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Abstract Background Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by accumulation of extracellular amyloid beta (Aβ) and intracellular neurofibrillary tangles, leading to chronic activation of astrocytes and microglia and persistent neuroinflammation. Aβ-linked activation of microglia and astrocytes leads to increased intracellular calcium and production of proinflammatory cytokines, impacting the progression of neurodegeneration. An N-terminal Aβ fragment (Aβ1–15) and a shorter hexapeptide core sequence within the N-Aβ fragment (N-Aβcore: Aβ10–15) have previously been shown to protect against Aβ-induced mitochondrial dysfunction, oxidative stress and apoptosis in neurons and rescue synaptic and spatial memory deficits in an APP/PSEN1 mouse model. Here, we hypothesized that the N-Aβ fragment and N-Aβcore are protective against Aβ-induced gliotoxicity, promoting a neuroprotective environment and potentially alleviating the characteristically persistent neuroinflammation present in AD. Methods We treated ex vivo organotypic brain slice cultures from an aged familial AD mouse model, 5xFAD, with the N-Aβcore and used immunocytochemistry to assess the impact on astrogliosis and microgliosis and alterations in synaptophysin-positive puncta engulfed by microglia. Isolated neuron/glia cultures, mixed glial cultures or a microglial cell line were treated with oligomeric human Aβ at concentrations mimicking the pathogenic concentrations (μM) observed in AD in the absence or presence of the non-toxic N-terminal Aβ fragments. Resultant changes in synaptic density, gliosis, oxidative stress, mitochondrial dysfunction, apoptosis, and the expression and release of proinflammatory markers were then determined. Results We demonstrate that the N-terminal Aβ fragments mitigated the phenotypic switch leading to astrogliosis and microgliosis induced by pathological concentrations of Aβ in mixed glial cultures and organotypic brain slice cultures from the transgenic 5xFAD mouse model, while protecting against Aβ-induced oxidative stress, mitochondrial dysfunction and apoptosis in isolated astrocytes and microglia. Moreover, the addition of the N-Aβcore attenuated the expression and release of proinflammatory mediators in microglial cells activated by Aβ and rescued microglia-mediated loss of synaptic elements induced by pathological levels of Aβ. Conclusions Together, these findings indicate the protective functions of the N-terminal Aβ fragments extend to reactive gliosis and gliotoxicity induced by Aβ, by preventing or reversing glial reactive states indicative of neuroinflammation and synaptic loss central to AD pathogenesis.

The ESX (or Type VII) secretion systems are protein export systems in mycobacteria and many Gram-positive bacteria that mediate a broad range of functions including virulence, conjugation, and metabolic regulation. These systems translocate folded dimers of WXG100-superfamily protein substrates across the cytoplasmic membrane. We report the cryo-electron microscopy structure of an ESX-3 system, purified using an epitope tag inserted with recombineering into the chromosome of the model organism Mycobacterium smegmatis. The structure reveals a stacked architecture that extends above and below the inner membrane of the bacterium. The ESX-3 protomer complex is assembled from a single copy of the EccB3, EccC3, and EccE3 and two copies of the EccD3 protein. In the structure, the protomers form a stable dimer that is consistent with assembly into a larger oligomer. The ESX-3 structure provides a framework for further study of these important bacterial transporters.

A novel data interpretation process that utilizes comprehensive particle-in-cell (PIC) simulations is developed for the new retarding field energy analyzer (RFEA) currently being constructed at DIII-D for the lower divertor using the Divertor Material Evaluation System. This probe is expected to survive a heat load of up to 100 MW/m2 for up to 5 s and reliably measure the main ion temperature (Ti) on the divertor target ranging from 10 to 200 eV. These extreme conditions posed significant engineering limitations on the probe geometry, thus extensive validation work has been performed. The conventional fitting method for the RFEA I–V characteristics is based on a simplified 1-D model without considering the ion space charge inside the probe cavity and may not be sufficient for probes designed for the DIII-D divertor environment. In this article, a more realistic description of the particle propagation process within the RFEA cavity is achieved by including both 3-D geometric effects and ion space charge in the PIC simulations, and the capability to reconstruct the ion energy distribution functions is demonstrated with reasonable consistency. [ABSTRACT FROM AUTHOR]

Large marine predators often aggregate seasonally in discrete locations to take advantage of optimal foraging conditions, leading to spatial and temporal variation in their exposure on other species. However, our understanding of the impacts this exposure may have on the behavior and physiology of prey is poor, especially in marine systems. Here, we evaluated the non-consumptive effects of potential exposure to large sharks (white sharks, Carcharodon carcharias (Linnaeus, 1758)) on the stress physiology of an economically important teleost, the striped bass (Morone saxatilis (Walbaum, 1792)), off Cape Cod, Massachusetts, USA. We sampled fish in habitats that varied significantly in shark exposure across 5 months and over 2 years, evaluating blood physiology stress indicators (i.e., cortisol, glucose, and lactate concentrations) and reflex impairment. None of the blood parameters were influenced by shark exposure, although we did observe subtle temperature and seasonal effects. One of the three reflex tests (the vertical orientation test) was negatively affected by shark exposure, although the mechanistic basis for this finding is unclear. This work supports the notion that predictable sources of predation pressure tend not to manifest in stress-related costs in free-ranging prey, which has implications for shaping our understanding of how large sharks influence ecosystems through non-consumptive effects. Key words: glucocorticoids, non-consumptive effects, predation, stress, striped bass, Morone saxatilis, white shark, Carcharodon carcharias. Les grands predateurs marins se regroupent souvent sur une base saisonniere dans des lieux precis pour profiter de conditions d'approvisionnement optimales, ce qui mene a des variations spatiales et temporelles de l'exposition d'autres especes a leur presence. La comprehension des impacts de cette exposition sur le comportement et la physiologie des proies demeure toutefois limitee, particulierement en ce qui concerne les systemes marins. Nous evaluons les effets non destructeurs de l'exposition potentielle a de grands requins (grands requins blancs, Carcharodon carcharias (Linnaeus, 1758)) sur la physiologie du stress d'un teleosteen d'importance commerciale, le bar d'Amerique (Morone saxatilis (Walbaum, 1792)), au large de Cape Cod (Massachusetts, Etats-Unis). Nous avons echantillonne des poissons dans des habitats presentant une diversite d'exposition aux requins sur une duree de cinq mois et sur deux annees, pour evaluer des indicateurs sanguins de la physiologie du stress (c.-a-d. concentrations de cortisol, glucose et lactate) et l'alteration des reflexes. Aucun des parametres sanguins n'etait influence par l'exposition aux requins, de subtils effets de la temperature et de la saison ayant toutefois ete observes. Un des trois tests des reflexes (le test d'orientation verticale) a releve un effet negatif de l'exposition aux requins, bien que le fondement mecaniste de cet effet demeure mal compris. Ces travaux appuient la notion voulant que les sources previsibles de pression de predation n'aient pas tendance a se refleter dans les couts associes au stress chez les proies en liberte, une constatation importante pour la comprehension de l'influence qu'exercent, par l'entremise d'effets non destructeurs, les grands requins sur les ecosystemes. [Traduit par la Redaction] Mots-cles : glucocorticoides, effets non destructeurs, predation, stress, bar d'Amerique, Morone saxatilis, grand requin blanc, Carcharodon carcharias., Introduction Predators can affect prey directly, through consumption, and indirectly, through non-consumptive effects, also known as 'risk effects' (Lima and Dill 1990; Laundre 2010). These non-consumptive effects can have strong [...]

As part of a population health-focused primary care transformation, in 2019 a health system in Minnesota developed a primary care team to exclusively care for high-cost high-need patients. Through its development and implementation, the team has discovered several key lessons in delivering care to complex patients. These lessons include the benefits of more integrative team-based care, the need and advantages of designated complex care team members, the importance of teamwork both within and outside of the complex care team, the need for frequent communication, and the importance of identifying mental health needs. In addition, there are several areas that require ongoing research and exploration, such as determining when a patient is able to graduate out of the program, how to enhance access to the complex care team, determining appropriate visit characteristics, and model feasibility. While addressing the needs of high cost high need patients is essential to improving quality of care and decreasing health care costs, there are several unique challenges and opportunities that come with caring for this patient population. Although this highly integrated model of care continues to evolve, the initial lessons learned may inform other health systems and care teams undertaking the care of complex patients.

Accurately predicting redeposition is vital for high-Z plasma-facing component (PFC) survivability in magnetic confinement fusion. In this study, we categorize high-Z redeposition into three mechanisms: geometric-driven (prompt), sheath-driven (local), and scrape-off-layer-driven (far) redeposition. To investigate these mechanisms, we employ Monte Carlo transport codes to simulate azimuthally symmetric tungsten source erosion and redeposition in a tokamak. By iteratively analyzing critical parameters, we evaluate redeposition scaling for each mechanism. Specifically, we investigate the impact of magnetic-field-to-PFC pitch angle assumptions on PFC losses into the scrape-off layer. Our findings reveal significant pitch angle sensitivity due to an asymmetric prompt vs local redeposition trade-off. These results enhance our understanding of redeposition phenomena in fusion plasma environments. [ABSTRACT FROM AUTHOR]

A novel impurity transport model that approximates SOL turbulence as a fluctuating poloidal electric field is shown to be an acceptable replacement for the traditional approach of assigning an arbitrary radial diffusion coefficient to the impurity ions. The model is implemented in the DIVIMP impurity transport code and applied to an L-Mode tungsten divertor experiment on DIII-D. The poloidal electric field is represented as fluctuating between ±1000 V m−1 based on previous measurements. The resulting intermittent vr = E θ × BT transport causes ions to transport both into the core as well as into the far-SOL. Simultaneous agreement with estimates of the W density just inside the separatrix as well as in the far-SOL is obtained (nW ∼ 1014 m−3 and nW ∼ 1012 m−3, respectively). Prompt re-deposition of the W ions was necessary to obtain agreement (fredep ∼ 99%). We conclude that simulating impurity transport using a physics-based approximation for turbulence in the SOL, versus arbitrarily assigning diffusion coefficients, may enable better reactor scale predictions of core impurity contamination. [ABSTRACT FROM AUTHOR]

The ESX (or Type VII) secretion systems are protein export systems in mycobacteria and many Gram-positive bacteria that mediate a broad range of functions including virulence, conjugation, and metabolic regulation. These systems translocate folded dimers of WXG100-superfamily protein substrates across the cytoplasmic membrane. We report the cryo-electron microscopy structure of an ESX-3 system, purified using an epitope tag inserted with recombineering into the chromosome of the model organism Mycobacterium smegmatis. The structure reveals a stacked architecture that extends above and below the inner membrane of the bacterium. The ESX-3 protomer complex is assembled from a single copy of the EccB3, EccC3, and EccE3 and two copies of the EccD3 protein. In the structure, the protomers form a stable dimer that is consistent with assembly into a larger oligomer. The ESX-3 structure provides a framework for further study of these important bacterial transporters.

The formation of neuron networks is a process of fundamental importance for understanding the development of the nervous system and for creating biomimetic devices for tissue engineering and neural repair. The basic process that controls the network formation is the growth of an axon from the cell body and its extension towards target neurons. Axonal growth is directed by environmental stimuli that include intercellular interactions, biochemical cues, and the mechanical and geometrical properties of the growth substrate. Despite significant recent progress, the steering of the growing axon remains poorly understood. In this paper, we develop a model of axonal motility, which incorporates substrate-geometry sensing. We combine experimental data with theoretical analysis to measure the parameters that describe axonal growth on micropatterned surfaces: diffusion (cell motility) coefficients, speed and angular distributions, and cell-substrate interactions. Experiments performed on neurons treated with inhibitors for microtubules (Taxol) and actin filaments (Y-27632) indicate that cytoskeletal dynamics play a critical role in the steering mechanism. Our results demonstrate that axons follow geometrical patterns through a contact-guidance mechanism, in which geometrical patterns impart high traction forces to the growth cone. These results have important implications for bioengineering novel substrates to guide neuronal growth and promote nerve repair.

Mutations within the Leucine-Rich Repeat Kinase 2 (LRRK2) gene are the most common genetic cause of autosomal and sporadic Parkinson’s disease (PD). LRRK2 is a large multidomain kinase that has reported interactions with several membrane proteins, including Rab and Endophilin, and has recently been proposed to function as a regulator of vesicular trafficking. It is unclear whether or how the spatiotemporal organization of the protein is altered due to LRRK2 activity. Therefore, we utilized fluctuation-based microscopy along with FLIM/FRET to examine the cellular properties and membrane recruitment of WT LRRK2-GFP (WT) and the PD mutant G2019S LRRK2-GFP (G2019S). We show that both variants can be separated into two distinct populations within the cytosol; a freely diffusing population associated with monomer/dimer species and a slower, likely vesicle-bound population. G2019S shows a significantly higher propensity to self-associate in both the cytosol and membrane regions when compared to WT. G2019S expression also resulted in increased hetero-interactions with Endophilin A1 (EndoA1), reduced cellular vesicles, and altered clathrin puncta dynamics associated with the plasma membrane. This finding was associated with a reduction in transferrin endocytosis in cells expressing G2019S, which indicates disruption of endocytic protein recruitment near the plasma membrane. Overall, this study uncovered multiple dynamic alterations to the LRRK2 protein as a result of the G2019S mutation—all of which could lead to neurodegeneration associated with PD.