Phenotypic analysis of hepatic T lymphocytes in a dog with chronic hepatitis

We evaluated hepatic T lymphocyte phenotypes in a dog with chronic hepatitis. Before treatment, numerous CD3+ lymphocytes were demonstrated in the liver, and the ratio of CD4+/CD8+ was remarkably high (2.96; reference range, 0.33+/-0.12). After treatment, CD3+ lymphocyte infiltration in the liver was reduced, and the ratio of CD4+/CD8+ decreased to 0.31. Therefore, hepatic T lymphocytes, especially CD4+ lymphocytes, might play a central role in the pathogenesis of this dog with chronic hepatitis.     

Suppression of Ralstonia solanacearum bacterial wilt disease by an organic hydroponic system

An organic hydroponic system that we developed has potential to control root diseases including bacterial wilt of hydroponically grown tomato. In inoculation tests with Ralstonia solanacearum during tomato plant cultivation in conventional inorganic hydroponics and in our organic system, many of the tomato seedlings in the conventional system wilted and died, but none of the seedlings in the organic hydroponics wilted or developed any symptoms, suggesting that the organic system can suppress this bacterial wilt disease. Interestingly, a rhizosphere biofilm, formed only on roots in the organic hydroponic system, may be responsible for the suppression of the bacterial wilt.     

Matrix metalloproteinases (MMPs) activity in cultured canine bone marrow stromal cells (BMSCs)

Autologous bone marrow stromal cells (BMSCs) infusion therapy improves the hepatic fibrosis. To investigate the mechanism of remission, we evaluated the matrix metalloproteinase (MMP)-2 and -9 activity in canine BMSCs and the effect of pro-inflammatory cytokines on their expression. The activity and the gene expression of MMPs were analyzed by gelatin zymography and quantitative RT-PCR, respectively. The specific gelatinase bands were indicative effect of MMP-2 and -9 in canine BMSCs. MMP-2 expression seemed to be increased by TNF-α and IL-1β while MMP-9 was enhanced by TNF-α and IL-6. These results suggested that remissive effect on liver fibrosis might be partly attributable to the MMP-2 and -9 activity in BMSCs under the inflammatory condition.     

Genomic gene encoding manganese peroxidase from a white-rot fungus Phanerochaete crassa WD1694

The gene encoding manganese peroxidase of a white-rot fungus Phanerochaete crassa WD1694 was cloned and sequenced. Four genomic clones were sequenced in which 3 clones were existed as alleles. The analysis of intron–exon structures divided the 4 clones into three subfamilies that corresponded to mnp2 and mnp3 of Phanerochaete chrysosporium, and a new subfamily possessing only five introns. The purified P. crassa WD1694 MnP consisted of 4 isozymes with same molecular weight, same N-terminal sequence, and different pI. N-terminal sequence of deduced protein of P. crassa mnpB3 gene was identical to those of 4 MnP isozymes; however, the other 3 mnp genes had different N-terminal sequence. The molecular weight of encoded mature protein of mnpB3 gene and purified MnP had a gap that could be difference between MnP proteins encoded by single gene. The results suggested that 4 MnP isozymes of P. crassa WD1694 arose from single gene.     

Artificial neural network modeling for predicting final moisture content of individual Sugi (Cryptomeria japonica) samples during air-drying

Sugi (Cryptomeria japonica D. Don) lumber is known to have a large variability in final moisture content (MC_f) and is difficult to dry. This study assessed the capability of artificial neural networks (ANNs) to predict the MC_f of individual wood samples. An ANN model was developed based on initial moisture content, basic density, annual ring orientation, annual ring width, heartwood ratio and lightness (L ^* in the CIE L ^* a ^* b ^* system). The performance of the ANN model was compared with a principal component regression (PCR) model. The ANN model showed good agreement with the experimentally measured MC_f with a higher correlation coefficient (r) and a lower root mean square error (RMSE) than the PCR model, demonstrating the importance of nonlinearity of the variables and the higher capability of the ANN model than the PCR model. By adding redness (a ^* ) and yellowness (b ^* ) and drying time to the input variables of ANNs, r and RMSE values were improved to 0.98 and 1.2 % for the training data set, and 0.85 and 2.2 % for the testing data set, respectively. Although the developed ANNs are available under the limited conditions of this study, our results suggest that the ANNs proposed offer reliable models and powerful prediction capability for the MC_f, even though wood properties vary considerably and their complex interrelations are not fully elucidated.     

Rapid discrimination of two cryptic species within Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae) by PCR–RFLP

Brontispa longissima (Coleoptera: Chrysomelidae) is a serious invasive pest of coconut palm (Cocos nucifera) supposedly originating from Indonesia and New Guinea. It has recently invaded Southeast and East Asia, where it has caused serious damage to coconut plants. Brontispa longissima as currently defined contains two cryptic species: we herein referred to one as the “Asian clade”, which is distributed over a wide area, including Asia and the Pacific region; and we referred to the other one as the “Pacific clade”, which is found in a limited area in the Pacific region. We developed a PCR–RFLP method for differentiating the two clades. Digestion of the PCR product of a 1,014-bp region within the mitochondrial cytochrome oxidase I gene (COI) with BslI, HpyCH4III, or NlaIV resulted in clade-specific patterns as estimated by the sequence data. We applied the method to specimens newly obtained from various locations to investigate the geographical distribution of B. longissima. Although B. longissima collected from Samoa in April 2003 had been placed in the Pacific clade, specimens collected from the same island in April 2010 were placed in the Asian clade, suggesting that the predominant clade may have been changing from the former to the latter. On Timor, specimens included both clades in apparently segregated habitats.     

Comparison of cholesterol levels among lipoprotein fractions separated by anion-exchange high-performance liquid chromatography in periparturient Holstein–Friesian dairy cows

Changes in lipoprotein profiles occur in dairy cows during the periparturient period and in cows with transition cow disease. Here, the lipoprotein profiles of Holstein–Friesian dairy cows during the periparturient period were obtained by anion-exchange, high-performance liquid chromatography to evaluate the usefulness of lipoprotein profile evaluation during the periparturient period and in cows with fatty liver and milk fever. Lipoprotein levels (including total and high- (HDL-C) and low-density lipoprotein (LDL-C) cholesterol) in 10 healthy cows were low 4 weeks prepartum, with the lowest values at calving or within 1 week of calving; the values increased at 8 weeks postpartum. The lipoprotein levels were measured in 16 cows diagnosed with fatty liver (n=10) or milk fever (n=6) and compared to 10 healthy dairy cows. A significant difference was observed in HDL-C between healthy cows (at calving and 1 week postpartum), and the fatty liver and milk fever cows. Cows with fatty liver and milk fever had a lower mean HDL-C than the 10 healthy dairy cows at calving and 1 week postpartum. HDL-C might be a good indicator of energy balance for differentiating healthy cows from those with transition cow disease.     

Comparative aspects on the role of polypyrimidine tract-binding protein in internal initiation of hepatitis C virus and picornavirus RNAs

We compared the effects of polypyrimidine tract-binding protein (PTB) on hepatitis C virus (HCV genotype IIa), encephalomyocarditis virus (EMCV) and poliovirus internal ribosome entry site (IRES) activities in vitro. It bound strongly to EMCV IRES, but weakly to PV and HCV RNAs. PV IRES showed the strongest dependency to PTB and it showed less than one-tenth of IRES activity after the immuno-depletion of PTB from HeLa S10 lysate with pre-coated anti-PTB IgG beads, comparing to the normal IgG beads-treated S10 lysate. EMCV IRES activity was approximately 40% of that of normal control after PTB depletion. Especially, HCV IRES activity was approximately 95%, and most weekly affected by the depletion of PTB. Repletion of PTB to depleted S10 lysate restored activities of PV and EMCV IRESs. The data suggest that PTB plays an important role in picornaviral IRESs, but not in HCV IRES.     

Nutritional evaluation of rotifers in rearing tanks without water exchange during seed production of amberjack <Emphasis Type="Italic">Seriola dumerili</Emphasis>

We evaluated the nutrient contents of rotifers sampled from larval-rearing tanks (tank rotifers) without water exchange during the seed production of amberjack Seriola dumerili at three facilities (Kamiura, Kagoshima, and Miyazaki) and compared them with the nutrient contents of freshly enriched rotifers. Compared to the enriched rotifers, the lipid contents, especially neutral lipids and proportion of 22:6n-3, tended to decrease in the tank rotifers. These trends were clearer at Miyazaki where the tank rotifers were sampled before daily supplementation of microalgae (Nannochloropsis). Crude protein content of the tank rotifers did not decrease markedly although the proportion of lysine tended to decrease. Vitamin C and E contents of the tank rotifers decreased significantly only at Miyazaki. Calcium content of the tank rotifers increased at Kamiura and Miyazaki, and the increases in iron and manganese contents of the tank rotifers at Miyazaki and zinc content at Kagoshima were pronounced. These results suggest that the nutritional value of rotifers in larval-rearing tanks without water exchange can be maintained by appropriate supplementation of microalgae. The effect of certain minerals that became high in tank rotifers on subsequent larval development requires further investigation.     

Synthesis and Structure–Activity Relationships of Miticidal 4,5-Dihydropyrazole-5-thiones

A series of novel 4,5-dihydropyrazole-5-thiones (DHPs) was synthesised by treating the corresponding dihydropyrazolones with ‘Lawesson’s reagent and evaluated for miticidal activity against two-spotted spider mites (Tetranychus urticae Koch). Of these, 3-(4-chlorophenyl)-4,4-dimethyl-1-phenyl-4,5-dihydropyrazole-5-thione, 3-(4-chlorophenyl)-4-ethyl-4-methyl-1-phenyl-4,5-dihydropyrazole-5-thione, 3-(4-chlorophenyl)-1-phenyl-4,5-dihydropyrazole-5-thione-4-spirocyclopentane and 4,4-dimethyl-1-phenyl-3-(4-trifluoromethyl-phenyl)-4,5-dihydropyrazole-5-thione were highly active (pEC₅₀>4·0) and were more effective than the miticide dicofol (pEC₅₀=3·879), which has traditionally been used for the control of phytophagous mites. Structure–activity relationship (SAR) studies were performed on each position of the pyrazole ring of DHPs. The results indicated that the unsubstituted phenyl, 4-substituted phenyl and thioxo groups on the 1-, 3- and 5-positions of DHPs respectively were required for activity. Quantitative SAR studies using physicochemical parameters of substituents and the capacity factor k′ as a hydrophobicity index suggested that: (a) the activities of all types of DHPs examined were mainly dominated by hydrophobicity, (b) the bulkiness of 4-substituents of the 3-phenyl ring favoured the activity and (c) the log k′ optimum for all DHPs was 1·675, equivalent to a log P_ow value of c. 5·0.