Gastric leiomyoma in a sea bass Dicentrarchus labrax broodfish.

This report describes a spontaneously arising non-infiltrative neoplasm of gastric tunica muscularis in a broodstock sea bass Dicentrarchus labrax from an Italian aquaculture farm. Microscopically, the mass was circumscribed and non-encapsulated and was composed of spindle cells arranged in parallel interlacing bundles or, occasionally, a whirling pattern. Cells had a small quantity of eosinophilic cytoplasm with distinct cell borders. Neoplastic cells were immuno-reactive with smooth muscle actin, vimentin and desmin; S100 was negative. The mucosal epithelium was intact with no neoplastic involvement. A gastric leiomyoma was diagnosed based on the findings. More efforts should be made to study the possible etiology of leiomyoma affecting fish from aquaculture.

Contagious agalactia: costs and control revisited.

The economic costs of contagious agalactia (CA) to the small ruminant dairy industry are not well known but include losses due to mortality, lowered milk production, spoiled products, abortions and animal welfare problems, as well as diagnosis and treatment. This paper reports financial estimates made in southern Europe, including a study on small- and large-scale farming systems in Italy, indicating that the financial losses are high and underestimated. Furthermore, the current control strategies, including chemotherapy and vaccination, in selected countries in Europe are described. In some countries, disease control is hampered by excessively strict veterinary legislation which discourages farmers and private veterinarians from notifying outbreaks because it leads to the prohibition of milk sales and can result in delays in lifting restrictions. In addition, new European Union legislation may downgrade the importance of CA, which will have implications for international research efforts. Finally, a series of recommendations are provided that cover the proper notification and handling of CA outbreaks, including movement control, current diagnostics, treatment, vaccination and disinfection.

Si le coût économique exact de l'agalaxie contagieuse pour le secteur ovin et caprin de production laitière n'est pas connu, on sait néanmoins qu'il recouvre les pertes dues à la mortalité dans les cheptels, à une chute de la production de lait, aux produits altérés, aux avortements et aux problèmes de bien-être animal, en plus des coûts du diagnostic et des traitements. Les auteurs font état d'estimations financières réalisées en Europe méridionale, dont une étude sur les exploitations familiales et les élevages de grande taille en Italie, qui coïncident dans le constat de pertes financières à la fois importantes et sous-estimées. Les auteurs décrivent également les stratégies de lutte mises en place actuellement par plusieurs pays d'Europe, en particulier l'antibiothérapie et la vaccination. Dans certains pays, les efforts de lutte sont entravés par une législation vétérinaire excessivement rigoureuse qui dissuade les éleveurs et les vétérinaires privés de notifier les foyers car cela entraîne l'interdiction de vendre le lait issu des troupeaux infectés et retarde la levée des mesures de restriction. En outre, la nouvelle réglementation de l'Union européenne risque d'abaisser l'importance de l'agalaxie contagieuse, ce qui aura des conséquences sur les efforts mobilisés par la recherche au niveau international. Pour conclure, les auteurs formulent plusieurs recommandations en vue d'une notification et gestion appropriées des foyers d'agalaxie contagieuse, notamment pour ce qui concerne le contrôle des mouvements d'animaux, les méthodes actuelles de diagnostic, le traitement, la vaccination et la désinfection.

Aunque no se conocen bien los costos económicos que la agalaxia contagiosa inflige a la industria lechera de pequeños rumiantes, se sabe que las pérdidas por mortalidad, mengua de la producción lechera, productos echados a perder, abortos y problemas de bienestar animal son un factor importante, sin olvidar los gastos de diagnóstico y tratamiento. Los autores dan cuenta de cálculos económicos realizados en Europa meridional, en particular a raíz de un estudio de pequeñas y grandes explotaciones ganaderas de Italia, que llevaron a la conclusión de que las pérdidas económicas son cuantiosas y están subestimadas. Además, los autores describen los métodos de lucha aplicados actualmente en determinados países de Europa, que incluyen tratamiento medicamentoso y vacunaciones. En algunos países la lucha contra la enfermedad se ve lastrada por una legislación veterinaria demasiado estricta, que no alienta a productores y veterinarios privados a notificar brotes porque ello conduce a la prohibición de las ventas de leche y puede demorar el levantamiento de las restricciones. Por otra parte, hay nuevos textos legislativos de la Unión Europea que quizá vengan a restar importancia a la agalaxia contagiosa, lo que repercutiría en las actividades internacionales de investigación. Por último, los autores formulan una serie de recomendaciones referidas a cuestiones que van desde la correcta notificación y gestión de los brotes de agalaxia contagiosa hasta el control de los desplazamientos, pasando por los procedimientos vigentes de diagnóstico o los métodos de tratamiento, vacunación y desinfección.

Immunohistochemical Study of Four Fish Tumors.

The present study supports the usefulness of ancillary techniques, such as immunohistochemistry, as a valid diagnostic tool in the field of fish oncology. The immunohistochemical patterns observed in four neoplasms on four individual teleosts belonging to different species are described. Cytokeratin, vimentin, actin, S100, calretinin, and Melan-A antibodies were used. Diagnoses of papilloma in a Bream Abramis brama, fibroma in a Sand Steenbras Lithognathus mormyrus, schwannoma in a Crucian Carp Carassius carassius, and melanoma in a spontaneously inbred Xiphophorus hybrid were made. Diagnosis of tumors in fish is not always easy to carry out, and the tool provided by antibodies used on mammalian tissue is essential for obtaining definitive, unambiguous, and inexpensive identification.

Electrospun PHEA-PLA/PCL Scaffold for Vascular Regeneration: A Preliminary in Vivo Evaluation.

BACKGROUND: There is increasing interest in the development of vessel substitutes, and many studies are currently focusing on the development of biodegradable scaffolds capable of fostering vascular regeneration. We tested a new biocompatible and biodegradable material with mechanical properties similar to those of blood vessels. METHODS: The material used comprises a mixture of α,ß-poly(N-2-hydroxyethyl)-d,l-aspartamide (PHEA) and polylactic acid (PLA), combined with polycaprolactone (PCL) by means of electrospinning technique. Low-molecular-weight heparin was also linked to the copolymer. A tubular PHEA-PLA/PCL sample was used to create an arteriovenous fistula in a pig model with the use of the external iliac vessels. The flow was assessed by means of Doppler ultrasound examination weekly, and 1 month after the implantation we removed the scaffold for histopathologic evaluation. RESULTS: The implants showed a perfect leak-proof seal and adequate elastic tension to blood pressure. About ∼3 weeks after the implantation, Doppler examination revealed thrombosis of the graft, so we proceeded to its removal. Histologic examination showed chronic inflammation, with the presence of foreign body cells and marked neovascularization. The material had been largely absorbed, leaving some isolated spot residues. CONCLUSIONS: The biocompatibility of PHEA-PLA/PCL and its physical properties make it suitable for the replacement of vessels. In the future, the possibility of functionalizing the material with a variety of molecules, to modulate the inflammatory and coagulative responses, will allow obtaining devices suitable for the replacement of native vessels.

NUPR1, a new target in liver cancer: implication in controlling cell growth, migration, invasion and sorafenib resistance.

Sorafenib, an oral multikinase inhibitor, is the only approved agent for the treatment of advanced hepatocellular carcinoma (HCC). However, its benefits are modest, and as its mechanisms of action remain elusive, a better understanding of its anticancer effects is needed. Based on our previous study results, we investigated here the implication of the nuclear protein 1 (NUPR1) in HCC and its role in sorafenib treatment. NUPR1 is a stress-inducible protein that is overexpressed in various malignancies, but its role in HCC is not yet fully understood. We found that NUPR1 expression was significantly higher in primary human HCC samples than in the normal liver. Knockdown of NUPR1 significantly increased cell sensitivity to sorafenib and inhibited the cell growth, migration and invasion of HCC cells, both in vitro and in vivo. Moreover, NUPR1 silencing influenced the expression of RELB and IER3 genes. Unsurprisingly, RELB and IER3 knockdown also inhibited HCC cell viability, growth and migration. Using gene expression profiling of HCC cells following stable NUPR1 knockdown, we found that genes functionally involved in cell death and survival, cellular response to therapies, lipid metabolism, cell growth and proliferation, molecular transport and cellular movement were mostly suppressed. Network analysis of dynamic gene expression identified NF-κB and ERK as downregulated gene nodes, and several HCC-related oncogenes were also suppressed. We identified Runt-related transcription factor 2 (RUNX2) gene as a NUPR1-regulated gene and demonstrated that RUNX2 gene silencing inhibits HCC cell viability, growth, migration and increased cell sensitivity to sorafenib. We propose that the NUPR1/RELB/IER3/RUNX2 pathway has a pivotal role in hepatocarcinogenesis. The identification of the NUPR1/RELB/IER3/RUNX2 pathway as a potential therapeutic target may contribute to the development of new treatment strategies for HCC management.

Parthenolide generates reactive oxygen species and autophagy in MDA-MB231 cells. A soluble parthenolide analogue inhibits tumour growth and metastasis in a xenograft model of breast cancer.

Triple-negative breast cancers (TNBCs) are clinically aggressive forms associated with a poor prognosis. We evaluated the cytotoxic effect exerted on triple-negative MDA-MB231 breast cancer cells both by parthenolide and its soluble analogue dimethylamino parthenolide (DMAPT) and explored the underlying molecular mechanism. The drugs induced a dose- and time-dependent decrement in cell viability, which was not prevented by the caspase inhibitor z-VAD-fmk. In particular in the first hours of treatment (1-3 h), parthenolide and DMAPT strongly stimulated reactive oxygen species (ROS) generation. The drugs induced production of superoxide anion by activating NADPH oxidase. ROS generation caused depletion of thiol groups and glutathione, activation of c-Jun N-terminal kinase (JNK) and downregulation of nuclear factor kB (NF-kB). During this first phase, parthenolide and DMAPT also stimulated autophagic process, as suggested by the enhanced expression of beclin-1, the conversion of microtubule-associated protein light chain 3-I (LC3-I) to LC3-II and the increase in the number of cells positive to monodansylcadaverine. Finally, the drugs increased RIP-1 expression. This effect was accompanied by a decrement of pro-caspase 8, while its cleaved form was not detected and the expression of c-FLIPS markedly increased. Prolonging the treatment (5-20 h) ROS generation favoured dissipation of mitochondrial membrane potential and the appearance of necrotic events, as suggested by the increased number of cells positive to propidium iodide staining. The administration of DMAPT in nude mice bearing xenografts of MDA-MB231 cells resulted in a significant inhibition of tumour growth, an increment of animal survival and a marked reduction of the lung area invaded by metastasis. Immunohistochemistry data revealed that treatment with DMAPT reduced the levels of NF-kB, metalloproteinase-2 and -9 and vascular endothelial growth factor, while induced upregulation of phosphorylated JNK. Taken together, our data suggest a possible use of parthenolide for the treatment of TNBCs.

Histologic effects of University of Wisconsin two-layer method preservation of rat pancreas.

Marginal donors represent a poorly utilized source of organs for transplantation despite their availability. The key is to reduce the ischemic damage in the effort to improve organ quality. This study investigated the histologic effects after in situ perfusion of preservation with a two-layer method compared with the classic University of Wisconsin preservation in term of tissue integrity and number of viable exocrine cells in the rat pancreas both after exsanguination and at 8 weeks of cryopreservation. Pancreata harvested from 60 rats were collected using 3 methods: two-layer method following University of Wisconsin perfusion; exsanguination; and classic University of Wisconsin perfusion/storage. In addition to histologic analysis of collected pancreata, we analyzed the number of CK19(+) cells and their viability using chi-square tests with values P < .05 considered to be significant. Rat pancreas histology showed as University of Wisconsin in situ perfusion and preservation by the two-layer method to be more effective to maintain the morphologic integrity of both exocrine and endocrine tissues. There were a larger number of CK19(+) cells with good viability. Moreover, the effects of oxygenation were visible in pancreas biopsies preserved after exsanguination. In situ University of Wisconsin perfusion and preservation for 240 minutes with the two-layer method yielded greater numbers and viability of CK19(+) cells even after 8 weeks of cryopreservation.

Expansion of intracellular IFN-γ positive lymphocytes during Mycoplasma agalactiae infection in sheep.

A method to assess the expansion of antigen-specific intracellular IFN-γ positive T cell subsets during the infection will be helpful for a better understanding of mycoplasmal infections physiopathology in the sheep. We analysed the percentage of antigen-specific lymphocytes positive for intracellular IFN-γ during the infection of sheep with Mycoplasma agalactiae by culturing peripheral blood mononuclear cells of infected or uninfected animals with irradiated M. agalactiae. The expansion of antigen-specific IFN-γ positive lymphocytes in infected sheep was initially sustained by CD4(+) T cells at day 15 after infection, when antigen specific IgG start to be detectable, followed by CD8/IFN-γ double positive cells. γδ T-cells were not expanded at any time point analysed. IFNγ(+) T cells disappear 60 days after infection, suggesting that antigen specific IFNγ(+) T cells, mainly detected in the early phase of the disease, could be useful to understand the role of cell-mediated immunity during M. agalactiae infection.

Embryonic and foetal Islet-1 positive cells in human hearts are also positive to c-Kit.

During embryogenesis, the mammalian heart develops from a primitive heart tube originating from two bilateral primary heart fields located in the lateral plate mesoderm. Cells belongings to the pre-cardiac mesoderm will differentiate into early cardiac progenitors, which express early transcription factors which are also common to the Isl-1 positive cardiac progenitor cells isolated from the developing pharyngeal mesoderm and the foetal and post-natal mice hearts. A second population of cardiac progenitor cells positive to c-Kit has been abundantly isolated from adult hearts. Until now, these two populations have been considered two different sets of progenitor cells present in the heart in different stages of an individual life. In the present study we collected embryonic, foetal and infant hearts, and we tested the hypotheses that c-Kit positive cells, usually isolated from the adult heart, are also present in the intra-uterine life and persist in the adult heart after birth, and that foetal Isl-1 positive cells are also positive to c-Kit. Using immunohistochemistry we studied the temporal distribution of Isl-1 positive and c-Kit/CD105 double positive cells, and by immunofluorescence and confocal analysis we studied the co-localization of c-Kit and Isl-1 positive cells. The results indicated that cardiomyocytes and interstitial cells were positive for c-Kit from the 9th to the 19th gestational week, that cells positive for both c-Kit and CD105 appeared in the interstitium at the 17th gestational week and persisted in the postnatal age, and that the Isl-1 positive cells were a subset of the c-Kit positive population.

Antimicrobial activity of the essential oil of Calamintha nepeta and its constituent pulegone against bacteria and fungi.

The chemical composition of the essential oil of Calamintha nepeta and its antimicrobial activity against Listeria monocytogenes, Bacillus cereus, Salmonella veneziana, S. paratyphi B. S. typhimurium, Fusarium moniliforme, Botrytis cinerea, Aspergillus niger and Pyricularia oryzae have been studied. Moreover the main constituents of the oil (limonene, menthone, pulegone, menthol) have been tested against the same microorganisms. Only pulegone showed antimicrobial activity, particularly against all the Salmonella species.