Search PubMed⌕ Search

Biomedical subjects

M Hasebe

Publications and source records attributed to M Hasebe.

At least 19 recordsLinked to original sources

Establishment of gene-trap and enhancer-trap systems in the moss Physcomitrella patens.

Because of its simple body plan and ease of gene knockout and allele replacement, the moss Physcomitrella patens is often used as a model system for studies in plant physiology and developmental biology. Gene-trap and enhancer-trap systems are useful techniques for cloning genes and enhancers that function in specific tissues or cells. Additionally, these systems are convenient for obtaining molecular markers specific for certain developmental processes. Elements for gene-trap and enhancer-trap systems were constructed using the uidA reporter gene with either a splice acceptor or a minimal promoter. Through a high rate of transformation conferred by a method utilizing homologous recombination, 235 gene-trap and 1073 enhancer-trap lines were obtained from 5637 and 3726 transgenic lines, respectively. The expression patterns of these trap lines in the moss gametophyte varied. The candidate gene trapped in a gene-trap line YH209, which shows rhizoid-specific expression, was obtained by 5' and 3' RACE. This gene was named PpGLU, and forms a clade with plant acidic alpha-glucosidase genes. Thus, these gene-trap and enhancer-trap systems should prove useful to identify tissue- and cell-specific genes in Physcomitrella.

Blotting, Northern↗

Isolation of homeodomain-leucine zipper genes from the moss Physcomitrella patens and the evolution of homeodomain-leucine zipper genes in land plants.

Homeobox genes encode transcription factors involved in many aspects of developmental processes. The homeodomain-leucine zipper (HD-Zip) genes, which are characterized by the presence of both a homeodomain and a leucine zipper motif, form a clade within the homeobox superfamily and were previously reported only from vascular plants. Here we report the isolation of 10 HD-Zip genes (named PPHB:1-PPHB:10) from the moss Physcomitrella patens. Based on a phylogenetic analysis of the 10 PPHB: genes and previously reported vascular plant HD-Zip genes, all of the PPHB: genes except Pphb3 belong to three of the four HD-Zip subfamilies (HD-Zip I, II, and III), indicating that these subfamilies originated before the divergence of the vascular plant and moss lineages. Pphb3 is sister to the HD-Zip II subfamily and has some distinctive characteristics, including the difference of the a(1) and d(1) sites of its leucine zipper motif, which are well conserved in each HD-Zip subfamily. Comparison of the genetic divergence of representative HD-Zip I and II genes showed that the evolutionary rate of HD-Zip I genes was faster than that of HD-Zip II genes.

Amino Acid Sequence↗

Tagged mutagenesis and gene-trap in the moss, Physcomitrella patens by shuttle mutagenesis.

The moss, Physcomitrella patens has been used as a useful material in many fields, because of its simple body plan, ease of gene targeting, and other reasons. Although many mutants have been reported, no method to isolate the corresponding genes was reported. We developed a gene tagging and gene-trap system in P. patens by using the shuttle mutagenesis technique, which has been used in the budding yeast. In 5264 tagged lines, 203 mutants with altered developmental or morphological phenotypes were obtained. In 129 of 4757 gene-trap lines, beta-glucuronidase (GUS) activity was detected in some tissue. Although multiple copies of a tag were detected in many tagged lines by Southern analyses, most copies are likely integrated at the same locus according to PCR analyses.

Blotting, Northern↗

Molecular phylogeny of Coriaria, with special emphasis on the disjunct distribution.

Coriaria, which has the most conspicuously disjunct distribution of the flowering plants, is distributed in four separate areas of the world. The phylogenetic relationships of 12 Coriaria species collected from the representative disjunct areas were inferred by comparing 2416 bp of the combined data set of rbcL (a large subunit of ribulose 1,5-bisphosphate carboxylase/oxygenase) and matK (maturase K) genes. The phylogenetic tree shows that the Chile-Papua New Guinea-New Zealand-Pacific islands species and the Central America-northern South America species form a sister group, and the Eurasian clade is more basal to them. The divergence time between the Eurasian group and the other species was estimated as 63 or 59 million years ago using rbcL and matK molecular clocks, respectively. These results do not support previously proposed hypotheses which explain the disjunct distribution on the basis of continental drift but suggest that the distribution pattern was formed by several geographical migrations and separations in the Cenozoic.

Cucurbitaceae↗

Phylogeny of the lady fern group, tribe Physematieae (Dryopteridaceae), based on chloroplast rbcL gene sequences.

Nucleotide sequences of the chloroplast gene rbcL from 42 species of the fern tribe Physematieae (Dryopteridaceae) were analyzed to gain insights into the inter- and intrageneric relationships and the generic circumscriptions in the tribe. The phylogenetic relationships were inferred using the neighbor-joining and maximum-parsimony methods, and both methods produced largely congruent trees. These trees reveal that: (1) Athyrium, Cornopteris, Pseudocystopteris, and Anisocampium form a clade and Athyrium is polyphyletic; (2) Deparia sensu lato is monophyletic and Dictyodroma formosana is included in the Deparia clade; (3) Diplaziopsis forms a clade with Homalosorus, which is isolated from the other genera of the Physematieae; (4) Monomelangium is included in the monophyletic Diplazium clade; and (5) Rhachidosorus is not closely related to either Athyrium or Diplazium.

Chloroplasts↗

Prevention of N-methylnitrosourea-induced colon carcinogenesis in rats by oxygenated carotenoid capsanthin and capsanthin-rich paprika juice.

Epidemiological and animal studies have provided evidence that dietary carotenoids may reduce the risk of certain types of cancer. An inhibitory activity of oxygenated carotenoid capsanthin, a potent antioxidant, and paprika juice rich in capsanthin (3.54 mg/100 ml) against colon carcinogenesis was investigated in F344 rats. In Experiment I (short-term assay), six rats each were given a gavage of 5 mg, 0.2 mg, or 0.008 mg capsanthin six times a week for Weeks 2-6 after receiving three intrarectal doses of 4 mg N-methylnitrosourea in Week 1. The number of colonic aberrant crypt foci, preneoplastic lesions, at Week 6 was significantly fewer (by 42%) in the 0.2 mg capsanthin group, but not in other groups, than the control group. In Experiment II (long-term assay), five groups of 30 or 25 rats each received an intrarectal dose of 2 mg N-methylnitrosourea three times a week for Weeks 1-3, and had either of 10 p.p.m. or 2 p.p.m. capsanthin solutions, 1:2.5 and 1:16.7 diluted solution of paprika juice (containing 10 p.p.m. or 2 p.p.m. capsanthin), and tap water (control fluid) as drinking fluid throughout the experiment. The experimental groups were fed 0.2 mg or 0.04 mg capsanthin/day/rat. The colon cancer incidence at Week 30 was significantly lower in the highly diluted paprika juice group (40%), but not in the moderately diluted paprika juice group (60%) and the capsanthin solution groups (68% and 68%) than the control group (83%). The results suggested that paprika juice may affect colon carcinogenesis. However, capsanthin alone failed to inhibit colon tumorigenesis, in spite of suppression of aberrant crypt foci formation in the short-term assay. Further studies are needed to explain this discrepancy.

Animals↗

Characterization of homeodomain-leucine zipper genes in the fern Ceratopteris richardii and the evolution of the homeodomain-leucine zipper gene family in vascular plants.

The homeodomain-leucine zipper (HD-Zip) genes encode transcription factors that are characterized by the presence of both a homeodomain and a leucine zipper motif. They belong to the homeobox gene superfamily and have been reported only from flowering plants. This article is the first report on the ferm HD-Zip genes (named Crhb1-Crhb11) isolated from the homosporous ferm Ceratopteris richardii. Phylogenetic analyses of the II Crhb genes with previously reported angiosperm HD-Zip genes show that the Crhb genes belong to three of the four different angiosperm HD-Zip subfamilies (HD-Zip I, II, and IV), indicating that these subfamilies of HD-Zip genes originated before the diversification of the ferm and seed plant lineages. The Crhb4-Crhb8 and Crhb11 genes belong to the HD-Zip I subfamily but differ from angiosperm HD-Zip I genes by the presence of a seven-amino-acid indel in the leucine zipper motif. By the northern analyses, Crhb1 and Crhb3 were expressed only in gametophyte tissue. Expression of Crhb2 and Crhb11 genes could not be detected in any tissue examined, while all other Crhb genes were expressed in most sporophytic and gametophytic tissues. Although the functions of the Crhb genes in Ceratopteris are unknown, their patterns of expression suggest that they regulate developmental or physiological processes common to both the gametophyte and the sporophyte generations of the fern. Differences in the expression of Crhb1 between male gametophytes and male-hermaphrodite mixed populations of gametophytes suggests that the Crhb1 gene is involved in gametophytic sex determination.

Amino Acid Sequence↗

[Advances in metabolic and nutritional management of patients in critical care].

Nutritional management often fails in critically ill patients such as trauma and sepsis because of their acutely progressive malnutrition and metabolic derangement being characteristic of such clinical settings. Nutritional strategies for these patients have been almost established by the methods of clinical epidemiology. A considerable amount of clinical evidences suggests that the following should be of benefit for the patients, enteral delivery of nutrition, a nutrition assessment including metabolic evaluation such as liver function or energy requirements, and supplementation of specialty nutrients. Our current data suggest that the metabolic evaluation should include measurement of energy expenditure, assessment of hepatic mitochondrial function, and measurement of the magnitude of body water deviations. Energy expenditure and hepatic mitochondrial function can be observed by the methods of indirect calorimetry and arterial ketone body measurement, respectively. These parameters are helpful to prevent overfeeding from the patients. We emphasize that application of a new method, body impedance spectrum analysis, is important and useful to evaluate the change of body water distributions resulting from metabolic derangement.

Critical Care↗

Characterization of MADS homeotic genes in the fern Ceratopteris richardii.

The MADS genes encode a family of transcription factors, some of which control the identities of floral organs in flowering plants. To understand the role of MADS genes in the evolution of floral organs, five MADS genes (CMADS1, 2, 3, 4, and 6) were cloned from the fern Ceratopteris richardii, a nonflowering plant. A gene tree of partial amino acid sequences of seed plant and fern MADS genes showed that the fern genes form three subfamilies. All members of one of the fern MADS subfamilies have additional amino-terminal amino acids, which is a synapomorphic character of the AGAMOUS subfamily of the flowering plant MADS genes. Their structural similarity indicates a sister relationship between the two subfamilies. The temporal and spatial patterns of expression of the five fern MADS genes were assessed by Northern blot analyses and in situ hybridizations. CMADS1, 2, 3, and 4 are expressed similarly in the meristematic regions and primordia of sporophyte shoots and roots, as well as in reproductive structures, including sporophylls and sporangial initials, although the amount of expression in each tissue is different in each gene. CMADS6 is expressed in gametophytic tissues but not in sporophytic tissues. The lack of organ-specific expression of MADS genes in the reproductive structures of the fern sporophyte may indicate that the restriction of MADS gene expression to specific reproductive organs and the specialization of MADS gene functions as homeotic selector genes in the flowering plant lineage were important in floral organ evolution.

Biological Evolution↗

Prevention of N-methylnitrosourea-induced colon carcinogenesis in F344 rats by lycopene and tomato juice rich in lycopene.

Epidemiological studies have suggested a protective effect of lycopene and lycopene-rich tomatoes against various cancers. Here, the inhibition of colon carcinogenesis by lycopene and tomato juice was investigated. Seven-week-old female F344/NSlc rats received an intrarectal dose of 2 mg (experiment I) or 4 mg (experiment II) of N-methylnitrosourea 3 times a week for 3 weeks, and had free access to one of 4 drinking fluids: plain water (control group), 17 ppm lycopene water solution (Ly group), and diluted tomato juice containing 17 ppm (Tj group) or 3.4 ppm (tj group) lycopene, throughout the experiments. The colon cancer incidence at week 35 was significantly lower in the Tj group, but not in the Ly group, than in the control group: 21% and 33% vs. 54%, in experiment I (24 rats in each group). It was significantly lower in the Tj group than in the tj and control groups, 40% vs. 72% and 84%, in experiment II (25 rats in each group). An appreciable amount of lycopene (0.02 microgram/g) was detected in the colon mucosa of rats in the Tj group, but not in the tj group. The results suggest that tomato juice rich in lycopene may have a protective effect against colon carcinogenesis.

Animals↗

[Nutrition strategies for postoperative infectious complications].

A considerable amount of data suggests that postoperative infectious complications result from malnutrition, organ impairment, and metabolic disorders. Since metabolism may become deranged once a complication occurs, appropriate pre- and postoperative nutritional support is very important for preventing postoperative infections. For patients who experience postoperative infections such as peritonitis, pyothorax, mediastinitis, or pneumonia, a special feeding formula for the metabolic derangement observed in sepsis and organ impairment should be administered. Branched-chain amino acids (BCAAs) are considered to reduce protein catabolism during the course of a septic insult. A BCAA-enriched parenteral nutrition formula is preferable for patients who cannot receive enteral feeding. Enteral nutrition should always be given the first priority in patients with a functional intestinal tract who are unable to consume adequate calories orally. Enteral formulas can include special nutrients such as dietary fiber and glutamine-analog which exert a trophic effect on the gut mucosa or enhance immunocompetence.

Humans↗

Transcriptional activation of H+/K+-ATPase genes by gastric GATA binding proteins.

H+/K+-ATPase (composed of alpha and beta subunits) and histamine H2 receptor are specifically expressed in gastric parietal cells. The GATA binding proteins (GATA-GT1 and GATA-GT2, also called GATA-6 and GATA-4, respectively) originally found in the gastric mucosa recognized a sequence motif [gastric motif, (G/C)PuPu(G/C)NGAT(A/T)PuPy] in the upstream regions of the ATPase genes [Tamura, S., Wang, X.-H., Maeda, M., and Futai, M. (1993) Proc. Natl. Acad. Sci. USA 90, 10876-10880]. These proteins activated the transcription of the reporter gene ligated downstream of the control region of the rat ATPase alpha or beta subunit gene but had no effect on the same reporter ligated downstream of the H2 receptor gene. Deletion analyses suggested that the upstream 249 (alpha gene) and 323 (beta gene) base pair sequences from the first letter of the initiation codon are sufficient for activation by the GATA proteins. Interestingly, two and three gastric motifs are located near the TATA-boxes of the alpha and beta genes, respectively. Mutagenesis studies demonstrated that the two motifs proximal to the TATA-box sequences of the ATPase alpha and beta subunit genes were essential for the activation. These results suggest that both the alpha and beta subunit genes are regulated similarly by the GATA binding proteins. The expression system established in this study is a useful system for analyzing the roles of GATA proteins in transcriptional regulation of the H+/K+-ATPase gene.

Animals↗

Identification of new chalcone synthase genes for flower pigmentation in the Japanese and common morning glories.

New cDNA sequences for chalcone synthase (CHS), a key enzyme in the flavonoid biosynthesis, were obtained from the Japanese and common morning glories; they are more closely related to other CHS sequences than the six previously described CHS genes from the same plants. The newly isolated CHS-D gene is abundantly expressed in the pigmented flower buds, while its expression is drastically reduced in the white flower buds. Thus CHS-D appears to produce major CHS transcripts for flower pigmentation.

Acyltransferases↗

Effect of dietary fiber on bowel mucosal integrity and bacterial translocation in burned rats.

The response of the bowel mucosa to enteral formula supplemented with dietary fiber was examined in rats with 30% full-thickness burns. The rats were fed a standard enteral formula without fiber or with one of two types of fiber (insoluble soy fiber or soluble guar gum fiber). Seventy-two hours after burn injury, the mesenteric lymph nodes were excised aseptically for bacterial culturing. Samples of the jejunum, ileum and cecum were also collected for histological examination. There were significantly fewer bacterial colonies in the lymph node cultures from rats given soy fiber compared to those from rats fed no fiber. In rats given soy fiber, the integrity of the bowel mucosa was maintained in the jejunum, ileum and cecum. In rats given guar gum fiber, however, the repair of mucosal erosions was observed in the jejunum and ileum as well as flattening of the cecal mucosa. These findings indicate that soy fiber is superior to guar gum fiber for maintaining bowel mucosal integrity and preventing bacterial translocation in burned rats receiving enteral feeding.

Animals↗