Acetylation of lysines on affinity-purification matrices to reduce co-digestion of bead-bound ligands v1

In mass-spectrometry-based interaction proteomics on-bead digestion protocols are commonly applied after affinity-enrichment due to their simplicity and high efficiency. However, on-bead digestion often leads to strong background signals due to co-digestion of the bead-bound ligands such as streptavidin or antibodies. We present an effective, rapid and low-cost method to specifically reduce the peptide signals from co-digested matrix ligands. A short pre-incubation of matrix beads with Sulfo-NHS-Acetate (S-NHS-Ac) leads to acetylation of free amines on lysine side-chains of the bead-bound ligands making them resistant to Lys-C-mediated proteolysis. After binding of bait proteins to the acetylated beads we employ a two-step digestion protocol with the sequential use of Lys-C protease for on-bead digestion followed by in-solution digestion with trypsin. The strong reduction of interfering ligand peptides improves signal strength and data quality for the peptides of interest in liquid chromatography mass spectrometry (LC-MS).

αVβ3 Integrin Expression Is Essential for Replication of Mosquito and Tick-Borne Flaviviruses in Murine Fibroblast Cells

The Flavivirus genus includes a number of important viruses that are pathogenic to humans and animals and are responsible for outbreaks across the globe. Integrins, a family of heterodimeric transmembrane molecules expressed in all nucleated cells mediate critical functions of cell physiology and cell cycle. Integrins were previously postulated to be involved in flavivirus entry and to modulate flavivirus replication efficiency. In the present study, mouse embryonic fibroblasts (MEF), lacking the expression of αVβ3 integrin (MEF-αVβ3−/−), were infected with four different flaviviruses, namely yellow fever virus (YFV), West Nile virus (WNV), Usutu virus (USUV) and Langat virus (LGTV). The effects of the αVβ3 integrin absence in double-knockout MEF-αVβ3−/− on flavivirus binding, internalization and replication were compared to the respective wild-type cells. Binding to the cell surface for all four flaviviruses was not affected by the ablation of αVβ3 integrin, whereas internalization of USUV and WNV was slightly affected by the loss of αVβ3 integrin expression. Most interestingly, the deletion of αVβ3 integrin strongly impaired replication of all flaviviruses with a reduction of up to 99% on virus yields and a strong reduction on flavivirus anti-genome RNA synthesis. In conclusion, our results demonstrate that αVβ3 integrin expression in flavivirus-susceptible cell lines enhances the flavivirus replication.

DARPin_9-29-Targeted Gold Nanorods Selectively Suppress HER2-Positive Tumor Growth in Mice

Near-infrared phototherapy has great therapeutic potential for cancer treatment. However, for efficient application, in vivo photothermal agents should demonstrate excellent stability in blood and targeted delivery to pathological tissue. Here, we demonstrated that stable bovine serum albumin-coated gold mini nanorods conjugated to a HER2-specific designed ankyrin repeat protein, DARPin_9-29, selectively accumulate in HER2-positive xenograft tumors in mice and lead to a strong reduction in the tumor size when being illuminated with near-infrared light. The results pave the way for the development of novel DARPin-based targeted photothermal therapy of cancer.

Vaccine Refusal in the Czech Republic Is Associated with Being Spiritual but Not Religiously Affiliated

A strong reduction in the deleterious effects of the COVID-19 pandemic can be achieved by vaccination. Religiosity and spirituality (R/S) may play an important role in vaccine acceptance. However, evidence is lacking for the associations with religious conspiracy theories (RCT) in a non-religious environment. This study investigated the associations between R/S and RCT about COVID-19 vaccination and the links of R/S with vaccine refusal and hesitancy. A sample of Czech adults (n = 459) participated in the survey. We measured R/S, RCT, religious fundamentalism, and COVID-19 vaccination intentions. We found spirituality to be significantly associated with RCT belief, with odds ratios (OR) of 2.12 (95% confidence interval [CI] 1.42–3.19). A combination of R/S groups revealed that spirituality with non-religious affiliation was associated with higher beliefs in RCT, with ORs from 3.51 to 7.17. Moreover, associations were found between spirituality with non-religious affiliation [OR 2.22(1.33–7.76)] with vaccine refusal. Our findings showed associations of spirituality and religious fundamentalism with RCT about COVID-19 vaccination. Furthermore, spirituality was linked to a higher possibility of vaccine refusal. Understanding these associations may help prevent the development of RCT and negative impact of spirituality on vaccine intentions and contribute to the effectiveness of the vaccination process.

Radiationless anapole states in on-chip photonics

High-index nanoparticles are known to support radiationless states called anapoles, where dipolar and toroidal moments interfere to inhibit scattering to the far field. In order to exploit the striking properties arising from these interference conditions in photonic integrated circuits, the particles must be driven in-plane via integrated waveguides. Here, we address the excitation of electric anapole states in silicon disks when excited on-chip at telecom wavelengths. In contrast to normal illumination, we find that the anapole condition—identified by a strong reduction of the scattering—does not overlap with the near-field energy maximum, an observation attributed to retardation effects. We experimentally verify the two distinct spectral regions in individual disks illuminated in-plane from closely placed waveguide terminations via far-field and near-field measurements. Our finding has important consequences concerning the use of anapole states and interference effects of other Mie-type resonances in high-index nanoparticles for building complex photonic integrated circuitry.

An internal expectation guides Drosophila egg-laying decisions

When presented with two egg-laying substrates, Drosophila lay most of their eggs on the option with higher relative value. How do flies make this relative-value decision? Might the strategy they use allow them to choose the best option even when they experience substrates with a more complex spatiotemporal profile than in canonical two-choice paradigms? We measured Drosophila egg-laying behavior in diverse substrate environments. In all cases, we found that flies dynamically increase or decrease their egg-laying rates as they explore substrates for a deposition site so as to target eggs to the best, recently visited option. Visiting the best option typically led to a strong reduction in egg laying on other substrates for several minutes, with this timescale varying across fly strains. Our data support a model in which flies compare the value of the current substrate with an internally constructed expectation on the value of available options to regulate the likelihood of laying an egg. We show that dopamine-neuron activity is critical for learning and/or expressing this expectation, similar to its role in certain tasks in vertebrates. Integrating sensory experiences over minutes to generate an internal sense of the quality of available options, i.e., forming an expectation, allows flies to use a dynamic reference point for judging the current substrate and might be a general way in which decisions are made, even beyond flies and egg laying.

Édition de gènes in vivo et thérapie génique

In vivo gene editing has been achieved in a phase I clinical trial and results in a strong reduction of the level of a pathogenic protein. While preliminary, these results open the way for many applications in gene therapy.

Lockdown: How the COVID-19 Pandemic Affected the Fishing Activities in the Adriatic Sea (Central Mediterranean Sea)

The coronavirus disease 2019 (COVID-19) has brought a global socio-economic crisis to almost all sectors including the fishery. To limit the infection, governments adopted several containment measures. In Italy, Croatia, and Slovenia, a lockdown period was imposed from March to May 2020, during which many activities, including restaurants had to close or limit their business. All of this caused a strong reduction in seafood requests and consequently, a decrease in fishing activities. The aim of this study is to investigate the effects of the COVID-19 in the Northern and Central Adriatic fleet, by comparing the fishing activities in three periods (before, during, and after the lockdown) of 2019 and 2020. The use of the Automatic Identification System (AIS) data allowed us to highlight the redistribution of the fishing grounds of the trawlers, mainly located near the coasts during the 2020 lockdown period, as well as a reduction of about 50% of fishing effort. This reduction resulted higher for the Chioggia trawlers (−80%) and, in terms of fishing effort decrease, the large bottom otter trawl was the fishing segment mainly affected by the COVID-19 event. Moreover, by analysing the landings of the Chioggia fleet and the Venice lagoon fleets, it was possible to point out a strong reduction both in landings and profits ranging from −30%, for the small-scale fishery operating at sea, to −85%, for the small bottom otter trawl.

Colour Planning in Town, a non profit colour project

Colour is much more than decoration. This medium definitely influences human beings –consciously and unconsciously. The IACC Chapter Europe, was invited in 2017 to create a new colour pallet for a City in Sothern Italy, maintaining the character and identity of the town. Eleven buildings of the centre were selected to be worked on. The collected 53 old colour samples were the base for the very careful analysis and the strong reduction to 21 nuances of the new pallet. Finally various colour collages were produced to show the community and the public of the city the change of impression - using just colour. Colour is definitely an absolute necessity for a balanced environment, indoors as well as outdoors. A medical statement says that more than 75% of all illnesses derive from stress: In this connection the stress diminishing property of colour is THE MOST EFFECTIVE one to take care of. El color es mucho más que decoración. Este medio influye definitivamente en los seres humanos -consciente e inconscientemente. El capítulo europeo de la IACC, fue invitado en 2017 a crear una nueva paleta de colores para una ciudad del sur de Italia, manteniendo el carácter y la identidad de la ciudad. Se seleccionaron once edificios del centro para trabajar en ellos. Las 53 muestras de color antiguas recogidas fueron la base para el análisis muy cuidadoso y la fuerte reducción a 21 matices de la nueva paleta. Por último, se elaboraron varios collages de colores para mostrar a la comunidad y al público de la ciudad el cambio de impresión, utilizando sólo el color. El color es definitivamente una necesidad absoluta para un entorno equilibrado, tanto en el interior como en el exterior. Una declaración médica dice que más del 75% de las enfermedades se derivan del estrés: En este sentido, la propiedad del color de reducir el estrés es la más eficaz.