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Eldercare workers experience higher levels of moral distress than other health and social care service workers. Moral distress is a psychological response to a morally challenging event. Very little is known about moral distress in the context of eldercare and about the mechanisms of preventing or mitigating moral distress. This qualitative study was conducted as part of the “Ensuring the availability of staff and the attractiveness of the sector in eldercareservices” project in Finland in 2021. The data were from 39 semi-structured interviews. This qualitative interview data were examined using two-stage content analysis. The key finding of this study, as reported by eldercare professionals, is that strategies to mitigate moral distress can be found at all organizational levels : organizational, workplace and individual. The tools that emerged from the interviews fell into four main categories: 1) organizational support and education 2) peer support 3) improving self-care and competence and 4) defending patients. The main identified categories confirmed the earlier findings but the qualitative, rich research interview data provided new insights into a little-studied topic: mitigating moral distress in eldercare. The main conclusion is that, in order to mitigate moral distress, ethical competence needs to be strengthened at all organizational levels.

Summary: The major lactiferous ducts of the human breast branch out and end at terminal ductal lobular units (TDLUs). Despite their functional and clinical importance, the three-dimensional (3D) architecture of TDLUs has remained undetermined. Our quantitative and volumetric imaging of healthy human breast tissue demonstrates that highly branched TDLUs, which exhibit increased proliferation, are uncommon in the resting tissue regardless of donor age, parity, or hormonal contraception. Overall, TDLUs have a consistent shape and branch parameters, and they contain a main subtree that dominates in bifurcation events and exhibits a more duct-like keratin expression pattern. Simulation of TDLU branching morphogenesis in three dimensions suggests that evolutionarily conserved mechanisms regulate mammary gland branching in humans and mice despite their anatomical differences. In all, our data provide structural insight into 3D anatomy and branching of the human breast and exemplify the power of volumetric imaging in gaining a deeper understanding of breast biology.

Abstract Background All patients with total hip arthroplasty (THA), especially those with metal-on-metal (MM) THA, are exposed to metallic particles and ions, which may cause total or site-specific mortality. We analyzed the causes of total and site-specific mortality among a cohort of patients with MM and with metal-on-polyethylene (MP) THA after a long follow-up time. Methods Standardized mortality ratios (SMR) of total and site-specific causes of death were calculated for 579 patients with MM (McKee-Farrar) and 1585 patients with MP (Brunswik, Lubinus) THA for primary osteoarthritis. Results Mean follow-up time was 17.9 years for patients with MM and 16.7 years for patients with MP. Overall SMR was 0.95 for the MM cohort and 0.90 for the MP cohort, as compared to the normal population. Both cohorts showed significantly decreased mortality for the first decade postoperatively, equal mortality over the next 10 years, and significantly increased mortality after 20 years. Patients with MM THA had higher cancer mortality (SMR 1.01) than those with MP THA (SMR 0.66) during the first 20 years postoperatively, but not thereafter. Conclusion Both MM and MP prostheses are safe based on total and site-specific mortality of recipients during the first 20 postoperative years in comparison with the general population.

Central to the success of functional precision medicine of solid tumors is to perform drug testing of patient-derived cancer cells (PDCs) in tumor-mimicking ex vivo conditions. While high throughput (HT) drug screening methods have been well-established for cells cultured in two-dimensional (2D) format, this approach may have limited value in predicting clinical responses. Here, we describe the results of the optimization of drug sensitivity and resistance testing (DSRT) in three-dimensional (3D) growth supporting matrices in a HT mode (3D-DSRT) using the hepatocyte cell line (HepG2) as an example. Supporting matrices included widely used animal-derived Matrigel and cellulose-based hydrogel, GrowDex, which has earlier been shown to support 3D growth of cell lines and stem cells. Further, the sensitivity of ovarian cancer PDCs, from two patients included in the functional precision medicine study, was tested for 52 drugs in 5 different concentrations using 3D-DSRT.Shortly, in the optimized protocol, the PDCs are embedded with matrices and seeded to 384-well plates to allow the formation of the spheroids prior to the addition of drugs in nanoliter volumes with acoustic dispenser. The sensitivity of spheroids to drug treatments is measured with cell viability readout (here, 72 h after addition of drugs). The quality control and data analysis are performed with openly available Breeze software. We show the usability of both matrices in established 3D-DSRT, and report 2D vs 3D growth condition dependent differences in sensitivities of ovarian cancer PDCs to MEK-inhibitors and cytotoxic drugs. This study provides a proof-of-concept for robust and fast screening of drug sensitivities of PDCs in 3D-DSRT, which is important not only for drug discovery but also for personalized ex vivo drug testing in functional precision medicine studies. These findings suggest that comparing results of 2D- and 3D-DSRT is essential for understanding drug mechanisms and for selecting the most effective treatment for the patient.

Abstract Background Co-morbidity is a powerful predictor of health care outcomes and costs, as well as an important cofounder in epidemiologic studies. The effect of co-morbidities is generally related to mortality or complications. This study evaluated the association between co-morbidity and health-related quality of life (HRQoL) in patients awaiting total joint replacement. Methods A total of 893 patients were recruited to the study between August 2002 and November 2003 in four Finnish hospitals. The effect of co-morbidity on HRQoL was measured by the generic 15D instrument and by a Visual Analog Scale (VAS). Comparative variance analysis of socio-demographic and clinical characteristics was described by using either an independent samples t-test or the Chi-square test. The differences in each of the 15D dimensions and the overall 15D single index score for patients were calculated. Two-sided p-values were calculated with the Levene Test for Equality of Variances. Results Patients with co-morbidity totaled 649; the incidence of co-morbidity was 73%. The mean number of co-morbidities among the patients was two. At baseline the 15D score in patients with and without co-morbidity was 0.778 vs 0.816, respectively. The difference of the score (0.038) was clinically and statistically significant (P < 0.001). The patients' scores with and without co-morbidity on the different 15D dimensions related to osteoarthritis-moving, sleeping, usual activities, discomfort and symptoms, vitality and sexual activity–were low in both groups. Patients with co-morbidity scored lower on the dimensions of moving, vitality and sexual activity compared to the patients without co-morbidity. Co-morbidity was significantly associated with a reduced HRQoL. Patients without co-morbidity had poorer VAS, arthritis had strong effect to their quality of life compared to the patients with co-morbidity. Conclusion Assessing co-morbidity in patients placed on the waiting list for joint replacement may be useful method to prioritization in medical decision-making for healthcare delivery. The assessment of co-morbidities during waiting time is important as well as evaluating how the co-morbidity may affect the final outcomes of the total joint replacement.

Abstract Background Several quality-of-life studies in patients awaiting major joint replacement have focused on the outcomes of surgery. Interest in examining patients on the elective waiting list has increased since the beginning of 2000. We assessed health-related quality of life (HRQoL) in patients waiting for total hip (THR) or knee (TKR) replacement in three Finnish hospitals, and compared patients' HRQoL with that of population controls. Methods A total of 133 patients awaiting major joint replacement due to osteoarthritis (OA) of the hip or knee joint were prospectively followed from the time the patient was placed on the waiting list to hospital admission. A sample of controls matched by age, gender, housing and home municipality was drawn from the computerised population register. HRQoL was measured by the generic 15D instrument. Differences between patients and the population controls were tested by the independent samples t-test and between the measurement points by the paired samples t-test. A linear regression model was used to explain the variance in the 15D score at admission. Results At baseline, 15D scores were significantly different between patients and the population controls. Compared with the population controls, patients were worse off on the dimensions of moving (P < 0.001), sleeping (P < 0.001), sexual activity (P < 0.001), vitality (P < 0.001), usual activities (P < 0.001) and discomfort and symptoms (P < 0.001). Further, psychological factors – depression (P < 0.001) and distress (P = 0.004) – were worse among patients than population controls. The patients showed statistically significantly improved average scores at admission on the dimensions of moving (P = 0.026), sleeping (P = 0.004) and discomfort and symptoms (P = 0.041), but not in the overall 15D score compared with the baseline. In patients, 15D score at baseline (P < 0.001) and body mass index (BMI) (P = 0.020) had an independent effect on patients' 15D score at hospital admission. Conclusion Although patients' HRQoL did not deteriorate while waiting, a consistently worse HRQoL was observed in patients waiting for major joint replacement compared with population controls.

Tutkimus käsittelee vanhustyössä työskentelevien hoitajien moraalista toimijuutta moraalisissa ristiriitatilanteissa. Moraalisen toimijuuden rajoittumisen on esitetty olevan keskeinen tekijä eettisen kuormituksen synnyssä. Eettinen kuormitus on yhdistetty esimerkiksi heikentyneeseen työhyvinvointiin ja hoidon laatuun sekä hoitoalan pito- ja vetovoimaisuuden vähenemiseen. Tutkimuskysymys on: miten vanhustyössä työskentelevien hoitajien kohtaamat moraaliset ristiriidat voivat rajoittaa heidän moraalista toimijuuttaan? Aineisto muodostui 40:n vanhustyössä työskentelevän hoitajan ja esihenkilön teemahaastatteluista. Aineisto analysoitiin teorialähtöisellä sisällönanalyysilla, hyödyntäen Hallamaan muotoilua moraalisesta toimijuudesta. Tutkimuksen päätulos on, että hoitajien moraalinen toimijuus näyttäytyi varsin rajoittuneena. Heidän mahdollisuutensa asettaa tavoitteita moraaliselle toiminnalle, tehdä rationaalisia moraalisia päätöksiä ja toimia moraalisesti oikein rajoittuivat. Johtopäätöksenä esitetään, että vanhustyössä tulee luoda riittävät edellytykset hoitajien moraaliselle toimijuudelle.

High-content screening prompted the development of software enabling discrete phenotypic analysis of single cells. Here, the authors show that supervised continuous machine learning can drive novel discoveries in diverse imaging experiments and present the Regression Plane module of Advanced Cell Classifier.

Changes in the dynamic architecture of podocytes, the glomerular epithelial cells, lead to kidney dysfunction. Previous studies on protein kinase C and casein kinase 2 substrates in neurons 2 (PACSIN2), a known regulator of endocytosis and cytoskeletal organization, reveal a connection between PACSIN2 and kidney pathogenesis. Here, we show that the phosphorylation of PACSIN2 at serine 313 (S313) is increased in the glomeruli of rats with diabetic kidney disease. We found that phosphorylation at S313 is associated with kidney dysfunction and increased free fatty acids rather than with high glucose and diabetes alone. Phosphorylation of PACSIN2 emerged as a dynamic process that fine-tunes cell morphology and cytoskeletal arrangement, in cooperation with the regulator of the actin cytoskeleton, Neural Wiskott–Aldrich syndrome protein (N-WASP). PACSIN2 phosphorylation decreased N-WASP degradation while N-WASP inhibition triggered PACSIN2 phosphorylation at S313. Functionally, pS313-PACSIN2 regulated actin cytoskeleton rearrangement depending on the type of cell injury and the signaling pathways involved. Collectively, this study indicates that N-WASP induces phosphorylation of PACSIN2 at S313, which serves as a mechanism whereby cells regulate active actin-related processes. The dynamic phosphorylation of S313 is needed to regulate cytoskeletal reorganization.

The Ethereum blockchain is one of the most popular permissionless blockchains. A consequence of its popularity has been the growth of processing and data storage requirements for any node participating in the Ethereum blockchain network. For constrained devices such requirements are often infeasible to meet. To cater for such nodes, a so-called light protocol has been proposed for Ethereum where the responsibility of maintaining a correct state representation is delegated to light protocol servers. Previous research has identified dependence on external state management as a potential security vulnerability that exclusively impacts light nodes. Although a simple mitigation strategy is available, it comes at the expense of increased latency. In this work, we propose a new Ethereum node type, which we call a subset node, as an extension of the light protocol. Our proposal allows subset nodes to gain a lower latency than a pure light node with comparable or even higher security assurances by tracking and evaluating only a subset of all of the transactions issued on the blockchain. We provide a formal proof on the correctness of the blockchain state used by the subset node under the proposed model. To evaluate the practical feasibility of the subset node model, we analyze one year of historical transaction data from Ethereum, and demonstrate that a subset node tracking the state of a single account can achieve a significant reduction in storage and computational requirements when compared to a full node.

Ethereum is a decentralized blockchain, known as being the second most popular public blockchain after Bitcoin. Since Ethereum is decentralised the canonical state is determined by the Ethereum network participants via a consensus mechanism without a centralized coordinator. The network participants are required to evaluate every transaction starting from the genesis block, which requires a large amount of network, computing, and storage resources. This is impractical for many devices with either limited computing resources or intermittent network connectivity. To overcome this drawback Ethereum defines a light client protocol where the light client fetches the blockchain state from a node operating as a light protocol server. Light clients are unable to maintain blockchain state internally, and as a consequence can only perform partial validation on blocks. Thus they rely on the light server for full block validation and to provide the updated blockchain state. Light clients connect to multiple light servers to mitigate the risk of relying on a single potentially dishonest server. Ethereum light clients are known to suffer from a probabilistic security model, but they are widely assumed to be secure under normal operating conditions. In fact, the implicit security assumptions of light clients have not been formally characterised in the literature. We present and analyse the probabilistic security guarantees under three different adversarial scenarios. The results show that for any adversary that is able to manipulate the network, the security assurances provided by the light protocol are severely impacted, and in some cases entirely lost. These results clearly demonstrate that the assumption of normal operating conditions is insufficient to justify the security assumptions of light clients. Our work also provides insight to the security of light clients under different security parameters, allowing light client implementers to more accurately understand the potential security trade-offs.

Hypervariable loops of HIV-1 Env protein gp120 are speculated to play roles in the conformational transition of Env to the receptor binding-induced metastable state. Structural analysis of full-length Env-based immunogens, containing the entire V2 loop, displayed tighter association between gp120 subunits, resulting in a smaller trimeric diameter than constructs lacking V2. A prominent basal quaternary location of V2 and V3' that challenges previous reports would facilitate gp41-independent gp120-gp120 interactions and suggests a quaternary mechanism of epitope occlusion facilitated by hypervariable loops. Deletion of V2 resulted in dramatic exposure of basal, membrane-proximal gp41 epitopes, consistent with its predicted basal location. The structural features of HIV-1 Env characterized here provide grounds for a paradigm shift in loop exposure and epitope occlusion, while providing substantive rationale for epitope display required for elicitation of broadly neutralizing antibodies, as well as substantiating previous pertinent literature disregarded in recent reports.

Cellular uptake of clustered α2β1-integrin induces the formation of membrane compartments that subsequently mature into a multivesicular body (MVB). Enhanced internalization mediated by clustered integrins was observed upon infection by the picornavirus echovirus 1 (EVI). We elucidated the structural features of virus-induced MVBs (vMVBs) in comparison to antibody-induced control MVBs (mock infection) by means of high-pressure cryo fixation of cells followed by immuno electron tomography during early entry of the virus. Three-dimensional tomograms revealed a marked increase in the size and complexity of these vMVBs and the intraluminal vesicles (ILVs) at 2 and 3.5 hours post infection (p.i.), in contrast to the control MVBs without virus. Breakages in the membranes of vMVBs were detected from tomograms after 2 and especially after 3.5 h suggesting that these breakages could facilitate the genome release to the cytoplasm. The in situ neutral-red labeling of viral genome showed that virus uncoating starts as early as 30 min p.i., while an increase of permeability was detected in the vMVBs between 1 and 3 hours p.i., based on a confocal microscopy assay. Altogether, the data show marked morphological changes in size and permeability of the endosomes in the infectious entry pathway of this non-enveloped enterovirus and suggest that the formed breakages facilitate the transfer of the genome to the cytoplasm for replication.

Electron tomography (ET) of biological samples is used to study the organization and the structure of the whole cell and subcellular complexes in great detail. However, projections cannot be acquired over full tilt angle range with biological samples in electron microscopy. ET image reconstruction can be considered an ill-posed problem because of this missing information. This results in artifacts, seen as the loss of three-dimensional (3D) resolution in the reconstructed images. The goal of this study was to achieve isotropic resolution with a statistical reconstruction method, sequential maximum a posteriori expectation maximization (sMAP-EM), using no prior morphological knowledge about the specimen. The missing wedge effects on sMAP-EM were examined with a synthetic cell phantom to assess the effects of noise. An experimental dataset of a multivesicular body was evaluated with a number of gold particles. An ellipsoid fitting based method was developed to realize the quantitative measures elongation and contrast in an automated, objective, and reliable way. The method statistically evaluates the sub-volumes containing gold particles randomly located in various parts of the whole volume, thus giving information about the robustness of the volume reconstruction. The quantitative results were also compared with reconstructions made with widely-used weighted backprojection and simultaneous iterative reconstruction technique methods. The results showed that the proposed sMAP-EM method significantly suppresses the effects of the missing information producing isotropic resolution. Furthermore, this method improves the contrast ratio, enhancing the applicability of further automatic and semi-automatic analysis. These improvements in ET reconstruction by sMAP-EM enable analysis of subcellular structures with higher three-dimensional resolution and contrast than conventional methods.

Collagen receptor integrins recycle between the plasma membrane and endosomes and facilitate formation and turnover of focal adhesions. In contrast, clustering of α2β1 integrin with antibodies or the human pathogen echovirus 1 (EV1) causes redistribution of α2 integrin to perinuclear multivesicular bodies, α2-MVBs. We show here that the internalized clustered α2 integrin remains in α2-MVBs and is not recycled back to the plasma membrane. Instead, receptor clustering and internalization lead to an accelerated down-regulation of α2β1 integrin compared to the slow turnover of unclustered α2 integrin. EV1 infection or integrin degradation is not associated with proteasomal or autophagosomal processes and shows no significant association with lysosomal pathway. In contrast, degradation is dependent on calpains, such that it is blocked by calpain inhibitors. We show that active calpain is present in α2-MVBs, internalized clustered α2β1 integrin coprecipitates with calpain-1, and calpain enzymes can degrade α2β1 integrin. In conclusion, we identified a novel virus- and clustering-specific pathway that diverts α2β1 integrin from its normal endo/exocytic traffic to a nonrecycling, calpain-dependent degradative endosomal route.

The isolation of single cells while retaining context is important for quantifying cellular heterogeneity but technically challenging. Here, the authors develop a high-throughput, scalable workflow for microscopy-based single cell isolation using machine-learning, high-throughput microscopy and laser capture microdissection.

Determining vesicle localization and association in live microscopy may be challenging due to non-simultaneous imaging of rapidly moving objects with two excitation channels. Besides errors due to movement of objects, imaging may also introduce shifting between the image channels, and traditional colocalization methods cannot handle such situations. Our approach to quantifying the association between tagged proteins is to use an object-based method where the exact match of object locations is not assumed. Point-pattern matching provides a measure of correspondence between two point-sets under various changes between the sets. Thus, it can be used for robust quantitative analysis of vesicle association between image channels. Results for a large set of synthetic images shows that the novel association method based on point-pattern matching demonstrates robust capability to detect association of closely located vesicles in live cell-microscopy where traditional colocalization methods fail to produce results. In addition, the method outperforms compared Iterated Closest Points registration method. Results for fixed and live experimental data shows the association method to perform comparably to traditional methods in colocalization studies for fixed cells and to perform favorably in association studies for live cells.

The article expands on Lewis and Maslin's "double two-step" historicization of the Anthropocene, with two major transitions in energy (agriculture and fossil fuels) and two in social organization (modernity and the Great Acceleration). Insofar as planetary impacts arise from "what we spend our time doing" - foraging, farming, feudal then waged labour, finally unsustainable consumption - such "doing" is understood as precisely 'performative' in the sense that its effects only arise from a massive social repetition that is confused with essential nature and thus concealed. Through a graphic model of such 'plural performativity,' four consecutive Anthropo(s)cenes are sketched: the Giving World of agriculture and state formation; the New World of colonial pillage and world trade; the Netherworld of wage labour and fossil capital; then 'All the World' but not with all of 'us' as players. Apart from environmental changes, the paper targets performances of power and inequality: normative histories of 'common sense' on the one hand, concealing 'people's histories' of conflict and opposition, on the other - the Anthropocene arising not simply from what the majority of people have been doing, but from what they have always been forced to do. [ABSTRACT FROM AUTHOR]

The Centennial of one of the cruelest of European civil wars fought in Finland between the Reds and the Whites from January to May 1918 has evoked a spectrum of theatre productions illustrating variations of styles and approaches on the events. The turn in the treatment of this cultural trauma occurred with the interpretations and narrative perspectives that were fixed in the 1960s, when an understanding for the defeated Red side was expressed in historiography, literature and theatre. Since that, the last six decades the Finnish theatre and public discourse on the Civil War have been dominated by the Red narrative as the memory of the 1918 Civil War provided an important part in the new identity politics for the 1969 generation. Since the 1980's the topic was mostly put aside so that before the 2018 revivals of the Civil War topic, the productions seem to have been reactions by the artists confronting the developments at the end of the Cold War. Some theatrical events can even be tied to the cultural trauma of the 1969 left evoked by the collapse of the socialist block. The Centennial productions repeated the Red narrative but they also provided more balanced interpretations on the tragic events. [ABSTRACT FROM AUTHOR]

The article presents the Trump presidency and the human-driven geological epoch of the Anthropocene as two arguable extremes among current notions of 'performativity': (1) a traditionally vertical model based on individual action and antagonism - where 'facts' matter less than 'making things great'; and (2) the more extended, horizontal human performance of things like global warming ("All the world's a stage"). Drawing freely on George Lakoff and Timothy Morton, it is argued that these models differ fundamentally in 'magnitude': where the one is direct, singular, vertical, and fast, the other is systemic, plural, horizontal, and slow beyond human perception. With Judith Butler and Naomi Klein, it is also argued that to actually confront the twin crises at issue, we need to acknowledge the kind of 'plural performativity' - of repetition, norms, and dissimulation - that brought them into being in the first place. [ABSTRACT FROM AUTHOR]