Metabolism of flupyrazofos in the isolated perfused rat liver

To investigate the hepatic metabolism of the new insecticide flupyrazofos [O,O-diethyl O-(1-phenyl-3-trifluoromethylpyrazol-5-yl) phosphorothioate], isolated rat liver was perfused with flupyrazofos under single-pass conditions. In outflow perfusate and bile, 1-phenyl-3-trifluoromethyl-5-hydroxyprazole (PTMHP), PTMHP-sulfate and PTMHP-glucuronide conjugates were identified as the metabolites of flupyrazofos. However, O,O-diethyl O-(1-phenyl-3-trifluoromethylpyrazol-5-yl) phosphate (flupyrazofos oxon) was not detected. A HPLC method with UV detection was used to investigate the hepatic disposition of flupyrazofos and its metabolite PTMHP. The concentrations of flupyrazofos, PTMHP and PTMHP conjugates in outflow perfusate reached steady-state levels within 20 min after commencing perfusion of 7.3 microM flupyrazofos. At steady state, the mean extraction ratio of flupyrazofos was 0.93 (+/- 0.01) and clearance was 26.1 (+/- 0.2) ml min-1 which nearly approached perfusate flow rate (28 ml min-1). PTMHP accounted for 55.7 (+/- 5.8)% of eliminated flupyrazofos and was recovered as unchanged PTMHP, PTMHP-sulfate and PTMHP-glucuronide in the bile as well as the outflow perfusate.     

Sporophytic photosynthesis and gametophytic growth of the kelp Ecklonia stolonifera affected by ocean acidification and warming

Juvenile sporophytes and gametophytes of Ecklonia stolonifera were incubated in combinations of three pCO₂ levels (360, 720 and 980 ppmv) and two temperatures (10 and 15°C for sporophytes; 15 and 20°C for gametophytes) to examine potential effects of climate change on photosynthesis and growth. Sporophytes had significantly higher maximum quantum yields (F_v/F_m) and maximum relative electron transport rates (rETR_max) at 720 ppmv than 360 and 980 ppmv. Also, these parameters were significantly lower at higher temperature of 15°C than at 10°C. Growth of female gametophytes was maximal at 360 ppmv rather than enriched pCO₂ levels. Female gametophytes had significantly lower growth at higher temperature of 20°C than at 15°C. These results indicate effects of elevated pCO₂ varied between generations: stimulating sporophytic photosynthesis and inhibiting gametophytic growth. Ocean acidification and warming would constitute a grave threat to seedling cultivation of E. stolonifera caused by growth inhibition of gametophytes at high pCO₂ levels and temperatures.     

Serologic and genetic characterization of North American H3N2 swine influenza A viruses

The H3N2 subtype of influenza A viruses isolated from pigs in the United States and Canada has shown both genetic and antigenic diversity. The objective of this study was to determine the serologic and genetic characteristics of contemporary strains of these viruses. Genetic analysis of 18 reference strains and 8 selected strains demonstrated differences in 1% to 9% of the nucleotides of the hemagglutinin (HA) gene. Phylogenetic analysis of the HA gene revealed 3 genetic clusters, as well as divergence of cluster III viruses from a cluster III prototype virus (A/Swine/Illinois/21587/99). By means of 1-way cross-hemagglutination inhibition with antiserum against 5 field isolates and 3 vaccine viruses, most of 97 isolates tested could be placed in 1 of 3 serogroups. The several isolates that did not react with any antiserum were in genetic cluster III, which suggests that continuous antigenic drift in cluster III may have resulted in virus variants. The efficacy of commercial vaccines against these virus variants should be evaluated with vaccination and challenge studies.     

Lysine restriction and realimentation affected growth,blood profiles and expression of genes related to protein and fat metabolism in weaned pigs

To investigate the effects of lysine restriction and subsequent realimentation on growth performance, blood profiles and gene expression of leptin and myostatin, 128 weaned pigs [initial body weight (BW) 6.96 ± 1.07 kg, 26 ± 2 days of age] were randomly allotted to four treatments. The starter diets during the first 2 weeks (P1) contained 100%, 80%, 70% or 60% of recommended lysine levels ( National Research Council, 1998 ). Then, common grower 1 and 2 diets were offered for 2 weeks (P2 and P3) each. During P1, average daily gain (ADG) was linearly reduced (p < 0.05) with the increasing levels of lysine restriction. Growth rate was greater in pigs previously fed lysine‐restricted diets than well‐fed pigs although it did not reach a significant level during realimentation. However, the final BW and overall ADG were the lowest (p < 0.05) and F/G was poor in pigs fed 60% lysine diet. Relative visceral organ weights and composition of skeletal muscle were similar (p > 0.05) among the treatment. Blood triglyceride and glucose levels were increased (p < 0.05) during P1, while blood urine nitrogen, total protein and albumin levels were decreased (p < 0.05) during P2 with the reduction in dietary lysine levels. The abundance of myostatin mRNA in skeletal muscle and leptin mRNA in subcutaneous adipose tissue were lower (p < 0.05) in lysine‐restricted pigs than in pigs fed non‐restricted diets. In conclusion, 80% and 70% lysine restriction of starter diets resulted in inferior growth and compensatory growth effect was noted during realimentation, while 60% lysine restriction had a negative influence on growth performance. Moreover, the changes in myostatin and leptin mRNA abundance caused by nutritional manipulations may be involved in the regulation of protein and fat deposition in young pigs.     

Isolation and characterization of porcine circovirus type-2 from sera of stillborn fetuses

In order to examine an association between porcine circovirus type-2 (PCV2) infection and reproductive failure in pigs, sera (n = 171) from stillborn fetuses were collected from 3 different farms with prolonged histories of reproductive problems. These sera were tested for the presence of antibodies to PCV2 using an immunoperoxidase monolayer assay. Of the 171 sera tested, 28 had PCV2 antibody titers of ≥ 1:16. When these 28 samples were tested by a polymerase chain reaction assay, 13 were found to contain PCV2 viral DNA. Of these 13 samples containing both PCV2 antibodies and viral DNA, 9 yielded PCV2 on virus isolation. Amino acid sequences comprising open reading frame 2 of PCV2 from 2 of these isolates were compared to PCV2 isolates from cases of post-weaning multi-systemic wasting syndrome (PMWS). The amino acid sequences of the 2 isolates from stillborn pigs were shown to be nearly identical to each other, as well as to other PCV2 isolates associated with reproductive failure. When compared with PMWS isolates, the isolates from the stillborn fetuses showed differences of at least 2 amino acids. These results confirm previous findings that transplacental infection of PCV2 occurs in the field and that stillbirths in pigs may be associated with PCV2 infections. At present, the significance of minor differences in amino acid sequences is not known.     

Aroma finishing of PET fabrics with PVAc nanoparticles containing lavender oil

A durable aroma finishing for PET fabric was carried out by adopting poly(vinyl acetate) (PVAc) nanoparticles containing lavender oil (LO) in core. Relatively small size of PVAc nanoparticles (ca. 244 nm of mean particle diameter) was expected to resist the frictional destruction of the particles, which is frequently observed in cases of microcapsules. PVAc nanoparticles containing LO in core were prepared by emulsification-diffusion method and their application as an aroma releasing agent for PET fabrics was assessed through the observation of releasing profiles of LO in ethanol for experimental acceleration. Melamine-formaldehyde (MF) microcapsules containing LO were also prepared and treated on fabrics for comparison. PVAc nanoparticles treated on PET fabric showed higher initial releasing amount, which was ascribed to the enhanced surface area. After 2 days of releasing, PET fabric treated with PVAc nanoparticles showed slower and more stable releasing profile and reached about 12 ppm of cumulative release after 16 days, which was under two thirds of that with MF microcapsules. PVAc nanoparticles can be used as an agent for durable aroma finishing of PET fabrics.     

Quantitative CT assessment of bone mineral density in dogs with hyperadrenocorticism

Canine hyperadrenocorticism (HAC) is one of the most common causes of general osteopenia. In this study, quantitative computed tomography (QCT) was used to compare the bone mineral densities (BMD) between 39 normal dogs and 8 dogs with HAC (6 pituitary-dependent hyperadrenocorticism [PDH]; pituitary dependent hyperadrenocorticism, 2 adrenal hyperadrenocorticism [ADH]; adrenal dependent hyperadrenocorticism) diagnosed through hormonal assay. A computed tomogaraphy scan of the 12th thoracic to 7th lumbar vertebra was performed and the region of interest was drawn in each trabecular and cortical bone. Mean Hounsfield unit values were converted to equivalent BMD with bone-density phantom by linear regression analysis. The converted mean trabecular BMDs were significantly lower than those of normal dogs. ADH dogs showed significantly lower BMDs at cortical bone than normal dogs. Mean trabecular BMDs of dogs with PDH using QCT were significantly lower than those of normal dogs, and both mean trabecular and cortical BMDs in dogs with ADH were significantly lower than those of normal dogs. Taken together, these findings indicate that QCT is useful to assess BMD in dogs with HAC.     

Growth hormone attenuates branchial HSP70 expression in silver sea bream

In this study the effects of growth hormone (GH) on silver sea bream branchial heat-shock protein 70 (HSP70) expression was investigated using in-vivo and in-vitro experiments. For in-vivo experiments, sea bream were administered recombinant bream GH or the GH secretagogue hexarelin. Pituitary levels of GH were unchanged in fish administered exogenous GH but decreased on hexarelin administration, in comparison with saline controls. Levels of HSP70 were measured using immunoanalysis and it was found that both GH and hexarelin administration caused a significant decrease in branchial HSP70 abundance. For in-vitro analysis, branchial filaments were exposed to a range of GH concentrations (1, 10, and 100 ng/ml) and it was found that HSP70 levels were significantly lowered in all cases. This study adds to the growing body of evidence surrounding the importance of hormones in regulating heat-shock protein expression in fish.     

Antiviral activity of Alpinia katsumadai extracts against rotaviruses

In vitro anti-rotavirus activity of Alpinia katsumadai (AK) extracts were evaluated against bovine G8P[7] and porcine G5P[7] rotaviruses in two different assay strategies, a mixed treatment assay and a post treatment assay. In the mixed treatment assay, six AK extracts [AK-1 (EtOH extract), AK-3 (H(2)O layer), AK-5 (40% methanol fraction), and AK-9-11 (H(2)O extract, polysaccharide fraction, supernatant fraction)] exhibited inhibitory activities against G5P[7] rotavirus with the EC(50) values ranging from 0.7±0.4 to 33.7±6.5 μg/mL. Extracts AK-1, AK-3, and AK-5 inhibited rotavirus infection against G8P[7] rotavirus, the with EC(50) values of 8.4±2.2 μg/mL, 6.5±0.8 μg/mL and 8.4±5.0 μg/mL, respectively. By hemagglutination inhibition (HI) assay, six AK extracts completely inhibited viral adsorption onto human RBCs in both strains of rotaviruses at less than 11 μg/mL. However, in the post treatment assay, there was no anti activity shown against both strains of rotaviruses. As a result, six AK extracts were attributed mainly to having a strong interaction with hemagglutinin protein on the outer surface of rotavirus, resulting to blockage of viral adsorption.     

Antimetastatic Effect of Halichondramide,a Trisoxazole Macrolide from the Marine Sponge Chondrosia corticata,on Human Prostate Cancer Cells via Modulation of Epithelial-to-Mesenchymal Transition

Halichondramide (HCA), a trisoxazole-containing macrolide isolated from the marine sponge Chondrosia corticata has been shown to exhibit cytotoxicity and antifungal activities. In our previous study, HCA was also found to exhibit antiproliferative activity against a variety of cancer cells. However, the precise mechanism of action of HCA in the antitumor activity remains to be elucidated. In the present study, we identified the antimetastatic activity of HCA in the highly metastatic PC3 human prostate cancer cells. HCA showed potent growth inhibitory activity of the PC3 cells with an IC₅₀ value of 0.81 µM. Further analysis revealed that HCA suppressed the expression of a potential metastatic biomarker, phosphatase of regenerating liver-3 (PRL-3), in PC3 cells. The suppression of PRL-3 by HCA sequentially down-regulates the expression of phosphoinositide 3-kinase (PI3K) subunits p85 and p110. The antimetastatic effect of HCA was also correlated with the down-regulation of matrix metalloproteases (MMPs) and the modulation of cadherin switches N-cadherin and E-cadherin. In addition, HCA also effectively suppressed the migration and invasion of PC3 cells. These findings suggest that halichondramide might serve as a potential inhibitor of tumor cell metastasis with the modulation of PRL-3.