Role of ERK1/2 in TNFα-induced internalization of Abcc2 in rat hepatocyte couplets

TNFα is a cytokine whose levels are increased in inflammatory pathologies that are associated with cholestasis. Endocytic internalization of Abcc2 (multidrug resistance-associated protein 2), a canalicular transporter of organic anions that is implicated in the clearance of clinically important drugs, is a phenomenon that occurs in inflammatory liver diseases, and it has been established that cytokines act as mediators. However, the intracellular mechanism involved in this effect remains unknown. The aim of the present work was to characterize the internalization of Abcc2 induced by TNFα and to study the role of ERK1/2 and reactive oxygen species as signaling mediators of transporter internalization. Using rat hepatocyte couplets, we found that TNFα (6.25 pg/ml) induced a decrease in Abcc2 activity estimated by the accumulation of the Abcc2 substrate glutathione methylfluorescein in the canalicular vacuole that was accompanied by internalization of Abcc2 from the canalicular membrane. Inhibition of MEK1/2 (upstream of ERK1/2) partially prevented TNFα effects on Abcc2 internalization and activity impairment. Reactive oxygen species (ROS) scavengers such as vitamin C and mannitol partially prevented both TNFα-induced decrease in Abcc2 activity and ERK1/2 phosphorylation. Apocynin, a NADPH oxidase inhibitor, prevented the increase in ROS and the phosphorylation of ERK1/2 produced by TNFα. Taken together, these results indicate that TNFα activates a pathway involving NADPH oxidase, ROS and MEK1/2-ERK1/2 that is partially responsible for the internalization of Abcc2. This internalization leads to an altered transport activity of Abcc2 that could impair drug disposal, enhancing drug toxicity in patients suffering from inflammatory liver diseases.Fil: Ciriaci, Nadia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Fisiología Experimental. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Fisiología Experimental; ArgentinaFil: Andermatten, Romina Belén. Universidad Nacional de Rosario; ArgentinaFil: Razori, María Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Fisiología Experimental. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Fisiología Experimental; ArgentinaFil: Schuck, Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Fisiología Experimental. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Fisiología Experimental; ArgentinaFil: Miszczuk, Gisel Sabrina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Fisiología Experimental. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Fisiología Experimental; ArgentinaFil: Medeot, Anabela Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Fisiología Experimental. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Fisiología Experimental; ArgentinaFil: Crocenzi, Fernando Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Fisiología Experimental. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Fisiología Experimental; ArgentinaFil: Roma, Marcelo Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Fisiología Experimental. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Fisiología Experimental; ArgentinaFil: Barosso, Ismael Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Fisiología Experimental. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Fisiología Experimental; ArgentinaFil: Ruiz, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Fisiología Experimental. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Fisiología Experimental; ArgentinaFil: Sanchez Pozzi, Enrique Juan. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Fisiología Experimental. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Fisiología Experimental; Argentin

Differential Expression and Distribution of α-, β-, and γ-Synuclein in the Developing Human Substantia Nigra

Although the functions of α-, β-, and γ-synuclein (αS, βS, γS, respectively) are unknown, these synaptic proteins are implicated in the pathogenesis of Parkinson's disease (PD) and related disorders. For example, αS forms Lewy bodies (LBs) in substantia nigra (SN) neurons of PD. However, since it is not known how these hallmark PD lesions contribute to the degeneration of SN neurons or what the normal function of αS is in SN neurons, we studied the developing human SN from 11 weeks gestational age (GA) to 16 years of age using immunohistochemistry and antibodies to αS, βS, γS, other synaptic proteins, and tyrosine hydoxylase (TH). SN neurons expressed TH at 11 weeks GA and αS, βS, and γS appeared initially at 15, 17, and 18 weeks GA, respectively. These synucleins first appeared in perikarya of SN neurons after synaptophysin, but about the same time as synaptotagmin and synaptobrevin. Redistribution of αS from perikarya to processes of SN neurons occurred by 18 weeks GA in parallel with synaptophysin, while βS and synaptotagmin were redistributed similarly between 20 and 28 weeks GA and this also occurred with γS and synaptobrevin between 33 weeks GA and 9 months postnatal. These data suggest that αS, βS, and γS may play a functional role in the development and maturation of SN neurons, but it remains to be determined how sequestration of αS as LBs in PD contributes to the degeneration of SN neurons

Fracture Shape and Orientation Contributions to P-Wave Velocity and Anisotropy of Alpine Fault Mylonites

P-wave anisotropy is significant in the mylonitic Alpine Fault shear zone. Mineral- and texture-induced anisotropy are dominant in these rocks but further complicated by the presence of fractures. Electron back-scattered diffraction and synchrotron X-ray microtomography (micro-CT) data are acquired on exhumed schist, protomylonite, mylonite, and ultramylonite samples to quantify mineral phases, crystal preferred orientations, microfractures, and porosity. The samples are composed of quartz, plagioclase, mica and accessory garnet, and contain 3–5% porosity. Based on the micro-CT data, the representative pore shape has an aspect ratio of 5:2:1. Two numerical models are compared to calculate the velocity of fractured rocks: a 2D wave propagation model, and a differential effective medium model (3D). The results from both models have comparable pore-free fast and slow velocities of 6.5 and 5.5 km/s, respectively. Introducing 5% fractures with 5:2:1 aspect ratio, oriented with the longest axes parallel to foliation decreases these velocities to 6.3 and 5.0 km/s, respectively. Adding both randomly oriented and foliation-parallel fractures hinders the anisotropy increase with fracture volume. The anisotropy becomes independent of porosity when 80% of fractures are randomly oriented. Modeled anisotropy in 2D and 3D are different for similar fracture aspect ratios, being 30 and 15%, respectively. This discrepancy is the result of the underlying assumptions and limitations. Our numerical results explain the effects that fracture orientations and shapes have on previously published field- and laboratory-based studies. Through this numerical study, we show how mica-dominated, pore-free P-wave anisotropy compares to that of fracture volume, shape and orientation for protolith and shear zone rocks of the Alpine Fault

Reliability and validity assessment of the observation of human-animal interaction for research (OHAIRE) behavior coding tool

The Observation of Human-Animal Interaction for Research (OHAIRE) is a coding tool developed to capture the behavior of children when interacting with social partners and animals in naturalistic settings. The OHAIRE behavioral categories of focus are emotional displays, social communication behaviors toward adults and peers, behaviors directed toward animals or experimental control objects, and interfering behaviors. To date, the OHAIRE has been used by 14 coders to code 2,732 min of video across four studies with a total of 201 participants ages 5 to 18 years (M = 10.1, SD = 2.5). Studies involved animal-assisted intervention with three species (i.e., dogs, horses, and guinea pigs) and three populations (i.e., autism spectrum disorder, attention-deficit hyperactivity disorder, and typically developing children) in a school, a therapeutic horseback riding program, a group therapy program, and the hospital setting. We explored the psychometric properties of the OHAIRE through analyses of its inter-rater reliability, intra-rater reliability, convergent and divergent validity, and internal structure, using data from these four human-animal interaction studies. The average inter-rater reliability was excellent (kappa = 0.81), with good reliability in most of the behavioral categories coded. Intra-rater reliability was consistently excellent (0.87 ≤ kappa ≤0.96). Internal structure analyses with Cronbach's alpha supported the exploratory use of subscales to measure social communication behaviors toward peers (α = 0.638) and adults (α = 0.605), and interactions experimental control objects (α = 0.589), and the use of a subscale to measure interactions with animals (α = 0.773). Correlation analyses with multiple questionnaires showed a convergence between positive emotional display and social behaviors as assessed by the OHAIRE and social skills as assessed by the Social Skills Rating System (SSRS) and the Social Communication Questionnaires (SCQ). Little concordance was found between the OHAIRE and the Social Responsiveness Scale (SRS) or the Aberrant Behavior Checklist-Community (ABC). The OHAIRE shows promise for wider use in the field of Human-Animal Interaction, with a need for generalization across more settings and ages

Controlled Assembly of Upconverting Nanoparticles for Low-Threshold Microlasers and Their Imaging in Scattering Media.

Micron-sized lasers fabricated from upconverting nanoparticles (UCNP) coupled to whispering gallery mode (WGM) microresonators can exhibit continuous-wave anti-Stokes lasing useful for tracking cells, environmental sensing, and coherent stimulation of biological activity. The integration of these microlasers into organisms and microelectronics requires even smaller diameters, however, which raises threshold pump powers beyond practical limits for biological applications. To meet the need for low lasing thresholds and high fidelity fabrication methods, we use correlative optical and electron microscopy to uncover the nanoparticle assembly process and structural factors that determine efficient upconverted lasing. We show that 5 μm microspheres with controlled submonolayer UCNP coatings exhibit, on average, 25-fold lower laser thresholds (1.7 ± 0.7 kW/cm2) compared to the mean values of the lowest threshold UCNP lasers, and variability is reduced 30-fold. WGMs are observed in the upconversion spectra for TiO2-coated microspheres as small as 3 μm, a size at which optical losses had previously prevented such observations. Finally, we demonstrate that the WGM signatures of these upconverting microlasers can be imaged and distinguished through tissue-mimicking phantoms. These advances will enable the fabrication of more efficient upconverting lasers for imaging, sensing, and actuation in optically complex environments

Additional file 1: of Sequential stages and distribution patterns of aging-related tau astrogliopathy (ARTAG) in the human brain

Summary of the statistical method and morphology of astroglial tau pathologies. (PDF 2229 kb

A Blended Learning Approach to Structural Field Mapping: Combining Local Geology, Virtual Geology, and Web-based Tools

In September 2020, the Corona crisis offered us an opportunity to develop and test a blended real and virtual interdisciplinary field mapping class, as well as revealing the need for, and stimulating development of new web-based tools for structural interpretation. Universität Mainz’ usual Master’s advanced field mapping, and Universität Tübingen’s usual Bachelor’s mapping classes were replaced with combinations of (i) virtual field mapping of Jurassic-Cretaceous sedimentary units at Molinos, Teruel Province, Spain, and (ii) field mapping of metamorphic rocks in the Mittelrhein Gorge and the Arh Valley, and outcrops of sedimentary rocks near Tübingen, Germany, which the students were mostly able to access on day trips using public transport or by bicycle. For the Molinos part of the exercise both groups were offered hand specimens containing distinctive fossils, linked to locations (and pseudo-locations) by google .kmz files, a variety of structural measurements also linked via .kmz files, and detailed satellite imagery within which mappable geological units display distinct characteristics. Introductions to the stratigraphy were made in three virtual outcrop sections examined in Google Street View from within Google Earth, and via web-based photogrammetric 3D outcrop models made available on the V3Geo virtual 3D geoscience platform. The students then extrapolated this stratigraphy based on the satellite imagery and .kmz file information. Our perception, validated by student feedback, is that the real parts of both field excursions were very important since they allowed us to teach and refine mapping and compass methodology and best demonstrate how to analyze 3D geometries of geological structures. Universität Mainz students particularly benefited from being able to visit locations where we had already made 3D outcrop models and offered a digital excursion, in the Ahr Valley (Rhenish Massif). They were able to compare real structural measurements with those derived from the precisely georeferenced 3D models, which enhanced their ability to subsequently obtain such information solely from the models. Although final student maps were of comparable quality to those produced in the field, structural interpretations were hampered by a lack of field measurements. In many cases, the Google Earth DEM is of too low resolution and ways should be found to include higher-resolution DEMs in web-based data sets. Overall, we think there were advantages compared to traditional field mapping, such as (i) enhanced evidence that methods like ‘structure contouring’ were used in all mapping, (ii) we were stimulated to teach the students to use digital methods to acquire field data, such as StraboSpot and Stereonet11 Apps. We observed these tools, and others we were unaware of, being used in combination with traditional paper and compass during the real mapping exercise. We hope to continue to employ this blended teaching approach even when the Corona crisis passes. This will be facilitated by our development of further 3D outcrop models, .kmz files with key information about outcrops in the Mittelrhein, and especially, web-based (rather than PC-based) tools to extract structural data such as plane and line orientations from 3D outcrop models and enable collaborative work on one data set

Additional file 4: of Sequential stages and distribution patterns of aging-related tau astrogliopathy (ARTAG) in the human brain

Hierarchical cluster analysis of astroglial tau pathologies in different disorders. (PDF 1463 kb

Additional file 2: of Sequential stages and distribution patterns of aging-related tau astrogliopathy (ARTAG) in the human brain

Pairwise conditional probability matrix and odds ratios of different ARTAG types. (PDF 876 kb

Additional file 3: of Sequential stages and distribution patterns of aging-related tau astrogliopathy (ARTAG) in the human brain

Pairwise conditional probability matrix and odds ratios of grey matter ARTAG and primary FTLD-tauopathy associated astroglial tau immunoreactivities. (PDF 437 kb