Simplified Monopalmitoyl Toll-like Receptor 2 Ligand Mini-UPam for Self-Adjuvanting Neoantigen-Based Synthetic Cancer Vaccines

Synthetic vaccines, based on antigenic peptides that comprise MHC−I and MHC-II T-cell epitopes expressed by tumors, show great promise for the immunotherapy of cancer. For optimal immunogenicity, the synthetic peptides (SPs) should be adjuvanted with suitable immunostimulatory additives. Previously, we have shown that improved immunogenicity in vivo is obtained with vaccine modalities in which an SP is covalently connected to an adjuvanting moiety, typically a ligand to Toll-like receptor 2 (TLR2). SPs were covalently attached to UPam, which is a derivative of the classic TLR2 ligand Pam₃CysSK₄. A disadvantage of the triply palmitoylated UPam is its high lipophilicity, which precludes universal adoption of this adjuvant for covalent modification of various antigenic peptides as it renders the synthetic vaccine insoluble in several cases. Here, we report a novel conjugatable TLR2 ligand, mini-UPam, which contains only one palmitoyl chain, rather than three, and therefore has less impact on the solubility and other physicochemical properties of a synthetic peptide. In this study, we used SPs that contain the clinically relevant neoepitopes identified in a melanoma patient who completely recovered after T-cell therapy. Homogeneous mini-UPam-SP conjugates have been prepared in good yields by stepwise solid-phase synthesis that employed a mini-UPam building block pre-prepared in solution and the standard set of Fmoc-amino acids. The immunogenicity of the novel mini-UPam-SP conjugates was demonstrated by using the cancer patient's T-cells.     

Emergency Door Capacity: Influence of Door Width, Population Composition and Stress Level

For the new version of the Dutch design guidelines for buildings, a threshold value for the capacity of emergency doors needs to be set. Innovative large-scale laboratory experiments have been performed to investigate the capacity of emergency doors during evacuation conditions. This paper focuses in particular on the relation between capacity and the independent variables doorway width, population composition, stress level and presence of an open door. It turned out that only the experiment with the widest doorway (275 cm) resulted in a capacity lower than the capacity from the current design guidelines (2.25 P/m/s). The average observed capacities are for all widths lowest for the lowest stress level and highest for the highest stress level. The population with a greater part of children has the highest capacity (on average 3.31 P/m/s). This is mainly due to the smaller physical size of children compared to adults and elderly, which makes it possible that more children can pass a door at the same time than adults. The lowest capacity (on average 2.02 P/m/s) is found for the experiment with 5% disabled participants. The presence of a door opened in the escape direction in an angle of 90° for a doorway of 85 cm results in a 20% capacity reduction compared to the reference experiment. The open door does not physically narrow the doorway, but it leads to interactions between participants reducing their speed and the corresponding outflow.     

Modelling collective decision making in groups and crowds: Integrating social contagion and interacting emotions, beliefs and intentions

Collective decision making involves on the one hand individual mental states such as beliefs, emotions and intentions, and on the other hand interaction with others with possibly different mental states. Achieving a satisfactory common group decision on which all agree requires that such mental states are adapted to each other by social interaction. Recent developments in social neuroscience have revealed neural mechanisms by which such mutual adaptation can be realised. These mechanisms not only enable intentions to converge to an emerging common decision, but at the same time enable to achieve shared underlying individual beliefs and emotions. This paper presents a computational model for such processes. As an application of the model, an agent-based analysis was made of patterns in crowd behaviour, in particular to simulate a real-life incident that took place on May 4, 2010 in Amsterdam. From available video material and witness reports, useful empirical data were extracted. Similar patterns were achieved in simulations, whereby some of the parameters of the model were tuned to the case addressed, and most parameters were assigned default values. The results show the inclusion of contagion of belief, emotion, and intention states of agents results in better reproduction of the incident than non-inclusion.     

Second Home Owner Perceptions of Their Environmental Impacts: The Case of Hartbeespoort

Second home tourism studies have found that there are environmental concerns which arise through the development and continued use of a second home. The majority of these studies, however, have been conducted in the Global North, and no research of this kind has yet been conducted in South Africa. This study, therefore, aims to investigate the environmental implications of second home tourism in Hartbeespoort, South Africa, with special reference to the perceptions second home owners hold with regards to their impacts on the environment. The perception held by majority of second home owner respondents is that neither they, nor their households, impact upon the environment. This perception brings the environmental sustainability of second home tourism in South Africa into question.     

Utilizing theory of mind for action selection applied in the domain of fighter pilot training

When developing intelligent agents, approaches that allow the anticipation of other agents is of utmost importance. For humans, this has also been shown to be crucial to establish good interactions. In this paper, a design for an agent that is equipped with theory of mind based reasoning capabilities is presented. The approach moves beyond the state of the art from several angles: first, it allows for the expression of certainties with respect to the predicted states of the other agents. Second, it allows the prediction during a substantial number of time steps in the future, thereby utilizing the theory of mind model multiple times. The approach has been applied to the domain of fighter pilots whereby intelligent opponents are developed to facilitate dedicated training for F16 fighter pilots.     

Activity-based profiling of retaining β-glucosidases: a comparative study

Activity-based protein profiling (ABPP) is a versatile strategy to report on enzyme activity in vitro, in situ, and in vivo. The development and use of ABPP tools and techniques has met with considerable success in monitoring physiological processes involving esterases and proteases. Activity-based profiling of glycosidases, on the other hand, has proven more difficult, and to date no broad-spectrum glycosidase activity-based probes (ABPs) have been reported. In a comparative study, we investigated both 2-deoxy-2-fluoroglycosides and cyclitol epoxides for their utility as a starting point towards retaining β-glucosidase ABP. We also investigated the merits of direct labeling and two-step bio-orthogonal labeling in reporting on glucosidase activity under various conditions. Our results demonstrate that 1) in general cyclitol epoxides are the superior glucosidase ABPs, 2) that direct labeling is the more efficient approach but it hinges on the ability of the glucosidase to be accommodated in the active site of the reporter (BODIPY) entity, and 3) that two-step bio-orthogonal labeling can be achieved on isolated enzymes but translating this protocol to cell extracts requires more investigation.     

Surface plasmon resonance evaluation of various aminoglycoside-RNA hairpin interactions reveals low degree of selectivity

Aminoglycoside antibiotics, which are able to selectively bind to RNA, are considered to be an important lead in RNA-targeting drug discovery. In this study, surface plasmon resonance (SPR) was employed to explore the interaction of aminoglycosides with known tobramycin-binding RNA hairpins (aptamers) and an unrelated RNA hairpin. It was established that aminoglycosides have multiple interactions with RNA hairpins. Unexpectedly, the different hairpins showed comparable affinity for a set of related aminoglycosides. The observed absence of selectivity presents an extra hurdle in the discovery of novel aminoglycosides as specific drugs that target defined RNA hairpins.     

Discovery of Selective Luteinizing Hormone Receptor Agonists Using the Bivalent Ligand Method

Two series of dimeric ligands for a G‐protein‐coupled receptor were prepared that differ by the interconnecting spacer system. Biological evaluation revealed that both dimeric series exhibit unique biological properties relative to their monomeric counterparts.

     

Numerical study of the Stratified smoke flow in a corridor: Full-scale calculations

The two-dimensional, steady-state, turbulent flow of hot air and smoke in a corridor with an adjacent room fire is calculated numerically. Turbulence is accounted for through the k-ε model. Buoyancy efects are included in both the vertical momentum and the turbulence equations. Convective and radiative heat losses are accounted for. The simple radiation model used assumes a well-mixed, spectrally gray and optically thick smoke layer.

A full-sized corridor of 75 m length and 2.5 m height is considered. Parameter variations include inflow temperature, radiative heat losses and net mass flow. Special attention is given to the stratification of the hot smoke layer. Strong stratification is found for all situations considered. However, the hot layer thickness increases significantly in some cases. Our calculations do not support the use of a critical overall Richardson number as a criterion for the occurrence of stratification.