The mechano-chemistry of cytoskeletal force generation

In this communication, we propose a model to study the non-equilibrium process by which actin stress fibers develop force in contractile cells. The emphasis here is on the non-equilibrium thermodynamics, which is necessary to address the mechanics as well as the chemistry of dynamic cell contractility. In this setting we are able to develop a framework that relates (a) the dynamics of force generation within the cell and (b) the cell response to external stimuli to the chemical processes occurring within the cell, as well as to the mechanics of linkage between the stress fibers, focal adhesions and extra-cellular matrix.Comment: 22 pages, 6 figures, 1 table, accepted in Biomechanics and Modeling in Mechanobiolog

Dietary Polyphenols and Their Effects on Cell Biochemistry and Pathophysiology 2014

Epidemiological studies suggest that high dietary intake of phytochemicals and in particular of polyphenols is associated with decreased risk of a multitude of diseases states including cardiovascular disease, cancer, and neurodegenerative diseases. As oxidative stress is involved in all these pathological conditions, the antioxidant properties of polyphenols and other natural compounds have attracted the interest of many authors

Redistribution of DNA topoisomerase II beta after in vitro stabilization of human erythroleukemic nuclei by heat or Cu++ revealed by confocal microscopy.

Using confocal laser scanning microscope and a monoclonal antibody we have examined by means of indirect immunofluorescence techniques the distribution of DNA topoisomerase II beta (the 180-kDa nucleolar isoform of topoisomerase II) following stabilization of isolated nuclei by exposure to moderate heat (37 degrees or 42 degrees C) or Cu++. In intact cells the antibody specifically decorated the nucleoli. The same pattern was maintained if nuclei were incubated at 0 degree C in a buffer containing spermine/spermidine/KCl or stabilized by means of 0.5 mM Cu++ for 10 minutes at 0 degree C in the same buffer. On the contrary, if stabilization was performed by incubating the nuclei either at 37 degrees or 42 degrees C, the immunoreactivity dispersed all over the nucleus, forming numerous speckles. This phenomenon was not detected if, in addition to spermine/spermidine/KCl, the incubation buffer also contained 5 mM Mg++ and the temperature was 37 degrees C. If the stabilization was performed at 42 degrees C, Mg++ failed to maintain the original distribution of DNA topoisomerase II beta, as seen in intact cells. The analysis on 2-D optical section showed the alteration of the nucleolar profile, particularly at 37 degrees C, even when the samples were treated with Mg++. The 3-D reconstruction figured out the irregularity of the surface at 37 degrees C and the variations of the volume occupied by the fluorescent figures. These were in close proximity to each other both in intact cells and in 0 degree C incubated nuclei; they showed a certain degree of shrinkage in 0 degree C plus Cu++ exposed samples (-20\% of the volume), and, on the contrary, the labeled structures were scattered in a volume increased two- or threefold when exposed to 37 degrees or 42 degrees C, respectively. The addition of Mg++ restored the original spatial relationship and volume at 37 degrees C, but not at 42 degrees C, where the volumetric analysis showed an increase of about 50\%. Our results demonstrate that heat stabilization of isolated nuclei in a buffer without Mg++ (i.e., a technique often employed to prepare the nuclear matrix or scaffold) cannot be considered an optimal procedure to maintain the original distribution of protein within the nucleus

La ricostruzione della balista di Vitruvio

Viene riportato il lavoro svolto durante le esercitazioni del primo anno dell’insegnamento Laboratorio di storia dell’ingegneria meccanica, tenuto presso la Facoltà di Ingegneria dell’Università di Bologna. La descrizione della balista di Vitruvio offre un esempio di progettazione modulare basato sul valore del diametro degli elementi elastici. La lettura critica del testo a noi pervenuto porta ad esaminare i disegni del Valturio, quelli di Fra Giocondo e le conclusioni del Barbaro, anche in relazione alle macchine scolpite sulla colonna Traiana. La funzione che interpola i dati di Vitruvio garantisce che macchine di diversa taglia, sollecitate nello stesso modo, a parità di alzo, abbiano la stessa gittata e quindi che il proietto abbia uguale velocità iniziale. Si trova anche che la palla da lanciare può essere usata come calibro per i fori per i quali passa la matassa elastica. Si tenta una argomentata ricostruzione della macchina, basandosi su calcoli, prove sui materiali e disegni. La struttura della macchina viene confrontata con quella di recenti ricostruzioni rintracciate in rete ed in letteratura. Si passa quindi allo “studio”, al disegno del complessivo e ai disegni costruttivi. In attesa della costruzione reale … si simulano le prove di lancio

Towards a regional ocean forecasting system for the IBI (Iberia-Biscay-Ireland area): developments and improvements within the ECOOP project framework

The regional ocean operational system remains a key element in downscaling from large scale (global or basin scale) systems to coastal ones. It enables the transition between systems in which the resolution and the resolved physics are quite different. Indeed, coastal applications need a system to predict local high frequency events (inferior to the day) such as storm surges, while deep sea applications need a system to predict large scale lower frequency ocean features. In the framework of the ECOOP project, a regional system for the Iberia-Biscay-Ireland area has been upgraded from an existing V0 version to a V2. This paper focuses on the improvements from the V1 system, for which the physics are close to a large scale basin system, to the V2 for which the physics are more adapted to shelf and coastal issues. Strong developments such as higher regional physics resolution in the NEMO Ocean General Circulation Model for tides, non linear free surface and adapted vertical mixing schemes among others have been implemented in the V2 version. Thus, regional thermal fronts due to tidal mixing now appear in the latest version solution and are quite well positioned. Moreover, simulation of the stratification in shelf areas is also improved in the V2

NADPH oxidase-4 and MATER expressions in granulosa cells: Relationships with ovarian aging

Aims Relevant roles in follicular development and ovulation are played by maternal antigen that embryos require (MATER), product of a maternal effect gene, and by reactive oxygen species (ROS), indispensable for the induction of ovulatory genes. At the moment, the relationship between these two biological systems and their involvement in the ovarian aging have not been still clarified. The aim of the current experimental study was to analyse the age-related changes of the MATER and NOX proteins. Materials and methods MATER and ROS homeostasis was studied in granulosa cells (GCs) and cumulus cells (CCs) of infertile patients who undergone oocyte retrieval for in vitro fertilization cycles using Western blot and confocal immunofluorescence analysis. Samples were obtained from subjects with age\ua0 65\ua040\ua0years (cases) and with age\ua0 64\ua037\ua0years (controls). Key findings The expression pattern of MATER and NOX observed in GCs was not different from that observed in CCs. High levels of both proteins were detected in the control samples. A significant lower expression of both MATER and NOX4 was observed in the case versus control samples. Significance The expression of MATER and NOX4 proteins are closely related to the follicular development and ovulation with particular regard for ovarian aging

Natural compounds as modulators of NADPH oxidases.

Reactive oxygen species (ROS) are cellular signals generated ubiquitously by all mammalian cells, but their relative unbalance triggers also diseases through intracellular damage to DNA, RNA, proteins, and lipids. NADPH oxidases (NOX) are the only known enzyme family with the sole function to produce ROS. The NOX physiological functions concern host defence, cellular signaling, regulation of gene expression, and cell differentiation. On the other hand, increased NOX activity contributes to a wide range of pathological processes, including cardiovascular diseases, neurodegeneration, organ failure, and cancer. Therefore targeting these enzymatic ROS sources by natural compounds, without affecting the physiological redox state, may be an important tool. This review summarizes the current state of knowledge of the role of NOX enzymes in physiology and pathology and provides an overview of the currently available NADPH oxidase inhibitors derived from natural extracts such as polyphenols

Human amniotic fluid-derived and dental pulp-derived stem cells seeded into collagen scaffold repair critical-size bone defects promoting vascularization

INTRODUCTION: The main aim of this study is to evaluate potential human stem cells, such as dental pulp stem cells and amniotic fluid stem cells, combined with collagen scaffold to reconstruct critical-size cranial bone defects in an animal model. METHODS: We performed two symmetric full-thickness cranial defects on each parietal region of rats and we replenished them with collagen scaffolds with or without stem cells already seeded into and addressed towards osteogenic lineage in vitro. After 4 and 8 weeks, cranial tissue samples were taken for histological and immunofluorescence analysis. RESULTS: We observed a new bone formation in all of the samples but the most relevant differences in defect correction were shown by stem cell–collagen samples 4 weeks after implant, suggesting a faster regeneration ability of the combined constructs. The presence of human cells in the newly formed bone was confirmed by confocal analysis with an antibody directed to a human mitochondrial protein. Furthermore, human cells were found to be an essential part of new vessel formation in the scaffold. CONCLUSION: These data confirmed the strong potential of bioengineered constructs of stem cell–collagen scaffold for correcting large cranial defects in an animal model and highlighting the role of stem cells in neovascularization during skeletal defect reconstruction

DEVELOPMENT OF A NOVEL METHOD FOR AMNIOTIC FLUID STEM CELL STORAGE

Background - Current procedures for collection of human Amniotic Fluid Stem Cells (hAFSCs) imply that amniotic fluid cells were cultured in flask for two weeks, than can be devoted to research purpose. However, hAFSCs could be retrieved directly from a small amount of amniotic fluid that can be obtained at the time of diagnostic amniocentesis. The aim of the study was to verify if a direct freezing of amniotic fluid cells is able to maintain and / or improve the potential of the sub-population of stem cells. Methods - We compared the potential of the hAFSCs depending on the moment in which they are frozen, cells obtained directly from amniotic fluid aspiration (D samples) and cells cultured in flask before freezing (C samples). Colony-forming-unit ability, proliferation, morphology, stemness-related marker expression, senescence, apoptosis, and differentiation potential of C and D samples were compared. Results - hAFSCs isolated from D samples expressed MSC markers until later passages, had a good proliferation rate, and exhibited differentiation capacity similar to hAFSCs of C samples. Interestingly, the direct freezing induce a higher concentration of cells positive for pluripotency stem cell markers, without teratoma formation in vivo. Conclusions - This study suggests that minimal processing may be adequate for the banking of amniotic fluid cells, avoiding in vitro passages before the storage and exposure to high oxygen concentration affecting stem cell properties. This technique might be a reasonable approach in terms of costs and for the process of accreditation in GMP for a stem cell bank