Rapamycin sharpens the sensitivity to cisplatin of cells isolated from ovine pulmonary adenocarcinoma

International audienc

Transformation mechanisms of type II alveolar cells in a virally-induced lung cancer

International audienc

Use of activity monitors for assessment of pruritus in an acute model of canine atopic dermatitis

BACKGROUND: We developed a canine model of acute atopic dermatitis to evaluate the potential of compounds to treat pruritus and skin lesions induced in Dermatophagoides farinae (Df)-sensitized dogs. HYPOTHESIS/OBJECTIVES: The aim was to investigate the effectiveness of long-term recording activity monitors to assess pruritus induced by allergen challenges. ANIMALS: Thirty-two Df-sensitized laboratory dogs. METHODS: In two blinded crossover studies, 28 Df-sensitized dogs were challenged on 3 days with a Df slurry applied to clipped abdominal skin. Dogs were treated with a positive control (prednisolone 1 mg/kg once daily for 5 days, starting 1 day before challenge) or left untreated; all were fitted with activity monitors. To confirm pruritus, a parallel study with four dogs was conducted, filming the dogs before and during challenge and assessing the film for pruritic behaviour. RESULTS: The activity of dogs treated with prednisolone was significantly lower between 00.00 and 03.00 h and between 03.00 and 06.00 h compared with untreated dogs (repeated-measures ANCOVA; P < 0.0001). To determine whether the recorded night-time activity corresponded to pruritic manifestations, we compared activity monitor and video recordings of four dogs for two periods (16.30-20.30 and 24.00-03.00 h) before and during a Df challenge. The correlation between night-time activity monitor activity and observed pruritic behaviour was highly significant (test of correlation coefficient versus zero: r = 0.57, P < 0.0001). CONCLUSIONS AND CLINICAL IMPORTANCE: Determination of night-time activity with activity monitors after allergen challenge appears to be an objective and practical way to assess pruritus in this experimental model of canine atopic dermatitis

High-level intracellular expression of heterologous proteins in <it>Brevibacillus choshinensis</it> SP3 under the control of a xylose inducible promoter

<p>Abstract</p> <p>Background</p> <p>In past years research has focused on the development of alternative Gram positive bacterial expression systems to produce industrially relevant proteins. <it>Brevibacillus choshinensis</it> is an easy to handle non-sporulating bacterium, lacking extracellular proteases, that has been already shown to provide a high level of recombinant protein expression. One major drawback, limiting the applicability of the <it>Brevibacillus</it> expression system, is the absence of expression vectors based on inducible promoters. Here we used the <it>PxylA</it> inducible promoter, commonly employed in other <it>Bacillae</it> expression systems, in <it>Brevibacillus</it>.</p> <p>Results</p> <p>Using GFP, α-amylase and TcdA-GT as model proteins, high level of intracellular protein expression (up to 250 mg/L for the GFP) was achieved <it>in Brevibacillus</it>, using the pHis1522 vector carrying the <it>B. megaterium</it> xylose-inducible promoter (<it>PxylA</it>)<it>.</it> The GFP expression yields were more than 25 fold higher than those reported for <it>B. megaterium</it> carrying the same vector. All the tested proteins show significant increment in their expression levels (2-10 folds) than those obtained using the available plasmids based on the P2 constitutive promoter.</p> <p>Conclusion</p> <p>Combining the components of two different commercially available Gram positive expression systems, such as <it>Brevibacillus</it> (from Takara Bio) and <it>B. megaterium</it> (from Mobitec), we demonstrate that vectors based on the <it>B. megaterium PxylA</it> xylose inducible promoter can be successfully used to induce high level of intracellular expression of heterologous proteins in <it>Brevibacillus</it>.</p

Viral load, organ distribution, histopathological lesions, and cytokine mRNA expression in goats infected with a molecular clone of the caprine arthritis encephalitis virus

Caprine arthritis encephalitis virus (CAEV) is a lentivirus of goats that causes persistent infection characterized by the appearance of inflammatory lesions in various organs. To define the sites of persistence, 5 goats were infected with a molecular clone of CAEV, and the viral load was monitored by real-time-PCR and RT-PCR in different sites 8 years after infection. The lymph nodes proved to be an important virus reservoir, with moderate virus replication relative to what is reported for lentiviruses of primates. Mammary gland and milk cells were preferred sites of viral replication. The viral load varied significantly between animals, which points to an important role of the genetic background. We found a clear association between occurrence of histopathological lesions and viral load in specific sites. The mRNA expression analysis of several cytokines did not reveal differences between animals that could explain the considerable individual variations in viral load observed

Transformation mechanisms of type II pneumocytes in a virally-induced lung cancer

National audienc

Analysis of the Antibody Response to an Immunodominant Epitope of the Envelope Glycoprotein of a Lentivirus and Its Diagnostic Potential

The envelope glycoprotein of small ruminant lentiviruses (SRLV) is a major target of the humoral immune response and contains several linear B-cell epitopes. We amplified and sequenced the genomic segment encoding the SU5 antigenic site of the envelope glycoprotein of several SRLV field isolates. With synthetic peptides based on the deduced amino acid sequences of SU5 in an enzyme-linked immunosorbent assay (ELISA), we have (i) proved the immunodominance of this region regardless of its high variability, (ii) defined the epitopes encompassed by SU5, (iii) illustrated the rapid and peculiar kinetics of seroconversion to this antigenic site, and (iv) shown the rapid and strong maturation of the avidity of the anti-SU5 antibody. Finally, we demonstrated the modular diagnostic potential of SU5 peptides. Under Swiss field conditions, the SU5 ELISA was shown to detect the majority of infected animals and, when applied in a molecular epidemiological context, to permit rapid phylogenetic classification of the infecting virus