Booster effect of canine distemper, canine parvovirus infection and infectious canine hepatitis combination vaccine in domesticated adult dogs

Domesticated adult dogs with antibody titer classified as below 'high' to one or more of canine distemper virus (CDV), canine parvovirus type-2 (CPV-2) and canine adenovirus type-1 (CAdV-1) were then given an additional inoculation, and the effectiveness of this booster evaluated 2 months later. Consequently, CDV and CAdV-1 antibody titer experienced a significant increase, but the same effect was not observed in the antibody titer of CPV-2. These findings suggest that with additional inoculation, a booster effect may be expected in increasing antibody titers for CDV and CAdV-1, but it is unlikely to give an increase in CPV-2 antibody titer.     

Effects of amino acid alterations on the transglycosylation reaction of endoglucanase I from Trichoderma viride HK-75

Endoglucanase I (EGI) from Trichoderma viride HK-75 catalyzes not only hydrolysis but also transglycosylation reactions of cellooligosaccharides. In order to characterize the important amino acid residues in transglycosylation of EGI, three Tyr, one Leu, and two Glu residues of EGI were replaced by Trp or Asp. The seven resulting EGI, except for L200W, had reduced activities toward carboxymethyl-cellulose compared to that of wild type EGI. The results from the mutations in the catalytic residues of E196 and E201 indicate that the space just around the catalytic residues is not directly related to the transglycosylation reactions of EGI. Analyses of the enzymes with mutations in the substrate-binding residues showed that Y146, Y170, and L200 of EGI are closely involved in the mode of transglycosylation and that several amino acid residues within the active site are involved in the transglycosylation reaction of EGI.     

Proteomic analysis of bone proteins adsorbed onto the surface of titanium dioxide

Osseointegration is the structural and functional connection between bone tissues and implants such as titanium dioxide (TiO₂). The bone-TiO₂ interface is thought to contain proteoglycans. However, exhaustive analysis of the proteins in this layer has not been performed. In this study, we evaluated the bone protein adhered on the surface of TiO₂ comprehensively. Pig bone protein was extracted by sequential elutions with guanidine, 0.1 M EDTA, and again with guanidine. The proteins obtained from these extractions were allowed to adhere to an HPLC column packed with TiO₂ and were eluted with 0.2 M NaOH. The eluted proteins were identified by LC/MS/MS and included not only proteoglycans but also other proteins such as extracellular matrix proteins, enzymes, and growth factors. Calcium depositions were observed on TiO₂ particles incubated with bone proteins, guanidine-extracted proteins adhered to TiO₂ displayed significantly high amounts of calcium depositions.     

Mutation of the plastidial alpha-glucan phosphorylase gene in rice affects the synthesis and structure of starch in the endosperm

Plastidial phosphorylase (Pho1) accounts for approximately 96% of the total phosphorylase activity in developing rice (Oryza sativa) seeds. From mutant stocks induced by N-methyl-N-nitrosourea treatment, we identified plants with mutations in the Pho1 gene that are deficient in Pho1. Strikingly, the size of mature seeds and the starch content in these mutants showed considerable variation, ranging from shrunken to pseudonormal. The loss of Pho1 caused smaller starch granules to accumulate and modified the amylopectin structure. Variation in the morphological and biochemical phenotype of individual seeds was common to all 15 pho1-independent homozygous mutant lines studied, indicating that this phenotype was caused solely by the genetic defect. The phenotype of the pho1 mutation was temperature dependent. While the mutant plants grown at 30 degrees C produced mainly plump seeds at maturity, most of the seeds from plants grown at 20 degrees C were shrunken, with a significant proportion showing severe reduction in starch accumulation. These results strongly suggest that Pho1 plays a crucial role in starch biosynthesis in rice endosperm at low temperatures and that one or more other factors can complement the function of Pho1 at high temperatures.     

Optimization of wastewater feeding for single-cell protein production in an anaerobic wastewater treatment process utilizing purple non-sulfur bacteria in mixed culture condition

Impacts of operation timing of feeding and withdrawal on anaerobic wastewater treatment utilizing purple non-sulfur bacteria have been investigated in mixed culture condition with acidogenic bacteria. Simulated wastewater containing glucose was treated in a laboratory-scale chemostat reactor, changing the timing of wastewater feeding and withdrawal. Rhodopseudomonas palustris, which does not utilize glucose as a substrate, was inoculated in the reactor. Rps. palustris was detected by a fluorescent in situ hybridization (FISH) technique using the specific Rpal686 probe. As a result, population ratios of Rps. palustris were over 20% through the operation. Rps. palustris could grow by utilizing metabolites of acidogenic bacteria that coexisted in the reactor. A morning feed was effective for a good growth of purple non-sulfur bacteria. A protein content of cultured bacteria was the highest when wastewater was fed in the morning. Dissolved organic carbon (DOC) removal was 94% independent of the timing control. Consequently, feeding in the morning is the optimum feed-timing control from the aspects of growth of purple non-sulfur bacteria and single-cell protein production.     

Subcellular Localization of PMES-2 Proteins Regulated by Their two Cytoskeleton-Associated Domains

1. PMES-2 is a protein, of which mRNA is translocated to the neurites of hippocampal neurons. Since the protein is present in the postsynaptic density, contributions to synaptic function have been predicted. 2. To elucidate the protein-protein interaction of PMES-2, yeast two-hybrid screening was performed with PMES-2 partial polypeptides as baits. We found that PMES-2 interacted with dynein light chain-2 (DLC-2), a light chain subunit of myosin-V and cytoplasmic dynein, via the C-terminal 20 amino acids. Exogenous PMES-2 colocalized with F-actin at the cell periphery, while a PMES-2 mutant lacking the DLC-binding site localized primarily in the nucleus. 3. This dual-targeting of PMES-2 constructs depends on an effector domain-like motif in the N-terminus. 4. These results indicate that PMES-2 links a motor complex to the membrane skeleton and that DLC-1/2 inhibits PMES-2 nuclear localization. PMES-2 possibly modifies the cytoskeletal architecture and protein transport at the synapse and/or regulates signal transduction from the synapse to the nucleus.     

Uncoupling of longevity and paraquat resistance in mutants of the nematode Caenorhabditis elegans

To analyze the relationship between resistance to oxidative stress and longevity, we isolated three novel paraquat-resistant mutants, mev-5, mev-6, and mev-7, from the nematode Caenorhabditis elegans. They all showed the Dyf (defective in dye filling) phenotype, but not always resistance to heat or UV. Life-span extension was observed only in the mev-5 mutant at 26 degrees C. These results indicate that longevity is uncoupled with the phenotype of paraquat resistance.     

Allene oxide cyclase is essential for theobroxide-induced jasmonic acid biosynthesis in Pharbitis nil

Theobroxide, a natural product, strongly stimulates the biosynthesis of jasmonic acid (JA) in Pharbitis nil. In this study, we investigated the accumulation of protein by the immunoblot analysis of lipoxygenase (LOX), allene oxide synthase (AOS), and allene oxide cyclase (AOC), key enzymes in JA biosynthesis, and how the endogenous levels of JA in P. nil are affected by theobroxide. The effect of JA on the accumulations of these proteins was monitored simultaneously. The results show that theobroxide treatment led to a high level accumulation of JA, which is due to high accumulations of LOX, AOS, and AOC proteins induced by theobroxide treatment both under short day (SD) and long day (LD) conditions. However, under SD conditions AOS and AOC proteins are not enhanced by JA treatment. Kinetic analysis of protein levels shows that a biphasic activation of AOC protein by theobroxide is displayed and the first activation of AOC protein together with elevated JA levels is observed within 30min after treatment. Meanwhile, AOS and LOX proteins are activated by theobroxide later than AOC protein, suggesting that AOC plays an essential role in the initial JA formation induced by theobroxide. Since theobroxide-increased JA levels also show a biphasic manner similar to AOC activation and AOS, LOX proteins are activated later than AOC, and thus we propose a positive JA feedback regulation. Interestingly, AOS protein, which is also the enzyme for the biosynthesis of 9,10-ketol-octadecadienoic acid (KODA, a flowering inducing factor), accumulates markedly due to the simultaneous involvement of theobroxide and SD conditions, suggesting that AOS probably plays a role in flower bud formation in P. nil.