Vitamin D receptor is necessary for mitochondrial function and cell health

Vitamin D receptor (VDR) mediates many genomic and non-genomic effects of vitamin D. Recently, the mitochondrial effects of vitamin D have been characterized in many cell types. In this article, we investigated the importance of VDR not only in mitochondrial activity and integrity but also in cell health. The silencing of the receptor in different healthy, non-transformed, and cancer cells initially decreased cell growth and modulated the cell cycle. We demonstrated that, in silenced cells, the increased respiratory activity was associated with elevated reactive oxygen species (ROS) production. In the long run, the absence of the receptor caused impairment of mitochondrial integrity and, finally, cell death. Our data reveal that VDR plays a central role in protecting cells from excessive respiration and production of ROS that leads to cell damage. Because we confirmed our observations in different models of both normal and cancer cells, we conclude that VDR is essential for the health of human tissues

Constructal thermodynamics combined with infrared experiments to evaluate temperature differences in cells

The aim of this work was to evaluate differences in energy flows between normal and immortalized cells when these distinct biological systems are exposed to environmental stimulation. These differences were considered using a constructal thermodynamic approach, and were subsequently verified experimentally. The application of constructal law to cell analysis led to the conclusion that temperature differences between cells with distinct behaviour can be amplified by interaction between cells and external fields. Experimental validation of the principle was carried out on two cellular models exposed to electromagnetic fields. By infrared thermography we were able to assess small changes in heat dissipation measured as a variation in cell internal energy. The experimental data thus obtained are in agreement with the theoretical calculation, because they show a different thermal dispersion pattern when normal and immortalized cells are exposed to electromagnetic fields. By using two methods that support and validate each other, we have demonstrated that the cell/environment interaction can be exploited to enhance cell behavior differences, in particular heat dissipation. We propose infrared thermography as a technique effective in discriminating distinct patterns of thermal dispersion and therefore able to distinguish a normal phenotype from a transformed one

Preparation and Characterization of a Novel Skin Substitute

Autologous epidermal cell cultures (CEA) represent a possibility to treat extensive burn lesions, since they allow a significative surface expansion which cannot be achieved with other surgical techniques based on autologous grafting. Moreover currently available CEA preparations are difficult to handle and their take rate is unpredictable. This study aimed at producing and evaluating a new cutaneous biosubstitute made up of alloplastic acellular glycerolized dermis (AAGD) and CEA to overcome these difficulties. A procedure that maintained an intact basement membrane was developed, so as to promote adhesion and growth of CEA on AAGD. Keratinocytes were seeded onto AAGD and cultured up to 21 days. Viability tests and immunohistochemical analysis with specific markers were carried out at 7, 14, and 21 days, to evaluate keratinocyte adhesion, growth, and maturation. Our results support the hypothesis that this newly formed skin substitute could allow its permanent engraftment in clinical application

Glycerolized reticular dermis as a new human acellular dermal matrix: An exploratory study

Human Acellular Dermal Matrices (HADM) are employed in various reconstructive surgery procedures as scaffolds for autologous tissue regeneration. The aim of this project was to develop a new type of HADM for clinical use, composed of glycerolized reticular dermis decellularized through incubation and tilting in Dulbecco's Modified Eagle's Medium (DMEM). This manufacturing method was compared with a decellularization procedure already described in the literature, based on the use of sodium hydroxide (NaOH), on samples from 28 donors. Cell viability was assessed using an MTT assay and microbiological monitoring was performed on all samples processed after each step. Two surgeons evaluated the biomechanical characteristics of grafts of increasing thickness. The effects of the different decellularization protocols were assessed by means of histological examination and immunohistochemistry, and residual DNA after decellularization was quantified using a real-time TaqMan MGB probe. Finally, we compared the results of DMEM based decellularization protocol on reticular dermis derived samples with the results of the same protocol applied on papillary dermis derived grafts. Our experimental results indicated that the use of glycerolized reticular dermis after 5 weeks of treatment with DMEM results in an HADM with good handling and biocompatibility properties

The cosmic ray primary composition in the "knee" region through the EAS electromagnetic and muon measurements at EAS-TOP

Abstract The evolution of the cosmic ray primary composition in the energy range 10 6 –10 7 GeV (i.e. the "knee" region) is studied by means of the e.m. and muon data of the Extensive Air Shower EAS-TOP array (Campo Imperatore, National Gran Sasso Laboratories). The measurement is performed through: (a) the correlated muon number ( N μ ) and shower size ( N e ) spectra, and (b) the evolution of the average muon numbers and their distributions as a function of the shower size. From analysis (a) the dominance of helium primaries at the knee, and therefore the possibility that the knee itself is due to a break in their energy spectrum (at E k He =(3.5±0.3)×10 6 GeV) are deduced. Concerning analysis (b), the measurement accuracies allow the classification in terms of three mass groups: light (p,He), intermediate (CNO), and heavy (Fe). At primary energies E 0 ≈10 6 GeV the results are consistent with the extrapolations of the data from direct experiments. In the knee region the obtained evolution of the energy spectra leads to: (i) an average steep spectrum of the light mass group ( γ p,He >3.1), (ii) a spectrum of the intermediate mass group harder than the one of the light component ( γ CNO ≃2.75, possibly bending at E k CNO ≈(6–7)×10 6 GeV), (iii) a constant slope for the spectrum of the heavy primaries ( γ Fe ≃2.3–2.7) consistent with the direct measurements. In the investigated energy range, the average primary mass increases from 〈ln A 〉=1.6–1.9 at E 0 ≃1.5×10 6 GeV to 〈ln A 〉=2.8–3.1 at E 0 ≃1.5×10 7 GeV. The result supports the standard acceleration and propagation models of galactic cosmic rays that predict rigidity dependent cut-offs for the primary spectra of the different nuclei. The uncertainties connected to the hadronic interaction model (QGSJET in CORSIKA) used for the interpretation are discussed

Activin A Induces Langerhans Cell Differentiation In Vitro and in Human Skin Explants

Langerhans cells (LC) represent a well characterized subset of dendritic cells located in the epidermis of skin and mucosae. In vivo, they originate from resident and blood-borne precursors in the presence of keratinocyte-derived TGFβ. Ιn vitro, LC can be generated from monocytes in the presence of GM-CSF, IL-4 and TGFβ. However, the signals that induce LC during an inflammatory reaction are not fully investigated. Here we report that Activin A, a TGFβ family member induced by pro-inflammatory cytokines and involved in skin morphogenesis and wound healing, induces the differentiation of human monocytes into LC in the absence of TGFβ. Activin A-induced LC are Langerin+, Birbeck granules+, E-cadherin+, CLA+ and CCR6+ and possess typical APC functions. In human skin explants, intradermal injection of Activin A increased the number of CD1a+ and Langerin+ cells in both the epidermis and dermis by promoting the differentiation of resident precursor cells. High levels of Activin A were present in the upper epidermal layers and in the dermis of Lichen Planus biopsies in association with a marked infiltration of CD1a+ and Langerin+ cells. This study reports that Activin A induces the differentiation of circulating CD14+ cells into LC. Since Activin A is abundantly produced during inflammatory conditions which are also characterized by increased numbers of LC, we propose that this cytokine represents a new pathway, alternative to TGFβ, responsible for LC differentiation during inflammatory/autoimmune conditions