Glycohistochemical characterization of histologically normal nasal mucosa and enzootic nasal tumor of sheep

Objective-To determine glycohistochemical characteristics of enzootic nasal tumors (ENTs) of sheep, compare results for ENT with those of histologically normal nasal mucosa of sheep, and identify the histologic origin of ENT. Sample-ENT and nasal mucosa samples obtained from cadavers of 5 adult Lacaune sheep with ENT and 5 Lacaune sheep unaffected by ENT, respectively. Procedures-Samples of ENT and nasal mucosa were collected from cadavers of sheep and sectioned. Conventional and lectin histochemical analyses were used to identify glycoconjugates in tissue sections on the basis of their principal chemical groups and principal terminal or internal oligosaccharidic glucidic residues, respectively. Results-ENTs had papillary and tubular portions. Cells in the papillary portion of ENTs had secretion and surface glycoconjugates, which included sulfated glycosaminoglycans and neutral and sialilated glycoproteins. Cells in the tubular portion of ENTs had surface glycoconjugates, which included neutral and sialilated glycoproteins. Both portions of ENTs had C(4)-acetylated sialoderivatives that were not detected in sections of histologically normal nasal mucosa. Conclusions and Clinical Relevance-The papillary portion of ENTs in sheep may originate from respiratory glands and goblet cells. The tubular portion of ENTs in sheep may originate from olfactory glands. Presence of C(4)-acetylated sialoderivatives in cells of ENTs could confer resistance against pathogens to those cells.     

Immunogenicity of eight Mycobacterium avium subsp. paratuberculosis specific antigens in DNA vaccinated and Map infected mice

Mycobacterium avium subsp. paratuberculosis (Map), the etiological agent of chronic enteritis of the small intestine in domestic and wild ruminants, causes substantial losses to livestock industry. Control of this disease is seriously hampered by the lack of adequate diagnostic tools and vaccines. Here we report on the immunogenicity of eight Map specific antigens, i.e. MAP1693c, Ag3, MAP2677c (identified by post-genomic and immunoproteomic analysis of Map secretome) and Ag5, Ag6, MAP1637c, MAP0388 and MAP3743 (identified by bioinformatic in silico screening of the Map genome). Strong, antigen-specific IFN-γ responses were induced in mice vaccinated with plasmid DNA encoding MAP1693c, MAP1637c, MAP0388 and MAP3743. In contrast, T cell responses in Map infected mice were directed preferentially against Ag5 and to a lesser extent against MAP3743. None of the tested DNA vaccines conferred protection against subsequent challenge with Map.     

Escherichia coli lipopolysaccharides and Staphylococcus aureus enterotoxin B differentially modulate inflammatory microRNAs in bovine monocytes

MicroRNAs (miRNAs) are a family of regulatory molecules involved in many physiological processes, including activation of cells of the immune system. This study investigated the effect of Escherichia coli lipopolysaccharide (LPS) and Staphylococcus aureus enterotoxin B (SEB) on the expression of five miRNAs involved in the inflammatory response, including miR-9, miR-125 b, miR-155, miR-146 a and miR-223, in bovine CD14(+) cells (monocytes). Incubation of monocytes with SEB induced down-regulation of miR-155, miR-223 and miR-125 b, but not the anti-inflammatory miRNA miR-146 a. Conversely, incubation with LPS upregulated both miR-155 and miR-146 a. In vitro incubation of isolated CD14(+) bovine monocytes with LPS and SEB elicited different and opposite expression of miRNAs reportedly involved in inflammatory reactions.     

Production of a bovine IL-12p40 probe and application using in situ hybridization on ruminant fixed tissues

Pro-inflammatory cytokines (particularly IL-12) are important for initiating protective T helper 1 (Th1)-type immune responses and hence vital for combating intracellular infections and tumours. In situ hybridization (ISH) provides a powerful diagnostic tool allowing the identification and localization of cells producing these mediators in fixed tissues. The objective of this work was to produce a bovine IL-12p40 probe that allows detection of IL-12p40 mRNA in fixed tissues from different ruminant species. The RNA probe sequence is 447 bp in length and from a region with high cross-species-sequence homology (>97.3% homology) to the ovine, cervine, caprine and bubaline IL-12p40 genes. ISH was carried out on paraformaldehyde fixed tissues collected from cattle, sheep and goats. The probe was efficient in identifying IL-12p40 expressing cells in fixed tissues from all these species. In conclusion, the IL-12p40 probe was efficient in identifying and localizing cells that express IL-12p40, and provides a good immuno-diagnostic technique to characterize immune responses in fixed tissues.     

Design and Synthesis of Pro-Apoptotic Compounds Inspired by Diatom Oxylipins

Oxylipins are a large and diverse family of fatty acid derivatives exhibiting different levels of oxidation of the carbon chain. They are involved in many biological functions in mammals, plants and diatoms. In this last group of organisms, they are suggested to play a role in the reproductive failure of copepod predators, showing clear pro-apoptotic effects on newborn nauplii. In this work, these compounds were tested for the ability to induce mitotic arrest in sea urchin embryos. We show for the first time that oxylipins have an increased efficacy in their corresponding methylated form. Natural oxylipins were also used as an inspiration for the rational design and synthesis of stable chemical analogs with apoptotic activity against tumor cell lines. This approach led to the synthesis of the linear C15-ketol (22) that was shown to induce apoptosis in human leukemia U-937 cells. These results are proof of the concept of the use of eco-physiological considerations as a platform to guide the search for novel drug candidates.     

In vitro adherence of Staphylococcus pseudintermedius to canine corneocytes is influenced by colonization status of corneocyte donors

The current knowledge of in vitro adherence of Staphylococcus pseudintermedius to canine corneocytes is limited to comparative analyses between strains, staphylococcal species or corneocytes collected from different breeds, body sites and hosts. However, the role played by colonization status of corneocyte donors remains unknown. The aim of this study was to evaluate the adherence properties of commensal S. pseudintermedius strains to corneocytes collected from dogs with different colonization status. For this purpose, corneocytes were collected from five dogs that were classified as persistently colonized (D1 and D2), intermittently colonized (D3 and D4) or non-colonized (D5) on the basis of the results of a previous longitudinal study. Adherence to corneocytes originating from each of the five dogs was assessed by an in vitro adhesion assay using four genetically unrelated strains isolated from the colonized dogs (S1 to S4). Irrespective of their host of origin, all strains adhered significantly better to corneocytes from D1 and D2 than to corneocytes from D3, D4 and D5 (P < 0.0001). The mean count of cells adhering to corneocytes from persistently colonized dogs was on average three times higher than the mean count using corneocytes from the other dogs. A significant difference between strains was only observed for one strain-corneocyte combination (S2-D4), indicating that S. pseudintermedius adherence to corneocytes is driven by host factors and only marginally influenced by strain factors. This finding has important implications for understanding and preventing S. pseudintermedius skin colonization and infection.