Design and synthesis of N-alkyl oxindolylidene acetic acids as a new class of potent Cdc25A inhibitors

The oxindolylidene acetic acids having long N-alkyl chains exhibited strong inhibitory activity toward dual specificity phosphatase Cdc25A.     

Analysis of sphingolipid classes and their contents in meals

Sphingolipids have attracted attention as physiologically functional lipids. We determined their class and content in Japanese meals that had been prepared by a nutritionist, mainly by using HPLC-ELSD. In all 12 meals tested, cerebroside and/or sphingomyelin were generally detected as the major sphingolipids. The total amounts of sphingolipids in typical high- and low-calorie meal samples over 2 days were 292 and 128 mg/day, and 81 and 45 mg/day, respectively.     

Phylogenetic relationships of ferns deduced from rbcL gene sequence

Part of the large subunit of the ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) gene (rbcL) was sequenced from three fern species: Adiantum capillus-veneris, Botrypus strictus, and Osmunda cinnamomea var. fokiensis. This region included 1,333 base pairs, about 90% of the gene. Maximum likelihood analysis of the deduced amino acid sequences indicated that (1) Botrypus (Ophioglossaceae) clustered monophyletically with other ferns (Adiantum, Angiopteris, Osmunda); the closest relative to Botrypus among the three species was Osmunda, which did not support the hypothesis that the Ophioglossaceae are linked to the progymnosperm-seed plant lineage. (2) Eusporangiate ferns containing Botrypus (Ophioglossaceae) and Angiopteris (Marattiaceae) were a paraphyletic group. (3) Seed plants and the four fern species examined formed a monophyletic group, but ferns and bryophytes (liverwort) did not. Variations in rates of substitution for synonymous and nonsynonymous codons were found in fern lineages.Correspondence to: M. Hasebe     

Phylogenetic relationships in gnetophyta deduced fromrbcL gene sequences

Part of the large subunit of the ribulose-1,5-bisphosphate carboxylase gene (rbcL) was sequenced (1333 base pairs) from three species of gymnosperms:Ephedra sinica Gnetum parvifolium, Welwitschia mirabilis. Phylogenetic trees inferred from the neighbor joining, Wagner parsimony and maximum likelihood methods showed thatGnetum andWelwitschia were more closely related to each other than either is toEphedra within Gnetophyta, and the result supports previous cladistical analysis of morphological data.     

A study on body temperature regulation and residential thermal environments of the elderly—I. whole country survey of residential thermal environment and evaluation methods: RTE-index

1. 1. As part of “research on environmental comfort,” that is, research which aims to make people's living environment more comfortable, we attempted to clarify the relation between the living environment of elderly people and physiological and psychological factors. We carried out a comprehensive study with a view to establishing comfort standards for the residential thermal environment, and for creating evaluation and control systems.

2. 2. The comfort of the living environment is closely related to the thermal environment and the temperature-regulation ability of the human body. This ability of the body to regulate temperature develops during childhood and recedes as the adult ages.

3. 3. We therefore carried out: (a) experiments on body-temperature regulation and on special characteristics of the body-temperature regulation of elderly people, (b) a nationwide survey of the actual residential thermal environments of elderly people and (c) a survey of the daily activities of elderly people, and how elderly people's physiological and psychological conditions change when they are engaged in these activities.

4. 4. As a result of these experiments and surveys, we were able to: (d) formulate standards for evaluation of the residential thermal environment and (e) numerically express the results of systematic evaluation of residential thermal environments of elderly people, by means of a RTE-index.

Expression of human tyrosine kinase, Lck, in yeast Saccharomyces cerevisiae: growth suppression and strategy for inhibitor screening

We report the successful expression and detection of a phosphorylated form of human T cell tyrosine kinase, Lck, in Saccharomyes cerevisiae, which leads to growth suppression of the yeast cells. Expression of an inactive Lck mutant resulted in no phosphorylation and no growth suppression, indicating that cell growth inhibition by Lck is due to the activity of the kinase, consistent with the observed tyrosine-phosphorylation of the Lck and yeast host cell proteins. The addition of a known inhibitor of Lck to the cell culture resulted in recovery of cell growth expressing the active Lck, suggesting that the growth inhibition by lck gene expression can be used to screen inhibitors for the gene product. We have extended such approach to Tob, another potential therapeutic target.     

Carbon and nitrogen content of Noctiluca scintillans in the Seto Inland Sea, Japan

The carbon and nitrogen content of Noctiluca scintillans cellsfrom the Seto Inland Sea, Japan was investigated in order toestimate its biomass in natural samples. The carbon contentof N.scintillans ranged from 123 to 627 ng C cell^–1 witha mean value of 353 ng C cell^–1, or 1.12 to 2.67 fg Cµm^–3 with a mean value of 1.98 fg C µm^–3.The nitrogen content ranged from 36.0 to 232 ng N cell^–1with a mean value of 131 ng N cell^–1, or 0.499 to 0.910fg N µm^–3 with a mean value of 0.694 fg N µm^–3.Total cell carbon and nitrogen increased but the carbon andnitrogen per cell volume decreased with increasing cell volume.The C/N ratio of the cells ranged from 2.3 to 4.4, which wasrelatively low compared with the Redfield ratio. The carbonand nitrogen content was extremely low (91.2 ng C cell^–1,41.8 ng N cell^–1) for starved cells, whereas it was extremelyhigh (528 ng C cell^–1, 205 ng N cell^–1) for cellswhich had ingested the large diatom, Coscinodiscus wailesii.Our results suggest that the carbon and nitrogen content ofN.scintillans varies depending on its physiological conditionand the type of food that it has recently consumed.     

Co-Overexpression of GEP100 and AMAP1 Proteins Correlates with Rapid Local Recurrence after Breast Conservative Therapy

A major problem of current cancer research and therapy is prediction of tumor recurrence after initial treatment, rather than the simple biological characterization of the malignancy and proliferative properties of tumors. Breast conservation therapy (BCT) is a well-approved, standard treatment for patients with early stages of breast cancer, which consists of lumpectomy and whole-breast irradiation. In spite of extensive studies, only ''age'' and ''Ki-67 positivity'' have been identified to be well correlated with local recurrence after BCT. An Arf6 pathway, activated by GEP100 under receptor tyrosine kinases (RTKs) and employs AMAP1 as its effector, is crucial for invasion and metastasis of some breast cancer cells. This pathway activates β1 integrins and perturbs E-cadherin-based adhesions, hence appears to be integral for epithelial-mesenchymal transdifferentiation (EMT). We here show that expression of the Arf6 pathway components statistically correlates with rapid local recurrence after BCT. We retrospectively analyzed four hundred seventy-nine patients who received BCT in Hokkaido University Hospital, and found 20 patients had local recurrence. We then analyzed pathological samples of patients who experienced local recurrence by use of Kaplan-Meier analysis, Stepwise regression analysis and the t-test, coupled with immunostaining, and found that co-overexpression of GEP100 and AMAP1 correlates with rapidity of the local recurrence. Their margin-status, node-positivity, and estrogen receptor (ER)- or progesterone receptor (PgR)-positivity did not correlated with the rapidity. This study is the first to show that expression of a certain set of proteins correlates with the rapidity of local recurrence. Our results are useful not only for prediction, but highlight the possibility of developing novel strategies to block local recurrence. We also discuss why mRNAs encoding these proteins have not been identified to correlate with local recurrence by previous conventional gene expression profiling analyses.     

A map-based cloning strategy employing a residual heterozygous line reveals that the GIGANTEA gene is involved in soybean maturity and flowering

Flowering is indicative of the transition from vegetative to reproductive phase, a critical event in the life cycle of plants. In soybean (Glycine max), a flowering quantitative trait locus, FT2, corresponding to the maturity locus E2, was detected in recombinant inbred lines (RILs) derived from the varieties "Misuzudaizu" (ft2/ft2; JP28856) and "Moshidou Gong 503" (FT2/FT2; JP27603). A map-based cloning strategy using the progeny of a residual heterozygous line (RHL) from the RIL was employed to isolate the gene responsible for this quantitative trait locus. A GIGANTEA ortholog, GmGIa (Glyma10g36600), was identified as a candidate gene. A common premature stop codon at the 10th exon was present in the Misuzudaizu allele and in other near isogenic lines (NILs) originating from Harosoy (e2/e2; PI548573). Furthermore, a mutant line harboring another premature stop codon showed an earlier flowering phenotype than the original variety, Bay (E2/E2; PI553043). The e2/e2 genotype exhibited elevated expression of GmFT2a, one of the florigen genes that leads to early flowering. The effects of the E2 allele on flowering time were similar among NILs and constant under high (43°N) and middle (36°N) latitudinal regions in Japan. These results indicate that GmGIa is the gene responsible for the E2 locus and that a null mutation in GmGIa may contribute to the geographic adaptation of soybean.