Investigation on the composition of agarose–collagen i blended hydrogels as matrices for the growth of spheroids from breast cancer cell lines

Three-dimensional (3D) cell culture systems mimic the structural complexity of the tissue microenvironment and are gaining increasing importance as they resemble the extracellular matrix (ECM)–cell and cell–cell physical interactions occurring in vivo. Several scaffold-based culture systems have been already proposed as valuable tools for large-scale production of spheroids, but they often suffer of poor reproducibility or high costs of production. In this work, we present a reliable 3D culture system based on collagen I-blended agarose hydrogels and show how the variation in the agarose percentage affects the physical and mechanical properties of the resulting hydrogel. The influence of the different physical and mechanical properties of the blended hydrogels on the growth, size, morphology, and cell motility of the spheroids obtained by culturing three different breast cancer cell lines (MCF-7, MDA-MB-361, and MDA-MB-231) was also evaluated. As proof of concept, the cisplatin penetration and its cytotoxic effect on the tumor spheroids as function of the hydrogel stiffness were also investigated. Noteworthily, the possibility to recover the spheroids from the hydrogels for further processing and other biological studies has been considered. This feature, in addition to the ease of preparation, the lack of cross-linking chemistry and the high re-producibility, makes this hydrogel a reliable biomimetic matrix for the growth of 3D cell structures

Oxidative stress and multi-organel damage induced by two novel phytocannabinoids, cbdb and cbdp, in breast cancer cells

Over the last few years, much attention has been paid to phytocannabinoids derived from Cannabis for their therapeutic potential. ∆9-tetrahydrocannabinol (∆9-THC) and cannabidiol (CBD) are the most abundant compounds of the Cannabis sativa L. plant. Recently, novel phytocannabinoids, such as cannabidibutol (CBDB) and cannabidiphorol (CBDP), have been discovered. These new molecules exhibit the same terpenophenolic core of CBD and differ only for the length of the alkyl side chain. Roles of CBD homologs in physiological and pathological processes are emerging but the exact molecular mechanisms remain to be fully elucidated. Here, we investigated the biological effects of the newly discovered CBDB or CBDP, compared to the well-known natural and synthetic CBD (nat CBD and syn CBD) in human breast carcinoma cells that express CB receptors. In detail, our data demonstrated that the treatment of cells with the novel phytocannabinoids affects cell viability, increases the production of reactive oxygen species (ROS) and activates cellular pathways related to ROS signaling, as already demonstrated for natural CBD. Moreover, we observed that the biological activity is significantly increased upon combining CBD homologs with drugs that inhibit the activity of enzymes involved in the metabolism of endocannabinoids, such as the monoacylglycerol lipase (MAGL) inhibitor, or with drugs that induces the activation of cellular stress pathways, such as the phorbol ester 12-myristate 13-acetate (PMA)

Effect of resveratrol on mitochondrial function: Implications in parkin-associated familiar Parkinson's disease

Mitochondrial dysfunction and oxidative stress occur in Parkinson's disease (PD), but the molecular mechanisms controlling these events are not completely understood. Peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α) is a transcriptional coactivator known as master regulator of mitochondrial functions and oxidative metabolism. Recent studies, including one from our group, have highlighted altered PGC-1α activity and transcriptional deregulation of its target genes in PD pathogenesis suggesting it as a new potential therapeutic target. Resveratrol, a natural polyphenolic compound proved to improve mitochondrial activity through the activation of several metabolic sensors resulting in PGC-1α activation. Here we have tested in vitro the effect of resveratrol treatment on primary fibroblast cultures from two patients with early-onset PD linked to different Park2 mutations. We show that resveratrol regulates energy homeostasis through activation of AMP-activated protein kinase (AMPK) and sirtuin 1 (SIRT1) and raise of mRNA expression of a number of PGC-1α's target genes resulting in enhanced mitochondrial oxidative function, likely related to a decrease of oxidative stress and to an increase of mitochondrial biogenesis. The functional impact of resveratrol treatment encompassed an increase of complex I and citrate synthase activities, basal oxygen consumption, and mitochondrial ATP production and a decrease in lactate content, thus supporting a switch from glycolytic to oxidative metabolism. Moreover, resveratrol treatment caused an enhanced macro-autophagic flux through activation of an LC3-independent pathway. Our results, obtained in early-onset PD fibroblasts, suggest that resveratrol may have potential clinical application in selected cases of PD-affected patients

Current Trends on MA Translation Courses in the UK: Changing Assessment Practices on Core Translation Modules

Assessment underpins all forms of translator training and is an essential element of any teaching and learning process. By looking at university assessment practices, we can gain an insight into current understandings about the nature of translation practice and what issues are foregrounded in translator education. This paper presents the findings of the second stage of our research into assessment practices on core translation modules of MA Translation courses offered in the UK, and follows on from a preliminary case study conducted at the University of Westminster in 2015 and 2016 with MA Translation tutors and students (n=16; n=53). The research presented in this paper was carried out via documentary research into all universities offering MA Translation courses (n=27) and via a survey which asked a representative of each UK university to fill out a questionnaire. The questionnaire was completed by 55% of universities (n=15). The areas explored include universities' learning outcomes, assessment instruments and criteria. In the light of the data gathered, our study aims to discover if and to what extent current assessment practices on the core translation modules reflect the competence-based understandings of the translation process and have adopted new forms of assessment

Short- and Long-Term Biomarkers for Bacterial Robustness: A Framework for Quantifying Correlations between Cellular Indicators and Adaptive Behavior

The ability of microorganisms to adapt to changing environments challenges the prediction of their history-dependent behavior. Cellular biomarkers that are quantitatively correlated to stress adaptive behavior will facilitate our ability to predict the impact of these adaptive traits. Here, we present a framework for identifying cellular biomarkers for mild stress induced enhanced microbial robustness towards lethal stresses. Several candidate-biomarkers were selected by comparing the genome-wide transcriptome profiles of our model-organism Bacillus cereus upon exposure to four mild stress conditions (mild heat, acid, salt and oxidative stress). These candidate-biomarkers—a transcriptional regulator (activating general stress responses), enzymes (removing reactive oxygen species), and chaperones and proteases (maintaining protein quality)—were quantitatively determined at transcript, protein and/or activity level upon exposure to mild heat, acid, salt and oxidative stress for various time intervals. Both unstressed and mild stress treated cells were also exposed to lethal stress conditions (severe heat, acid and oxidative stress) to quantify the robustness advantage provided by mild stress pretreatment. To evaluate whether the candidate-biomarkers could predict the robustness enhancement towards lethal stress elicited by mild stress pretreatment, the biomarker responses upon mild stress treatment were correlated to mild stress induced robustness towards lethal stress. Both short- and long-term biomarkers could be identified of which their induction levels were correlated to mild stress induced enhanced robustness towards lethal heat, acid and/or oxidative stress, respectively, and are therefore predictive cellular indicators for mild stress induced enhanced robustness. The identified biomarkers are among the most consistently induced cellular components in stress responses and ubiquitous in biology, supporting extrapolation to other microorganisms than B. cereus. Our quantitative, systematic approach provides a framework to search for these biomarkers and to evaluate their predictive quality in order to select promising biomarkers that can serve to early detect and predict adaptive traits

English in Translation Studies: Methodological perspectives.

This volume contains a selection of papers on the use and teaching of English in translation presented at the 9th ESSE (European Society for the Study of English) Conference, held in Aarhus, Denmark, on 22-26 August 2008. The volume builds on the notion of English language and translation competence to discuss methodological issues and teaching methods in the field of translation training. The eight contributions raise issues concerned, among others, with the academic-vocational dichotomy in specialized language training, the issue of translator identity, the psychological implications of resorting to creativity in translation, the assessment in translation teaching and the effects of the spreading of English as a Lingua Franca. The volume also offers an integrated model of translation competence which gives prominence to the translation process on the background of interrelated competencies, and which can be used to assess the acquisition of translation competence from both the quantitative and the qualitative viewpoint. The book is directed to an audience of translation scholars and practitioners interested in the evolving field of English used as a language of translation

Lhermitte-Duclos disease: MR diffusion and spectroscopy

AIM: To describe two cases of Lhermitte-Duclos disease studied with Diffusion MRI and MRi Spectroscopy. MATERIALS AND METHODS: Two patients showing a space-occupying lesion in the cerebellar hemisphere were studied by conventional MRI associated with Diffusion MRI and Spectroscopy. RESULTS: Conventional MRI showed the typical morphological pattern of Lhermitte-Duclos disease with a slow-growing cerebellar mass mainly involving the cortex and non-enhancing thickened cerebellar folia. The mass exerted mild compression of the IV ventricle causing hydrocephalus On Diffusion imaging showed the lesion to be hyperintense due to residual T2-activity and hypointense on the ADC map. Spectroscopy was typical with an increased lactate peak and a reduced choline peak, indicating abnormal anaerobic glycolysis and demyelination, respectively. DISCUSSION: Lhermitte-Duclos disease is a neurological condition which presents several signs and symptoms related to the cerebellar mass. Cowden's syndrome, an autosomal dominant neoplastic disorder with malignant features is associated in 40% of cases. MRI associated with functional examinations such as Diffusion Imaging and Spectroscopy is the imaging modality of choice in LDD patients. They allow a definite diagnosis as the neuroradiological findings correlate well with molecular biology theories regarding the malformative nature of dysplastic cerebellar gangliocytoma

MULTIMODAL DIAGNOSTIC APPROACH IN ISCHEMIC STROKE

Aim In our study, we explain the application of the new multimodal neuroradiological protocol for acute ischemic stroke. Material and methods Diffusion weighted Imaging, Perfusion weighted Imaging and Magnetic Resonance Angiography have become an important tool for the identification of early ischemic Injury. Results DWI lesions present irreversible damaged tissue – the infarct core. PWI maps demonstrate the area of minor perfusion in patients with acute vessel occlusion. Combined DWI and PWI has the potential to be more powerful than either study alone in predicting infarct outcome apart from CT and MRI. Discussion and conclusions In acute ischemic stroke multimodal MRI diagnostic approach is fundamental for a very early diagnosis and for the trombolytic therapy with recanalization of the occluded cerebral vessel