[Successful preparation of mitoxantrone emulsion containing non-ionic contrast medium].

Mitoxantrone, a new anti-cancer agent, was successfully prepared for Lipiodol emulsion. The mixture of Mitoxantrone and non-ionic contrast medium, Omnipaque 300, was combined with Lipiodol at the ratio of 1:2. When the ratio of Mitoxantrone and Lipiodol was 1:4, microscopic study revealed stabilized water in oil emulsion, which could release the anti-cancer agent slowly. We applied it for a case of hepatocellular carcinoma with good result. Intra-arterial infusion of this emulsion might be considered effective for treatment of hepatocellular carcinoma.

Intrinsic colossal magnetoresistance effect in thin-film Pr0.5Sr0.5MnO3 through dimensionality switching.

A homogeneous colossal magnetoresistance (CMR) effect at low temperatures has been found in a thin-film perovskite manganite Pr0.5Sr0.5MnO3. The transition is driven not by the spin alignment as in usual CMR in bulk samples but by the localization-delocalization transition switched by the change in the effective dimensionality. Two-dimensional (x2-y2)-orbital ordering enhanced by the substrate strain is essential for the stabilization of the insulating localized state, which is on the verge of the first-order transition to the three-dimensional metallic ferromangetic state.

Persistent and reversible all-optical phase control in a manganite thin film.

Persistent and reversible optical phase control has been achieved in a manganite thin film through a careful choice of the composition of Pr1-x(Ca1-ySr(y))xMnO3 near a multicritical point. Pulsed laser light brings the lower temperature metallic phase out of the higher temperature charge-ordered insulator, while a cw light reverses the effect by heating. We clearly demonstrate the two competing roles played by light, heating, and excitation across the charge gap, which are important in both the application and the understanding of the physics of electron correlation.

Isolation and characterization of a laccase-derepressed mutant of Neurospora crassa.

Laccase from the ascomycete Neurospora crassa is an inducible secretory enzyme. Production of this enzyme is repressed in vegetative cultures but can be induced by treatment with low concentrations of cycloheximide. Isolation and characterization of a derepressed mutant, the lah-1 mutant, that is capable of producing laccase in vegetative cultures without induction by cycloheximide are described. The lah-1 mutation is mapped between nit-2 and leu-3 on linkage group I, and it behaved as a recessive mutation in a forced heterokaryon. No differences were detected biochemically or immunologically between the laccase protein produced by the lah-1 mutant in the absence of cycloheximide and that induced with cycloheximide in the wild-type strain. This suggests that both laccases (66 kilodaltons) are products of the same structural gene. Relative amounts of laccase in the culture filtrate of the lah-1 mutant were much higher than those induced with cycloheximide in the wild-type strain, demonstrating high efficiency of the lah-1 mutant in production and secretion of laccase. The time course of laccase production by the lah-1 mutant revealed that expression of 66-kilodalton laccase was repressed in conidia and derepressed during vegetative mycelial growth. This suggests that a multiple regulatory mechanism is involved in the production and/or maturation of Neurospora laccase. The lah-1 mutant may be useful for identifying genes that regulate expression of the laccase gene in N. crassa.

Microsphere resonators strongly coupled to a plane dielectric substrate: coupling via the optical near field.

A model is proposed that describes the essential optical process in the recently observed resonant light scattering from a microsphere resonator that is strongly coupled to the substrate. The experimentally observed field patterns across the resonance can be reproduced quite well by a numerical calculation taking into account only a few vector spherical waves that are converted from nonpropagating to propagating waves at the substrate surface. Explicit consideration of the multiple-reflection effect is not necessary to reproduce the experimental results. Comparison of the experiment and the calculation suggests the splitting of degenerate resonance modes that have different azimuthal mode numbers within a single broad resonance line. These results are discussed on the basis of the strongly coupled nature of the system.

Observation of a modulation effect caused by a microsphere resonator strongly coupled to a dielectric substrate.

The effect of modulation caused by a microsphere resonator is experimentally investigated with a model system consisting of a microsphere resonator and a plane substrate. We used total internal reflection microscopy (TIRM), which is a combination of conventional optical microscopy and the total internal reflection method, and observed the intensity distribution under the resonator in the evanescent-wave incidence condition. The TIRM patterns drastically change when the wavelength of the incident beam is scanned across a resonance. The response of the system is discussed on the basis of a recent proposal of traveling-wave resonance.

A histone H3 methyltransferase controls DNA methylation in Neurospora crassa.

DNA methylation is involved in epigenetic processes such as X-chromosome inactivation, imprinting and silencing of transposons. We have demonstrated previously that dim-2 encodes a DNA methyltransferase that is responsible for all known cytosine methylation in Neurospora crassa. Here we report that another Neurospora gene, dim-5, is required for DNA methylation, as well as for normal growth and full fertility. We mapped dim-5 and identified it by transformation with a candidate gene. The mutant has a nonsense mutation in a SET domain of a gene related to histone methyltransferases that are involved in heterochromatin formation in other organisms. Transformation of a wild-type strain with a segment of dim-5 reactivated a silenced hph gene, apparently by 'quelling' of dim-5. We demonstrate that recombinant DIM-5 protein specifically methylates histone H3 and that replacement of lysine 9 in histone H3 with either a leucine or an arginine phenocopies the dim-5 mutation. We conclude that DNA methylation depends on histone methylation.

Isolation and characterization of mutants defective in production of laccase in Neurospora crassa.

A protein synthesis inhibitor, cycloheximide, induces excretion of laccase in Neurospora crassa. The lah-1 mutation results in excretion of a large amount of laccase even in the absence of cycloheximide. Ten mutations were induced that suppress derepressed excretion of laccase in the lah-1 mutant. Of these, seven second-site mutations were found to confer a laccase-noninducible phenotype, and were classified into two different complementation groups. Four mutations define a locus designated lni-1, found to be closely linked to ylo-1 on linkage group VI. The other three mutations were mapped to second locus, designated lni-2, that lies between nic-3 and thi-3 on linkage group VII. The lni-2 locus was shown to encode laccase by RFLP mapping of the DNA fragment encoding laccase and by transformation of the lni-2 mutant with plasmid pBL1 carrying the laccase gene (the locus encoding laccas is hereafter described as lacc). All lacc mutants examined (whether mutagen-induced or inactivated by repeat-induced point mutation) appeared to exhibit no phenotypic deficiency during both asexual and sexual cycles, suggesting that the laccase gene is dispensable in N. crassa. Northern analysis of total cellular RNA from the four lni-1 mutants demonstrated that the lni-1 mutations abolish increased transcription of the laccase gene under inducing conditions. Consequently, the lni-1 locus is inferred to encode a trans-acting positive regulator required for transcriptional activation of the laccase gene in response to cycloheximide. Possible functions of the lah-1 gene are also described.

Transcriptional activation of a cycloheximide-inducible gene encoding laccase is mediated by cpc-1, the cross-pathway control gene, in Neurospora crassa.

Expression of the laccase gene (lacc) of Neurospora crassa is transcriptionally inducible by the protein synthesis inhibitor cycloheximide. A lni-1 mutation, conferring the laccase non-inducible phenotype, was found to be a cpc-1 allele. Northern blots probed with plasmid pLA1, which carries the lacc gene revealed that the cpc-1 mutation abolishes the induced transcription of the lacc gene, indicating requirement of the cpc-1 gene for transcriptional activation of the lacc gene. In Northern blots probed with plasmid pAB1, which bears arg-2 a gene whose transcription is under the control of CPC1, the level of the arg-2 transcript was shown to increase several-fold in wild-type mycelia but remained low in cpc-1 mycelia, after treatment with cycloheximide. This suggests that inhibition of protein synthesis with cycloheximide, as well as amino acid limitation, elicits the CPC1-mediated cross-pathway control. Characterization of the lacc upstream region using a series of 5'-deletion plasmids led to the identification of a 170 bp DNA region required for the induced lacc expression. Sequence analysis of this DNA region demonstrated that it includes a 9 bp sequence with dyad symmetry, ATGAATCAT, which differs only by a central base pair from ATGA(C/G)TCAT, the recognition sequence characteristic of CPC1 and GCN4 binding sites. Possible mechanisms by which CPC1 mediates transcriptional activation of the lacc gene are discussed.