Engineered Perineural Vascular Plexus for Modeling Developmental Toxicity.

There is a vital need to develop in vitro models of the developing human brain to recapitulate the biological effects that toxic compounds have on the brain. To model perineural vascular plexus (PNVP) in vitro, which is a key stage in embryonic development, human embryonic stem cells (hESC)-derived endothelial cells (ECs), neural progenitor cells, and microglia (MG) with primary pericytes (PCs) in synthetic hydrogels in a custom-designed microfluidics device are cocultured. The formation of a vascular plexus that includes networks of ECs (CD31+, VE-cadherin+), MG (IBA1+), and PCs (PDGFRß+), and an overlying neuronal layer that includes differentiated neuronal cells (ßIII Tubulin+, GFAP+) and radial glia (Nestin+, Notch2NL+), are characterized. Increased brain-derived neurotrophic factor secretion and differential metabolite secretion by the vascular plexus and the neuronal cells over time are consistent with PNVP functionality. Multiple concentrations of developmental toxicants (teratogens, microglial disruptor, and vascular network disruptors) significantly reduce the migration of ECs and MG toward the neuronal layer, inhibit formation of the vascular network, and decrease vascular endothelial growth factor A (VEGFA) secretion. By quantifying 3D cell migration, metabolic activity, vascular network disruption, and cytotoxicity, the PNVP model may be a useful tool to make physiologically relevant predictions of developmental toxicity.

Customized hydrogel substrates for serum-free expansion of functional hMSCs.

We describe a screening approach to identify customized substrates for serum-free human mesenchymal stromal cell (hMSC) culture. In particular, we combine a biomaterials screening approach with design of experiments (DOE) and multivariate analysis (MVA) to understand the effects of substrate stiffness, substrate adhesivity, and media composition on hMSC behavior in vitro. This approach enabled identification of poly(ethylene glycol)-based and integrin binding hydrogel substrate compositions that supported functional hMSC expansion in multiple serum-containing and serum-free media, as well as the expansion of MSCs from multiple, distinct sources. The identified substrates were compatible with standard thaw, seed, and harvest protocols. Finally, we used MVA on the screening data to reveal the importance of serum and substrate stiffness on hMSC expansion, highlighting the need for customized cell culture substrates in optimal hMSC biomanufacturing processes.

3-D culture and endothelial cells improve maturity of human pluripotent stem cell-derived hepatocytes.

Human-induced pluripotent stem cell (hiPSC)-derived hepatocytes (iHEP) offer an attractive alternative to primary human hepatocytes (PHH) for drug toxicity studies, as PHHs are limited in supply, vary in their metabolic activity between donors, and rapidly lose their functionality in vitro. However, one of the major drawbacks with iHEP cells in drug safety studies is their decreased phenotypic maturity, with lower liver specific enzyme activity compared with that of PHH. Here we evaluated the effects of 3D culture and non-parenchymal cells on the maturation of iHEPs. We describe a serum-free, chemically defined 3D in vitro model using iHEP cells, which is compatible with automation and conventional assay plates. The iHEP cells cultured in this model form polarized aggregates with functional bile canaliculi and strongly increased expression of albumin, urea and genes encoding phase I and II drug metabolism enzymes and bile transporters. Cytochrome P450-mediated metabolism is significantly higher in 3D iHEP aggregates compared to 2D iHEP culture. Furthermore, addition of human liver sinusoidal endothelial cells (sECs) and iPS-derived endothelial cells (iECs) improved mature hepatocyte function and CYP450 enzyme activity. Also, ECs formed endothelial networks within the hepatic 3D cultures, mimicking aspects of an in vivo architecture. Collectively, these results suggest that the iHEP/EC aggregates described here may have the potential to be used for many applications, including as an in vitro model to study liver diseases associated with sinusoidal endothelial cells. STATEMENT OF SIGNIFICANCE: iPS-derived hepatocytes provide an inexhaustible source of cells for drug screening, toxicology studies and cell-based therapies, but lack mature phenotype of adult primary human hepatocytes (PHH). Herein, we show that 3D culture of iPS-derived hepatocytes and their co-culture with human sinusoidal endothelial cells (sECs) to improve their maturity.

Electrophysiological correlates of visual singleton detection.

Identifying a fixed-feature singleton that pops out from an otherwise uniform array of distractors elicits an ERP component called the N2pc over the posterior scalp. The N2pc has been used to track attention with millisecond accuracy, inform theories of visual selection, and test for specific attention deficits in clinical populations, yet it is still unclear what neuro-cognitive process gives rise to the component. One hypothesis is that the N2pc reflects a spatial filtering process that suppresses irrelevant distractors. In support of this hypothesis, Luck and Hillyard (1994a) showed that the N2pc is eliminated when the features of the target and distractors switch unpredictably across trials (so that participants cannot prepare to filter out irrelevant items). The present study aimed to replicate Luck and Hillyard's singleton detection experiment but with modifications to enhance the N2pc signal and to gain statistical power. We show that orientation singletons do, in fact, elicit the N2pc as well as an earlier-onset and longer-lasting singleton detection positivity over the occipital scalp when the target and distractor orientations swap randomly across trials. We conclude that spatial filtering might not play a major role in the generation of the N2pc and that the selection processes required to search for fixed-feature targets (in feature-search mode) are also engaged in the detection of variable-feature singletons (in singleton detection mode).

Dynamic, Bioresponsive Hydrogels via Changes in DNA Aptamer Conformation.

DNA aptamers are integrated into synthetic hydrogel networks with the aim of creating hydrogels that undergo volume changes when exposed to target molecules. Specifically, single-stranded DNA aptamers in cDNA-bound, extended state are incorporated into hydrogel networks as cross-links, so that the nanoscale conformational change of DNA aptamers upon binding to target molecules will induce macroscopic volume decreases of hydrogels. Hydrogels incorporating adenosine triphosphate (ATP)-binding aptamers undergo controllable volume decreases of up to 40.3 ± 4.6% when exposed to ATP, depending on the concentration of DNA aptamers incorporated in the hydrogel network, temperature, and target molecule concentration. Importantly, this approach can be generalized to aptamer sequences with distinct binding targets, as demonstrated here that hydrogels incorporating an insulin-binding aptamer undergo volume changes in response to soluble insulin. This work provides an example of bioinspired hydrogels that undergo macroscopic volume changes that stem from conformational shifts in resident DNA-based cross-links.

Competitive action video game players display rightward error bias during on-line video game play.

Research in asymmetrical visuospatial attention has identified a leftward bias in the general population across a variety of measures including visual attention and line-bisection tasks. In addition, increases in rightward collisions, or bumping, during visuospatial navigation tasks have been demonstrated in real world and virtual environments. However, little research has investigated these biases beyond the laboratory. The present study uses a semi-naturalistic approach and the online video game streaming service Twitch to examine navigational errors and assaults as skilled action video game players (n = 60) compete in Counter Strike: Global Offensive. This study showed a significant rightward bias in both fatal assaults and navigational errors. Analysis using the in-game ranking system as a measure of skill failed to show a relationship between bias and skill. These results suggest that a leftward visuospatial bias may exist in skilled players during online video game play. However, the present study was unable to account for some factors such as environmental symmetry and player handedness. In conclusion, video game streaming is a promising method for behavioural research in the future, however further study is required before one can determine whether these results are an artefact of the method applied, or representative of a genuine rightward bias.

Materials and toxicological approaches to study metal and metal-oxide nanoparticles in the model organism Caenorhabditis elegans.

Understanding the in vivo fate and transport of nanoparticles (NPs) is challenging, but critical. We review recent studies of metal and metal oxide NPs using the model organism Caenorhabditis elegans, summarizing major findings to date. In a joint transdisciplinary effort, we highlight underutilized opportunities offered by powerful techniques lying at the intersection of mechanistic toxicology and materials science,. To this end, we firstly summarize the influence of exposure conditions (media, duration, C. elegans lifestage) and NP physicochemical properties (size, coating, composition) on the response of C. elegans to NP treatment. Next, we focus on the techniques employed to study NP entrance route, uptake, biodistribution and fate, emphasizing the potential of extending the toolkit available with novel and powerful techniques. Next, we review findings on several NP-induced biological responses, namely transport routes and altered molecular pathways, and illustrate the molecular biology and genetic strategies applied, critically reviewing their strengths and weaknesses. Finally, we advocate the incorporation of a set of minimal materials and toxicological science experiments that will permit meta-analysis and synthesis of multiple studies in the future. We believe this review will facilitate coordinated integration of both well-established and underutilized approaches in mechanistic toxicology and materials science by the nanomaterials research community.

Take your seats: leftward asymmetry in classroom seating choice.

Despite an overall body symmetry, human behavior is full of examples of asymmetry, from writing or gesturing to kissing and cradling. Prior research has revealed that theatre patrons show a bias towards sitting on the right side of a movie theatre. Two competing theories have attempted to explain this seating asymmetry: one posits that expectation of processing demand drives the bias; the other posits that basic motor asymmetries drive the bias. To test these theories we assessed the real-world classroom seating choices of university students using photographs. A bias for students to choose seats on the left side of the classroom was observed, in contrast to the right side bias observed in theatre seating studies. These results provide evidence in support of a processing-expectation bias.

Dynamic stimuli: accentuating aesthetic preference biases.

Despite humans' preference for symmetry, artwork often portrays asymmetrical characteristics that influence the viewer's aesthetic preference for the image. When presented with asymmetrical images, aesthetic preference is often given to images whose content flows from left-to-right and whose mass is located on the right of the image. Cerebral lateralization has been suggested to account for the left-to-right directionality bias; however, the influence of cultural factors, such as scanning habits, on aesthetic preference biases is debated. The current research investigates aesthetic preference for mobile objects and landscapes, as previous research has found contrasting preference for the two image types. Additionally, the current experiment examines the effects of dynamic movement on directionality preference to test the assumption that static images are perceived as aesthetically equivalent to dynamic images. After viewing mirror-imaged pairs of pictures and videos, right-to-left readers failed to show a preference bias, whereas left-to-right readers preferred stimuli with left-to-right directionality regardless of the location of the mass. The directionality bias in both reading groups was accentuated by the videos, but the bias was significantly stronger in left-to-right readers. The findings suggest that scanning habits moderate the leftward bias resulting from hemispheric specialization and that dynamic stimuli further fluent visual processing.

Lateral bias in theatre-seat choice.

Examples of behavioural asymmetries are common in the range of human behaviour; even when faced with a symmetrical environment people demonstrate reliable asymmetries in behaviours like gesturing, cradling, and even seating. One such asymmetry is the observation that participants tend to choose seats to the right of the screen when asked to select their preferred seating location in a movie theatre. However, these results are based on seat selection using a seating chart rather than examining real seat choice behaviour in the theatre context. This study investigated the real-world seating patterns of theatre patrons during actual film screenings. Analysis of bias scores calculated using photographs of theatre patrons revealed a significant bias to choose seats on the right side of the theatre. These findings are consistent with the prior research in the area and confirm that the seating bias observed when seats are selected from a chart accurately reflects real-world seating behaviour.