Molecular characterization of coral sulfate transporter homolog that is up-regulated by the presence of symbiotic algae

Reef-building corals are in symbiosis with the dinoflagellates Symbiodinium spp. In our previous study, the expression of two mRNAs (AtSym-01 and 02) was up-regulated by the presence of Symbiodinium cells (strain PL-TS-1) in juveniles of the reef-building coral Acropora tenuis. In this study, the AtSym-01 mRNA was found to encode a cnidarian ortholog of the vertebrate SLC26A11 sulfate transporter. The AtSym-01 and human SLC26A11 proteins exhibited 46% identity over 542 amino acids. Real-time polymerase chain reaction analysis showed that the expression level of the AtSym-01 mRNA was also increased by the presence of Symbiodinium strain CCMP2467 cells. Immunohistochemical analysis was performed using polyclonal antibodies against the AtSym-01 protein, in order to study the distribution of the protein in A. tenuis tissues. Cell-specific immunoreaction was observed in diverse tissues in juvenile and adult specimens. Notable immunoreaction was observed in mucocytes (mucus cells) in the outer epithelium of juveniles, and gastrodermal cells located between the coelenteron and skeleton of the adult colony. These observations suggest the possibility that the AtSym-01 protein is involved in uptake of sulfate ion for synthesis of sulfated macromolecules that are contained in the mucus and organic matrix of the calcified skeleton.     

A plant vacuolar protease, VPE, mediates virus-induced hypersensitive cell death

Programmed cell death (PCD) in animals depends on caspase protease activity. Plants also exhibit PCD, for example as a response to pathogens, although a plant caspase remains elusive. Here we show that vacuolar processing enzyme (VPE) is a protease essential for a virus-induced hypersensitive response that involves PCD. VPE deficiency prevented virus-induced hypersensitive cell death in tobacco plants. VPE is structurally unrelated to caspases, although VPE has a caspase-1 activity. Thus, plants have evolved a regulated cellular suicide strategy that, unlike PCD of animals, is mediated by VPE and the cellular vacuole.     

An Improved Method for Isolating Violet Root Rot Fungus, Helicobasidium mompa, from Basidiocarps

Basidiocarps of the violet root rot fungus, Helicobasidium mompa, are less frequently used for isolation than are mycelial strands and sclerotia even though the basidiocarps are conspicuously produced at the trunk base of diseased plants. Basidiocarps are also more suitable for storage. This paper describes an improved method for obtaining pure cultures from basidiocarps using microcentrifuge tubes to facilitate the awkward steps of rinsing fungal materials under a dissecting microscope. Received 28 June 2000/ Accepted in revised form 4 August 2000     

PCR detection of Porcine circovirus type 2 DNA in whole blood,serum, oropharyngeal swab,nasal swab,and feces from experimentally infected pigs and field cases

Porcine circovirus type 2 (PCV2) shedding patterns were investigated by polymerase chain reaction (PCR) for the detection of PCV2 DNA, and the diagnostic suitability of a sample for the PCR was examined by using different types of samples. In the experimental infection, sixteen pigs were inoculated intranasally with PCV2. The samples, including oropharyngeal and nasal swabs, feces, whole blood and serum became positive for PCV2 DNA by PCR immediately after the inoculation, and almost all samples remained positive during the observation period, post-inoculation-day 70. Field samples were collected from 313 pigs in five different age groups. The overall percentages of positive samples in the whole blood, nasal swabs, and feces detected by PCR were 30.4%, 19.2%, and 20.4%, respectively. The frequency of positive samples increased after the nursery stages and reached a peak in the 3 to 4-month-old pigs. These results indicate that PCV2 infection may occur after weaning, that PCV2 DNA may be present in whole blood for a long period after infection, and that whole blood and serum are the most suitable sample types for the PCR analysis of PCV2.     

Assignment of the 1H and 13C NMR spectra of 2-aminobenzamide-labeled galacto- and arabinooligosaccharides

1,4-Linked β-d-galactooligosaccharides with a degree of polymerization (DP) between 1 and 7 and 1,5-linked α-l-arabinooligosaccharides with a DP between 1 and 8 were labeled at their reducing ends with 2-aminobenzamide (2AB) in the presence of sodium cyanoborohydride. The 2AB-labeled oligosaccharides were shown to be homogeneous using high-performance anion-exchange chromatography (HPAEC) and by electrospray ionization mass spectrometry (ESI-MS). The signals in the ¹H and ¹³C nuclear magnetic resonance (NMR) spectra of the 2AB-labeled oligosaccharides were then assigned using one- and two-dimensional NMR spectroscopy. These NMR data will be useful for the structural analysis of enzymatically synthesized galactan and arabinan side chains derived from rhamnogalacturonan I.     

Prediction of bending stiffness and moment carrying capacity of sugi cross-laminated timber

Cross-laminated timber (CLT) panels consist of several layers of lumber stacked crosswise and glued together on their faces. Prototype sugi CLT floor panels were manufactured and bending tests were carried out under the different parameters of lumber modulus of elasticity (MOE), number of layers, thickness of lumber and thickness of CLT panels. On the basis of above tests, bending stiffness and moment carrying capacity were predicted by Monte Carlo method. MOE of lumber was measured by using grading machine and tensile strength of lumber was assumed to be 60 % of bending strength based on the obtained bending test. Bending stiffness EI of CLT panels could be estimated by adopting composite theory and equivalent section area. Experimental moment carrying capacity showed 12 % higher value than the calculated moment carrying capacity by average lumber failure method, and also showed 45 % higher value than the calculated moment carrying capacity by minimum lumber failure method due to the reinforcement of the outer layer by the neighboring cross layer.     

Protection from pseudorabies virus challenge in mice by a combination of purified gII, gIII and gVI antigens.

The antigens gII, gIII and gVI were purified from the lysates of the pseudorabies virus (PRV) -infected HmLu-1 cells using Sepharose 4 B coupled with MAbs against these antigens. Mice immunized with either gII, gIII, gVI antigen or a mixture of them were challenged intraperitoneally with 8.5 x 10(3) plaque forming units of PRV. All the mice immunized with 1.5 and 4.5 micrograms of the mixture and 4.5 micrograms of the gIII antigen survived. The sera of mice immunized with the mixture had virus neutralizing activity which was independent of complement, hemagglutination inhibition activity, and recognized major (93 kilodaltons) and minor (129, 74, 68 and 50 kilodaltons) PRV proteins under reducing conditions in western blotting. The serological activity levels in the lower survival group were not different from those in the complete survival. These results indicate that the mixture of each glycoprotein is more effective for eliciting of protective immunities in mice and that serological activity do not always correlate with protection.