The influence of a stressful microenvironment on tumor exosomes: a focus on the DNA cargo

Exosomes secreted by tumor cells, through the transport of bioactive molecules, reprogram the surroundings, building a microenvironment to support the development of the tumor. The discovery that exosomes carry genomic DNA reflecting that of the tumor cell of origin has encouraged studies to use them as non-invasive biomarkers. The exosome-mediated transfer of oncogenes suggested a new mechanism of malignant transformation that could play a role in the formation of metastases. Several studies have examined the role of tumor exosomes on the modulation of the tumor microenvironment, but relatively few have been directed to assess how stressful stimuli can influence their production and cargo. Understanding the changes in exosome loads and the production pattern of the stressed tumor cell may uncover actionable mechanisms responsible for tumor progression

Mandibular osteosarcoma in a goat

Introduction-A few large surveys on tumour prevalence in goats indicate that tumours in general are quite common in this species. Reviews of neoplastic diseases in goats indicate a prevalence ranging from 0,8 to 11%. However, osteogenic tumours arising from facial bones in goats are rare. Regarding oral localization only a few mesenchymal tumours have been described arising from the gengiva. Case presentation-A 4-year old, female crossbred goat was referred with a history of dysorexia and a slow growing painful mass on the face. On physical examination the animal showed poor body condition and the left side of the face was deformed by a voluminous mass which, at the inspection of the oral cavity, displaced the maxillary teeth. Differential diagnoses included os-teomyelitis and benign (osteoma, chondroma, ossifying and non ossifying fibroma, odontogenic tumours) as well as malignant (osteosarcoma, chondrosarcoma) mesenchymal tumours arising from either the connective tissue and bone. The goat was euthanized because of the extension of the lesion and a complete necropsy was performed. Grossly, the face was deformed by the presence of a hard mass arising from the branch of the left mandible. Histologically the oral mass was composed of heterogeneous proliferation of malignant osteoblasts intermigled with brightly eosinophilic strands or island of osteoid matrix. Neoplastic cells, interpreted as malignant osteoblasts, were characterized by plump to round or spindle-shape morphology, with moderate basophilic cytoplasm and an eccentrically located voluminous nucleus containing a large prominent nucleolus. Mitotic figures were found and were either bipolar and atypical. At necropsy no metastases were found and the final diagnosis was non-metastasizing mandibular osteoblastic osteosarcoma. Conclusion-In conclusion, regardless the type of tumour, the goat was euthanized because of the extension and the severity of the lesion. Necropsy and histological examination were necessary to correctly classify the tumour as a non-metastasizing mandibular osteosarcoma

Mitochondrial F0F1-ATP synthase is a molecular target of 3-iodothyronamine, an endogenous metabolite of thyroid hormone

Background & Purpose:\u2002 T1AM is a thyronamine derivative of thyroid hormone acting as a signalling molecule via non-genomic effectors and can reach intracellular targets. In light of the importance of F(0) F(1) -ATPsynthase as a target in drug development, T1AM interaction with the enzyme is demonstrated by its effects on the activity and a model of binding locations is depicted. Experimental Approach:\u2002 Kinetic analyses were performed on F(0) F(1) -ATPsynthase in sub-mitochondrial particles and soluble F(1) -ATPase. Activity assays and immunodetection of the inhibitor protein IF(1) were used and combined with molecular docking analyses. In situ respirometric analysis of T1AM effect was investigated on H9c2 cardiomyocytes. Key Results:\u2002 T1AM is a non-competitive inhibitor of F(0) F(1) -ATPsynthase whose binding is mutually exclusive with that of the inhibitors IF(1) and aurovertin B. Distinct T1AM binding sites are consistent with results from both kinetic and docking analyses: at low nanomolar concentrations, T1AM binds to a high affinity-region likely located within the IF(1) binding site, causing IF(1) release; at higher concentrations, T1AM binds to a low affinity-region likely located within the aurovertin binding cavity and inhibits enzyme activity. Low nanomolar concentrations of T1AM elicit in cardiomyocytes an increase in ADP-stimulated mitochondrial respiration indicative for an activation of F(0) F(1) -ATPsynthase consistent with displacement of endogenous IF(1, ) thereby reinforcing the in vitro results. Conclusions & Implications:\u2002 The T1AM effects upon F(0) F(1) -ATPsynthase are twofold: IF(1) displacement and enzyme inhibition. By targeting F(0) F(1) -ATPsynthase within mitochondria T1AM might affect cell bioenergetics with a positive effect on mitochondrial energy production at low endogenous concentration. T1AM putative binding locations overlapping with IF(1) and aurovertin binding sites are depicted

Skeletal muscle oxidative function in vivo and ex vivo in athletes with marked hypertrophy from resistance training

Oxidative function during exercise was evaluated in 11 young athletes with marked skeletal muscle hypertrophy induced by long-term resistance training (RTA, body mass 102.67.3 kg, meanSD) and 11 controls (CTRL, body mass 77.86.0). Pulmonary O2 uptake (V\u27O2) and vastus lateralis muscle fractional O2 extraction (by near-infrared spectroscopy) were determined during an incremental cycle ergometer (CE) and one-leg knee-extension (KE) exercise. Mitochondrial respiration was evaluated ex vivo by high-resolution respirometry in permeabilized vastus lateralis fibers obtained by biopsy. Quadriceps femoris muscle cross sectional area, volume (determined by magnetic resonance imaging) and strength were greater in RTA vs. CTRL (by ~40%, ~33% and ~20%, respectively). V\u27O2peak during CE was higher in RTA vs. CTRL (4.050.64 L min-1 vs. 3.560.30)no difference between groups was observed during KE. The O2 cost of CE exercise was not different between groups. When divided per muscle mass (for CE) or quadriceps muscle mass (for KE) V\u27O2peak was lower (by 15-20%) in RTA vs. CTRL. Vastus lateralis fractional O2 extraction was lower in RTA vs. CTRL at all work rates, both during CE and KE. RTA had higher ADP-stimulated mitochondrial respiration (56.723.7 pmolO2s-1mg-1 ww) vs. CTRL (35.710.2), and a tighter coupling of oxidative phosphorylation. In RTA the greater muscle mass and maximal force, and the enhanced mitochondrial respiration seem to compensate for the hypertrophy-induced impaired peripheral O2 diffusion. The net results are an enhanced whole body oxidative function at peak exercise, and unchanged efficiency and O2 cost at submaximal exercise, despite a much greater body mas

A human neuronal model of Niemann Pick C disease developed from stem cells isolated from patient's skin.

Niemann Pick C (NPC) disease is a neurovisceral lysosomal storage disorder due to mutations in NPC1 or NPC2 genes, characterized by the accumulation of endocytosed unesterified cholesterol, gangliosides and other lipids within the lysosomes/late endosomes. Even if the neurodegeneration is the main feature of the disease, the analysis of the molecular pathways linking the lipid accumulation and cellular damage in the brain has been challenging due to the limited availability of human neuronal models.The aim of this study was to develop a human neuronal model of NPC disease by inducing neuronal differentiation of multipotent adult stem cells (MASC) isolated from NPC patients.Stem cells were isolated from 3 NPC patients and 3 controls both from skin biopsies and previously established skin fibroblast cultures. Cells were induced to differentiate along a neuronal fate adapting methods previously described by Beltrami et al, 2007. The surface immunophenotype of stem cells was analyzed by FACS. Stem cell and neuronal markers expression were evaluated by immunofluorescence. Intracellular accumulation of cholesterol and gangliosides were assessed by filipin staining and immunofluorescence, respectively. A morphometric analysis was performed using a Neurite outgrowth image program.After 3 passages in selective medium, MASC isolated either from skin biopsies or previously established skin fibroblast cultures displayed an antigenic pattern characteristic of mesenchymal stem cells and expressed the stem cell markers Oct-4, Nanog, Sox-2 and nestin. A massive lysosomal accumulation of cholesterol was observed only in cells isolated from NPC patients. After the induction of neural differentiation, remarkable morphologic changes were observed and cells became positive to markers of the neuronal lineage NeuN and MAP2. Differentiated cells from NPC patients displayed characteristic features of NPC disease, they showed intracellular accumulation of unesterified cholesterol and GM2 ganglioside and presented morphological differences with respect to cells derived from healthy donors.In conclusion, we generated a human neuronal model of NPC disease through the induction of differentiation of stem cells obtained from patient's easily accessible sources. The strategy described here may be applied to easily generate human neuronal models of other neurodegenerative diseases

Tuberculosis in roe deer from Spain and Italy

TUBERCULOSIS (TB) is a chronic infectious disease caused by bacteria of the genus Mycobacterium (Grange and others 1990). The detection of wildlife reservoirs of disease is important, particularly in areas where there is a relatively low incidence of the disease in domestic animals. Tuberculosis cases in roe deer (Capreolus capreolus) are reported only sporadically, despite the wide distribution and the abundance of this cervid. Roe deer with TB have been reported in Germany (Schmidt 1938), Switzerland (Bouvier 1963), France (Zanella and others 2008) and the UK (Gunning 1985, Delahay and others 2007). This short communication is the first report of TB in roe deer in Spain and Italy, and discusses the implications of these findings for wildlife and livestock disease control. The prevalence of mycobacterial infections, such as TB and paratuberculosis, seems to be increasing in Spain. Wildlife species may act as disease reservoirs, so this short communication also elucidates the epidemiology of mycobacterial infections in species such as roe deer