Analysis of differential accumulation of winged bean Kunitz chymotrypsin inhibitor mRNA species by a sequence-specific termination method

Winged bean Kunitz chymotrypsin inhibitor (WCI) is encoded by a multigene family and accumulation of its mRNA is restricted in mid-maturation stage seeds and tuberous roots. In this paper, we analyzed the accumulation of mRNA derived from each WCI gene using a novel method: sequence-specific termination analysis. The results demonstrated that the accumulation of each WCI mRNA was differentially regulated in winged bean plants.     

Interinstitutional patient transfers between rapid chemotherapy cycles were feasible to utilize proton beam therapy for pediatric Ewing sarcoma family of tumors

Aim

To assess the feasibility of transferring to the University of Tsukuba Hospital for proton beam therapy (PBT) during intensive chemotherapy in children with Ewing sarcoma family of tumors (ESFT) who had been diagnosed and started their first-line treatment at prefectural or regional centers for pediatric oncology.

PAR‐3 controls endothelial planar polarity and vascular inflammation under laminar flow

Impaired cell polarity is a hallmark of diseased tissue. In the cardiovascular system, laminar blood flow induces endothelial planar cell polarity, represented by elongated cell shape and asymmetric distribution of intracellular organelles along the axis of blood flow. Disrupted endothelial planar polarity is considered to be pro‐inflammatory, suggesting that the establishment of endothelial polarity elicits an anti‐inflammatory response. However, a causative relationship between polarity and inflammatory responses has not been firmly established. Here, we find that a cell polarity protein, PAR‐3, is an essential gatekeeper of GSK3β activity in response to laminar blood flow. We show that flow‐induced spatial distribution of PAR‐3/aPKCλ and aPKCλ/GSK3β complexes controls local GSK3β activity and thereby regulates endothelial planar polarity. The spatial information for GSK3β activation is essential for flow‐dependent polarity to the flow axis, but is not necessary for flow‐induced anti‐inflammatory response. Our results shed light on a novel relationship between endothelial polarity and vascular homeostasis highlighting avenues for novel therapeutic strategies.     

Increased Levels of Monodehydroascorbate Radical in UV-B-Irradiated Broad Bean Leaves

Light-induced monodehydroascorbate (MDA) radical productionwas not detectable by EPR spectroscopy in untreated broad beanleaves, but it was observed after exposing the leaves to UV-B(280–320 nm) irradiation. After this pre-treatment, alow level of MDA radicals was also detectable without illumination.Light-induced MDA, MDA production in the dark, oxygen evolution,quantum yield of PSII as measured by Chi fluorescence, MDA producingand reducing enzyme activities were determined and comparedin leaves after irradiation with various doses of UV-B. Ourresults suggest that UV-B irradiation disturbs the balance ofMDA radical production and reduction, resulting in increasedlight induced MDA signal. The enhancement of ascorbate photooxidationat the UV-B damaged donor side of PSII appears as a major factorin this process. (Received November 22, 1996; Accepted March 25, 1997)     

Duplication of the LDH gene loci by polyploidization in the fish order Clupeiformes

Summary Within the order Clupeiformes, chromosome analysis and DNA measurements have indicated a diploid-tetraploid relationship among closely related species. To confirm the presence of polyploidization at single gene loci we studied the LDH isoenzyme system. The results obtained are in agreement with the hypothesis of polyploidization. While the diploid species show two gene loci for LDH, the tetraploid species exhibit four separate gene loci.In Duarte, supported in part by CA-05138 from the National Cancer Institute, U.S. Public Health Service.     

Enzyme enhancement activity of N-octyl-beta-valienamine on beta-glucosidase mutants associated with Gaucher disease

Gaucher disease (GD), caused by a defect of beta-glucosidase (beta-Glu), is the most common form of sphingolipidosis. We have previously shown that a carbohydrate mimic N-octyl-beta-valienamine (NOV), an inhibitor of beta-Glu, could increase the protein level and enzyme activity of F213I mutant beta-Glu in cultured GD fibroblasts, suggesting that NOV acted as a pharmacological chaperone to accelerate transport and maturation of this mutant enzyme. In the current study, NOV effects were evaluated in GD fibroblasts with various beta-Glu mutations and in COS cells transiently expressing recombinant mutant proteins. In addition to F213I, NOV was effective on N188S, G202R and N370S mutant forms of beta-Glu, whereas it was ineffective on G193W, D409H and L444P mutants. When expressed in COS cells, the mutant proteins as well as the wild-type protein were localized predominantly in the endoplasmic reticulum and were sensitive to Endo-H treatment. NOV did not alter this localization or Endo-H sensitivity, suggesting that it acted in the endoplasmic reticulum. Profiling of N-alkyl-beta-valienamines with various lengths of the acyl chain showed that N-dodecyl-beta-valienamine was as effective as NOV. These results suggest a potential therapeutic value of NOV and related compounds for GD with a broad range of beta-Glu mutations.