Comparative Susceptibility of Sheep of Different Origins, Breeds and PRNP Genotypes to Challenge with Bovine Spongiform Encephalopathy and Scrapie

Sheep are natural hosts of the prion disease, scrapie. They are also susceptible to experimental challenge with various scrapie strains and with bovine spongiform encephalopathy (BSE), which affects cattle and has been accidentally transmitted to a range of other species, including man. Incidence and incubation period of clinical disease in sheep following inoculation is controlled by the PRNP gene, which has different alleles defined on the basis of polymorphisms, particularly at codons 136, 154 and 171, although other codons are associated with survival time, and the exact responses of the sheep may be influenced by other breed-related differences. Here we report the results of a long term single study of experimental scrapie and BSE susceptibility of sheep of Cheviot, Poll Dorset and Suffolk breeds, originating from New Zealand and of a wide range of susceptible and resistant PRNP genotypes. Responses were compared with those of sheep from a closed Cheviot flock of UK origin (Roslin Cheviot flock). The unusually long observation period (6-8 years for most, but up to 12 years for others) allows us to draw robust conclusions about rates of survival of animals previously regarded as resistant to infection, particularly PRNP heterozygotes, and is the most comprehensive such study reported to date. BSE inoculation by an intracerebral route produced disease in all genotype groups with differing incubation periods, although M112T and L141F polymorphisms seemed to give some protection. Scrapie isolate SSBP/1, which has the shortest incubation period in sheep with at least one VRQ PRNP allele, also produced disease following sub-cutaneous inoculation in ARQ/ARQ animals of New Zealand origin, but ARQ/ARQ sheep from the Roslin flock survived the challenge. Our results demonstrate that the links between PRNP genotype and clinical prion disease in sheep are much less secure than previously thought, and may break down when, for example, a different breed of sheep is moved into a new flock

Enzymes immobilized in Langmuir-Blodgett films: Why determining the surface properties in Langmuir monolayer is important?

ABSTRACT In this review we discuss about the immobilization of enzymes in Langmuir-Blodgett films in order to determine the catalytic properties of these biomacromolecules when adsorbed on solid supports. Usually, the conformation of enzymes depends on the environmental conditions imposed to them, including the chemical composition of the matrix, and the morphology and thickness of the film. In this review, we show an outline of manuscripts that report the immobilization of enzymes as LB films since the 1980’s, and also some examples of how the surface properties of the floating monolayer prepared previously to the transfer to the solid support are important to determine the efficiency of the resulting device

Secondary structure analysis of HIV-1-gp41 in solution and adsorbed to aluminium hydroxide by Fourier transform infrared spectroscopy

International audienc

Transmission and characterization of bovine spongiform encephalopathy sources in two ovine transgenic mouse lines (TgOvPrP4 and TgOvPrP59).

International audienc

SEM3De: image restoration for FIB-SEM.

FIB-SEM (Focused Ion Beam-Scanning Electron Microscopy) is a technique to generate 3D images of samples up to several microns in depth. The principle is based on the alternate use of SEM to image the surface of the sample (a few nanometers thickness) and of FIB to mill the surface of the sample a few nanometers at the time. In this way, huge stacks of images can thus be acquired.Although this technique has proven useful in imaging biological systems, the presence of some visual artifacts (stripes due to sample milling, detector saturation, charge effects, focus or sample drift, etc.) still raises some challenges for image interpretation and analyses. With the aim of meeting these challenges, we developed a freeware (SEM3De) that either corrects artifacts with state-of-the-art approaches or, when artifacts are impossible to correct, enables the replacement of artifactual slices by an in-painted image created from adjacent non-artifactual slices. Thus, SEM3De improves the overall usability of FIB-SEM acquisitions. SEM3De can be downloaded from https://sourceforge.net/projects/sem3de/ as a plugin for ImageJ

Primary and metastatic intraaxial brain tumors: prospective comparison of multivoxel 2D chemical-shift imaging (CSI) proton MR spectroscopy, perfusion MRI, and histopathological findings in a group of 159 patients

This study aims to assess the diagnostic value of multivoxel 2D chemical-shift imaging (CSI) proton magnetic resonance (MR) spectroscopy combined with perfusion magnetic resonance imaging (MRI) in the differential diagnosis and grading of brain tumors by comparing neuroimaging data with histopathological findings obtained after resection or biopsy. A total of 159 patients with a previous brain tumor diagnosis underwent multivoxel 2D CSI proton MR spectroscopy and perfusion MRI. MR spectroscopy multivoxel 2D CSI was performed with an echo time of 30, TR 1,500, FOV 160 mm, acquisition time 7 min 34 s. rCBV maps were evaluated during postprocessing. Statistical analysis was performed on the examination of distributive normality, with logarithmic transformations, Fisher's test, and Bonferroni's test. We used the Pearson's test to compare percentages. In the differential diagnosis between GBM and metastases, MR spectroscopy multivoxel 2D CSI, combined with dynamic contrast enhanced MRI (DCE-MRI) perfusion, reached high sensibility and specificity (p < 0.000001). In brain tumor grading, the same method reached high sensibility and specificity (p < 0.000001) in distinguishing grade III-IV gliomas but encountered difficulty in determining grades within the two main groups of primary brain tumors, especially where mixed gliomas were involved. The systematic use of CSI spectroscopy and perfusion imaging has shown a high potential in the differential diagnosis and grading of brain tumors. Further exploration into diagnostic procedures that can significantly distinguish between grade III-IV and grade II tumors is needed