Pancreatic microexons regulate islet function and glucose homeostasis.

Pancreatic islets control glucose homeostasis by the balanced secretion of insulin and other hormones, and their abnormal function causes diabetes or hypoglycaemia. Here we uncover a conserved programme of alternative microexons included in mRNAs of islet cells, particularly in genes involved in vesicle transport and exocytosis. Islet microexons (IsletMICs) are regulated by the RNA binding protein SRRM3 and represent a subset of the larger neural programme that are particularly sensitive to SRRM3 levels. Both SRRM3 and IsletMICs are induced by elevated glucose levels, and depletion of SRRM3 in human and rat beta cell lines and mouse islets, or repression of particular IsletMICs using antisense oligonucleotides, leads to inappropriate insulin secretion. Consistently, mice harbouring mutations in Srrm3 display defects in islet cell identity and function, leading to hyperinsulinaemic hypoglycaemia. Importantly, human genetic variants that influence SRRM3 expression and IsletMIC inclusion in islets are associated with fasting glucose variation and type 2 diabetes risk. Taken together, our data identify a conserved microexon programme that regulates glucose homeostasis.

A developmentally programmed splicing failure contributes to DNA damage response attenuation during mammalian zygotic genome activation.

Transition from maternal to embryonic transcriptional control is crucial for embryogenesis. However, alternative splicing regulation during this process remains understudied. Using transcriptomic data from human, mouse, and cow preimplantation development, we show that the stage of zygotic genome activation (ZGA) exhibits the highest levels of exon skipping diversity reported for any cell or tissue type. Much of this exon skipping is temporary, leads to disruptive noncanonical isoforms, and occurs in genes enriched for DNA damage response in the three species. Two core spliceosomal components, Snrpb and Snrpd2, regulate these patterns. These genes have low maternal expression at ZGA and increase sharply thereafter. Microinjection of Snrpb/d2 messenger RNA into mouse zygotes reduces the levels of exon skipping at ZGA and leads to increased p53-mediated DNA damage response. We propose that mammalian embryos undergo an evolutionarily conserved, developmentally programmed splicing failure at ZGA that contributes to the attenuation of cellular responses to DNA damage.

Cooperative epithelial phagocytosis enables error correction in the early embryo.

Errors in early embryogenesis are a cause of sporadic cell death and developmental failure1,2. Phagocytic activity has a central role in scavenging apoptotic cells in differentiated tissues3-6. However, how apoptotic cells are cleared in the blastula embryo in the absence of specialized immune cells remains unknown. Here we show that the surface epithelium of zebrafish and mouse embryos, which is the first tissue formed during vertebrate development, performs efficient phagocytic clearance of apoptotic cells through phosphatidylserine-mediated target recognition. Quantitative four-dimensional in vivo imaging analyses reveal a collective epithelial clearance mechanism that is based on mechanical cooperation by two types of Rac1-dependent basal epithelial protrusions. The first type of protrusion, phagocytic cups, mediates apoptotic target uptake. The second, a previously undescribed type of fast and extended actin-based protrusion that we call 'epithelial arms', promotes the rapid dispersal of apoptotic targets through Arp2/3-dependent mechanical pushing. On the basis of experimental data and modelling, we show that mechanical load-sharing enables the long-range cooperative uptake of apoptotic cells by multiple epithelial cells. This optimizes the efficiency of tissue clearance by extending the limited spatial exploration range and local uptake capacity of non-motile epithelial cells. Our findings show that epithelial tissue clearance facilitates error correction that is relevant to the developmental robustness and survival of the embryo, revealing the presence of an innate immune function in the earliest stages of embryonic development.

Welfare technology, ethics and well-being a qualitative study about the implementation of welfare technology within areas of social services in a Swedish municipality.

Purpose: Digitalization and e-health have potential to generate good quality, equal health, well-being and to develop and strengthen individuals' resources with the goal of increased independence and participation in society. The implementation of welfare technology requires knowledge of digitalization, as well as an awareness of its meaning in terms of ethical principles and ethical analysis. The purpose of this study was to describe ethical analysis concerning the implementation of welfare technology, in terms of both strategies and tools, within areas of social services in a Swedish municipality. Method: We followed a working model that focused on increased knowledge and experience in the implementation of welfare technology from an ethical perspective. In the data collection were observations, a questionnaire with open-ended questions and focus group discussions used. Results: The analysis showed that when welfare technology was introduced and implemented within the area of social services in a municipality, ethical awareness resulting from the conflicts between various interests and values had to be addressed. Conclusions: The ethical analysis improved implementation of strategies and tools in terms of facts and values, and invisible underlying values to the concept of well-being.

Production of Mannosylerythritol Lipids (MELs) to be Used as Antimicrobial Agents Against S. aureus ATCC 6538.

Antimicrobial resistance (AMR) is a current major health issue, both for the high rates of resistance observed in bacteria that cause common infections and for the complexity of the consequences of AMR. Pathogens like Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Mycobacterium tuberculosis among others are clear examples of antibiotic-resistant threats. Biosurfactants have recently emerged as a potential new generation of anti-adhesive and anti-biofilm agents; mannosylerythritol lipids (MELs) are biosurfactants produced by a range of fungi. A range of structural variants of MELs can be formed and the proportion of each isomer in the fermentation depends on the yeast used, the carbon substrate used for growth and the duration of the fermentation. In order to allow assessment of the possible functions of MELs as antimicrobial molecules, small quantities of MEL were produced by controlled fermentation. Fermentations of the yeast Pseudozyma aphidis using rapeseed oil as a carbon source yielded up to 165 gMELs/kgSubstrate. The MELs formed by this strain was a mixture of MEL-A, MEL-B, MEL-C and MEL-D. The MELs produced were tested against S. aureus ATCC 6538 on pre-formed biofilm and on co-incubation biofilm experiments on silicone discs; showing a disruption of biomass, reduction of the biofilm metabolic activity and a bacteriostatic/bactericidal effect confirmed by a release of oxygen uptake [Formula: see text], the reduction of citrate synthase activity and scanning electron microscopy. The results show that MELs are promising antimicrobial molecules for biomedical technological applications that could be studied in detail in large-scale systems and in conjunction with animal tissue models.

Intersectional perspective in elderly care.

Earlier research has shown that power relationships at workplaces are constructed by power structures. Processes related to power always influence the working conditions for (in this study in elderly care) the working groups involved. Power structures are central for intersectional analysis, in the sense that the intersectional perspective highlights aspects such as gender and ethnicity (subjective dimensions) and interrelates them to processes of power (objective dimension). This qualitative study aims to explore in what way an intersectional perspective could contribute to increased knowledge of power structures in a nursing home where the employees were mostly immigrants from different countries. By using reflexive dialogues related to an intersectional perspective, new knowledge which contributes to the employees' well-being could develop. Narrative analysis was the method used to conduct this study. Through a multi-stage focus group on six occasions over 6 months, the staff were engaged in intersectional and critical reflections about power relationship with the researchers, by identifying patterns in their professional activities that could be connected to their subjectivities (gender, ethnicity, etc.). The result of this study presents three themes that express the staff's experiences and connect these experiences to structural discrimination. 1) Intersectionality, knowledge, and experiences of professionalism; 2) Intersectionality, knowledge, and experiences of collaboration; and 3) Intersectionality, knowledge, and experiences of discrimination. The result demonstrates that an intersectional perspective reinforces the involved abilities, during the conversations, into being clear about, for example, their experiences of discrimination, and consequently developing a better understanding of their professionalism and collaboration. Such deeper reflections became possible through a process of consciousness raising, strengthening the employee's self-confidence, in a positive way.

Versatile approach for the fabrication of functional wrinkled polymer surfaces.

A simple and versatile approach to obtaining patterned surfaces via wrinkle formation with variable dimensions and functionality is described. The method consists of the simultaneous heating and irradiation with UV light of a photosensitive monomer solution confined between two substrates with variable spacer thicknesses. Under these conditions, the system is photo-cross-linked, producing a rapid volume contraction while capillary forces attempt to maintain the contact between the monomer mixture and the cover. As a result of these two interacting forces, surface wrinkles were formed. Several parameters play a key role in the formation and final characteristics (amplitude and period) of the wrinkles generated, including the formulation of the photosensitive solution (e.g., the composition of the monomer mixture) and preparation conditions (e.g., temperature employed, irradiation time, and film thickness). Finally, in addition, the possibility of modifying the surface chemical composition of these wrinkled surfaces was investigated. For this purpose, either hydrophilic or hydrophobic comonomers were included in the photosensitive mixture. The resulting surface chemical composition could be finely tuned as was demonstrated by significant variations in the wettability of the structured surfaces, between 56° and 104°, when hydrophilic and hydrophobic monomers were incorporated, respectively.

Versatile functional microstructured polystyrene-based platforms for protein patterning and recognition.

We report the preparation of different functional surface patterns based on the optimization of the photo-cross-linking/degradation kinetics of polystyrene (PS) upon exposure to UV-light. We employed a PS-b-PGA (polystyrene-block-poly(l-glutamic acid)) block copolymer that will, in addition to the surface pattern, provide functionality. By using short irradiation times, PS can be initially cross-linked, whereas an excess of the exposure time provokes the degradation of the material. As a result of the optimization of time of exposure, the use of an appropriate cover, or the incorporation of an appropriate amount of absorbing active species (photoinitiator), different tailor-made surface patterns can be obtained, from boxes to needles. Moreover, in addition to the surface pattern, we introduced changes on the chemical composition of the polystyrene using an amphiphilic block copolymer (for instance, we employ PS-b-PGA) that will provide functional surfaces with major advantages. In particular, the presence of carboxylic functional groups provides a unique opportunity to anchor, for instance polypeptide sequences. We describe the immobilization of polypeptide sequences in precise surface positions that allows the use of the surfaces for protein recognition purposes. The immobilization of the proteins evidence the success of the recognition and opens a new alternative for protein patterning on surfaces for many biotechnological and biomedical applications.

Risk factors for a low linezolid trough plasma concentration in acute infections.

Linezolid is an antibiotic with time-dependent activity, and both the percentage of time that plasma concentrations exceed the MIC and the area under the concentration-time curve over 24 h in the steady state divided by the MIC (AUC24/MIC ratio) are associated with clinical response. The aim of this study was to analyze the linezolid trough plasma concentration (C(min)) and to determine factors associated with a C(min) < 2 mg/liter and other clinically relevant thresholds. Characteristics of 78 patients receiving 600 mg/12 h of linezolid with a C(min) determination at the steady state and within the first 10 days of treatment were retrospectively reviewed. Concentrations were measured using high-pressure liquid chromatography. Univariate and multivariate analysis were performed to identify risk factors of low C(min). A total of 29.5% of patients had a C(min) < 2 mg/liter. The percentage was significantly higher in patients with an estimated glomerular filtration (eGF) > 80 ml/min, in intensive care unit (ICU) patients, and in patients with an infection due to Staphylococcus aureus. The independent predictors of C(min) < 2 mg/liter were an eGF > 80 ml/min (odds ratio [OR], 10; 95% confidence interval [CI], 2.732 to 37.037; P = 0.001) and infection due to S. aureus (OR, 5.906; 95% CI, 1.651 to 21.126; P = 0.006). A linezolid C(min) of <2 mg/liter was found in 29.5% of cases, and the risk was significantly higher among those with an eGF > 80 ml/min and in infections due to S. aureus. In patients with severe sepsis, a loading dose or continuous infusion and drug monitoring could improve the pharmacodynamic parameters associated with linezolid efficacy.

Decreased serum linezolid concentrations in two patients receiving linezolid and rifampicin due to bone infections.

Methicillin-resistant Staphylococcus is a common cause of orthopaedic implant infections. In such cases, rifampicin is the antibiotic of choice, but it should not be administered alone to avoid the selection of resistant mutants. Linezolid has activity against resistant staphylococci and a high oral bioavailability; therefore, it could be a good option for combining with rifampicin. We describe 2 patients admitted to our hospital due to orthopaedic implant infections, who received combination therapy with linezolid and rifampicin. In both cases, the trough serum concentration of linezolid during rifampicin treatment was below the minimum inhibitory concentration required to inhibit the growth of 90% of organisms (MIC(90)) for staphylococci, but increased after rifampicin withdrawal. This finding suggests an interaction between rifampicin and linezolid, and a possible explanation is discussed.

Controlling optical properties and function of BODIPY by using asymmetric substitution effects.

Asymmetrically substituted BODIPY analogues of the dye PM567 have been synthesised from 2-acylpyrroles and pyrroles that bear indene, fluorene or difluorene units. The type of linkage between the fluorene and the BODIPY core plays an important role in the photophysics of the BODIPY chromophore. Indeed, an aliphatic bridge gives rise to an energy-transfer process between the chromophores, whereas a vinyl spacer allows an electronic interaction between them, leading to a large red shift of the spectral bands. The laser action of the new dyes has been analysed under transversal pumping at 10 Hz repetition rate, in both liquid phase and incorporated into solid polymeric matrices. Lasing efficiencies of up to 40% were reached with high photostabilities with the laser output remaining at the initial level after 1×10(5) pump pulses in the same position of the sample. The laser action of the new dyes outperforms the laser behaviour of commercial dyes that emit in the same spectral region. The replacement of fluorene by indene quenches the fluorescence and laser emission, but allows the development of an iron cation fluorescent sensor.

Can drug resistance mutations influence the measurement of plasma HIV-RNA by different viral load techniques?

Guidelines for the use of antiretroviral agents consider that viral load values may differ using the branched DNA (bDNA) and polymerase chain reaction (PCR) methods. Whereas the most commonly used PCR assay (Amplicor v1.5, Roche) targets HIV gag sequences, the bDNA (Quantiplex v3.0, Bayer) is based on the recognition of HIV pol sequences. The possibility that accumulation of drug-resistance mutations at the reverse transcriptase and/or protease genes could influence viral load measurements using bDNA was examined in a case control study. Plasma samples collected from 46 HIV-infected individuals receiving antiretroviral therapy and carrying different number of drug resistance mutations were analyzed. The performance of both bDNA and PCR assays was found to be comparable irrespective of the presence or absence of drug resistance mutations.