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Toxoplasma gondii holds significant therapeutic potential; however, its nonspecific invasiveness results in off-target effects. The purpose of this study is to evaluate whether T. gondii specificity can be improved by surface display of scFv directed against dendritic cells’ endocytic receptor, DEC205, and immune checkpoint PD-L1. Anti-DEC205 scFv was anchored to the T. gondii surface either directly via glycosylphosphatidylinositol (GPI) or by fusion with the SAG1 protein. Both constructs were successfully expressed, but the binding results suggested that the anti-DEC-SAG1 scFv had more reliable functionality towards recombinant DEC protein and DEC205-expressing MutuDC cells. Two anti-PD-L1 scFv constructs were developed that differed in the localization of the HA tag. Both constructs were adequately expressed, but the localization of the HA tag determined the functionality by binding to PD-L1 protein. Co-incubation of T. gondii displaying anti-PD-L1 scFv with tumor cells expressing/displaying different levels of PD-L1 showed strong binding depending on the level of available biomarker. Neutralization assays confirmed that binding was due to the specific interaction between anti-PD-L1 scFv and its ligand. A mixed-cell assay showed that T. gondii expressing anti-PD-L1 scFv predominately targets the PD-L1-positive cells, with negligible off-target binding. The recombinant RH-PD-L1-C strain showed increased killing ability on PD-L1+ tumor cell lines compared to the parental strain. Moreover, a co-culture assay of target tumor cells and effector CD8+ T cells showed that our model could inhibit PD1/PD-L1 interaction and potentiate T-cell immune response. These findings highlight surface display of antibody fragments as a promising strategy of targeting replicative T. gondii strains while minimizing nonspecific binding.

Pelagic stingray (Pteroplatytrygon violacea) is the only species of stingray (Dasyatidae) that utilizes both pelagic and demersal habitats. It is the main bycatch species in pelagic longline fisheries targeting bluefin tuna (Thunnus thynnus) in the Gulf of Lions. In the Mediterranean Sea, their stock structure, behavioural ecology and movements are unknown. For the first time in the Mediterranean, 17 individuals (39–60 cm disc width) were tagged with pop-up satellite archival transmitting tags using a novel method of tag attachment to investigate horizontal and vertical movements. The tags were attached for between two and 60 days. Between the months of July and October, pelagic stingray occupied a temperature range of 12.5–26.6 °C, and a depth range extending from the surface to 480 m. Monthly trends in catch-per-unit-effort (CPUE) of pelagic stingray peaked in August and decreased by late autumn. Pelagic stingray may aggregate on the continental shelf during summer and move southwards in early autumn, and this movement pattern is considered in relation to the reproductive cycle and overwintering. At-vessel mortality was low, but there was varying evidence of post-release mortality, indicating the need for further work. Future work and options for bycatch mitigation are also discussed.

Because they are safe and easy to use, collaborative robots are revolutionizing many sectors, including industry, medicine, and agriculture. Controlling their dynamics, movements, and postures are key points in this evolution. Inverse kinematics is then crucial for robot motion planning. In 6R serial robots, achieving a desired pose is possible with different joint combinations. In this paper, our focus lies in studying forward and, mainly, inverse kinematics of the FANUC CRX-10iA cobot, a 6R cobotic arm with a non-spherical wrist. Its specific structural parameters implies that no analytical solutions exist except for some particular situations. FANUC does not provide the complete set of inverse kinematic solutions, even when 16 solutions are possible, only 8 of them are provided in Roboguide software. Furthermore, the existing literature on joints-to-workspace mapping for CRX cobots is currently very limited. It either lacks or provides partial or inconsistent inverse kinematics analysis. We present and detail a novel fully geometric method for numerically solving inverse kinematics meeting the requirement of high precision and a fast response. This approach provides both the exact number of inverse kinematics solutions and the sets of joint angles even for singular configuration. Its effectiveness was verified through simulations using the Roboguide Software and experimentation on the actual CRX-10iA cobot. Several examples (8, 12, or 16 inverse kinematic solutions) have enabled us to validate and prove the robustness and reliability of this geometric approach.

Tactical urbanism is a modality of intervention resulting from citizen initiatives, involving temporary, reversible and inexpensive material transformation, whose objective is to introduce rapid changes in the use and appropriation of urban public space. During or shortly after the spring 2020 lockdown due to the COVID-19 pandemic, tactical urbanism proved to be a simple, quick and inexpensive way to respond to the health emergency of social distancing. In particular, bicycle lanes and paths, referred to as pop-up infrastructures or in France as “coronapists,” were created in order to avoid a modal shift to the car by people afraid of using public transport. Based on research conducted with interviews and video elicitation in five French cities–Grenoble, Lyon, Montpellier, Rennes and Saint-Étienne–with experienced and novice cyclists, the article analyzes how these pop-up infrastructures have been perceived and used according to the context, the type of infrastructure and the profile of cyclists. Building on the discussion of the role of infrastructure, the aim here is to understand how the contingencies arising from the layout of transitory infrastructure produce a different relationship to space and mobility. In doing so, we examine the potential spillover effect of the coronapists on a population that did not practice utilitarian cycling before the pandemic and the continued practice of those who were already utilitarian cyclists. The purpose is to examine how these new infrastructures may have acted as a facilitator of modal shift or as a learning facilitator, and what particular role coronapists played in the practice of utilitarian cycling. The article first examines the reception of the infrastructure and then questions the uses related to an assessment of the cyclability in terms of comfort, safety and security. The article ends with a discussion of the interests and limitations of the tactical dimension in the implementation of coronapists. The diversity of the territory surveyed has shown the political force of the infrastructures and the effect that these transitional infrastructures have had as a complement to other measures. They promote new uses and encourage new people, including children, to adopt cycling on a daily or more regular basis. This result invites the pursuit of this proactive policy. These promising results encourage the development of these infrastructures in sparsely populated, suburban or rural areas. In addition, we have shown that ridership is highly dependent on cyclists’ assessment of the comfort, safety, and security of the path. This perception varies greatly depending on the context, the type of facility and the experience of the cyclists. In most cases, the coronapists have helped to stimulate more cycling by providing greater coverage and safer or more direct routes. The expectancies and dissatisfaction generated by transitional facilities, which relate to perceived bikeability, inform us of the importance of considering the diversity of uses in their design as well as their physical quality and maintenance. The results of our survey encourage the design of readable, wide, physically separated, continuous and maintained facilities to improve cyclists’ sense of safety. Finally, the results also show the importance of thinking about cycling policy by considering the full range of possible uses and experience levels from the design stage onwards in order to attract new cyclists.

Background Data and Safety Monitoring is integral to quality assurance of clinical trials. Although monitoring is a core legal component of regulated clinical trials, non-regulated trials are not mandated to incorporate monitoring. Consequently, the monitoring process has been underutilised and underreported in this setting. This research report outlines the development and plans for implementing a bespoke Clinical Monitoring Strategy within the ‘Rehabilitation Strategies Following Oesophagogastric and Hepatopancreaticobiliary Cancer (ReStOre II) Trial’, a non-regulated trial comparing a 12-week multidisciplinary programme of rehabilitation to standard care in a cohort of 120 cancer survivors. Methods This research note provides a detailed overview of the ReStOre II Clinical Monitoring Strategy and describes the development of the strategy pre and post awarding of the grant. The strategy consists of the establishment and implementation of a comprehensive trial governance structure, inclusive of a Trial Management Group, Trial Steering Committee Meeting, and Independent Data Monitoring Committee. In addition, external trial monitoring by the Clinical Research Facility at St James’s Hospital. Three monitoring visits will be conducted during the trial; i) site initiation visit, ii) interim monitoring visit, and iii) close our visit. Results The Clinical Monitoring Strategy has been finalised and is currently being implemented within the ReStOre II Trial. Two site initiation visits and one interim monitoring visit have been completed to date. Conclusion This research note provides a template for implementation of a Clinical Monitoring Strategy in a non-regulated clinical trial. Registration ReStOre II Trial: https://clinicaltrials.gov/ct2/show/NCT03958019

PurposeLamb-Shaffer syndrome (LAMSHF) is a neurodevelopmental disorder described in just over two dozen patients with heterozygous genetic alterations involving SOX5, a gene encoding a transcription factor regulating cell fate and differentiation in neurogenesis and other discrete developmental processes. The genetic alterations described so far are mainly microdeletions. The present study was aimed at increasing our understanding of LAMSHF, its clinical and genetic spectrum, and the pathophysiological mechanisms involved.MethodsClinical and genetic data were collected through GeneMatcher and clinical or genetic networks for 41 novel patients harboring various types ofSOX5 alterations. Functional consequences of selected substitutions were investigated.ResultsMicrodeletions and truncating variants occurred throughout SOX5. In contrast, most missense variants clustered in the pivotal SOX-specific high-mobility-group domain. The latter variants prevented SOX5 from binding DNA and promoting transactivation in vitro, whereas missense variants located outside the high-mobility-group domain did not. Clinical manifestations and severity varied among patients. No clear genotype-phenotype correlations were found, except that missense variants outside the high-mobility-group domain were generally better tolerated.ConclusionsThis study extends the clinical and genetic spectrum associated with LAMSHF and consolidates evidence that SOX5 haploinsufficiency leads to variable degrees of intellectual disability, language delay, and other clinical features.

Inflammatory gut disorders, including inflammatory bowel disease (IBD), can be impacted by dietary, environmental, and genetic factors. While the incidence of IBD is increasing worldwide, we still lack a complete understanding of the gene-by-environment interactions underlying inflammation and IBD. Here, we profiled the colon transcriptome of 52 BXD mouse strains fed with a chow or high-fat diet (HFD) and identified a subset of BXD strains that exhibit an IBD-like transcriptome signature on HFD, indicating that an interplay of genetics and diet can significantly affect intestinal inflammation. Using gene co-expression analyses, we identified modules that are enriched for IBD-dysregulated genes and found that these IBD-related modules share cis-regulatory elements that are responsive to the STAT2, SMAD3, and REL transcription factors. We used module quantitative trait locus analyses to identify genetic loci associated with the expression of these modules. Through a prioritization scheme involving systems genetics in the mouse and integration with external human datasets, we identified Muc4 and Epha6 as the top candidates mediating differences in HFD-driven intestinal inflammation. This work provides insights into the contribution of genetics and diet to IBD risk and identifies two candidate genes, MUC4 and EPHA6, that may mediate IBD susceptibility in humans.

Chlamydia trachomatis (Ct) infections are the most common sexually transmitted infection (STI). Despite effective antibiotics for Ct, undetected infections or delayed treatment can lead to infertility, ectopic pregnancies, and chronic pelvic pain. Besides humans, chlamydia poses similar health challenges in animals such as C. suis (Cs) in pigs. Based on the similarities between humans and pigs, as well as their chlamydia species, we use pigs as a large biomedical animal model for chlamydia research. In this study, we used the pig model to develop a vaccine candidate against Ct. The vaccine candidate consists of TriAdj-adjuvanted chlamydial-protease-like activity factor (CPAF) protein. We tested two weekly administration options—twice intranasal (IN) followed by twice intramuscular (IM) and twice IM followed by twice IN. We assessed the humoral immune response in both serum using CPAF-specific IgG (including antibody avidity determination) and also in cervical and rectal swabs using CPAF-specific IgG and IgA ELISAs. The systemic T-cell response was analyzed following in vitro CPAF restimulation via IFN-γ and IL-17 ELISpots, as well as intracellular cytokine staining flow cytometry. Our data demonstrate that while the IN/IM vaccination mainly led to non-significant systemic immune responses, the vaccine candidate is highly immunogenic if administered IM/IN. This vaccination strategy induced high serum anti-CPAF IgG levels with strong avidity, as well as high IgA and IgG levels in vaginal and rectal swabs and in uterine horn flushes. In addition, this vaccination strategy prompted a pronounced cellular immune response. Besides inducing IL-17 production, the vaccine candidate induced a strong IFN-γ response with CD4 T cells. In IM/IN-vaccinated pigs, these cells also significantly downregulated their CCR7 expression, a sign of differentiation into peripheral-tissue-homing effector/memory cells. Conclusively, this study demonstrates the strong immunogenicity of the IM/IN-administered TriAdj-adjuvanted Ct CPAF vaccine candidate. Future studies will test the vaccine efficacy of this promising Ct vaccine candidate. In addition, this project demonstrates the suitability of the Cs pre-exposed outbred pig model for Ct vaccine development. Thereby, we aim to open the bottleneck of large animal models to facilitate the progression of Ct vaccine candidates into clinical trials.