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Abstract: A bovine herpesvirus 4 was isolated from the milk cell fraction of a healthy cow and his full genome cloned as a bacterial artificial chromosome. So cloned viral genome was used as a vector platform to deliver in vitro and in vivo an optimized secreted chimeric peptide obtained by the fusion of the bovine viral diarrhoea virus glycoprotein E2 ectodomain with the bovine herpesvirus 1 glycoprotein D ectodomain. Recombinant virus infected cells robustly expressed and secreted the chimeric peptide into the culture medium and inoculated animals with the recombinant virus successfully responded toward antigens, gE2 and gD. Thus, this work has implications for the development of safe and effective polyvalent vaccines. [Copyright &y& Elsevier]

A project dedicated to stereotactic body radiotherapy (SBRT) dosimetric aspects started in the framework of the Italian Association of Medical Physics (AIFM) SBRT working group. Its main objectives are manifold but sharing the knowledge between different clinics is the one of the most important. More than 30 centres, equipped with Varian, Elekta, Siemens or CyberKnife linac were enrolled. The project was divided in several work-packages, the first one evaluated the relative measurements with detectors routinely used by individual centers and in the following different detectors run in various centre. Each centre in the same work-package performed dose profile of field size ranging from 0.6 × 0.6 cm2 to 5.0 × 5.0 cm2, and relative output factors (ROF) measurements with a diamond or silicon diode or scintillator detector. In workpackage-1 ROF values measured in the first phase were compared with the ones measured with a microdiamond showing a higher inter-center consistency with this dosimeter compared to routine detectors. In workpackage-2 a silicon diode of new generation was used also to develop a mathematical relation from multicentric experimental data, which describes and predicts the ROF as a function of effective field size for TrueBeam Varian and Elekta linacs. Workpackage-3 used a plastic scintillator for TPR20, 10 and ROF; the latter showed a greater variability. Workpackage-4 used diamond and plastic scintillator with Cyberknife beams, to evaluate if microDiamond could be a suitable alternative to silicon diodes for OF determination and to validate the feasibility of using the scintillator as a reference detector in consideration of the ČerenkovLightRatio and experimental uncertainties. The results of the study for all the detectors emphasized the usefulness of a multi-center validation over a single center approach. [ABSTRACT FROM AUTHOR]

Abstract: Many immunological aspects of pregnancy, such as the role played by gamma interferon (IFN-γ) in abortion, are not well understood. Neospora caninum is an intracellular protozoan considered to be among the main causes of abortion in cattle worldwide. The present study analyzes the interaction between IFN-γ production and N. caninum infection in naturally infected pregnant cows. Data were obtained from 126 pregnant cows: 86 seropositive and 40 seronegative for the parasite. Pregnancy diagnosis and blood sample collection were performed on days 40, 90, 120, 150, 180 and 210 post-insemination or until the time of abortion detection. Plasma was tested for antibodies against N. caninum and IFN-γ. Interferon-gamma was detected at some point along the pregnancy in 16 (19%) of the 86 Neospora-seropositive cows yet was undetectable in the 40 seronegative animals. Of the 126 pregnancies examined, 22 (17.5%) ended in abortion. Abortion occurred in 24.4% of seropositive cows (21/86) and in 2.5% of seronegative animals (1/40). Significant (P <0.0001) interaction was observed between Neospora-seropositivity and IFN-γ production. Based on the odds ratio, the risk of abortion was 15.6 times higher in seropositive cows not producing IFN-γ than in seronegative animals, whereas neosporosis had no effect in seropositive cows with IFN-γ production. A significant (P =0.001) negative effect of IFN-γ production on the Neospora titer was furthermore observed in the 65 non-aborting seropositive animals. These results indicate that IFN-γ production affords protection against abortion in Neospora-infected cows and also point to a reduced humoral immune response to N. caninum during gestation in cows producing IFN-γ. [Copyright &y& Elsevier]

Abstract: VP2 is the outermost Bluetongue virus (BTV) antigenic protein, forming triskelion motifs on the virion surface. Although VP2 has been expressed successfully through many systems, its paracrine expression as a soluble form by mammalian cells represents a difficult task. In the present paper two fragments of VP2 have been expressed successfully into the medium of transiently transfected mammalian cells through a fusion peptides strategy. The crude conditioned medium containing the secreted peptide could be employed for immunodiagnostic assay development or vaccine purposes. [ABSTRACT FROM AUTHOR]

A monoclonal antibody (antik-B) against an oligopeptide of 23 AA corresponding to the region 131-153 of bovine κ-casein (κ-CN) B was generated using the Human Combinatorial Antibody Library (HuCAL) technology. Both AA substitutions distinguishing κ-CN A and B are located in that region (positions 136 and 148). In this study, the reactivity of antik-B to milk samples collected from cows previously genotyped as CSN3*AA, CSN3*AB, and CSN3*BB was tested. According to Western blot results, antik-B recognized κ-CN B and it showed no cross-reactivity toward κ-CN A and other milk proteins. Furthermore, a modified Western blot method, urea-PAGE Western blot, was set up to assess the reactivity of antik-B toward all isoforms of κ-CN B. In conclusion, antik-B was specific to κ-CN B in milk and it seemed to be reactive toward all its isoforms. [ABSTRACT FROM AUTHOR]