A simple and quick method for loading proteins in extracellular vesicles

Extracellular vesicles (EVs) mediate intercellular transport of biomolecular cargo in the body, making them promising delivery vehicles for bioactive compounds. Genetic engineering of producer cells has enabled encapsulation of therapeutic proteins in EVs. However, genetic engineering approaches can be expensive, time-consuming, and incompatible with certain EV sources, such as human plasma and bovine milk. The goal of this study was to develop a quick, versatile, and simple method for loading proteins in EVs post-isolation. Proteins, including CRISPR associated protein 9 (Cas9), were bound to cationic lipids that were further complexed with MDA-MB-231 cell-derived EVs through passive incubation. Size-exclusion chromatography was used to remove components that were not complexed with EVs. The ability of EVs to mediate intracellular delivery of proteins was compared to conventional methods, such as electroporation and commercial protein transfection reagents. The results indicate that EVs retain native features following protein-loading and obtain similar levels of intracellular protein delivery as conventional methods, but display less toxicity. This method opens up opportunities for rapid exploration of EVs for protein delivery

PEDOT:PSS Morphostructure and ion-to-electron transduction and amplification mechanisms in organic electrochemical transistors

Organic electrochemical transistors (OECTs) represent a powerful and versatile type of organic-based device, widely used in biosensing and bioelectronics due to potential advantages in terms of cost, sensitivity, and system integration. The benchmark organic semiconductor they are based on is poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS), the electrical properties of which are reported to be strongly dependent on film morphology and structure. In particular, the literature demonstrates that film processing induces morphostructural changes in terms of conformational rearrangements in the PEDOT:PSS in-plane phase segregation and out-of-plane vertical separation between adjacent PEDOT-rich domains. Here, taking into account these indications, we show the thickness-dependent operation of OECTs, contextualizing it in terms of the role played by PEDOT:PSS film thickness in promoting film microstructure tuning upon controlled-atmosphere long-lasting thermal annealing (LTA). To do this, we compared the LTA-OECT response to that of OECTs with comparable channel thicknesses that were exposed to a rapid thermal annealing (RTA). We show that the LTA process on thicker films provided OECTs with an enhanced amplification capability. Conversely, on lower thicknesses, the LTA process induced a higher charge carrier modulation when the device was operated in sensing mode. The provided experimental characterization also shows how to optimize the OECT response by combining the control of the microstructure via solution processing and the effect of postdeposition processing

Nanomolar detection of the antitumor drug tamoxifen by flexible organic electrochemical devices

Organic Electrochemical Transistors (OECTs) represent a versatile tool successfully exploited in the field of Bioelectronics. In particular, OECTs have been used for the detection of a wide set of bioanalytes, often showing superior performance compared to that of commonly used sensors. In this study, we propose a flexible, disposable OECT, based on poly (3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS) channels and few layers graphene (FLG) sheets gate electrodes, for the detection of Tamoxifen (TAM), an important antitumor drug widely used in breast cancer therapy. The optimal device operation conditions in terms of sensitivity and limit of detection (LOD) have been investigated too

Aerosol Jet Printed Organic Memristive Microdevices Based on a Chitosan:PANI Composite Conductive Channel

In this study we show a chitosan:polyaniline (CPA)-based ink, responding to eco-biofriendly criteria, specifically developed for the manufacturing of the first organic memristive device (OMD) with an aerosol jet printed conductive channel. Our contribution is in the context of bioelectronics, where there is an increasing interest in emulating neuro-morphic functions. In this framework, memristive devices and systems have been shown to be well suited. In particular organic-based devices are envisaged as very promising in some applications, such as brain-machine interfacing, owing to specific properties of organics (e.g., biocompatibility, mixed ionic-electronic conduction). On the other hand, the research activities on flexible organic (bio)electronic devices and direct writing (DW) noncontact techniques increasingly overlap in the effort of achieving reliable applications benefiting from the rapid prototyping to accomplish a fast device optimization. In this context, ink-based techniques, such as aerosol jet printing (AJP), although particularly well suited to implement 3D-printed electronics due to advantages it offers in terms of a wide set of allowed printable materials, still require research efforts aimed at conferring printability to the desired precursors. The developed CPA composite was characterized by FTIR, DLS, and MALDI-TOF techniques, while the related aerosol jet printed films were studied by SEM and profilometry. Taking advantage of the intrinsic and stable electrical conductivity of CPA films, which do not necessarily require any acidic treatment to promote a sustained charge carrier conduction, 10 mu m short-channel OMDs were hence manufactured by interfacing the printed CPA layers with a solid polyelectrolyte (SPE). We accordingly demonstrated prototypes of stable and best performing OMD devices with downscaled features, showing well-defined counterclockwise hysteresis/rectification and an enhanced durability. These properties pave the way to further improving performance, as well as to realizing a direct integration of the devices into hardware neural networks by in-line fabrication routes

Interfacing aptamers, nanoparticles and graphene in a hierarchical structure for highly selective detection of biomolecules in OECT devices

In several biomedical applications, the detection of biomarkers demands high sensitivity, selectivity and easy-to-use devices. Organic electrochemical transistors (OECTs) represent a promising class of devices combining a minimal invasiveness and good signal transduction. However, OECTs lack of intrinsic selectivity that should be implemented by specific approaches to make them well suitable for biomedical applications. Here, we report on a biosensor in which selectivity and a high sensitivity are achieved by interfacing, in an OECT architecture, a novel gate electrode based on aptamers, Au nanoparticles and graphene hierarchically organized to optimize the final response. The fabricated biosensor performs state of the art limit of detection monitoring biomolecules, such as thrombin-with a limit of detection in the picomolar range (≤ 5 pM) and a very good selectivity even in presence of supraphysiological concentrations of Bovine Serum Albumin (BSA-1mM). These accomplishments are the final result of the gate hierarchic structure that reduces sterich indrance that could contrast the recognition events and minimizes false positive, because of the low affinity of graphene towards the physiological environment. Since our approach can be easily applied to a large variety of different biomarkers, we envisage a relevant potential for a large series of different biomedical applications

Coherent X-ray diffraction imaging of nanoengineered polymeric capsules

© 2017, Pleiades Publishing, Inc. For the first time, nanoengineered polymeric capsules and their architecture have been studied with coherent X-ray diffraction imaging technique. The use of coherent X-ray diffraction imaging technique allowed us to analyze the samples immersed in a liquid. We report about the significant difference between polymeric capsule architectures under dry and liquid conditions

Regulating financial conglomerates

We investigate the optimal regulation of financial conglomerates which combine a bank and a non-bank financial institution. The conglomerate’s risk-taking incentives depend upon the level of market discipline it faces, which in turn is determined by the conglomerate’s liability structure. We examine optimal capital requirements for stand-alone institutions, for integrated financial conglomerates, and for financial conglomerates that are structured as holding companies. For a given risk profile, integrated conglomerates have a lower probability of failure than either their stand-alone or decentralized equivalent. However, when risk profiles are endogenously selected, conglomeration may extend the reach of the deposit insurance safety net and hence provide incentives for increased risk-taking. As a result, integrated conglomerates may optimally attract higher capital requirements. In contrast, decentralised conglomerates are able to hold assets in the socially most efficient place. Their optimal capital requirements encourage this. Hence, the practice of “regulatory arbitrage”, or of transferring assets from one balance sheet to another, is welfare-increasing. We discuss the policy implications of our finding in the context not only of the present debate on the regulation of financial conglomerates but also in the light of existing US bank holding company regulation

3D printed masks for powders and viruses safety protection using food grade polymers: Empirical tests

The production of 3D printed safety protection devices (SPD) requires particular attention to the material selection and to the evaluation of mechanical resistance, biological safety and surface roughness related to the accumulation of bacteria and viruses. We explored the possibility to adopt additive manufacturing technologies for the production of respirator masks, responding to the sudden demand of SPDs caused by the emergency scenario of the pandemic spread of SARS‐COV‐ 2. In this study, we developed different prototypes of masks, exclusively applying basic additive manufacturing technologies like fused deposition modeling (FDM) and droplet‐based precision extrusion deposition (db‐PED) to common food packaging materials. We analyzed the resulting mechanical characteristics, biological safety (cell adhesion and viability), surface roughness and resistance to dissolution, before and after the cleaning and disinfection phases. We showed that masks 3D printed with home‐grade printing equipment have similar performances compared to the industrial‐grade ones, and furthermore we obtained a perfect face fit by customizing their shape. Finally, we developed novel approaches to the additive manufacturing post‐processing phases essential to assure human safety in the production of 3D printed custom medical devices

‘We are getting there slowly’: lesbian teacher experiences in the post-Section 28 environment

Prior to the subtraction of Section 28 from the 1988 Local Government Act in 2003, a substantial amount of research was published that specifically examined the experiences of lesbian physical education (PE) teachers. This article contributes to the existing academic literature by exploring the lives of lesbian, gay, bisexual and transsexual teachers working in a post-Section 28 school environment. Drawing on life history interviews of two lesbian PE teachers, we offer insights into how the abolition of Section 28 has affected their lives. Comparable to previous studies, both women reported feeling fearful of the consequences of identifying as lesbian and employed various strategies in order to maintain a divide between their public and private lives so as to conceal their sexual identities from colleagues, pupils and parents. However, in contrast to much of the previous literature, we found that the teachers in this study also identified with narratives of resistance. Despite being fearful of coming out at work, they nevertheless remained committed to coming out when the context is appropriate, to challenging the heteronormative symbolic order configured around the heterosexual/homosexual binary and to more proactively promoting sexual diversity and tolerance in schools