Epithelial‐to‐mesenchymal transition: immunohistochemical investigation of related molecules in canine cutaneous epithelial tumours

Background –  Epithelial‐to‐mesenchymal transition (EMT) is a multistep process, important in tumour invasion and metastasis, characterized by loss of epithelial markers, redistribution of β‐catenin and gain of mesenchymal markers. Hyposthesis/Objectives –  Our aim was to investigate the immunohistochemical aberrant expression of cytokeratin, vimentin, survivin and heat shock protein 72 (Hsp72) in canine cutaneous epithelial tumours, to understand the association of expression of these molecules with features of malignancy and their role in the EMT phenotype. Methods –  Ten canine squamous cell carcinomas (SCCs; one with lymph node metastasis), 30 canine hair follicle tumours (six pilomatricomas, eight infundibular keratinizing acanthomas, six trichoepitheliomas and 10 trichoblastomas) and five normal skin samples were investigated by immunohistochemistry using specific anti‐vimentin, ‐cytokeratin, ‐survivin and ‐Hsp72 antibodies. A semi‐quantitative method was used to analyse the results, as follows: 0 to <5%; ≥5 to <10%; ≥10 to <25%; and ≥25% of positive cells. Immunofluorescence was performed to investigate survivin–vimentin and survivin–Hsp72 colocalization in selected SCCs. Results –  In malignant hair follicle tumours and SCCs, a reduced intensity of cytokeratin and increased survivin and Hsp72 expression were observed. In SCCs, loss of cytokeratin expression and vimentin immunolabelling, suggestive of the EMT phenotype, were evident in <5% of neoplastic cells in the front of tumour invasion. In the same areas, strong nuclear survivin and cytoplasmic Hsp72 staining was evident, often colocalizing. Only a few neoplastic cells in the front of tumour invasion showed vimentin–survivin colocalization. Conclusions and clinical importance –  A possible simultaneous involvement of survivin and Hsp72 in tumour invasion and the multistep process of EMT of cutaneous epithelial tumours of dogs is suggested.     

Location and stability of a recombinant ovine prion protein in synthetic humic-like mineral complexes

Location and stability of a recombinant prion protein (recPrP) and its interaction with humic-like complexes were investigated by low-temperature ashing (LTA), thermal gravimetric (TG), and scanning electron microscopy (SEM) analyses. Humic-like complexes were obtained by abiotic polymerization of catechol, one of the possible precursors of soil humic matter, through the catalysis of birnessite, a manganese oxide common in soil environment. The recPrP was immobilized in organomineral complexes via sorption or entrapment. Complexes were treated by LTA, allowing the controlled removal of organic matter layer by layer, from the external to the internal side, with minimal disturbance of mineral constituents. Thermal gravimetric and SEM analyses were performed on specimens before and after LTA treatment. Entrapped recPrP, compared with sorbed, resulted less easily accessible to LTA treatment and showed a higher thermal stability by TGA analyses. On the basis of these findings, we hypothesize that the processes leading to newly formed organic complexes can enhance prion stability in soil and thus influence the environmental diffusion of infectivity.     

BAX and BAK regulation of endoplasmic reticulum Ca2+: a control point for apoptosis

BAX and BAK are "multidomain" proapoptotic proteins that initiate mitochondrial dysfunction but also localize to the endoplasmic reticulum (ER). Mouse embryonic fibroblasts deficient for BAX and BAK (DKO cells) were found to have a reduced resting concentration of calcium in the ER ([Ca2+]er) that results in decreased uptake of Ca2+ by mitochondria after Ca2+ release from the ER. Expression of SERCA (sarcoplasmic-endoplasmic reticulum Ca2+ adenosine triphosphatase) corrected [Ca2+]er and mitochondrial Ca2+ uptake in DKO cells, restoring apoptotic death in response to agents that release Ca2+ from intracellular stores (such as arachidonic acid, C2-ceramide, and oxidative stress). In contrast, targeting of BAX to mitochondria selectively restored apoptosis to "BH3-only" signals. A third set of stimuli, including many intrinsic signals, required both ER-released Ca2+ and the presence of mitochondrial BAX or BAK to fully restore apoptosis. Thus, BAX and BAK operate in both the ER and mitochondria as an essential gateway for selected apoptotic signals.     

Assessment of Eco-Physiological Performance of Quercus ilex L. Leaves in Urban Area by an Integrated Approach

Bioretention cells, also known as raingardens, are increasingly being constructed as a means to collect, infiltrate, and treat stormwater runoff. There are concerns, however, about how stormwater management practices might function in terms of infiltration and pollutant removal as they age. Saturated hydraulic conductivity (K _sat) values were obtained for eight cells in 2006 and again for three of those cells in 2010 using an infiltrometer. A strong positive correlation of mean K _sat with service time was observed (slope = 10.2?±?2.4 cm/h per year, R ² = 0.67). Results from metals analyses of bioretention media cores collected from six bioretention cells showed the expected trend of Cu and Zn enrichment at the surface while Cd was detected only in one out of 72 media samples analyzed. Sorption isotherms from batch testing of field media samples (T = 22.5 °C, pH = 7.2) were used to estimate metal sorption capacities based on representative stormwater Cd and Zn concentrations. Cu was not considered, as very little of the metal is dissolved under these conditions (22.8?±?7.1 %). The mean equilibrium sorption capacities for Cd (10.2?±?3.1 mg/kg) and Zn (294.9?±?14.9 mg/kg) far exceeded observed levels in the bioretention media such that the remaining sorption capacity was ≥83 % for Zn and ≥90 % for Cd for the cells. Overall, the results of this investigation suggest that bioretention cells can provide many years of effective infiltration (>6 years) and metals removal performance (>25 years).     

Gamma-ray flares from the Crab Nebula

A young and energetic pulsar powers the well-known Crab Nebula. Here, we describe two separate gamma-ray (photon energy greater than 100 mega-electron volts) flares from this source detected by the Large Area Telescope on board the Fermi Gamma-ray Space Telescope. The first flare occurred in February 2009 and lasted approximately 16 days. The second flare was detected in September 2010 and lasted approximately 4 days. During these outbursts, the gamma-ray flux from the nebula increased by factors of four and six, respectively. The brevity of the flares implies that the gamma rays were emitted via synchrotron radiation from peta-electron-volt (10(15) electron volts) electrons in a region smaller than 1.4 × 10(-2) parsecs. These are the highest-energy particles that can be associated with a discrete astronomical source, and they pose challenges to particle acceleration theory.     

Fifty Years on: Long‐term Patterns of Land Sensitivity to Desertification in Italy

The Mediterranean region has been regarded as a critical hotspot for desertification due to the impact of soil degradation, the land‐use changes and the climate variations. Few large‐scale studies have been devoted to analyse trends in land sensitivity to desertification in the northern Mediterranean basin. The present paper contributes to this deserving issue by quantifying the level of land sensitivity to desertification in Italy at seven points between 1960 and 2010 at a fine spatial scale. The approach used followed the Environmentally Sensitive Area scheme that assesses changes in four key themes (climate, soil, vegetation and land management) related to land degradation processes. Italian land was classified into four levels of sensitivity to desertification (non‐affected, potentially affected, fragile and critical) according to the Environmentally Sensitive Area framework. Interestingly, although land surface area classified as ‘fragile’ and ‘critical’ grew homogeneously in Italy between 1960 and 1990, the increase observed in the most recent time period was spatially clustered and contributed to reverse the polarisation in ‘structurally vulnerable’ and ‘non‐affected’ regions observed in Italy. The paper discussed these trends in the light of socioeconomic changes that occurred in Italy after World War II. Copyright © 2013 John Wiley & Sons, Ltd.