Human iPSC-Derived Astrocytes: A Powerful Tool to Study Primary Astrocyte Dysfunction in the Pathogenesis of Rare Leukodystrophies

Astrocytes are very versatile cells, endowed with multitasking capacities to ensure brain homeostasis maintenance from brain development to adult life. It has become increasingly evident that astrocytes play a central role in many central nervous system pathologies, not only as regulators of defensive responses against brain insults but also as primary culprits of the disease onset and progression. This is particularly evident in some rare leukodystrophies (LDs) where white matter/myelin deterioration is due to primary astrocyte dysfunctions. Understanding the molecular defects causing these LDs may help clarify astrocyte contribution to myelin formation/maintenance and favor the identification of possible therapeutic targets for LDs and other CNS demyelinating diseases. To date, the pathogenic mechanisms of these LDs are poorly known due to the rarity of the pathological tissue and the failure of the animal models to fully recapitulate the human diseases. Thus, the development of human induced pluripotent stem cells (hiPSC) from patient fibroblasts and their differentiation into astrocytes is a promising approach to overcome these issues. In this review, we discuss the primary role of astrocytes in LD pathogenesis, the experimental models currently available and the advantages, future evolutions, perspectives, and limitations of hiPSC to study pathologies implying astrocyte dysfunctions.     

Aquaporins Are One of the Critical Factors in the Disruption of the Skin Barrier in Inflammatory Skin Diseases

The skin is the largest organ of the human body, serving as an effective mechanical barrier between the internal milieu and the external environment. The skin is widely considered the first-line defence of the body, with an essential function in rejecting pathogens and preventing mechanical, chemical, and physical damages. Keratinocytes are the predominant cells of the outer skin layer, the epidermis, which acts as a mechanical and water-permeability barrier. The epidermis is a permanently renewed tissue where undifferentiated keratinocytes located at the basal layer proliferate and migrate to the overlying layers. During this migration process, keratinocytes undertake a differentiation program known as keratinization process. Dysregulation of this differentiation process can result in a series of skin disorders. In this context, aquaporins (AQPs), a family of membrane channel proteins allowing the movement of water and small neutral solutes, are emerging as important players in skin physiology and skin diseases. Here, we review the role of AQPs in skin keratinization, hydration, keratinocytes proliferation, water retention, barrier repair, wound healing, and immune response activation. We also discuss the dysregulated involvement of AQPs in some common inflammatory dermatological diseases characterised by skin barrier disruption.     

Probabilistic seismic damage assessment of reinforced concrete buildings considering directionality effects

Most of buildings and structures are usually projected according to two main axes. However, the geographical position of these buildings varies randomly. Such random distributions of the azimuthal positions of structures, in most of the cities, generally, are not accounted for when assessing their seismic risk; certainly, the direction of the seismic loads is another highly random variable. Moreover, an additional important source of uncertainty is related to the structural response, mainly due to the random character of the mechanical properties. There is a consensus that uncertainties must be considered for adequately assessing the seismic risk of structures, but these directionality effects have not been deeply explored so far. In this article, the influence of the high uncertainty involved in these input variables on the expected seismic damage is analysed. Thus, an actual earthquake, which affected the southern part of Spain, is studied. Notably, damages on a group of affected buildings, located close to the epicentre, are analysed and discussed in detail. The results show that the influence of the random azimuthal position of structures is an important source of uncertainty and that it should be taken into account when estimating the expected seismic risk in urban areas.     

Dissolved trace element concentrations in the East River-Long Island Sound system: relative importance of autochthonous versus allochthonous sources

Dissolved trace metal (Ag, Cd, Cu, Fe, Ni, Pb, and Zn), inorganic nutrient (NO3, NH4, PO4, H4SiO4), and DOC concentrations were measured at 43 stations during low (July 2000) and high (April 2001) river discharge conditions in surface waters of Long Island Sound (LIS). To evaluate the impact of fluvial sources to the total metal budget of the sound, samples were collected from major tributaries discharging into LIS (Thames, Quinnipiac, Housatonic, Connecticut, and East Rivers). To compare LIS with other coastal embayments, samples were also collected from five LIS coastal embayments (Manhassett Bay, Huntington Harbor, Oyster Bay, Hempstead Harbor, and Port Jefferson Harbor), which are monitored by the U.S. National Status and Trends Program. Metal and nutrient distributions identified two biogeochemical regimes within LIS: an area of relatively high nutrient and metal concentrations in the East River/Narrows region in western LIS and an area in the eastern region of the sound that had comparatively lower concentrations. Mass balance estimates indicated that, during low flow conditions, the East River was the dominant allochthonous source of most trace metals (Ag, Cd, Cu, Ni, Zn) and inorganic nutrients (NO3 and PO4); during high flow conditions, the most influential source of these constituents was the Connecticut River. Mass balance estimates also evidenced a large autochthonous source of Cu, Ni, and Zn, as their spatial distributions displayed elevated concentrations away from point sources such as the East River. Principal component analysis suggested that metal and nutrient distributions in the LIS system were influenced by different seasonal processes: remobilization from contaminated sediments, anthropogenic inputs from sewage discharges and phytoplankton scavenging during the spring freshet, and benthic remobilization during summer conditions.     

Preliminary Investigation on the Involvement of Cytoskeleton-Related Proteins,DAAM1 and PREP,in Human Testicular Disorders

Herein, for the first time, the potential relationships between the cytoskeleton-associated proteins DAAM1 and PREP with different testicular disorders, such as classic seminoma (CS), Leydig cell tumor (LCT), and Sertoli cell-only syndrome (SOS), were evaluated. Six CS, two LCT, and two SOS tissue samples were obtained during inguinal exploration in patients with a suspect testis tumor based on clinical examination and ultrasonography. DAAM1 and PREP protein levels and immunofluorescent localization were analyzed. An increased DAAM1 protein level in CS and SOS as compared to non-pathological (NP) tissue was observed, while LCT showed no significant differences. Conversely, PREP protein level increased in LCT, while it decreased in CS and SOS compared to NP tissue. These results were strongly supported by the immunofluorescence staining, revealing an altered localization and signal intensity of DAAM1 and PREP in the analyzed samples, highlighting a perturbed cytoarchitecture. Interestingly, in LCT spermatogonia, a specific DAAM1 nuclear localization was found, probably due to an enhanced testosterone production, as confirmed by the increased protein levels of steroidogenic enzymes. Finally, although further studies are needed to verify the involvement of other formins and microtubule-associated proteins, this report raised the opportunity to indicate DAAM1 and PREP as new potential markers, supporting the cytoskeleton dynamics changes occurring during normal and/or pathological cell differentiation.     

Assessment of k–ε models using tetrahedral grids to describe the turbulent flow field of a PBT impeller and validation through the PIV technique

In turbulence modeling, the RNG and Realizable models have important improvements in the turbulent production and dissipation terms in comparison to the Standard. The selection of the appropriate turbulence model has an impact on the convergence and solution in STRs, and they are used in mixing, multiphase modeling or as starting solution of transient models as DES and LES. Although there are several studies with the pitched blade turbine(PBT) impeller, most of them used the Standard model as representative of all k–ε models, using structured hexahedral grids composed of low number of cells, and in some cases under axial symmetry assumptions.Accordingly, in this work the assessment of the Standard, RNG and Realizable models to describe the turbulent flow field of this impeller, using the Multiple Reference Frame(MRF) and Sliding Mesh(SM) approaches with tetrahedral domains in dense grids, is presented. This kind of cell elements is especially suitable to reproduce complex geometries. Flow velocities and turbulent parameters were verified experimentally by PIV and torque measurements. The three models were capable of predicting fairly the pumping number, the power number based on torque, and velocities. Although the RNG improved the predictions of the turbulent kinetic energy and dissipation rate, the Realizable model presented better performance for both approaches. All models failed in the prediction of the total dissipation rate, and a dependence of its value on the number of cells for the MRF was found.