Ultrastructural anatomy of nodes of Ranvier in the peripheral nervous system as revealed by STED microscopy.

We used stimulated emission depletion (STED) superresolution microscopy to analyze the nanoscale organization of 12 glial and axonal proteins at the nodes of Ranvier of teased sciatic nerve fibers. Cytoskeletal proteins of the axon (betaIV spectrin, ankyrin G) exhibit a high degree of one-dimensional longitudinal order at nodal gaps. In contrast, axonal and glial nodal adhesion molecules [neurofascin-186, neuron glial-related cell adhesion molecule (NrCAM)] can arrange in a more complex, 2D hexagonal-like lattice but still feature a ∼190-nm periodicity. Such a lattice-like organization is also found for glial actin. Sodium and potassium channels exhibit a one-dimensional periodicity, with the Nav channels appearing to have a lower degree of organization. At paranodes, both axonal proteins (betaII spectrin, Caspr) and glial proteins (neurofascin-155, ankyrin B) form periodic quasi–one-dimensional arrangements, with a high degree of interdependence between the position of the axonal and the glial proteins. The results indicate the presence of mechanisms that finely align the cytoskeleton of the axon with the one of the Schwann cells, both at paranodal junctions (with myelin loops) and at nodal gaps (with microvilli). Taken together, our observations reveal the importance of the lateral organization of proteins at the nodes of Ranvier and pave the way for deeper investigations of the molecular ultrastructural mechanisms involved in action potential propagation, the formation of the nodes, axon–glia interactions, and demyelination diseases

Subcortical cytoskeleton periodicity throughout the nervous system

Superresolution fluorescence microscopy recently revealed a ~190 nm periodic cytoskeleton lattice consisting of actin, spectrin, and other proteins underneath the membrane of cultured hippocampal neurons. Whether the periodic cytoskeleton lattice is a structural feature of all neurons and how it is modified when axons are ensheathed by myelin forming glial cells is not known. Here, STED nanoscopy is used to demonstrate that this structure is a commonplace of virtually all neuron types in vitro. To check how the subcortical meshwork is modified during myelination, we studied sciatic nerve fibers from adult mice. Periodicity of both actin and spectrin was uncovered at the internodes, indicating no substantial differences between unmyelinated and myelinated axons. Remarkably, the actin/spectrin pattern was also detected in glial cells such as cultured oligodendrocyte precursor cells. Altogether our work shows that the periodic subcortical cytoskeletal meshwork is a fundamental characteristic of cells in the nervous system and is not a distinctive feature of neurons, as previously thought

Historical data and modern methods reveal insights in measles epidemiology: a retrospective closed cohort study

OBJECTIVES Measles was endemic in England during the early 1800s; however, it did not arrive in Australia until 1850 whereas other infectious diseases were known to have arrived much earlier-many with the First Fleet in 1788-leading to the question of why there was a difference. DESIGN Ships surgeons' logbooks from historical archives, 1829-1882, were retrospectively reviewed for measles outbreak data. Infectious disease modelling techniques were applied to determine whether ships would reach Australia with infectious measles cases. SETTING Historical ship surgeon logbooks of measles outbreaks occurring on journeys from Britain to Australia were examined to provide new insights into measles epidemiology. PRIMARY AND SECONDARY OUTCOME MEASURES Serial intervals and basic reproduction numbers (R(0)), immunity, outbreak generations, age-distribution, within-family transmission and outbreak lengths for measles within these closed cohorts. RESULTS Five measles outbreaks were identified (163 cases). The mean serial interval (101 cases) was 12.3 days (95% CI 12.1 to 12.5). Measles R(0) (95 cases) ranged from 7.7-10.9. Immunity to measles was lowest among children ≤10 years old (range 37-42%), whereas 94-97% of adults appeared immune. Outbreaks ranged from 4-6 generations and, before 1850, were 41 and 38 days in duration. Two outbreaks after 1850 lasted longer than 70 days and one lasted 32 days. CONCLUSIONS Measles syndrome reporting in a ship surgeon's logs provided remarkable detail on prevaccination measles epidemiology in the closed environment of ship voyages. This study found lower measles R(0) and a shorter mean clinical serial interval than is generally reported. Archival ship surgeon log books indicate it was unlikely that measles was introduced into Australia before 1850, owing to high levels of pre-existing immunity in ship passengers, low numbers of travelling children and the journey's length from England to Australia.g BP was supported by a Master of Applied Epidemiology scholarship from the Australian Government and a Hunter Medical Research Institute Research Fellowshi

Multicomponent Hydrogels for the Formation of Vascularized Bone-like Constructs In Vitro.

The native extracellular matrix (ECM) is a complex gel-like system with a broad range of structural features and biomolecular signals. Hydrogel platforms that can recapitulate the complexity and signaling properties of this ECM would have enormous impact in fields ranging from tissue engineering to drug discovery. Here, we report on the design, synthesis, and proof-of-concept validation of a microporous and nanofibrous hydrogel exhibiting multiple bioactive epitopes designed to recreate key features of the bone ECM. The material platform integrates self-assembly with orthogonal enzymatic cross-linking to create a supramolecular environment comprising hyaluronic acid modified with tyramine (HA-Tyr) and peptides amphiphiles (PAs) designed to promote cell adhesion (RGDS-PA), osteogenesis (Osteo-PA), and angiogenesis (Angio-PA). Through individual and co-cultures of human adipose derived mesenchymal stem cells (hAMSCs) and human umbilical vascular endothelial cells (HUVECs), we confirmed the capacity of the HA-Tyr/RGDS-PA/Osteo-PA/Angio-PA hydrogel to promote cell adhesion as well as osteogenic and angiogenic differentiation in both 2D and 3D setups. Furthermore, using immunofluorescent staining and reverse transcription-quantitative polymerase chain reaction (RT-qPCR), we demonstrated co-differentiation and organization of hAMSCs and HUVECs into 3D aggregates resembling vascularized bone-like constructs

Uncovering current pyroregions in Italy using wildfire metrics

Background: Pyrogeography is a major field of investigation in wildfire science because of its capacity to describe the spatial and temporal variations of fire disturbance. We propose a systematic pyrogeographic analytical approach to cluster regions on the basis of their pyrosimilarities. We employed the Affinity Propagation algorithm to cluster pyroregions using Italian landscape as a test bed and its current wildfire metrics in terms of density, seasonality and stand replacing fire ratio. A discussion follows on how pyrogeography varies according to differences in the human, biophysical, socioeconomic, and climatic spheres. Results: The algorithm identified seven different pyroregion clusters. Two main gradients were identified that partly explain the variability of wildfire metrics observed in the current pyroregions. First, a gradient characterized by increasing temperatures and exposure to droughts, which coincides with a decreasing latitude, and second, a human pressure gradient displaying increasing population density in areas at lower elevation. These drivers exerted a major influence on wildfire density, burnt area over available fuels and stand replacing, which were associated to warm-dry climate and high human pressure. The study statistically highlighted the importance of a North–South gradient, which represents one of the most important drivers of wildfire regimes resulting from the variations in climatic conditions but showing collinearity with socioeconomic aspects as well. Conclusion: Our fully replicable analytical approach can be applied at multiple scales and used for the entire European continent to uncover new and larger pyroregions. This could create a basis for the European Commission to promote innovative and collaborative funding programs between regions that demonstrate pyrosimilarities

In Vitro Hair Growth Promoting Effect of a Noncrosslinked Hyaluronic Acid in Human Dermal Papilla Cells

Dermal papilla cells (DPCs) are a source of nutrients and growth factors, which support the proliferation and growth of keratinocytes as well as promoting the induction of new hair follicles and maintenance of hair growth. The protection from reactive oxygen species (ROS) and the promotion of angiogenesis are considered two of the basal mechanisms to preserve the growth of the hair follicle. In this study, a noncrosslinked hyaluronic acid (HA) filler (HYDRO DELUXE BIO, Matex Lab S.p.A.) containing several amino acids was tested with in vitro assays on human follicle dermal papilla cells (HFDPCs). The experiments were carried out to investigate the possible protection against oxidative stress and the ability to increase the vascular endothelial growth factor (VEGF) release. The results demonstrated the restoration of cell viability against UVB-induced cytotoxicity and an increase in the VEGF secretion. These data demonstrate the capability of the product to modulate human dermal papilla cells, suggesting a future use in mesotherapy, a minimally invasive local intradermal therapy (LIT), after further clinical investigations

Neuronal chemotaxis by optically manipulated liposomes

We probe chemotaxis of single neurons, induced by signalling molecules which were optically delivered from liposomes in the neighbourhood of the cells. We implemented an optical tweezers setup combined with a micro-dissection system on an inverted microscope platform. Molecules of Netrin-1 protein were encapsulated into micron-sized liposomes and manipulated to micrometric distances from a specific growth cone of a hippocampal neuron by the IR optical tweezers. The molecules were then released broken the liposomes with UV laser pulses. Chemotaxis induced by the delivered molecules was confirmed by the migration of the growth cone toward the liposome position. Since the delivery can be manipulated with high temporal and spatial resolution and the number of molecules released can be controlled quite precisely by tuning the liposome size and the solution concentration, this technique opens new opportunities to investigate the effect of physiological active compounds as Netrin-1 to neuronal signalling and guidance, which represents an important issue in neurobiology

Evaluation of anti-photoaging effects of a novel cosmeceutical containing a retinoids mixture using in vitro cell models

11noopenPhysiological ageing due to the passing of time and prolonged exposure to harmful sun rays generate wrinkles and reduce skin elasticity. These visible and clinical signs can be prevented or reversed by known strategies, such as the daily use of cosmetic products with antioxidant combina-tions or retinoids. A new dermocosmetic formulation enriched with a complex of retinoids, called RETINOIDS SERUM, was investigated through in vitro assays using human skin cells. The experiments were carried out to assess the anti-ageing activity in normal human dermal fibroblasts (NHDF) and epidermal keratinocytes (HaCaT). After the preliminary MTT assay, the proliferation together with the synthesis of collagen and elastin fibers was performed on NHDF cells after 24 h treatment with the two non-cytotoxic concentrations. Using UVB-irradiated HaCaT cells, the measurement of matrix metalloproteinase-1 (MMP-1) levels was also investigated. In vitro studies show that the dermocosmetic product improves collagen and elastin synthesis and the renewal of dermal fibrob-lasts. Moreover, a reduction in the MMP-1 secretion was also highlighted in UVB-irradiated HaCaT cells. These results suggest that the cosmetic formulation containing functional compounds such as retinoids can be useful to prevent the natural sign of ageing.openZerbinati N.; Sommatis S.; Maccario C.; Capillo M.C.; Di Francesco S.; Liga E.; Rauso R.; D'este E.; Dalla Gasperina D.; Guida S.; Mocchi R.Zerbinati, N.; Sommatis, S.; Maccario, C.; Capillo, M. C.; Di Francesco, S.; Liga, E.; Rauso, R.; D'Este, E.; Dalla Gasperina, D.; Guida, S.; Mocchi, R