Search

Abstract Chassis strains, derived from Streptomyces coelicolor M145, deleted for one or more of its four main specialized metabolites biosynthetic pathways (CPK, CDA, RED and ACT), in various combinations, were constructed for the heterologous expression of specialized metabolites biosynthetic pathways of various types and origins. To determine consequences of these deletions on the metabolism of the deleted strains comparative lipidomic and metabolomic analyses of these strains and of the original strain were carried out. These studies unexpectedly revealed that the deletion of the peptidic clusters, RED and/or CDA, in a strain deleted for the ACT cluster, resulted into a great increase in the triacylglycerol (TAG) content, whereas the deletion of polyketide clusters, ACT and CPK had no impact on TAG content. Low or high TAG content of the deleted strains was correlated with abundance or paucity in amino acids, respectively, reflecting high or low activity of oxidative metabolism. Hypotheses based on what is known on the bio‐activity and the nature of the precursors of these specialized metabolites are proposed to explain the unexpected consequences of the deletion of these pathways on the metabolism of the bacteria and on the efficiency of the deleted strains as chassis strains.

Since nitrogen limitation is known to be an important trigger of triacylglycerol (TAG) accumulation in most microorganisms, we first assessed the global lipid content of 21 strains derived from Streptomyces coelicolor M145 deleted for genes involved in nitrogen metabolism. Seven of these strains deleted for genes encoding proteins involved in polyamine (GlnA2/SCO2241, GlnA3/SCO6962, GlnA4/SCO1613), or protein (Pup/SCO1646) degradation, in the regulation of nitrogen metabolism (GlnE/SCO2234 and GlnK/SCO5584), or the global regulator DasR/SCO5231 that controls negatively the degradation of N-acetylglucosamine, a constituent of peptidoglycan, had a higher TAG content than the original strain, whereas five of these strains (except the glnA2 and pup mutants) had a lower cardiolipin (CL) content. The production of the blue polyketide actinorhodin (ACT) was totally abolished in the dasR mutant in both Pi conditions, whereas the deletion of pup, glnA2, glnA3, and glnA4 was correlated with a significant increase in total ACT production, but mainly in Pi limitation. Unexpectedly, ACT production was strongly reduced in the glnA3 mutant in Pi proficiency. Altogether, our data suggest that high TAG and ACT biosynthesis and low CL biosynthesis might all contribute to the lowering of oxidative stress resulting from nitrogen limitation or from other causes.

We present an analytical framework aimed at predicting the local brain activity in uncontrolled experimental conditions based on multimodal recordings of participants' behavior, and its application to a corpus of participants having conversations with another human or a conversational humanoid robot. The framework consists in extracting high-level features from the raw behavioral recordings and applying a dynamic prediction of binarized fMRI-recorded local brain activity using these behavioral features. The objective is to identify behavioral features required for this prediction, and their relative weights, depending on the brain area under investigation and the experimental condition. In order to validate our framework, we use a corpus of uncontrolled conversations of participants with a human or a robotic agent, focusing on brain regions involved in speech processing, and more generally in social interactions. The framework not only predicts local brain activity significantly better than random, it also quantifies the weights of behavioral features required for this prediction, depending on the brain area under investigation and on the nature of the conversational partner. In the left Superior Temporal Sulcus, perceived speech is the most important behavioral feature for predicting brain activity, regardless of the agent, while several features, which differ between the human and robot interlocutors, contribute to the prediction in regions involved in social cognition, such as the TemporoParietal Junction. This framework therefore allows us to study how multiple behavioral signals from different modalities are integrated in individual brain regions during complex social interactions.

Abstract Human–human interactions (HHI) and human–robot interactions (HRI) are compared to identify differences between cognitive processes reflecting bonding in social interactions with natural and artificial agents. We capitalize on a unique corpus of neuroimaging data (fMRI) recorded while participants freely discussed with another human or a conversational robotic head, in order to study a crucial parameter of human social cognition, namely that social interactions are adaptive bidirectional processes that evolve over time. We used linear statistics to identify regions of the brain where activity changes differently when participants carry out twelve one-minute conversations, alternating between a human and a robotic interlocutor. Results show that activity in the posterior cingulate cortex, a key region associated with social cognition, increases over time in HHI but not in HRI. These results are interpreted as reflecting a process of strengthening social bonding during repeated exchanges when the interacting agent is a human, but not a robot.

Emotional contagion, in particular of happiness, is essential to creating social bonds. The somatic marker hypothesis posits that embodied physiological changes associated with emotions and relayed to the brain by the autonomous nervous system influence behavior. Perceiving others’ positive emotions should thus be associated with activity in brain regions relaying information from and to the autonomic nervous system. Here, we address this question using a unique corpus of brain activity recorded during unconstrained conversations between participants and a human or a humanoid robot. fMRI recordings are used to test whether activity in key brain regions of the autonomic system, the amygdala, hypothalamus, and insula, is differentially affected by the level of happiness expressed by the human and robot agents. Results indicate that for the hypothalamus and the insula, in particular the anterior agranular region strongly involved in processing social emotions, activity in the right hemisphere increases with the level of happiness expressed by the human but not the robot. Perceiving positive emotions in social interactions induces local brain responses predicted by the contagion of somatic markers of emotions only when the interacting agent is a fellow human.

Abstract Human behaviors from toolmaking to language are thought to rely on a uniquely evolved capacity for hierarchical action sequencing. Testing this idea will require objective, generalizable methods for measuring the structural complexity of real-world behavior. Here we present a data-driven approach for extracting action grammars from basic ethograms, exemplified with respect to the evolutionarily relevant behavior of stone toolmaking. We analyzed sequences from the experimental replication of ~ 2.5 Mya Oldowan vs. ~ 0.5 Mya Acheulean tools, finding that, while using the same “alphabet” of elementary actions, Acheulean sequences are quantifiably more complex and Oldowan grammars are a subset of Acheulean grammars. We illustrate the utility of our complexity measures by re-analyzing data from an fMRI study of stone toolmaking to identify brain responses to structural complexity. Beyond specific implications regarding the co-evolution of language and technology, this exercise illustrates the general applicability of our method to investigate naturalistic human behavior and cognition.

In so-called ethorobotics and robot-supported social cognitive neurosciences, robots are used as scientific tools to study animal behavior and cognition. Building on previous epistemological analyses of biorobotics, in this article it is argued that these two research fields, widely differing from one another in the kinds of robots involved and in the research questions addressed, share a common methodology, which significantly differs from the “synthetic method” that, until recently, dominated biorobotics. The methodological novelty of this strategy, the research opportunities that it opens, and the theoretical and technological challenges that it gives rise to, will be discussed with reference to the peculiarities of the two research fields. Some broad methodological issues related to the generalization of results concerning robot-animal interaction to theoretical conclusions on animal-animal interaction will be identified and discussed.

Streptomyces coelicolor M145 is a model strain extensively studied to elucidate the regulation of antibiotic biosynthesis in Streptomyces species. This strain abundantly produces the blue polyketide antibiotic, actinorhodin (ACT), and has a low lipid content. In a process designed to delete the gene encoding the isocitrate lyase (sco0982) of the glyoxylate cycle, an unexpected variant of S. coelicolor was obtained besides bona fide sco0982 deletion mutants. This variant produces 7- to 15-fold less ACT and has a 3-fold higher triacylglycerol and phosphatidylethanolamine content than the original strain. The genome of this variant was sequenced and revealed that 704 genes were deleted (9% of total number of genes) through deletions of various sizes accompanied by the massive loss of mobile genetic elements. Some deletions include genes whose absence could be related to the high total lipid content of this variant such as those encoding enzymes of the TCA and glyoxylate cycles, enzymes involved in nitrogen assimilation as well as enzymes belonging to some polyketide and possibly trehalose biosynthetic pathways. The characteristics of this deleted variant of S. coelicolor are consistent with the existence of the previously reported negative correlation existing between lipid content and antibiotic production in Streptomyces species.

This article investigates the differences in cognitive and neural mechanisms between human-human and human-virtual agent interaction using a dataset recorded in an ecologically realistic environment. We use Convergent Cross Mapping (CCM) to investigate functional connectivity between pairs of regions involved in the framework of social cognitive neuroscience, namely the fusiform gyrus, superior temporal sulcus (STS), temporoparietal junction (TPJ), and the dorsolateral prefrontal cortex (DLPFC)—taken as prefrontal asymmetry. Our approach is a compromise between investigating local activation in specific regions and investigating connectivity networks that may form part of larger networks. In addition to concording with previous studies, our results suggest that the right TPJ is one of the most reliable areas for assessing processes occurring during human-virtual agent interactions, both in a static and dynamic sense.

Yarrowia lipolytica is a promising oleaginous yeast for producing unusual lipids, such as odd-chain fatty acids (OCFA). Their diverse applications and low natural production make OCFA particularly interesting. In recent studies, inhibiting the catabolic pathway of precursor, boosting precursor pools, and optimizing substrate combination greatly improved the production of OCFA in Y. lipolytica. We explored the lipid readjustment of OCFA in engineered Y. lipolytica strains. NPLC-Corona-CAD® evidenced a time-dependent overproduction of free fatty acids, diglycerides, and phosphatidylcholine (PC) in obese LP compared to obese L. Phosphatidylethanolamine (PE) and phosphatidylinositol, largely overproduced in obese LP at 72 h compared to obese L, vanished at 216 h. The fatty acyls (FAs) composition of glycero- and glycerophospholipids was determined by NPLC-APPI+-HRMS from in-source generated monoacylglycerol-like fragment ions. C18:1 and C17:1 were predominant acylglycerols in obese L and obese LP, respectively. Phosphatidic acid, PE, and PC exhibited similar FAs composition but differed in their molecular species distributions. Cardiolipin (CL) is known to contain mostly C18:2 FAs corresponding to the composition in obese L, 50% of C18:2, and 35% of C18:1. In obese LP, both FAs dropped to drop to 20%, and C17:1 were predominant, reaching 55%. We hypothesize that CL-modified composition in obese LPs may alter mitochondrial function and limit lipid production.

In this issue we demonstrated that the phospholipid content of Streptomyces lividans varies greatly with Pi availability being was much lower in Pi limitation than in Pi proficiency whereas that of Streptomyces coelicolor varied little with Pi availability. In contrast the content in phosphate free ornithine lipids was enhanced in both strains in condition of phosphate limitation. Ornithine lipids biosynthesis starts with the N-acylation of ornithine to form lyso-ornithine that is then O-acylated to yield ornithine lipid. The operon sco1222-23 was proposed to be involved in the conversion of specific amino acids into ornithine in condition of phosphate limitation whereas the sco0921-20 operon encoding N- and O-acyltransferase, respectively, was shown to be involved in the biosynthesis of these lipids. The expression of these two operons was shown to be under the positive control of the two components system PhoR/PhoP and thus induced in phosphate limitation. The expression of phoR/phoP being weak in S. coelicolor, the poor expression of these operons resulted into a fivefold lower ornithine lipids content in this strain compared to S. lividans. In the deletion mutant of the sco0921-20 operon of S. lividans, lyso-ornithine and ornithine lipids were barely detectable and TAG content was enhanced. The complementation of this mutant by the sco0921-20 operon or by sco0920 alone restored ornithine lipids and TAG content to wild type level and was correlated with a twofold increase in the cardiolipin content. This suggested that SCO0920 bears, besides its broad O-acyltransferase activity, an N-acyltransferase activity and this was confirmed by the detection of lyso-ornithine in this strain. In contrast, the complementation of the mutant by sco0921 alone had no impact on ornithine lipids, TAG nor cardiolipin content but was correlated with a high lyso-ornithine content. This confirmed that SCO0921 is a strict N-acyltransferase. However, interestingly, the over-expression of the sco0921-20 operon or of sco0921 alone in S. coelicolor, led to an almost total disappearance of phosphatidylinositol that was correlated with an enhanced DAG and TAG content. This suggested that SCO0921 also acts as a phospholipase C, degrading phosphatidylinositol to indirectly supply of phosphate in condition of phosphate limitation.

Retroviruses package two copies of their genomic RNA (gRNA) as non-covalently linked dimers. Many studies suggest that the retroviral nucleocapsid protein (NC) plays an important role in gRNA dimerization. The upper part of the L3 RNA stem-loop in the 5′ leader of the avian leukosis virus (ALV) is converted to the extended dimer by ALV NC. The L3 hairpin contains three stems and two internal loops. To investigate the roles of internal loops and stems in the NC-mediated extended dimer formation, we performed site-directed mutagenesis, gel electrophoresis, and analysis of thermostability of dimeric RNAs. We showed that the internal loops are necessary for efficient extended dimer formation. Destabilization of the lower stem of L3 is necessary for RNA dimerization, although it is not involved in the linkage structure of the extended dimer. We found that NCs from ALV, human immunodeficiency virus type 1 (HIV-1), and Moloney murine leukemia virus (M-MuLV) cannot promote the formation of the extended dimer when the apical stem contains ten consecutive base pairs. Five base pairs correspond to the maximum length for efficient L3 dimerization induced by the three NCs. L3 dimerization was less efficient with M-MuLV NC than with ALV NC and HIV-1 NC.

In condition of over-expression, SCO3201, a regulator of the TetR family was previously shown to strongly inhibit antibiotic production and morphological differentiation in Streptomyces coelicolor M145. In order to elucidate the molecular processes underlying this interesting, but poorly understood phenomenon, a comparative analysis of the lipidomes and transcriptomes of the strain over-expressing sco3201 and of the control strain containing the empty plasmid, was carried out. This study revealed that the strain over-expressing sco3201 had a higher triacylglycerol content and a lower phospholipids content than the control strain. This was correlated with up- and down- regulation of some genes involved in fatty acids biosynthesis (fab) and degradation (fad) respectively, indicating a direct or indirect control of the expression of these genes by SCO3201. In some instances, indirect control might involve TetR regulators, whose encoding genes present in close vicinity of genes involved in lipid metabolism, were shown to be differentially expressed in the two strains. Direct interaction of purified His6-SCO3201 with the promoter regions of four of such TetR regulators encoding genes (sco0116, sco0430, sco4167, and sco6792) was demonstrated. Furthermore, fasR (sco2386), encoding the activator of the main fatty acid biosynthetic operon, sco2386-sco2390, has been shown to be an illegitimate positive regulatory target of SCO3201. Altogether our data demonstrated that the sco3201 over-expressing strain accumulates TAG and suggested that degradation of fatty acids was reduced in this strain. This is expected to result into a reduced acetyl-CoA availability that would impair antibiotic biosynthesis either directly or indirectly.

Background: Recent studies suggest that Posttraumatic Stress Disorder (PTSD) might be associated with dysfunctional reward circuitry. However, further research is needed to understand the key role of the reward system in PTSD symptomatology. Methods: Twenty participants with PTSD and 21 Trauma-Exposed matched Controls (TECs) completed the Monetary Incentive Delay (MID) task during an MRI session. Reaction times (RTs) and hit rates were recorded. Brain activity was investigated during the anticipation and the outcome of monetary gains and losses. Results: During the anticipation of monetary loss, PTSD participants had higher RTs than TECs. However, the groups did not differ at the neurofunctional level. During successful avoidance of monetary loss, PTSD patients showed higher activation than TECs in the left caudate nucleus. During the anticipation of monetary gains, no differences in RTs were found between groups. PTSD patients had specific activations in the right amygdala, nucleus accumbens, putamen, and middle frontal gyrus (p

Multivariate pattern analysis (MVPA) has become vastly popular for analyzing functional neuroimaging data. At the group level, two main strategies are used in the literature. The standard one is hierarchical, combining the outcomes of within-subject decoding results in a second-level analysis. The alternative one, inter-subject pattern analysis, directly works at the group-level by using, e.g. a leave-one-subject-out cross-validation. This study provides a thorough comparison of these two group-level decoding schemes, using both a large number of artificial datasets where the size of the multivariate effect and the amount of inter-individual variability are parametrically controlled, as well as two real fMRI datasets comprising 15 and 39 subjects, respectively. We show that these two strategies uncover distinct significant regions with partial overlap, and that inter-subject pattern analysis is able to detect smaller effects and to facilitate the interpretation. The core source code and data are openly available, allowing to fully reproduce most of these results.

Objective: Neurobiological models of Posttraumatic Stress Disorder (PTSD) implicate fear processing impairments in the maintenance of the disorder. Eye Movement Desensitization and Reprocessing (EMDR) is one of the most efficient psychotherapies to treat PTSD. We aimed at exploring the brain mechanisms of the fear circuitry involved in PTSD patients’ symptom remission after EMDR therapy. Method: Thirty-six PTSD participants were randomly assigned to either EMDR group receiving EMDR therapy or Wait-List (WL) group receiving supportive therapy. Participants underwent a behavioural fear conditioning and extinction paradigm during functional magnetic resonance (fMRI). In the EMDR group, patients were scanned at baseline, before EMDR and one week after remission. In the WL group, patients were scanned at baseline and within the same time interval as the EMDR group. Results: In the EMDR group after treatment, fear responses in the late extinction were significantly lower than before therapy. In parallel, significant functional activity and connectivity changes were found in the EMDR group versus the WL during the late extinction. These changes involve the fear circuit (amygdalae, left hippocampus), the right inferior frontal gyrus, the right frontal eye field and insula (pFWE < .05). Conclusion: These functional modifications underlie a significant improvement of fear extinction learning in PTSD patients after EMDR therapy.

The anthropomorphic bias describes the finding that the perceived naturalness of a biological motion decreases as the human-likeness of a computer-animated agent increases. To investigate the anthropomorphic bias in autistic children, human or cartoon characters were presented with biological and artificial motions side by side on a touchscreen. Children were required to touch one that would grow while the other would disappear, implicitly rewarding their choice. Only typically developing controls depicted the expected preference for biological motion when rendered with human, but not cartoon, characters. Despite performing the task to report a preference, children with autism depicted neither normal nor reversed anthropomorphic bias, suggesting that they are not sensitive to the congruence of form and motion information when observing computer-animated agents' actions.

Cyclosporin A (CsA) is a molecule with well-known immunosuppressive properties. As it also acts on the opening of mitochondrial permeability transition pore (mPTP), CsA has been evaluated for ischemic heart diseases (IHD). However, its distribution throughout the body and its physicochemical characteristics strongly limit the use of CsA for intravenous administration. In this context, nanoparticles (NPs) have emerged as an opportunity to circumvent the above-mentioned limitations. We have developed in our laboratory an innovative nanoformulation based on the covalent bond between squalene (Sq) and cyclosporin A to avoid burst release phenomena and increase drug loading. After a thorough characterization of the bioconjugate, we proceeded with a nanoprecipitation in aqueous medium in order to obtain SqCsA NPs of well-defined size. The SqCsA NPs were further characterized using dynamic light scattering (DLS), cryogenic transmission electron microscopy (cryoTEM), and high-performance liquid chromatography (HPLC), and their cytotoxicity was evaluated. As the goal is to employ them for IHD, we evaluated the cardioprotective capacity on two cardiac cell lines. A strong cardioprotective effect was observed on cardiomyoblasts subjected to experimental hypoxia/reoxygenation. Further research is needed in order to understand the mechanisms of action of SqCsA NPs in cells. This new formulation of CsA could pave the way for possible medical application.

The COVID-19 pandemic led to many European countries imposing lockdown measures and limiting people’s movement during spring 2020. During the summer 2020, these strict lockdown measures were gradually lifted while in autumn 2020, local restrictions started to be re-introduced as a second wave emerged. After initial restrictions on visitors accessing many Nature Protected Areas (PAs) in Europe, management authorities have had to introduce measures so that all users can safely visit these protected landscapes. In this paper, we examine the challenges that emerged due to COVID-19 for PAs and their deeper causes. By considering the impact on and response of 14 popular European National and Nature Parks, we propose tentative longer-term solutions going beyond the current short-term measures that have been implemented. The most important challenges identified in our study were overcrowding, a new profile of visitors, problematic behavior, and conflicts between different user groups. A number of new measures have been introduced to tackle these challenges including information campaigns, traffic management, and establishing one-way systems on trail paths. However, measures to safeguard public health are often in conflict with other PA management measures aiming to minimize disturbance of wildlife and ecosystems. We highlight three areas in which management of PAs can learn from the experience of this pandemic: managing visitor numbers in order to avoid overcrowding through careful spatial planning, introducing educational campaigns, particularly targeting a new profile of visitors, and promoting sustainable tourism models, which do not rely on large visitor numbers.

Streptomycetes are well known antibiotic producers and are among the rare prokaryotes able to store carbon as lipids. Previous comparative studies of the weak antibiotic producer Streptomyces lividans with its ppk mutant and with Streptomyces coelicolor, which both produce antibiotics, suggested the existence of a negative correlation between total lipid content and the ability to produce antibiotics. To determine whether such a negative correlation can be generalized to other Streptomyces species, fifty-four strains were picked randomly and grown on modified R2YE medium, limited in phosphate, with glucose or glycerol as the main carbon source. The total lipid content and antibiotic activity against Micrococcus luteus were assessed for each strain. This study revealed that the ability to accumulate lipids was not evenly distributed among strains and that glycerol was more lipogenic than glucose and had a negative impact on antibiotic biosynthesis. Furthermore, a statistically significant negative Pearson correlation between lipid content and antibiotic activity could be established for most strains, but a few strains escape this general law. These exceptions are likely due to limits and biases linked to the type of test used to determine antibiotic activity, which relies exclusively on Micrococcus luteus sensitivity. They are characterized either by high lipid content and high antibiotic activity or by low lipid content and undetectable antibiotic activity against Micrococcus luteus. Lastly, the comparative genomic analysis of two strains with contrasting lipid content, and both named Streptomyces antibioticus (DSM 41,481 and DSM 40,868, which we found to be phylogenetically related to Streptomyces lavenduligriseus), indicated that some genetic differences in various pathways related to the generation/consumption of acetylCoA could be responsible for such a difference.

Recent studies are unraveling the impact of microorganisms from the roots and rhizosphere on interactions between plants and herbivorous insects and are gradually changing our perception of the microorganisms' capacity to affect plant defenses, but the reverse effect has seldom been investigated. Our study aimed at determining how plant herbivory influences the dynamics of root and rhizosphere microbial community assemblages and whether potential changes in root metabolites and chemical elements produced during herbivory can be related to microbial community diversity. We conducted our study on oilseed rape (Brassica napus) and its major belowground herbivore, the cabbage root fly (Delia radicum). We further assessed the influence of initial soil microbial diversity on these interactions. Different microbial diversities based on a common soil matrix were obtained through a removal-recolonization method. Root and rhizosphere sampling targeted different stages of the herbivore development corresponding to different perturbation intensities. Root bacterial communities were more affected by herbivory than some rhizosphere bacterial phyla and fungal communities, which seemed more resistant to this perturbation. Root herbivory enhanced the phylum of γ-Proteobacteria in the roots and rhizosphere, as well as the phylum of Firmicutes in the rhizosphere. Herbivory tended to decrease most root amino acids and sugars, and it increased trehalose, indolyl glucosinolates, and sulfur. Higher abundances of four bacterial genera (Bacillus, Paenibacillus, Pseudomonas, and Stenotrophomonas) were associated following herbivory to the increase of trehalose and some sulfur-containing compounds. Further research would help to identify the biological functions of the microbial genera impacted by plant infestation and their potential implications in plant defense.

We investigated the consequences of premature birth on the functional neuroanatomy of the dorsal stream of visual processing. fMRI was recorded while sixteen healthy participants, 8 (two men) adults (19 years 6 months old, SD 10 months) born premature (mean gestational age 30 weeks), referred to as Premas, and 8 (two men) matched controls (20 years 1 month old, SD 13 months), performed a 1-back memory task of Object or Grip information using a hand grasping a drinking vessel as stimulus. While history of prematurity did not significantly affect task performance, Group by Task analysis of variance in regions of interest spanning the occipital, temporal and parietal lobes revealed main effects of Task and interactions between the two factors. Object processing activated the left inferior occipital cortex and bilateral ventral temporal regions, belonging to the ventral stream, with no effect of Group. Grip processing across groups activated the early visual cortex and the left supramarginal gyrus belonging to the dorsal stream. Group effect on the brain activity during Grip suggested that Controls represented the actions' goal while Premas relied more on low-level visual information. This shift from higher- to lower-order visual processing between Controls and Premas may reflect a more general trend, in which Premas inadequately recruit higher-order visual functions for dorsal stream task performance, and rely more on lower-level functions. (Contains 2 tables and 2 figures.)

The adaptive threat-detection advantage takes the form of a preferential orienting of attention to threatening scenes. In this study, we compared attention to social scenes in 15 high-functioning individuals with autism (ASD) and matched typically developing (TD) individuals. Eye-tracking was recorded while participants were presented with pairs of scenes, either emotional positive-neutral, emotional negative-neutral or neutral-neutral scenes. Early allocation of attention, the first image fixated in each pair, differed between groups: contrary to TD individuals who showed the typical threat-detection advantage towards negative images, the ASD group failed to show a bias toward threat-related scenes. Later processing of stimuli, indicated by the total fixation to the images during the 3-s presentation, was found unaffected in the ASD group. These results support the hypothesis of an early atypical allocation of attention towards natural social scenes in ASD, that is compensated in later stages of visual processing.

Stone tools provide some of the most abundant, continuous, and high resolution evidence of behavioral change over human evolution, but their implications for cognitive evolution have remained unclear. We investigated the neurophysiological demands of stone toolmaking by training modern subjects in known Paleolithic methods ("Oldowan", "Acheulean") and collecting structural and functional brain imaging data as they made technical judgments (outcome prediction, strategic appropriateness) about planned actions on partially completed tools. Results show that this task affected neural activity and functional connectivity in dorsal prefrontal cortex, that effect magnitude correlated with the frequency of correct strategic judgments, and that the frequency of correct strategic judgments was predictive of success in Acheulean, but not Oldowan, toolmaking. This corroborates hypothesized cognitive control demands of Acheulean toolmaking, specifically including information monitoring and manipulation functions attributed to the "central executive" of working memory. More broadly, it develops empirical methods for assessing the differential cognitive demands of Paleolithic technologies, and expands the scope of evolutionary hypotheses that can be tested using the available archaeological record.

The High Energy Stereoscopic System (H.E.S.S.) is an array of imaging atmospheric Cherenkov telescopes (IACTs) located in Namibia. It was built to detect Very High Energy (VHE, >100 GeV) cosmic gamma rays, and consists of four 12 m diameter Cherenkov telescopes (CT1-4), built in 2003, and a larger 28 m telescope (CT5), built in 2012. The larger mirror surface of CT5 permits to lower the energy threshold of the array down to 30 GeV. The cameras of CT1-4 are currently undergoing an extensive upgrade, with the goals of reducing their failure rate, reducing their readout dead time and improving the overall performance of the array. The entire camera electronics has been renewed from ground-up, as well as the power, ventilation and pneumatics systems, and the control and data acquisition software. Technical solutions forseen for the next-generation Cherenkov Telescope Array (CTA) observatory have been introduced, most notably the readout is based on the NECTAr analog memory chip. The camera control subsystems and the control software framework also pursue an innovative design, increasing the camera performance, robustness and flexibility. The CT1 camera has been upgraded in July 2015 and is currently taking data; CT2-4 will upgraded in Fall 2016. Together they will assure continuous operation of H.E.S.S at its full sensitivity until and possibly beyond the advent of CTA. This contribution describes the design, the testing and the in-lab and on-site performance of all components of the newly upgraded H.E.S.S. camera.