The usefulness of anti-fucosylated antigen antibody YB-2 for diagnosis of hepatocellular carcinoma

Levels of fucosylated antigens in sera from patients with liver diseases were examined by a newly developed sandwich-type enzyme immuno assay with the aid of anti-fucosylated antigen antibody, YB-2 which reacts simultaneously with Y, Leb and H type 2 antigens. When the cut-off value was set arbitrarily at mean [3 SD values of normal, 30 (69.8%) of the 43 patients with HCC, 14 (53.8%) of the 26 patients with liver cirrhosis (LC) and 24 (45.3%) of the 53 patients with chronic hepatitis (CH) were found to be positive, whereas all of the 30 samples from healthy controls were negative. The levels of -fetoprotein (AFP) and protein induced by vitamin K absence or antagonist-II (PIVKA-II) in HCC were not correlated with those of YB-2 antigens. The positive rates of the combination YB-2 and AFP assay and YB-2 and PIVKA-II assay in HCC were significantly higher (83.7 and 86.0%, respectively) than that of the AFP and PIVKA-II combination (65.1%) which had been reported to be the best combination up to this time.     

Frontal cortical oxygenation changes during gravity-induced loss of consciousness in humans: a near-infrared spatially resolved spectroscopic study.

Gravity (G)-induced loss of consciousness (G-LOC), which is presumably caused by a reduction of cerebral blood flow resulting in a decreased oxygen supply to the brain, is a major threat to pilots of high-performance fighter aircraft. The application of cerebral near-infrared spectroscopy (NIRS) to monitor gravity-induced cerebral oxygenation debt has generated concern over potential sources of extracranial contamination. The recently developed NIR spatially resolved spectroscopy (SRS-NIRS) has been confirmed to provide frontal cortical tissue hemoglobin saturation [tissue oxygenation index (TOI)]. In this study, we monitored the TOI and the standard NIRS measured chromophore concentration changes of oxygenated hemoglobin and deoxygenated hemoglobin in 141 healthy male pilots during various levels of +G(z) (head-to-foot inertial forces) exposure to identify the differences between subjects who lose consciousness and those who do not during high +G(z) exposure. Subjects were exposed to seven centrifuge profiles, with +G(z) levels from 4 to 8 G(z) and an onset rate from 0.1 to 6.0 G(z)/s. The SRS-NIRS revealed an approximately 15% decrease in the TOI in G-LOC. The present study also demonstrated the TOI to be a useful variable to evaluate the effect of the anti-G protection system. However, there was no significant difference found between conditions with and without G-LOC in subjects with terminated G exposure. Further studies that elucidate the mechanism(s) behind the wide variety of individual differences may be needed for a method of G-LOC prediction to be effectively realized.     

Production of mouse monoclonal antibody with galactose-extended sugar chain by suspension cultured tobacco BY2 cells expressing human beta(1,4)-galactosyltransferase

Previously, we developed a transgenic tobacco BY2 cell line (GT6) in which glycosylation was modified by expressing human beta(1,4)-galactosyltransferase (betaGalT). In this study, we produced a mouse monoclonal antibody in GT6 cells, and determined the sugar chain structures of plant-produced antibodies. Galactose-extended N-linked glycans comprised 16.7%, and high-mannose-type and complex-type glycans comprised 38.5% and 35.0% of the total number of glycans, respectively. N-linked glycans with the plant-specific sugars beta(1,2)-xylose and alpha(1,3)-fucose comprised 9.8%. The introduction of human betaGalT into suspension cultured tobacco cells resulted in the production of recombinant proteins with galactose-extended glycans and decreased contents of beta(1,2)-xylose and alpha(1,3)-fucose.     

RNA-binding protein hoip accelerates polyQ-induced neurodegeneration in Drosophila

Abnormal polyglutamine (polyQ) expansion in the N-terminal domain of the human androgen receptor (hAR) is known to cause spinobulbar muscular atrophy (SBMA), a hereditary human neurodegenerative disorder. To explore the molecular mechanisms of neurodegeneration in SBMA, we genetically screened modulators of neurodegeneration in a Drosophila SBMA experimental model system. We identified hoip as an accelerator of polyQ-induced neurodegeneration. We found that hoip forms a complex with 18s rRNA together nop56 and nop5 proteins, whose human homologs are known to form a snoRNP complex involved in ribosomal RNA processing. Significantly, the levels of mutant polyQ-hAR were up-regulated in a mutant line overexpressing hoip. Consistently, severe neurodegeneration phenotype (rough eye) was also observed in both nop56 and nop5 overexpression mutant lines. These findings suggest that the process of neurodegeneration induced by abnormal polyQ expansion in the hAR may be regulated by the activity of snoRNP complex.     

Cold-active halophilic bacteria from the ice-sealed Lake Vida,Antarctica

Lake Vida is a large, permanently ice-covered lake in the Victoria Valley of the McMurdo Dry Valleys, Antarctica and is unique among Dry Valley lakes because it is ice-sealed, with an ice-cover of nearly 19 m. Enrichment cultures of melt-water from Lake Vida 15.9 m ice yielded five pure cultures of aerobic, heterotrophic bacteria. Of these, one strain grew at −8°C and the four others at −4°C. All isolates were either halotolerant or halophilic, with two strains capable of growth at 15% NaCl. Phylogenetic analysis revealed the Lake Vida isolates to be Gammaproteobacteria, related to species of Psychrobacter and Marinobacter. This is the first report of pure cultures of bacteria from Lake Vida, and the isolates displayed a phenotype consistent with life in a cold hypersaline environment.     

Wickerhamomyces edaphicus sp. nov. and Pichia jaroonii sp. nov., two ascomycetous yeast species isolated from forest soil in Thailand

Four yeast strains were isolated from soil in a mixed deciduous forest in Amphoe Wang Nam Khiao, Nakhon Ratchasima province, Thailand. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics, and the sequence analyses of the D1/D2 domain of the large-subunit (LSU) rRNA gene, small-subunit rRNA gene and internal transcribed spacer (ITS) region, the three strains (S-29, S-63 and S-80) were found to represent a single species of the genus Wickerhamomyces , which were named Wickerhamomyces edaphicus sp. nov. The type strain is S-29^T (BCC 21231^T=NBRC 101969^T=CBS 10408^T). Strain S-75 represented a novel species of the genus Pichia on the basis of morphological, biochemical, physiological and chemotaxonomic characteristics, and the sequence analyses of the D1/D2 domain of the LSU rRNA gene, for which the name Pichia jaroonii sp. nov. is proposed. The type strain is S-75^T (BCC 23061^T, NRBC 102180^T=CBS 10930^T).     

Cloning of Lactobacillus plantarum IAM 12477 lysine biosynthetic genes encoding functional aspartate semialdehyde dehydrogenase, dihydrodipicolinate synthase, and dihydrodipicolinate reductase

Summary The lysine biosynthetic genes asd, dapA, and dapB, encoding aspartate semialdehyde dehydrogenase (ASADH), dihydrodipicolinate synthase (DHPS), and dihydrodipicolinate reductase (DHPR), respectively, have been cloned from Lactobacillus plantarum IAM 12477 by heterologous complementation to Escherichia coli mutants. The amino acid sequences of the cloned genes showed considerable similarities to the corresponding protein from other gram-positive bacteria. We identified the amino acids that correspond to key catalytic residues of ASADH, DHPS, and DHPR and found them to be conserved in the protein from L. plantarum. ASADH, DHPS, and DHPR activity was detected in the cell extracts of E. coli mutant harboring each gene, indicating that the cloned genes were functionally expressed in E. coli. The regulation of ASADH, DHPS, and DHPR were studied in the cell extracts of both the E.␣coli mutant harboring the gene and L. plantarum; however, those enzymes were found not to be regulated by the end products of the pathway. This paper represents a portion of the thesis submitted by M. N. Cahyanto to Osaka University as partial fulfillment of the requirements for the PhD degree.     

Nuphar alkaloids with immediately apoptosis-inducing activity from Nuphar pumilum and their structural requirements for the activity

The methanolic extract and its alkaloid fraction from the rhizomes of Nuphar pumilum showed cytotoxic effects on human leukemia cell (U937), mouse melanoma cell (B16F10), and human fibroblast (HT1080). Dimeric sesquiterpene thioalkaloids with the 6-hydroxyl group (6-hydroxythiobinupharidine, 6,6'-dihydroxythiobinupharidine, 6-hydroxythionuphlutine B) showed substantial cytotoxic activity at a concentration of 10 microM, but dimeric sesquiterpene thioalkaloids lacking the 6-hydroxyl group (thiobinupharidine, thionuphlutine B, 6'-hydroxythionuphlutine B, neothiobinupharidine, thionuphlutine B beta-sulfoxide, and neothiobinupharidine beta-sulfoxide) and monomeric sesquiterpene alkaloids (nupharidine, 7-epideoxynupharidine, and nupharolutine) showed weak activity. Next, apoptosis-inducing activity of a principal active constituent, 6-hydroxythiobinupharidine, on U937 was examined using morphological observation and DNA fragmentation assay (TUNEL method). Apoptosis of U937 was immediately observed within 1 h after treatment of 6-hydroxythiobinupharidine at 2.5-10 microM.