Experiments as Code and its application to VR studies in human-building interaction.

Experiments as Code (ExaC) is a concept for reproducible, auditable, debuggable, reusable, & scalable experiments. Experiments are a crucial tool to understand Human-Building Interactions (HBI) and build a coherent theory around it. However, a common concern for experiments is their auditability and reproducibility. Experiments are usually designed, provisioned, managed, and analyzed by diverse teams of specialists (e.g., researchers, technicians, engineers) and may require many resources (e.g., cloud infrastructure, specialized equipment). Although researchers strive to document experiments accurately, this process is often lacking. Consequently, it is difficult to reproduce these experiments. Moreover, when it is necessary to create a similar experiment, the "wheel is very often reinvented". It appears easier to start from scratch than trying to reuse existing work. Thus valuable embedded best practices and previous experiences are lost. In behavioral studies, such as in HBI, this has contributed to the reproducibility crisis. To tackle these challenges, we propose the ExaC paradigm, which not only documents the whole experiment, but additionally provides the automation code to provision, deploy, manage, and analyze the experiment. To this end, we define the ExaC concept, provide a taxonomy for the components of a practical implementation, and provide a proof of concept with an HBI desktop VR experiment that demonstrates the benefits of its "as code" representation, that is, reproducibility, auditability, debuggability, reusability, & scalability.

The role of strategic visibility in shaping wayfinding behavior in multilevel buildings.

In this paper, we explore the mutual effect of prior background expectations and visibility afforded by the 3D configuration of the physical environment on wayfinding efficiency and strategy in multilevel buildings. We perform new analyses on data from 149 participants who performed six unaided and directed wayfinding tasks in virtual buildings with varying degrees of visibility. Our findings reveal that the interaction between visibility and prior background expectations significantly affects wayfinding efficiency and strategy during between-floor wayfinding tasks. We termed this interaction effect strategic visibility, which emphasizes the importance of the strategic allocation of visibility towards actionable building elements in promoting efficient wayfinding and shaping wayfinding strategy. Our study highlights the significance of strategic visibility in promoting inclusive and accessible built environments for neurodiversity. Finally, we provide an open-source dataset that can be used to develop and test new wayfinding theories and models to advance research in the emerging field of human-building interaction.

Whole cell hydride Meisenheimer complex biotransformation guided optimization of antimycobacterial benzothiazinones.

Nitrobenzothiazinones (BTZs) are potent active substances against Mycobacterium tuberculosis with currently two investigational drugs in clinical development for the treatment of tuberculosis. BTZs are the first examples for which a metabolic pathway towards transient hydride Meisenheimer complexes (HMC) has been shown in mammals, including humans. In this study, lead optimization efforts on BTZs are guided by the systematic evaluation of the HMC formation propensity combined with multiparameter assessment. For this purpose, a novel cell-based assay was specifically developed and fully implemented, and a library of 5- and 7-substituted BTZs was prepared to study substituent effects on the HMC formation. The multiparameter optimization revealed 5-methylated BTZs as the most preferred scaffolds, demonstrating a reduced HMC formation propensity combined with potent activity and good microsomal stability in vitro. In vivo experiments showed good systemic exposure upon oral administration and efficacy in a murine M. tuberculosis infection model. This study reports a qualified in vitro HMC assay, which not only enabled the selection of next-generation BTZs with improved pharmacokinetic properties but also allowed forecasting their in vivo metabolism.

The influence of upward social comparison on retail trading behaviour.

Online investing is often facilitated by digital platforms, where the information of peer top performers can be widely accessible and distributed. However, the influence of such information on retail investors' psychology, their trading behaviour and potential risks they may be prone to is poorly understood. We investigate the impact of upward social comparison on risk-taking, trading activity and investor satisfaction using a tailored experiment with 807 experienced retail investors trading on a dynamically evolving simulated stock market, designed to systematically measure various facets of trading activity. We find that investors presented with an upward social comparison take more risk and trade more actively, and they report significantly lower satisfaction with their own performance. Our findings demonstrate the pitfalls of modern investment platforms with peer information and social trading. The broad implications of this study also provide guidelines for improving retail investor satisfaction and protection.

Motivation moderates gender differences in navigation performance.

Gender differences in navigation performance are a recurrent and controversial topic. Previous research suggests that men outperform women in navigation tasks and that men and women exhibit different navigation strategies. Here, we investigate whether motivation to complete the task moderates the relationship between navigation performance and gender. Participants learned the locations of landmarks in a novel virtual city. During learning, participants could trigger a top-down map that depicted their current position and the locations of the landmarks. During testing, participants were divided into control and treatment groups and were not allowed to consult the map. All participants were given 16 minutes to navigate to the landmarks, but those in the treatment group were monetarily penalized for every second they spent completing the task. Results revealed a negative relationship between physiological arousal and the time required to locate the landmarks. In addition, gender differences in strategy were found during learning, with women spending more time with the map and taking 40% longer than men to locate the landmarks. Interestingly, an interaction between gender and treatment group revealed that women in the control group required more time than men and women in the treatment group to retrieve the landmarks. During testing, women in the control group also took more circuitous routes compared to men in the control group and women in the treatment group. These results suggest that a concurrent and relevant stressor can motivate women to perform similarly to men, helping to diminish pervasive gender differences found in the navigation literature.

Cognitive Path Planning With Spatial Memory Distortion.

Human path-planning operates differently from deterministic AI-based path-planning algorithms due to the decay and distortion in a human's spatial memory and the lack of complete scene knowledge. Here, we present a cognitive model of path-planning that simulates human-like learning of unfamiliar environments, supports systematic degradation in spatial memory, and distorts spatial recall during path-planning. We propose a Dynamic Hierarchical Cognitive Graph (DHCG) representation to encode the environment structure by incorporating two critical spatial memory biases during exploration: categorical adjustment and sequence order effect. We then extend the "Fine-To-Coarse" (FTC), the most prevalent path-planning heuristic, to incorporate spatial uncertainty during recall through the DHCG. We conducted a lab-based Virtual Reality (VR) experiment to validate the proposed cognitive path-planning model and made three observations: (1) a statistically significant impact of sequence order effect on participants' route-choices, (2) approximately three hierarchical levels in the DHCG according to participants' recall data, and (3) similar trajectories and significantly similar wayfinding performances between participants and simulated cognitive agents on identical path-planning tasks. Furthermore, we performed two detailed simulation experiments with different FTC variants on a Manhattan-style grid. Experimental results demonstrate that the proposed cognitive path-planning model successfully produces human-like paths and can capture human wayfinding's complex and dynamic nature, which traditional AI-based path-planning algorithms cannot capture.

A mobile EEG study on the psychophysiological effects of walking and crowding in indoor and outdoor urban environments.

Environmental psychologists have established multiple psychological benefits of interaction with natural, compared to urban, environments on emotion, cognition, and attention. Yet, given the increasing urbanisation worldwide, it is equally important to understand how differences within different urban environments influence human psychological experience. We developed a laboratory experiment to examine the psychophysiological effects of the physical (outdoor or indoor) and social (crowded versus uncrowded) environment in healthy young adults, and to validate the use of mobile electroencephalography (EEG) and electrodermal activity (EDA) measurements during active walking. Participants (N = 42) were randomly assigned into a walking or a standing group, and watched six 1-min walk-through videos of green, urban indoor and urban outdoor environments, depicting high or low levels of social density. Self-reported emotional states show that green spaces is perceived as more calm and positive, and reduce attentional demands. Further, the outdoor urban space is perceived more positively than the indoor environment. These findings are consistent with earlier studies on the psychological benefits of nature and confirm the effectiveness of our paradigm and stimuli. In addition, we hypothesised that even short-term exposure to crowded scenes would have negative psychological effects. We found that crowded scenes evoked higher self-reported arousal, more negative self-reported valence, and recruited more cognitive and attentional resources. However, in walking participants, they evoked higher frontal alpha asymmetry, suggesting more positive affective responses. Furthermore, we found that using recent signal-processing methods, the EEG data produced a comparable signal-to-noise ratio between walking and standing, and that despite differences between walking and standing, skin-conductance also captured effectively psychophysiological responses to stimuli. These results suggest that emotional responses to visually presented stimuli can be measured effectively using mobile EEG and EDA in ambulatory settings, and that there is complex interaction between active walking, the social density of urban spaces, and direct and indirect affective responses to such environments.

High-Dose Mycobacterium tuberculosis H37rv Infection in IL-17A- and IL-17A/F-Deficient Mice.

During experimental tuberculosis (TB), interleukin (IL)-17A appears to be involved in the formation of lung granulomas, possibly through the attraction of neutrophils to the sites of infection. However, the protective impact of cytokine appears to depend on the degree of its induction. Hence, robust production of IL-17A in mice infected with the hypervirulent isolate Mycobacterium tuberculosis (Mtb) HN878 mediates protection, while the cytokine is dispensable for protective immune responses against low-dose infection with the less virulent strain H37rv. Here, we show that after experimental infection with high doses of Mtb H37rv, IL-17A-deficient (-/-) mice exhibited high susceptibility to the infection, which was mediated by the strong accumulation of neutrophils in the infected lung tissue. Accordingly, we observed nearly unrestricted bacterial replication within the neutrophils, indicating that they may serve as a survival niche for Mtb. By use of IL-17A/IL-17F-double-deficient mice, we demonstrated that the susceptibility in the absence of IL-17A is mediated by a compensatory expression of IL-17F, which, however, appeared not to be dependent on neutrophils. Together, our results illustrate the compensatory potential of the Th17-secreted cytokines IL-17A and IL-17F in the context of experimental TB and once again emphasize the detrimental effect of excessive neutrophil infiltration in response to Mtb.

Tuberculostearic Acid-Containing Phosphatidylinositols as Markers of Bacterial Burden in Tuberculosis.

One-fourth of the global human population is estimated to be infected with strains of the Mycobacterium tuberculosis complex (MTBC), the causative agent of tuberculosis (TB). Using lipidomic approaches, we show that tuberculostearic acid (TSA)-containing phosphatidylinositols (PIs) are molecular markers for infection with clinically relevant MTBC strains and signify bacterial burden. For the most abundant lipid marker, detection limits of ∼102 colony forming units (CFUs) and ∼103 CFUs for bacterial and cell culture systems were determined, respectively. We developed a targeted lipid assay, which can be performed within a day including sample preparation─roughly 30-fold faster than in conventional methods based on bacterial culture. This indirect and culture-free detection approach allowed us to determine pathogen loads in infected murine macrophages, human neutrophils, and murine lung tissue. These marker lipids inferred from mycobacterial PIs were found in higher levels in peripheral blood mononuclear cells of TB patients compared to healthy individuals. Moreover, in a small cohort of drug-susceptible TB patients, elevated levels of these molecular markers were detected at the start of therapy and declined upon successful anti-TB treatment. Thus, the concentration of TSA-containing PIs can be used as a correlate for the mycobacterial burden in experimental models and in vitro systems and may prospectively also provide a clinically relevant tool to monitor TB severity.

Do Anti-tuberculosis Drugs Reach Their Target?─High-Resolution Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging Provides Information on Drug Penetration into Necrotic Granulomas.

Tuberculosis (TB) is characterized by mycobacteria-harboring centrally necrotizing granulomas. The efficacy of anti-TB drugs depends on their ability to reach the bacteria in the center of these lesions. Therefore, we developed a mass spectrometry (MS) imaging workflow to evaluate drug penetration in tissue. We employed a specific mouse model that─in contrast to regular inbred mice─strongly resembles human TB pathology. Mycobacterium tuberculosis was inactivated in lung sections of these mice by γ-irradiation using a protocol that was optimized to be compatible with high spatial resolution MS imaging. Different distributions in necrotic granulomas could be observed for the anti-TB drugs clofazimine, pyrazinamide, and rifampicin at a pixel size of 30 µm. Clofazimine, imaged here for the first time in necrotic granulomas of mice, showed higher intensities in the surrounding tissue than in necrotic granulomas, confirming data observed in TB patients. Using high spatial resolution drug and lipid imaging (5 µm pixel size) in combination with a newly developed data analysis tool, we found that clofazimine does penetrate to some extent into necrotic granulomas and accumulates in the macrophages inside the granulomas. These results demonstrate that our imaging platform improves the predictive power of preclinical animal models. Our workflow is currently being applied in preclinical studies for novel anti-TB drugs within the German Center for Infection Research (DZIF). It can also be extended to other applications in drug development and beyond. In particular, our data analysis approach can be used to investigate diffusion processes by MS imaging in general.

Interleukin-13-Overexpressing Mice Represent an Advanced Preclinical Model for Detecting the Distribution of Antimycobacterial Drugs within Centrally Necrotizing Granulomas.

The Mycobacterium tuberculosis-harboring granuloma with a necrotic center surrounded by a fibrous capsule is the hallmark of tuberculosis (TB). For a successful treatment, antibiotics need to penetrate these complex structures to reach their bacterial targets. Hence, animal models reflecting the pulmonary pathology of TB patients are of particular importance to improve the preclinical validation of novel drug candidates. M. tuberculosis-infected interleukin-13-overexpressing (IL-13tg) mice develop a TB pathology very similar to patients and, in contrast to other mouse models, also share pathogenetic mechanisms. Accordingly, IL-13tg animals represent an ideal model for analyzing the penetration of novel anti-TB drugs into various compartments of necrotic granulomas by matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MS imaging). In the present study, we evaluated the suitability of BALB/c IL-13tg mice for determining the antibiotic distribution within necrotizing lesions. To this end, we established a workflow based on the inactivation of M. tuberculosis by gamma irradiation while preserving lung tissue integrity and drug distribution, which is essential for correlating drug penetration with lesion pathology. MALDI-MS imaging analysis of clofazimine, pyrazinamide, and rifampicin revealed a drug-specific distribution within different lesion types, including cellular granulomas, developing in BALB/c wild-type mice, and necrotic granulomas in BALB/c IL-13tg animals, emphasizing the necessity of preclinical models reflecting human pathology. Most importantly, our study demonstrates that BALB/c IL-13tg mice recapitulate the penetration of antibiotics into human lesions. Therefore, our workflow in combination with the IL-13tg mouse model provides an improved and accelerated evaluation of novel anti-TB drugs and new regimens in the preclinical stage.

Gasdermin D mediates host cell death but not interleukin-1ß secretion in Mycobacterium tuberculosis-infected macrophages.

Necrotic cell death represents a major pathogenic mechanism of Mycobacterium tuberculosis (Mtb) infection. It is increasingly evident that Mtb induces several types of regulated necrosis but how these are interconnected and linked to the release of pro-inflammatory cytokines remains unknown. Exploiting a clinical cohort of tuberculosis patients, we show here that the number and size of necrotic lesions correlates with IL-1ß plasma levels as a strong indicator of inflammasome activation. Our mechanistic studies reveal that Mtb triggers mitochondrial permeability transition (mPT) and subsequently extensive macrophage necrosis, which requires activation of the NLRP3 inflammasome. NLRP3-driven mitochondrial damage is dependent on proteolytic activation of the pore-forming effector protein gasdermin D (GSDMD), which links two distinct cell death machineries. Intriguingly, GSDMD, but not the membranolytic mycobacterial ESX-1 secretion system, is dispensable for IL-1ß secretion from Mtb-infected macrophages. Thus, our study dissects a novel mechanism of pathogen-induced regulated necrosis by identifying mitochondria as central regulatory hubs capable of delineating cytokine secretion and lytic cell death.

Visibility matters during wayfinding in the vertical.

Visibility is the degree to which different parts of the environment can be observed from a given vantage point. In the absence of previous familiarity or signage, the visibility of key elements in a multilevel environment (e.g., the entrance, exit, or the destination itself) becomes a primary input to make wayfinding decisions and avoid getting lost. Previous research has focused on memory-based wayfinding and mental representation of 3D space, but few studies have investigated the direct effects of visibility on wayfinding. Moreover, to our knowledge, there are no studies that have explicitly observed the interaction between visibility and wayfinding under uncertainty in a multilevel environment. To bridge this gap, we studied how the visibility of destinations, as well as the continuity of sight-lines along the vertical dimension, affects unaided and goal-directed wayfinding behavior in a multilevel desktop Virtual Reality (VR) study. We obtained results from a total of 69 participants. Each participant performed a total of 24 wayfinding trials in a multilevel environment. Results showcase a significant and nonlinear correlation between the visibility of destinations and wayfinding behavioral characteristics. Specifically, once the destination was in sight, regardless of whether it was highly or barely visible, participants made an instantaneous decision to switch floors and move up towards the destination. In contrast, if the destination was out-of-sight, participants performed 'visual exploration', indicated by an increase in vertical head movements and greater time taken to switch floors. To demonstrate the direct applicability of this fundamental wayfinding behavioral pattern, we formalize these results by modeling a visibility-based cognitive agent. Our results show that by modeling the transition between exploration and exploitation as a function of visibility, cognitive agents were able to replicate human wayfinding patterns observed in the desktop VR study. This simple demonstration shows the potential of extending our main findings concerning the nonlinear relationship between visibility and wayfinding to inform the modeling of human cognition.

Interleukin-23 instructs protective multifunctional CD4 T cell responses after immunization with the Mycobacterium tuberculosis subunit vaccine H1 DDA/TDB independently of interleukin-17A.

Interleukin (IL)-17A-producing T helper (Th)17 cells are increasingly being acknowledged to be associated with protective immunity to Mycobacterium tuberculosis (Mtb). Subunit vaccines potently promote protective immune responses against Mtb infection that correlate with an expansion of IL-23-dependent Th17 cells. Previous studies revealed that after vaccination, IL-23 is required for protection against challenge with Mtb but the underlying IL-23-dependent-and possibly IL-17A-mediated-mechanisms remain elusive. Therefore, we here analyzed the early outcome of Mtb infection in C57BL/6, IL-23p19-deficient (-/-), and IL-17A-/- mice after vaccination with the subunit vaccine H1-DDA/TDB to investigate the role of the IL-23-Th17 immune axis for the instruction of vaccine-induced protection. While in IL-23p19-/- mice the protective effect was reduced, protection after vaccination was maintained in IL-17A-/- animals for the course of infection of 6 weeks, indicating that after vaccination with H1-DDA/TDB early protection against Mtb is-although dependent on IL-23-not mediated by IL-17A. In contrast, IL-17A deficiency appears to have an impact on maintaining long-term protection. In fact, IL-23 instructed the vaccine-induced memory immunity in the lung, in particular the sustained expansion of tumor necrosis factor (TNF)+IL-2+ multifunctional T cells, independently of IL-17A. Altogether, a targeted induction of IL-23 during vaccination against Mtb might improve the magnitude and quality of vaccine-induced memory immune responses. KEY MESSAGES: After subunit Mtb vaccination with H1-DDA/TDB, IL-23 but not IL-17A contributes to vaccine-induced early protection against infection with Mtb. IL-17F does not compensate for IL-17A deficiency in terms of H1-DDA/TDB-induced protection against Mtb infection. IL 23 promotes the H1-DDA/TDB-induced accumulation of effector memory T cells independently of IL 17A. IL-23 arbitrates the induction of H1-specific IFN-γ-TNF+IL-2+ double-positive multifunctional CD4 T cells after subunit Mtb vaccination in an IL-17A-independent manner.

WNT6/ACC2-induced storage of triacylglycerols in macrophages is exploited by Mycobacterium tuberculosis.

In view of emerging drug-resistant tuberculosis (TB), host-directed adjunct therapies are urgently needed to improve treatment outcomes with currently available anti-TB therapies. One approach is to interfere with the formation of lipid-laden "foamy" macrophages in the host, as they provide a nutrient-rich host cell environment for Mycobacterium tuberculosis (Mtb). Here, we provide evidence that Wnt family member 6 (WNT6), a ligand of the evolutionarily conserved Wingless/Integrase 1 (WNT) signaling pathway, promotes foam cell formation by regulating key lipid metabolic genes including acetyl-CoA carboxylase 2 (ACC2) during pulmonary TB. Using genetic and pharmacological approaches, we demonstrated that lack of functional WNT6 or ACC2 significantly reduced intracellular triacylglycerol (TAG) levels and Mtb survival in macrophages. Moreover, treatment of Mtb-infected mice with a combination of a pharmacological ACC2 inhibitor and the anti-TB drug isoniazid (INH) reduced lung TAG and cytokine levels, as well as lung weights, compared with treatment with INH alone. This combination also reduced Mtb bacterial numbers and the size of mononuclear cell infiltrates in livers of infected mice. In summary, our findings demonstrate that Mtb exploits WNT6/ACC2-induced storage of TAGs in macrophages to facilitate its intracellular survival, a finding that opens new perspectives for host-directed adjunctive treatment of pulmonary TB.

Monocyte progenitors give rise to multinucleated giant cells.

The immune response to mycobacteria is characterized by granuloma formation, which features multinucleated giant cells as a unique macrophage type. We previously found that multinucleated giant cells result from Toll-like receptor-induced DNA damage and cell autonomous cell cycle modifications. However, the giant cell progenitor identity remained unclear. Here, we show that the giant cell-forming potential is a particular trait of monocyte progenitors. Common monocyte progenitors potently produce cytokines in response to mycobacteria and their immune-active molecules. In addition, common monocyte progenitors accumulate cholesterol and lipids, which are prerequisites for giant cell transformation. Inducible monocyte progenitors are so far undescribed circulating common monocyte progenitor descendants with high giant cell-forming potential. Monocyte progenitors are induced in mycobacterial infections and localize to granulomas. Accordingly, they exhibit important immunological functions in mycobacterial infections. Moreover, their signature trait of high cholesterol metabolism may be piggy-backed by mycobacteria to create a permissive niche.

DGCR8 deficiency impairs macrophage growth and unleashes the interferon response to mycobacteria.

The mycobacterial cell wall glycolipid trehalose-6,6-dimycolate (TDM) activates macrophages through the C-type lectin receptor MINCLE. Regulation of innate immune cells relies on miRNAs, which may be exploited by mycobacteria to survive and replicate in macrophages. Here, we have used macrophages deficient in the microprocessor component DGCR8 to investigate the impact of miRNA on the response to TDM. Deletion of DGCR8 in bone marrow progenitors reduced macrophage yield, but did not block macrophage differentiation. DGCR8-deficient macrophages showed reduced constitutive and TDM-inducible miRNA expression. RNAseq analysis revealed that they accumulated primary miRNA transcripts and displayed a modest type I IFN signature at baseline. Stimulation with TDM in the absence of DGCR8 induced overshooting expression of IFNß and IFN-induced genes, which was blocked by antibodies to type I IFN. In contrast, signaling and transcriptional responses to recombinant IFNß were unaltered. Infection with live Mycobacterium bovis Bacille Calmette-Guerin replicated the enhanced IFN response. Together, our results reveal an essential role for DGCR8 in curbing IFNß expression macrophage reprogramming by mycobacteria.

The effect of navigation method and visual display on distance perception in a large-scale virtual building.

Immersive virtual reality (VR) technology has become a popular method for fundamental and applied spatial cognition research. One challenge researchers face is emulating walking in a large-scale virtual space although the user is in fact in a small physical space. To address this, a variety of movement interfaces in VR have been proposed, from traditional joysticks to teleportation and omnidirectional treadmills. These movement methods tap into different mental processes of spatial learning during navigation, but their impacts on distance perception remain unclear. In this paper, we investigated the role of visual display, proprioception, and optic flow on distance perception in a large-scale building by manipulating four different movement methods. Eighty participants either walked in a real building, or moved through its virtual replica using one of three movement methods: VR-treadmill, VR-touchpad, and VR-teleportation. Results revealed that, first, visual display played a major role in both perceived and traversed distance estimates but did not impact environmental distance estimates. Second, proprioception and optic flow did not impact the overall accuracy of distance perception, but having only an intermittent optic flow (in the VR-teleportation movement method) impaired the precision of traversed distance estimates. In conclusion, movement method plays a significant role in distance perception but does not impact the configurational knowledge learned in a large-scale real and virtual building, and the VR-touchpad movement method provides an effective interface for navigation in VR.