray-assisted IP steps, and it is suggested that the increased water permeability have been caused by the increased interfacial surface area of the 2-step PA membranes.

We have identified and characterized a Coprinopsis cinerea mutant defective in stipe elongation during fruiting body development. In the wild-type, stipe cells elongate at the maturation stage of fruiting, resulting in very slender cells. In the mutant, the stipe cells fail to elongate, but become rather globular at the maturation stage. We found that the mutant phenotype is rescued by a gene encoding a homolog of Saccharomyces cerevisiae CDC3 septin, Cc.Cdc3. The C. cinerea genome includes 6 septin genes, 5 of which, including Cc.cdc3, are highly transcribed during stipe elongation in the wild type. In the mutant, the level of Cc.cdc3 transcription in the stipe cells remains the same as that in the mycelium, and the level of Cc.cdc10 transcription is approximately 100 times lower than that in the wild-type stipe cells. No increase in transcription of Cc.cdc3 in the mutant may be due to the fact that the Cc.cdc3 gene has a 4-base pair insertion in its promoter and/or that the promoter region is methylated in the mutant. Overexpressed EGFP-Cc.Cdc3 fusion protein rescues the stipe elongation in the transformants, localizes to the cell cortex and assembles into abundant thin filaments in the elongating stipe cells. In contrast, in vegetative hyphae, EGFP-Cc.Cdc3 is localized to the hyphal tips of the apical cells of hyphae. Cellular defects in the mutant, combined with the localization of EGFP-Cc.Cdc3, suggest that septin filaments in the cell cortex provide the localized rigidity to the plasma membrane and allow cells to elongate cylindrically.

Focal cortical dysplasia (FCD) is an important cause of intractable epilepsy. Previous rat studies have utilized freeze lesioning of neonatal animals to model FCD; however, such models are unable to demonstrate spontaneous seizures without seizure-provoking events. Therefore, we created an animal model with multiple FCD, produced during embryonic development, and observed whether spontaneous seizures occurred. Furthermore, we examined the relationship between FCD and epileptogenesis using immunohistochemistry. At 18 days postconception, a frozen metal probe was placed bilaterally on the scalps of Sprague-Dawley rat embryos through the uterus wall to produce multiple FCD. Electroencephalogram (EEG) and video recording were performed from postnatal day (P) 35 to P77. Brain tissues were examined immunohistochemically at P28 and P78 using semiquantitative densitometry. Eleven of 16 rats (68.8%) showed spontaneous seizures arising in the hippocampus from P47. Movement cessation followed by sniffing and mastication, culminating in wet-dog shaking, was seen during the hippocampal EEG discharges. FCD was observed in the bilateral frontoparietal lobes. The expression levels of N-methyl-d-aspartate receptor (NMDAR) subunits 1, 2A, 2B, the glutamate/aspartate transporter and the glial glutamate transporter 1 (GLT1) at FCD sites were increased at P28 and P78. There were no major histological abnormalities in the hippocampi compared with those in the cortex. However, the expression levels of NMDAR 2A and 2B were increased at P28. Levels of NMDAR1, 2A and 2B, the glutamate/aspartate transporter and GLT1 were also increased at P78. We created an animal model showing spontaneous seizures without a provoking event except for the existence of cortical dysplasia, and without a genetic or general systematic cause like MAM injection or irradiation. The seizures resembled human temporal lobe epilepsy both clinically and on EEG. Alterations in the levels of glutamatergic and GABAergic receptors were investigated during growth. This model should enable better clarification of the mechanisms underlying the development of human epilepsy.

We characterized a Coprinopsis cinerea mutant strain, Spe20, defective in fruiting initiation, which was isolated after restriction enzyme-mediated integration (REMI) mutagenesis of a homokaryotic fruiting strain, 326. A plasmid rescue followed by complementation experiments, RACE, and cDNA analyses revealed that the gene, a mutation of which is responsible for the phenotype, is predicted to encode a protein that exhibits a high similarity to yeast Snf5p, a key component of the chromatin remodeling complex SWI/SNF, and named Cc.snf5. Cc.Snf5 is, however, different from Snf5p in that the former has, in addition to an Snf5 domain comprising N-terminal repeat1 (rp1) and C-terminal repeat2 (rp2) subdomains in a middle region, a GATA Zn-finger domain in a C-terminal region. In strain Spe20, plasmid pPHT1 used for REMI is inserted in the ORF encoding rp2. This raised the possibility that in strain Spe20, the disrupted Cc.Snf5 is functionally active albeit incompletely because it retains rp1. Thus, we disrupted the whole SNF5 domain and its downstream peptide and found that the disruption results in inhibition of not only fruiting initiation but also dikaryon development, a prerequisite for fruiting. We also found that specific disruption of the Zn-finger domain results in inhibition of fruiting initiation. These results indicate that Cc.Snf5 plays an essential role in sexual development of C. cinerea.

This study introduces a promising strategy, called co-solvent assisted interfacial polymerization (CAIP), for the synthesis of thin polyamide membranes by interfacial polymerization of 1,3-phenylenediamine (MPD) with 1,3,5-benzenetricarbonyl trichloride (TMC) on polysulfone ultrafiltration supports. A synergistic co-solvent added in the hexane phase was used to control the polymerization reaction zone and modify the membrane network structure. The resultant membrane exhibited selective molecular sieving of small molecules from larger ones. The addition of larger amounts of acetone as a co-solvent to the hexane solution increased both pore size and water flux, which was determined by analysis of the membrane permeation properties. In the permselectivity test of a 500 ppm glucose aqueous solution, the best-performing membrane was prepared by 2 wt% acetone addition. It showed a rejection of more than 99.4% and a high water transport at a rate of more than 1 × 10 −11 m 3 /(m 2 Pa s), which was more than 4-fold higher than the membrane prepared without acetone. The effects of reaction conditions, including co-solvent content and interfacial polymerization time, were studied.

A promising strategy is reported for the synthesis of inorganic-polyamide nanocomposite membranes on an ultrafiltration polysulfone support via metal-alkoxide-assisted interfacial polymerization. Three types of nanocomposite membranes were prepared using three different metal alkoxides. The metal alkoxides used here were titanium tetraisopropoxide, bis(triethoxysilyl)ethane and phenyltriethoxysilane. The as-prepared nanocomposite membranes exhibited performance superior to that of the pure polyamide membrane. Water flux and salt rejection were observed for each of the nanocomposite membranes. Addition of greater amounts of metal alkoxide to the hexane solution increased both pore size and water flux, which were determined by analysis of the membrane permeation data using aqueous solutions of sodium chloride and organic solutes at a pressure of 1.5 MPa and a temperature of 25 °C. The best nanocomposite membrane that was prepared with phenyltriethoxysilane showed water flux that was increased approximately 2-fold compared with the pure polyamide membrane with negligible rejection loss.

The agaricomycete (homobasidiomycete) Coprinopsis cinerea has been used as a model to study the molecular mechanism for photomorphogenesis. Molecular genetic analyses of mutants defective in fruiting body (mushroom) photomorphogenesis of C. cinerea identified two genes, dst1 and dst2. dst1 encodes a homolog of WC-1, a fungal blue-light photoreceptor first identified in Neurospora crassa, while dst2 encodes a novel protein with a putative flavin adenine dinucleotide (FAD)-binding-4 domain. In addition, reverse genetic analysis revealed that disruption of a C. cinerea gene encoding a WC-2 homolog, the partner of WC-1, causes the same blind phenotype. Searches on the genome data show that both WC-1 and WC-2 homologs are present in some agaricomycetes other than C. cinerea. Furthermore, in an agaricomycete, Lentinula edodes, it has been shown in vitro that the WC-1 and WC-2 homologs interact with each other. These findings suggest that the presumptive mechanism for blue-light sensing in agaricomycetes is fundamentally similar to that in Neurospora crassa, in which the WC-1/WC-2 complex plays a central role. Since the WC-1/WC-2 complex operates as a photoreceptor and a transcription factor, future studies will include identification of the targets of the WC-1/WC-2 complex that regulate photomorphogenesis in agaricomycetes. Another future challenge will be elucidation of the role of the newly identified photomorphogenetic protein, Dst2, in the blue-light-sensing mechanism.

The fruiting-body primordium of Coprinopsis cinerea exhibits remarkable photomorphogenesis. Under a 12-h light/12-h dark regime, the primordium proceeds to the fruiting-body maturation phase in which the primordium successively undergoes basidiospore formation, stipe elongation and pileus expansion, resulting in the mature fruiting-body. In continuous darkness, however, the primordium never proceeds to the maturation phase: the pileus and stipe tissues at the upper part of the primordium remain rudimentary while the basal part of the primordium elongates, producing the etiolated "dark stipe" phenotype. In our previous studies, blind mutants, which produce dark stipes under light conditions that promote fruiting-body maturation in the wild-type, have been isolated, and two genes, dst1 and dst2, responsible for the mutant phenotype have been identified. In this study we show that the dst2-1 mutant exhibits a blind phenotype during asexual spore production in addition to that in fruiting-body photomorphogenesis. We also reveal that dst2 is predicted to encode a protein with a putative flavin adenine dinucleotide (FAD)-binding-4 domain. The two blind phenotypes, together with the existence of an FAD-binding domain in Dst2, suggest that Dst2 may play a role in perceiving blue light.

The homobasidiomycete Coprinopsis cinerea is a member of the fungi known as inky cap mushrooms, and its fruiting-body pileus autolyzes soon after completion of the development. During the last 3h of the development, the pileus exhibits umbrella-like expansion: the pileal tissue is cracked at the base of each gill and then each gill tissue is split to form a V-shape, as seen in a cross section. We identified two C. cinerea mutants defective in both pileus expansion and autolysis. The defects in both mutants are due to recessive mutations in a single gene, designated exp1. The exp1 gene is predicted to encode an HMG1/2-like protein with two HMG domains. The transcription of exp1 is strongly induced in the pileus 3h before pileus expansion. This result, together with the fact that the exp1 mutations cause a specific developmental phenotype, suggest that Exp1 is a novel, transcriptional regulator controlling the final phase of fruiting-body morphogenesis.

We identified and characterized elongationless3 (eln3-1), a restriction enzyme-mediated integration (REMI) mutation affecting fruiting body morphogenesis in Coprinus cinereus. The mutant produces an aberrant fruiting body in which the stipe hardly elongates during fruiting body maturation. In the wild type, cylindrical stipe cells, elongation growth of which is responsible for stipe elongation, make side-by-side contact with one another and run parallel to the stipe axis, whereas in the mutant, the organization of the stipe tissue is disturbed and much space is produced between stipe cells. This disorganization of the stipe tissue, together with reduced elongation of the stipe cells, causes the mutant stipe short and bulgy. After a plasmid rescue, the eln3 gene was identified as a DNA fragment that complements the eln3-1 mutation. The eln3 ORF is predicted to encode a protein of 927 amino acids with a general glycosyltransferase domain and to be located in the plasma membrane. Transcription of the eln3 gene is specifically activated in rapidly elongating stipes. Possible involvement of the putative Eln3 enzyme in cell-to-cell connection is discussed. Index Descriptors: Homobasidiomycete; Coprinus cinereus; Fruiting body morphogenesis; REMI mutant; Stipe elongation; Cell-to-cell connection; Membrane protein; Glycosyltransferase

To remove nucleic acids from cellular products as drugs, cross-linked N,N-dimethylaminopropylacrylamide (DMP) particles with cationic functional groups were prepared. The particle's hydrophobicity and its anion-exchange capacity were easily adjusted by changing the cross-linking agent and the DMP ratio in the cross-linking, respectively. When divinylbenzene (DVB) was used as a cross-linking agent and the DMP ratio (in the cross-linking) was adjusted to 90 mol%, the particles (DMP-DVB, 90:10) showed the highest adsorbing activity of DNA (salmon spermary). Its adsorption capacity was 54 mg/ml adsorbent. On the other hand, the adsorption of bovine serum albumin (BSA) to the DMP-DVB extremely increased with increase in the adsorbent's pore size (molecular mass exclusions; M(lim)) from 2 x 10(3) to 1 x 10(4), but decreased with increase in the buffer's ionic strength (mu) to 0.2 or stronger. As a result, when the DMP-DVB (80:20) with M(lim) 2 x 10(3) was used as adsorbent by a column method at pH, 7.2 and mu = 0.17, it only selectively removed DNA from a BSA solution, including 1000 microg/ml of BSA and 10 microg/ml of DNA. The adsorbent decreased the concentration of DNA in the BSA solution to < 10 ng/ml, and the recovery rate of BSA was more 98%.

A genetic linkage map of the basidiomycete Coprinus cinereus was constructed on the basis of the segregation of 219 RAPD markers, 28 RFLP markers and the A and B mating-type loci among 40 random basidiospore progeny from a single cross between a wild-type homokaryon, KF3#2, and an AmutBmut strain, #326. Thirteen linkage groups covering a total of 1346 cM were identified and correlated to the 13 chromosomes of this fungus by hybridization of RFLP and RAPD marker probes to CHEF blots. These probes also revealed chromosome length polymorphisms (CLP), which could be associated with haplotype plots of the progeny. The average kb/cM ratio in this cross was approximately 27.9 kb/cM. The AmutBmut strain undergoes sexual development without mating, because of mutations in both A and B mating-type loci, and has been used to identify mutations affecting developmental processes such as dikaryosis, fruit body morphogenesis, and meiosis. The markers in the map, especially the RAPD ones, would facilitate mapping of genes responsible for such mutations induced in the AmutBmut strain.

We constructed a common-AB diploid strain of Coprinus cinereus and mated this to a compatible haploid strain to construct a diploid-haploid dikaryon. We examined the positions of the diploid and haploid nuclei in the apical and subapical cells of the dikaryon by fluorescence microscopy and microfluorometry. In 60% of apical cells the leading nucleus (the nucleus proximal to the hyphal apex) was diploid and the second nucleus (the nucleus distal to the apex) was haploid, whereas in the remaining 40% of apical cells the order of the two nuclei was reversed. It was also observed that in 97% of hyphae examined the order of the diploid and haploid nuclei was reversed between the apical cell and the subapical cell. Based on these observations, we conclude that the two nuclei alternate in taking the leading and second positions in the apical cell at almost every conjugate division in the dikaryon. Copyright 1998 Academic Press.

We used fluorescence microscopy to examine the positioning of the two nuclei, the configuration of the mitotic apparatus, and the site of septation during conjugate division in eight dikaryons homozygous for either one α-or one of the seven β-tubulin mutations as well as the parental wild-type dikaryon of Coprinus cinereus . In the wild-type dikaryon, more than 90% of the mitotic apparatus had distinct asters, whereas in the mutants the frequency of the apparatus with distinct asters was more or less lowered. In one of the β-tubulin mutants, BEN 193, the defect of astral microtubules was most remarkable and 75% of the mitotic apparatus lacked astral microtubules, although spindle formation and elongation occurred normally. In BEN 193, the two nuclei were positioned normally during conjugate division in the majority of hyphal cells examined, despite the defect of astral microtubules; the leading nucleus divided in the clamp and the second nucleus in the main hypha just beneath the clamp. This result provided evidence against a direct involvement of astral microtubules in the positioning of the two nuclei during conjugate division. In BEN 193, however, septation in the clamp was disturbed in spite of the fact that a mitotic nucleus was positioned normally in the clamp. In the mutant, the actin ring, which forms prior to septation, was also disturbed in its angle in the clamp. These results strongly suggested that astral microtubules control septation in the clamp.