Application of nox-restriction fragment length polymorphism for the differentiation of Brachyspira intestinal spirochetes isolated from pigs and poultry in Australia.

Sixty-nine intestinal spirochetes isolated from pigs and poultry in eastern Australia were selected to evaluate the effectiveness of a species-specific PCR-based restriction fragment length polymorphism (RFLP) analysis of the Brachyspira nox gene. For comparative purposes, all isolates were subjected to species-specific PCRs for the pathogenic species Brachyspira hyodysenteriae and Brachyspira pilosicoli, and selected isolates were examined further by sequence analysis of the nox and 16S ribosomal RNA genes. Modifications to the original nox-RFLP method included direct inoculation of bacterial cells into the amplification mixture and purification of the PCR product, which further optimized the nox-RFLP for use in a veterinary diagnostic laboratory, producing sufficient product for both species identification and future comparisons. Although some novel profiles that prevented definitive identification were observed, the nox-RFLP method successfully classified 45 of 51 (88%) porcine and 15 of 18 (83%) avian isolates into 5 of the 6 recognized species of Brachyspira. This protocol represents a significant improvement over conventional methods currently used in veterinary diagnostic laboratories for rapid specific identification of Brachyspira spp. isolated from both pigs and poultry.     

Temperature effects on the action of acylurea insecticides against tobacco hornworm (Manduca sexta) larvae

The toxic effects of six acylurea insecticides on larvae of the tobacco hornworm were investigated at each of four environmental temperatures (20, 25, 30 and 35°C). This spans the range of temperatures which the insects can tolerate. For all the acylureas tested, mortality increased with temperature when either newly hatched or fourth-instar larvae were given insecticide in their food. Sub-lethal growth inhibition also became more pronounced at progressively higher environmental temperatures. This temperature dependence of acylurea action was not due to altered uptake of the insecticide, since there was no significant variation with temperature in the amount of [¹⁴C]flufenoxuron taken up by fifth-instar larvae when given a single meal containing labelled insecticide. Additionally, mortality of fourth-instar larvae given a single intra-haemocoelic injection of flufenoxuron was significantly greater at higher temperatures, implying that temperature affects a process that occurs after insecticide uptake. The intrinsic ability of acylureas to inhibit chitin synthesis is temperaturesensitive, since flufenoxuron inhibited the incorporation of [¹⁴C]N-acetylalucosamine into chitin by proleg epidermis in vitro significantly less well at 20°C than at the higher temperatures tested. However, there was no significant variation between the effectiveness of in-vitro chitin synthesis inhibition at 25, 30 and 35°C. These data show that the effectiveness of acylurea insecticides is subject to strong temperature effects in the range of temperatures likely to be experienced in the field.     

EFFECTS OF FLUID AND COMPUTED TOMOGRAPHIC TECHNICAL FACTORS ON CONSPICUITY OF CANINE AND FELINE NASAL TURBINATES

Turbinate destruction is an important diagnostic criterion in canine and feline nasal computed tomography (CT). However decreased turbinate visibility may also be caused by technical CT settings and nasal fluid. The purpose of this experimental, crossover study was to determine whether fluid reduces conspicuity of canine and feline nasal turbinates in CT and if so, whether CT settings can maximize conspicuity. Three canine and three feline cadaver heads were used. Nasal slabs were CT‐scanned before and after submerging them in a water bath; using sequential, helical, and ultrahigh resolution modes; with images in low, medium, and high frequency image reconstruction kernels; and with application of additional posterior fossa optimization and high contrast enhancing filters. Visible turbinate length was measured by a single observer using manual tracing. Nasal density heterogeneity was measured using the standard deviation (SD) of mean nasal density from a region of interest in each nasal cavity. Linear mixed‐effect models using the R package ‘nlme’, multivariable models and standard post hoc Tukey pair‐wise comparisons were performed to investigate the effect of several variables (nasal content, scanning mode, image reconstruction kernel, application of post reconstruction filters) on measured visible total turbinate length and SD of mean nasal density. All canine and feline water‐filled nasal slabs showed significantly decreased visibility of nasal turbinates (P < 0.001). High frequency kernels provided the best turbinate visibility and highest SD of aerated nasal slabs, whereas medium frequency kernels were optimal for water‐filled nasal slabs. Scanning mode and filter application had no effect on turbinate visibility.     

Breast cancer prevention by green tea catechins and black tea theaflavins in the C3(1) SV40 T,t antigen transgenic mouse model is accompanied by increased apoptosis and a decrease in oxidative DNA adducts

Tea consumption is associated with a reduced risk of mammary cancer as reflected by epidemiological studies and experiments in carcinogen-induced rodent models of mammary carcinogenesis. We tested the hypothesis that green tea catechins (GTC) or theaflavins from black tea (BTT) interfere with mammary carcinogenesis in C3(1) SV40 T,t antigen transgenic multiple mammary adenocarcinoma (TAg) mice and that GTC/BTT affect tumor survival or oxidation status. TAg mice received GTC/BTT (0.05%) in drinking water for their lifetime. As compared to control mice, they survived longer and had smaller tumors. On microscopic inspection, the size of the largest tumor per mouse was decreased by 40-42% (p<0.01). GTC (0.01%) and BTT (0.05%) increased levels of cleaved caspase 3 in tumor tissue by 67 and 38%, respectively (p<0.05), intimating increased apoptosis. Tumor levels of the malondialdehyde-DNA adduct M1dG in mice receiving GTC or BTT (0.05%) were reduced by 78 (p<0.001) or 63% (p<0.05), respectively, as compared to controls. The results render the exploration of the breast cancer chemopreventive properties of tea preparations in humans worthwhile.