Canine cutaneous epitheliotropic T-cell lymphoma with vesiculobullous lesions resembling human bullous mycosis fungoides

The broad spectrum of clinical signs in canine cutaneous epitheliotropic T-cell lymphoma mimics many inflammatory skin diseases and is a diagnostic challenge. A 13-year-old-male castrated golden retriever crossbred dog presented with multifocal flaccid bullae evolving into deep erosions. A shearing force applied to the skin at the periphery of the erosions caused the epidermis to further slide off the dermis suggesting intraepidermal or subepidermal separation. Systemic signs consisted of profound weight loss and marked respiratory distress. Histologically, the superficial and deep dermis were infiltrated by large, CD3-positive neoplastic lymphocytes and mild epitheliotropism involved the deep epidermis, hair follicle walls and epitrichial sweat glands. There was partial loss of the stratum basale. Bullous lesions consisted of large dermoepidermal and intraepidermal clefts that contained loose accumulations of neutrophils mixed with fewer neoplastic cells in proteinaceous fluid. The lifted epidermis was often devitalized and bordered by hydropic degeneration and partial epidermal collapse. Similar neoplastic lymphocytes formed small masses in the lungs associated with broncho-invasion. Clonal rearrangement analysis of antigen receptor genes in samples from skin and lung lesions using primers specific for canine T-cell receptor gamma (TCRγ) produced a single-sized amplicon of identical sequence, indicating that both lesions resulted from the expansion of the same neoplastic T-cell population. Macroscopic vesiculobullous lesions with devitalization of the lesional epidermis should be included in the broad spectrum of clinical signs presented by canine cutaneous epitheliotropic T-cell lymphoma.     

Identification of CD3+ T lymphocytes in the green turtle Chelonia mydas

To understand the role of the immune system with respect to disease in reptiles, there is the need to develop tools to assess the host's immune response. An important tool is the development of molecular markers to identify immune cells, and these are limited for reptiles. We developed a technique for the cryopreservation of peripheral blood mononuclear cells and showed that a commercially available anti-CD3 epsilon chain antibody detects a subpopulation of CD3 positive peripheral blood lymphocytes in the marine turtle Chelonia mydas. In the thymus and in skin inoculated with phytohemagglutinin, the same antibody showed the classical staining pattern observed in mammals and birds. For Western blot, the anti-CD3 antibodies identified a 17.6 kDa band in membrane proteins of peripheral blood mononuclear cell compatible in weight to previously described CD3 molecules. This is the first demostration of CD3+ cells in reptiles using specific antibodies.     

Specific proteins allow classification of pigs according to sire breed, rearing environment and gender

The present study used a proteomic data set obtained from a 2 × 2 × 2 factorial experiment on longissimus muscle of 24 pigs to identify single or couples of proteins allowing correct classification of pigs according to gender, sire breed, or rearing environment. An actin isoform, a myosin light chain 2 isoform and cytochrome Bc1 allowed each, correct classification of all pigs according to gender. Peroxiredoxin 6 allowed correct classification of 23 pigs according to indoor or outdoor rearing environment, but only if gender was also taken into account. Heat shock protein 73 allowed correct classification of 21 pigs according to sire breed, Duroc or Large White. Results show that proteins may be used as biomarkers for traceability or genetic research. Relationships between treatment factors and intracellular regulatory mechanisms associated with these proteins are discussed.     

Effect of tapping activity on the dynamics of radial growth of Hevea brasiliensis trees

Rubber tree (Hevea brasiliensis Müll. Arg.) radial growth dynamics were monitored with displacement sensors, together with latex production, to investigate three aspects of the dual production of latex and wood: (1) the usefulness of fine-scale dendrometric measurements as a physiological tool to detect water shortage through radial growth; (2) the dynamic aspects, both at the seasonal and at the multi-year scale, of the competition between latex and wood production; and (3) the spatial distribution of radial growth rates around the tapping cut. Radial growth of untapped control trees started with the onset of the rainy season and lasted until the onset of the dry season, ceasing completely during the driest period. Displacement sensors provided a sensitive means of detecting water shortage, with a clear correlation between diameter variations and changes in water availability (both daily evapotranspiration and monthly rainfall) over the whole annual cycle. However, the correlation was significantly disturbed in tapped trees. After resumption of tapping, the radial growth rate dropped sharply within two weeks and the effect persisted throughout the whole season, so that the cumulative growth of tapped trees was about half that of untapped trees, with the cumulative growth deficit reaching 80% for the period from mid-June to November. This long-known negative impact of tapping on growth was much stronger in the second year of tapping than in the first, whereas latex production increased significantly between the first and second year of tapping. The increased latex production, which could not be ascribed to climatic conditions, shows that the establishment of an artificial latex sink is a progressive, long-term process likely involving many aspects of metabolism. As expected, ethylene significantly increased latex production in both years; however, ethylene had no effect on the growth rates of tapped trees. Radial growth was differentially affected at different locations around the tapping cut, with growth rates significantly lower in the tapped panel than in the untapped panel, and higher above the cut than below the cut. Thus, caution is needed when deriving whole stem wood production from girth measurements at one location on the stem, especially from girth measurements made close to the tapping cut. This also provides new evidence for the location of the latex regeneration area in the tapped panel, below the cut.     

“Aberrant” plants in cauliflower: 2. Aneuploidy and global DNA methylation

Aberrant phenotypes of cauliflower were detected throughout the cultivation period and in any variety type. The rate of these phenotypes in the field has recently increased. We reported previously on the first part of our results which showed that (1) the rate of aberrant plants varied with genotype and cultivation area, (2) the aberrant phenotypes can evolve or reverse to normality during the plant cycle and (3) the capacity to express a variant phenotype can be transmitted to the progeny. An epigenetic hypothesis has been proposed to explain the determinism of the phenomenon. Further investigation on the “aberrant” character focussed on the flow cytometric estimation of ploidy levels and on the parallel observation of meiosis. Only a fraction of aberrant plants did show aneuploidy and various ploïdy levels were found for the same phenotype. Indeed, aneuploidy could not be related to the aberrant phenotype although it could probably be a consequence of the aberration phenomenon. HPLC analysis of global DNA methylation rates showed that DNA hypermethylation occurred in plants which exhibited an evolution of their phenotype during vegetative cycle. The epigenetic origin of aberrant phenotypes in cauliflower is discussed with reference to epigenetic diseases described in human beings.     

Production and isolation of azaspiracid-1 and -2 from Azadinium spinosum culture in pilot scale photobioreactors

Azaspiracid (AZA) poisoning has been reported following consumption of contaminated shellfish, and is of human health concern. Hence, it is important to have sustainable amounts of the causative toxins available for toxicological studies and for instrument calibration in monitoring programs, without having to rely on natural toxin events. Continuous pilot scale culturing was carried out to evaluate the feasibility of AZA production using Azadinium spinosum cultures. Algae were harvested using tangential flow filtration or continuous centrifugation. AZAs were extracted using solid phase extraction (SPE) procedures, and subsequently purified. When coupling two stirred photobioreactors in series, cell concentrations reached 190,000 and 210,000 cell · mL(-1) at steady state in bioreactors 1 and 2, respectively. The AZA cell quota decreased as the dilution rate increased from 0.15 to 0.3 day(-1), with optimum toxin production at 0.25 day(-1). After optimization, SPE procedures allowed for the recovery of 79 ± 9% of AZAs. The preparative isolation procedure previously developed for shellfish was optimized for algal extracts, such that only four steps were necessary to obtain purified AZA1 and -2. A purification efficiency of more than 70% was achieved, and isolation from 1200 L of culture yielded 9.3 mg of AZA1 and 2.2 mg of AZA2 of >95% purity. This work demonstrated the feasibility of sustainably producing AZA1 and -2 from A. spinosum cultures.     

Evaluation of thermomechanical pretreatment for enzymatic hydrolysis of pure microcrystalline cellulose and cellulose from Brewers’ spent grain

Microcrystalline cellulose (Avicel PH102) and Brewers’ spent grain (BSG) were subjected to Détente Instantanée Contrôlée (DIC) thermomechanical pre-treatment before exposure to cellulases (Celluclast 1.5 L). In a first part, we showed that the addition of β-glucosidase (Novozym-188) increased the hydrolysis yield of Avicel. A maximal theoretical yield (100%), was obtained for 5 and 10 g/L of Avicel using a mixture of Celluclast 1.5 L/Novozym-188. After DIC pre-treatments, the initial rate and final yield of hydrolysis decreased in comparison with those from untreated microcrystalline cellulose. This phenomenon may be due to the modification of the crystallinity of pure cellulose and the formation of inhibitors during the pre-treatment. In a second part, BSG was thermomechanically pre-treated and hydrolyzed. The results showed that the hydrolysis yield of BSG treated at pressure levels between 2 and 7 bar during 15 min was strongly improved compared to hydrolysis yield of untreated BSG. The optimized hydrolysis process, under intensive DIC conditions, achieved a glucose yield corresponding to 100% of the theoretical cellulose value. The morphology of BSG samples was studied with Scanning Electronic Microscopy (SEM) and highlighted that the structure of pre-treated BSG showed an important disruption compared to the rigid structure of untreated BSG.     

A systematic review of the evidence of reduced allergenicity and clinical benefit of food hydrolysates in dogs with cutaneous adverse food reactions

Several hydrolysate-based diets have been commercialized for helping diagnose or treat dogs with cutaneous adverse food reactions (CAFR). This systematic review was performed to examine the evidence in favour of reduced immunological and clinical allergenicity of hydrolysates in dogs with CAFR. Citation databases, meeting abstracts and article bibliographies were scanned for relevant citations, and companies were contacted to provide unpublished reports. Eleven studies relevant to this study were identified. Some evidence of reduced serum IgE binding to a soy hydrolysate (1 study) and decreased intradermal test reactivity to hydrolysed proteins (three studies) was found. In four reports, the feeding of dogs suspected of having CAFR with hydrolysate-based diets reduced or eliminated clinical signs in a variable proportion of subjects. The percentage of dogs with CAFR that still reacted to these hydrolysate-based diets could not be assessed, however. Importantly, up to 50% of dogs with CAFR enrolled in three controlled studies exhibited increases in clinical signs after ingesting partial hydrolysates derived from foods to which they were hypersensitive. In conclusion, the limited number of studies undertaken point to reduced – but not eliminated – immunological and clinical allergenicity of hydrolysate-based commercial diets. A variable proportion of dogs with CAFR will exhibit a worsening of clinical signs when fed partial hydrolysates. Clinicians must weigh the clinical benefit of these diets versus their high cost and low risk of reduced appetence or gastrointestinal sign development. At this time, hydrolysate-containing diets are probably best used in dogs suspected not to be hypersensitive to their individual components.