Western blot detection of PrP Sc in archived paraffin-embedded brainstem from scrapie-affected sheep

Scrapie is a naturally occurring fatal neurodegenerative disease of adult sheep and goats, one of a group of mammalian diseases known as transmissible spongiform encephalopathies (TSE) or prion diseases. Immunoassays that identify disease-associated prion protein (PrP Sc) are integral to the diagnosis of scrapie and other prion diseases. Results obtained by either immunohistochemistry (IHC) or Western blot (WB) assay are generally adequate for the definitive diagnosis. Approved or accepted methods for WB diagnosis of TSEs requires the use of fresh or frozen nonfixed tissue samples, whereas formalin-fixed, paraffin-embedded tissue is required for the localization of PrP Sc by IHC. Because disparate processing methods are used for these accepted diagnostic techniques, separate tissue samples are collected from the same animal. Occasions arise in which there is either insufficient quantity of tissue available to complete analysis by both techniques or initial tissue processing is incompatible with one of the assays. Also, results between the assays may differ because of the vagaries of sampling, especially in case material that contains moderate-to-low levels of PrP Sc. The present article describes a method to conduct a WB assay from the same paraffin-embedded brainstem sample used for the IHC diagnosis of experimentally induced sheep scrapie.     

Choline: high-affinity uptake by rat brain synaptosomes

Synaptosomes from rat brain accumulate choline by two kinetically distinct processes, a high-affinity uptake system [Michaelis constant (K(m)) = 1 x 10(-6)M], and a low-affinity system (K(m) = 9 x 10(-5)M). The high-affinity uptake system requires sodium, and is associated with considerable formation of acetylcholine. The low-affinity uptake system is less dependent on sodium, and does not appear to be associated with a marked degree of acetylcholine formation. The high-affinity choline uptake appears to represent selective choline accumulation by cholinergic neurons.     

Nitrogen-fixing anabaena: physiological adaptations instrumental in maintaining surface blooms

Both laboratory and in situ studies indicate that the nitrogen-fixing blue-green nuisance algae Anabaena spp. have developed adaptive means of dominating surface lake waters. During the dramatic diurnal shifts in surface light intensity and oxygen saturation accompanying blooms, Anabaena can overcome oxygen toxicity by sequential optimization of carbon dioxide and nitrogen fixation and by pigment alteration. These mechanisms allow optimal utilization of the radiant energy while minimizing competition for photoreductant between two main energy-demanding processes.     

Effects of dietary protein level and cold exposure on tissue responsiveness and sensitivity to insulin in sheep

The effects of dietary crude protein (CP) level and cold exposure on tissue responsiveness and sensitivity to insulin were studied in sheep. Nine rams were assigned to one of three isoenergetic diets which contained 70, 100, and 140% of CP for maintenance. They were exposed from a thermoneutral environment (20 degrees C) to a cold environment (0 degrees C) for 7 days. A hyperinsulinemic euglycemic clamp approach was applied for the determination of tissue responsiveness to insulin (the maximal glucose infusion rate, GIRmax) and tissue sensitivity to insulin (the plasma insulin concentration at half maximal glucose infusion rate, ED50). Dietary CP level influenced digestibilities of dry matter and CP (P=0.002 and P=0.001, respectively), and cold exposure decreased (P=0.01) CP digestibility. The GIRmax and ED50 tended to be influenced (P=0.08) by dietary CP level. The GIRmax was enhanced (P=0.0001) during cold exposure. Significant interactions between diet and environment were found for the GIRmax (P=0.04), but not for ED50 (P=0.07). It is concluded that in sheep dietary CP level can modify insulin action in response to cold exposure.     

Adherence of Actinobacillus pleuropneumoniae to primary cultures of porcine lung epithelial cells

To study adherence of Actinobacillus pleuropneumoniae to porcine lower respiratory epithelium, a cell culture model was developed using primary cultures of porcine lung epithelial cells (LEC). Adherence assays were performed and results were compared with data obtained with swine kidney cells (SK6). A. pleuropneumoniae efficiently adhered to LEC with up to 62 bacteria per cell after 2h of incubation. Reference strain of serotype 3 (R3) adhered better to LEC than reference strains of serotypes 1 (R1), 7 (R7) and 8 (R8). Overall the adherence to LEC was more rapid and up to 30-fold more efficient than adherence to SK6 cells. In search for the mechanism involved in the adherence event, we tested the effect of LPS which has previously been demonstrated to cause adherence of the pathogen to upper respiratory epithelium. Adherence assays with LPS transposon mutants demonstrated unaltered (mutant with modification in core/lipid A moiety) or even three-fold more adherence (mutants lacking O antigen) compared to the parent micro-organisms. Purified LPS of strains R1, R3, R7 and R8 did not inhibit adherence of R8 to LEC either, suggesting that LPS and particularly the O-antigen are not essential for adherence of A. pleuropneumoniae to LEC. The efficient, LPS-independent adherence of A. pleuropneumoniae to LEC cells indicates that A. pleuropneumoniae may carry different, cell type-specific adhesins and that primary cultures of lower respiratory epithelium are valuable infection models in studying A. pleuropneumoniae pathogenesis.     

The physical appearance of the vulva in ureaplasma infected ewes

The vulvar labia of ewes naturally infected with ureaplasma were significantly more swollen and red than those of uninfected ewes. Similar changes were observed following experimental infection of previously uninfected ewes. These differences, although statistically significant, were not sufficiently marked to be useful in diagnosing infection by clinical examination.     

Imbalanced lysine to arginine ratios reduced performance in juvenile cobia (Rachycentron canadum) fed high plant protein diets

Cobia (8.4 ± 0.1 g body weight) were fed to satiation with three test diets of high plant protein‐based ingredients and different lysine to arginine ratios, and one commercial diet (currently used for cobia rearing in Vietnam as a control for growth) for 6 weeks. The test diets contained 206 g marine ingredients kg^?1, including fishmeal, krill meal and fish protein concentrate (in order of high to low inclusion), while the rest of the dietary protein was a blend of soya and pea protein concentrate, wheat protein and sunflower meal. Crystalline lysine and arginine were added in the test diets to produce either a balanced lysine to arginine ratio (BL/A; 1.1) and a high or low lysine to arginine ratio (HL/A; 1.8 and LL/A; 0.8, respectively). There were no significant differences in final body weight, weight gain, feed conversion ratio or protein gain between cobia fed BL/A‐ and commercial control diet (CCT). Cobia fed BL/A diet performed better than fish fed either HL/A‐ or LL/A diet. This was partly due to a higher feed intake and protein and lipid gain in cobia fed BL/A diet as compared to HL/A‐ and LL/A diet.     

Band gap fluorescence from individual single-walled carbon nanotubes

Fluorescence has been observed directly across the band gap of semiconducting carbon nanotubes. We obtained individual nanotubes, each encased in a cylindrical micelle, by ultrasonically agitating an aqueous dispersion of raw single-walled carbon nanotubes in sodium dodecyl sulfate and then centrifuging to remove tube bundles, ropes, and residual catalyst. Aggregation of nanotubes into bundles otherwise quenches the fluorescence through interactions with metallic tubes and substantially broadens the absorption spectra. At pH less than 5, the absorption and emission spectra of individual nanotubes show evidence of band gap-selective protonation of the side walls of the tube. This protonation is readily reversed by treatment with base or ultraviolet light.