Identification of psychedelic new psychoactive substances (NPS) showing biased agonism at the 5-HT2AR through simultaneous use of beta-arrestin 2 and miniG alpha(q) bioassays

Psychedelic new psychoactive substances (NPS), compounds exerting their main pharmacological effects through the activation of the serotonin 2A receptor (5-HT2AR), continuously comprise a substantial portion of the reported NPS. However, these substances and their exact mechanism of action, differentiating them from nonpsychedelic 5-HT2AR agonists, require further characterization. One potentially relevant phenomenon is the occurrence of biased agonism, in which (a) certain signaling pathway(s) is preferentially activated over the other (s). To this end, a new bioassay was developed, monitoring the recruitment of an engineered miniGαq protein to the activated 5-HT2AR. The setup was designed to be analogous to that of a previously developed bioassay monitoring β-arrestin 2 recruitment through the NanoBiT system, enabling estimation of the potential preference of a substance to trigger recruitment of one protein over the other. This approach yielded several statistically significantly biased agonists within the group of phenylalkylamine psychedelics, more specifically the N-benzyl substituted 25H analogues 25H-NBF, 25H-NBMD, 25H-NBOH and 25H-NBOMe. All four compounds show a statistically significant preference towards the recruitment of β-arrestin 2 over miniGαq, as compared to the reference psychedelic substance LSD. We identified markedly different responses for Bromo-DragonFLY in the two bioassays, suggesting biased agonism, though the calculated bias factor equalled out to approximately 0. This demonstrates that the accurate assessment of biased agonism requires both the consideration of the observed trends in addition to the numerical value of the bias factor. A second panel of structural (I-substituted) analogues of the former group of phenylalkylamines showed a similar trend in the ranking order of the bias factors, resulting in one additional compound (25I-NBF) being statistically significantly biased

Localizer:fast, accurate, open-source, and modular software package for superresolution microscopy

We present Localizer, a freely available and open source software package that implements the computational data processing inherent to several types of superresolution fluorescence imaging, such as localization (PALM/STORM/GSDIM) and fluctuation imaging (SOFI/pcSOFI). Localizer delivers high accuracy and performance and comes with a fully featured and easy-to-use graphical user interface but is also designed to be integrated in higher-level analysis environments. Due to its modular design, Localizer can be readily extended with new algorithms as they become available, while maintaining the same interface and performance. We provide front-ends for running Localizer from Igor Pro, Matlab, or as a stand-alone program. We show that Localizer performs favorably when compared with two existing superresolution packages, and to our knowledge is the only freely available implementation of SOFI/pcSOFI microscopy. By dramatically improving the analysis performance and ensuring the easy addition of current and future enhancements, Localizer strongly improves the usability of superresolution imaging in a variety of biomedical studies

Network virtualization as an integrated solution for emergency communication

In this paper the Virtual Private Ad Hoc Networking (VPAN) platform is introduced as an integrated networking solution for many applications that require secure transparent continuous connectivity using heterogeneous devices and network technologies. This is done by creating a virtual logical self-organizing network on top of existing network technologies reducing complexity and maintaining session continuity right from the start. One of the most interesting applications relies in the field of emergency communication with its specific needs which will be discussed in this paper and matched in detail against the architecture and features of the VPAN platform. The concept and dynamics are demonstrated and evaluated with measurements done on real hardware

Sensitive and specific detection of E. coli using biomimetic receptors in combination with a modified heat-transfer method

We report on a novel biomimetic sensor that allows sensitive and specific detection of Escherichia colt (E. coli) bacteria in a broad concentration range from 10(2) up to 10(6) CFU/mL in both buffer fluids and relevant food samples (i.e. apple juice). The receptors are surface-imprinted polyurethane layers deposited on stainless-steel chips. Regarding the transducer principle, the sensor measures the increase in thermal resistance between the chip and the liquid due to the presence of bacteria captured on the receptor surface. The low noise level that enables the low detection limit originates from a planar meander element that serves as both a heater and a temperature sensor. Furthermore, the experiments show that the presence of bacteria in a liquid enhances the thermal conductivity of the liquid itself. Reference tests with a set of other representative species of Enterobacteriaceae, closely related to E. coli, indicate a very low cross-sensitivity with a sensor response at or below the noise level

Complementarity of PALM and SOFI for super-resolution live cell imaging of focal adhesions

Live cell imaging of focal adhesions requires a sufficiently high temporal resolution, which remains a challenging task for super-resolution microscopy. We have addressed this important issue by combining photo-activated localization microscopy (PALM) with super-resolution optical fluctuation imaging (SOFI). Using simulations and fixed cell focal adhesion images, we investigated the complementarity between PALM and SOFI in terms of spatial and temporal resolution. This PALM-SOFI framework was used to image focal adhesions in living cells, while obtaining a temporal resolution below 10 s. We visualized the dynamics of focal adhesions, and revealed local mean velocities around 190 nm per minute. The complementarity of PALM and SOFI was assessed in detail with a methodology that integrates a quantitative resolution and signal-to-noise metric. This PALM and SOFI concept provides an enlarged quantitative imaging framework, allowing unprecedented functional exploration of focal adhesions through the estimation of molecular parameters such as the fluorophore density and the photo-activation and photo-switching rates