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Twórczość Leo Lipskiego stanowi przykład pisarstwa, w którym struktury tekstowo-językowe pozostają w nierozerwalnym splocie ze strukturami egzystencji. Dzieło Lipskiego, w perspektywie radykalnego ujęcia formy pisania jako pracy w obrębie cielesności, świadomości, pamięci staje się szczególną metodą uobecniania, reprezentacji, mediacji i moderacji własnego doświadczenia. W tym sensie pisanie – twórczy akt – Lipskiego jawi się jako faktyczna formacja życia. W artykule zostaje przedstawiona próba wydobycia wskazanych kwestii z dzieła autora Niespokojnych poddanego lekturze mikrologicznej, meletycznej, zorientowanej na pisanie.

Artykuł stanowi polemiczne i krytyczne omówienie kilku przykładowych tłumaczeń gier słownych w powieści Jamesa Joyce’a Ulisses w nowym przekładzie Macieja Świerkockiego. Aby pokazać inne możliwości translatoryczne, autor artykułu przytacza także tłumaczenia Macieja Słomczyńskiego i Aloysa Skoumala.

Cel pracyCelem pracy była ocena zachowań prozdrowotnych jako działań ochronnych przed objawami lęku i depresji w grupie pracowników służby zdrowia, w okresie pierwszej fali pandemii wirusa SARS-CoV-2.MetodaW badanu udział wzięło 114 osób, w tym 46 lekarzy w wieku 41,10±11,89 lat i 68 pielęgniarek w wieku 48,16±8,54 lat. Metody: Do badań użyto: Inwentarza Zachowań Zdrowotnych (IZZ), Inwentarza Objawów Depresyjnych (BDI) oraz Inwentarza Stanu i Cechy Lęku (STAI).WynikiW grupie badanej w IZZ otrzymano średni wynik 79,61±13,08 pkt. W kwestionariuszu BDI badani uzyskali średnio 3,7±4,65pkt. W kwestionariuszu STAI, w części dotyczącej lęku-stanu wynik średni w grupie badanej wynosił 38,08±9,46 pkt., a w odniesieniu do lęku-cechy 38,35±8,44pkt. Biorąc pod uwagę składowe IZZ jedynie wyniki uzyskane w podskalach: pozytywne nastawienie psychiczne (PNP) oraz praktyki zdrowotne (PZ) korelowały ujemnie z wynikami uzyskanymi w skalach STAI i BDI. Ponadto wykazano prozdrowotny wpływ PNP na objawy lęku i depresji.WnioskiNie zaobserwowano znacznego nasilenia objawów lękowych i depresyjnych wśród personelu medycznego w okresie pierwszej fali pandemii. Zachowania prozdrowotne, a zwłaszcza pozytywne nastawienia psychiczne mogą pełnić ochronna funkcję w odniesieniu do objawów lęku i depresji w sytuacji stresowej.

Increased parental monitoring is protective against cannabis use (CU) for justice involved youth, although discrepancies across parent/adolescent reports of monitoring may confer risk. Baseline data were drawn from two randomized clinical trials (152 adolescents; Mage= 15.9; 68% male). Adolescents reported on past 60-day CU and adolescents and parents completed a measure of parental knowledge, parental solicitation, parental control, and child disclosure. Multiple regression models that varied operationalization of discrepancies were performed, in which CU was predicted from each monitoring construct. Inclusion of main effects of parent and adolescent reports improved prediction of CU, particularly parental knowledge and child disclosure. When operationalized categorically, discrepancies improved prediction of CU for parental knowledge. Discrepancies did not improve prediction of CU for the other aspects of parental monitoring. Findings diverge from previous research on adolescent alcohol use; explanations of findings and implications for treatment are discussed.

Paraoxonases (PON) 1-3 are lactonases with antioxidant and atheroprotective properties. The best known single nucleotide polymorphisms (SNPs) within the PON family, include: Q192R (rs662), L55M (rs854560) in the PON1 gene and C311S (rs7493) in the PON2 gene. Their influence on the occurrence and course of coronary artery disease (CAD) is unclear. The aim of this study was to assess the association between the most common PON1 and PON2 genetic variants with the presence of CAD, as well as their relation to coronary lesion complexity in accordance with the ACC/AHA standard.We included 1027 individuals: 367 CAD patients qualified for coronary angiography and 660 healthy volunteers as controls. We extracted DNA from circulating blood leukocytes, amplified the PON1 and PON2 genetic sequence and used restriction enzymes to identify the SNPs. Patients with CAD underwent coronary angiography and were assigned to two groups based on lesion severity: patients with at least one type C lesion and without a type C lesion. The former where categorized into those with a significant narrowing (≥50% diameter stenosis) and those without one.We found no association between the analyzed SNPs and symptomatic CAD. However, in patients with diagnosed CAD, the PON311S allele was independently associated with the risk of the most complex type C coronary lesion occurrence.Our study is the first report of an association between PON2 311S SNP and the type of coronary atherosclerotic lesions in humans.

The glycocalyx is a soft material composed of glycosylated proteins and lipids decorating the plasma membrane of cells. Although the glycocalyx is a mechanical conduit between the plasma membrane and the cell surroundings, the coupled dynamics of the glycocalyx and membrane are poorly understood in most cell processes. Here, we construct a dynamic model to predict the coupled mechanical behaviors of the glycocalyx and membrane due to internal cytosolic pressure in a cell interacting with an external substrate. We report how the glycocalyx constituents bear cytosolic loads, physically rearrange, and shape the cell membrane. Simulations of the model predict that highly flexible, polymeric elements in the glycocalyx are uniformly compressed in the cell-substrate contact zone. However, more rigid, fiber-like glycocalyx constituents dynamically rearrange over timescales on the order of 10 milliseconds, concentrating into clusters. This spatial heterogeneity allows close apposition of the plasma membrane with the substrate, a geometry that would be favorable for subsequent adhesive bond formation. Clustered glycocalyx constituents imprint nanotopographical features on the plasma membrane with longitudinal dimensions of 200 – 300 nm. Analysis of the membrane topographies reveals curvatures that could be sufficient to elicit biological responses through curvature sensing proteins, such as BAR-domain proteins. Together, our simulations suggest how instabilities in the compressed glycocalyx could mediate downstream adhesive and signaling processes in the bleb-substrate interface.

Cells respond to inflammatory stimuli such as cytokines by activation of the nuclear factor-κB (NF-κB) signalling pathway, resulting in oscillatory translocation of the transcription factor p65 between nucleus and cytoplasm in some cell types. We investigate the relationship between p65 and inhibitor-κB⍺ (IκBα) protein levels and dynamic properties of the system, and how this interaction impacts on the expression of key inflammatory genes. Using bacterial artificial chromosomes, we developed new cell models of IκB⍺-eGFP protein overexpression in a pseudo-native genomic context. We find that cells with high levels of the negative regulator IκBα remain responsive to inflammatory stimuli and maintain dynamics for both p65 and IκBα. In contrast, canonical target gene expression is dramatically reduced by overexpression of IκBα, but can be partially rescued by overexpression of p65. Treatment with leptomycin B to promote nuclear accumulation of IκB⍺ also suppresses canonical target gene expression, suggesting a mechanism in which nuclear IκB⍺ accumulation prevents productive p65 interaction with promoter binding sites. This causes reduced target promoter binding and gene transcription, which we validate by chromatin immunoprecipitation and in primary cells. Overall, we show how inflammatory gene transcription is modulated by the expression levels of both IκB⍺ and p65. This results in an anti-inflammatory effect on transcription, demonstrating a broad mechanism to modulate the strength of inflammatory response.

This is a single pdf file that contains: (1) supplementary methods; (2) supplementary table 1 (microarray glycans), supplementary table 2 (ADCs), and supplementary table 3 (primers); (3) supplementary figure 1 (ch735 Ab expression/purification), supplementary figure 2 (mo735 mAb ELISA), supplementary figure 3 (mo735 glycan microarray profiling), supplementary figure 4 (binding affinity data), supplementary figure 5 (histograms of Ab-stained cell lines), supplementary figure 6 (cell surface binding of cancer cell lines), supplementary figure 7 (CRISPR-Cas knockout cell lines), supplementary figure 8 (internalization of mo735), supplementary figure 9 (colocalization of mo735), and supplementary figure 10 (synthesis of ADC).

The specific targeting of differentially expressed glycans in malignant cells has emerged as an attractive anticancer strategy. One such target is the oncodevelopmental antigen polysialic acid (polySia), a polymer of α2,8-linked sialic acid residues that is largely absent during postnatal development but is re-expressed during progression of several malignant human tumors, including small-cell and non–small cell lung carcinomas, glioma, neuroblastoma, and pancreatic carcinoma. In these cancers, expression of polySia correlates with tumor progression and poor prognosis and appears to modulate cancer cell adhesion, invasiveness, and metastasis. To evaluate the potential of PolySia as a target for anticancer therapy, we developed a chimeric human polySia-specific mAb that retained low nanomolar (nmol/L) target affinity and exhibited exquisite selectivity for polySia structures. The engineered chimeric mAb recognized several polySia-positive tumor cell lines in vitro and induced rapid endocytosis of polySia antigens. To determine whether this internalization could be exploited for delivery of conjugated cytotoxic drugs, we generated an antibody–drug conjugate (ADC) by covalently linking the chimeric human mAb to the tubulin-binding maytansinoid DM1 using a bioorthogonal chemical reaction scheme. The resulting polySia-directed ADC demonstrated potent target-dependent cytotoxicity against polySia-positive tumor cells in vitro. Collectively, these results establish polySia as a valid cell-surface, cancer-specific target for glycan-directed ADC and contribute to a growing body of evidence that the tumor glycocalyx is a promising target for synthetic immunotherapies.Significance:These findings describe a glycan-specific antibody-drug conjugate that establishes polySia as a viable cell surface target within the tumor glycocalyx.

Supplemental Methods, References, and Figure Legends

Vinculin is localized to cell-ECM borders in invasive breast cancer and adhesome proteins are upregulated across human cancers.

Scanning angle interference microscopy - validation experiments.

MECs remodel 3D collagen gels for cell invasion. Individual collagen fibers are stiff and >500nm in diameter.

Vinculin nanoscale localization reflects altered molecular activity.

List of Adhesome, Oncogene and Control Genes used in Bioinformatics Analysis.

ECM cues, malignant transformation, and integrin signaling, promote cell spreading and invasion.

List of Cancer Tissue with Unbiased Matched Normal Tissue used in Bioinformatics Analysis.

Cells respond to inflammatory stimuli such as cytokines by activation of the nuclear factor-κB (NF-κB) signalling pathway, resulting in oscillatory translocation of the transcription factor p65 between nucleus and cytoplasm to mediate immune response. We investigate the relationship between p65 and inhibitor-κBα (IκBα) protein levels and dynamic properties of the system, and how this interaction impacts on the expression of key inflammatory genes. Using bacterial artificial chromosomes, we developed new cell models of IκBα-eGFP protein overexpression in a native genomic context. We find that cells with high levels of the negative regulator IκBα remain responsive to inflammatory stimuli and maintain dynamics for both p65 and IκBα. In contrast, canonical target gene expression is dramatically reduced by overexpression of IκBα, but can be partially rescued by overexpression of p65. Treatment with leptomycin B to promote nuclear accumulation of IκBα also suppresses canonical target gene expression, suggesting a mechanism in which nuclear IκBα accumulation prevents productive p65 interaction with promoter binding sites. This causes reduced target promoter binding and gene transcription, which we validate by chromatin immune precipitation and in primary cells. Overall, we show how inflammatory gene transcription is modulated by the expression levels of both IκBα and p65, and that transcription can be partially decoupled from p65 protein dynamics. This results in an anti-inflammatory effect on transcription, demonstrating a broad mechanism to modulate the strength of inflammatory response.

Aberrant DNA methylation is an important mechanism by which the normal patterns of microRNA expression are disrupted in human cancers including B-cell precursor acute lymphoblastic leukemia (BCP ALL), the most common pediatric malignancy.To characterize the methylation profile landscape of microRNA genes in BCP ALL patients.We employed Infinium® MethylationEPIC BeadChip Arrays to measure the methylation of microRNA genes from bone marrow samples of children with BCP ALL (n = 38) and controls without neoplasms (n = 4).This analysis revealed differential methylation of the microRNA genes in the pediatric BCP ALL when compared to the control. A subcluster amongst BCP ALL patients with TCF3-PBX1 genetic subtype was also observed. No other differences were observed in association with age, gender or risk group. Several interesting leukemia-related phenotypes are enriched by the genes with hyperand hypomethylated sites located in promoters as well as gene bodies. The top 3 miRNA genes, promoters of which were the most statistically significantly hypermethylated in BCP ALL were MIR1273G, MIR1304 and MIR663, and the top 3 hypomethylated were MIR4442, MIR155 and MIR3909.In this study, a different microRNA genes methylation landscape was shown in pediatric BCP ALL compared to children without neoplasms. A visible subcluster among BCP ALL samples consisted of individuals with TCF3-PBX1 genetic subtype. No other differences were observed in association with age, gender or risk group. Several interesting leukemia-connected phenotypes were found, associated with genes with hyperand hypomethylated sites located on promoters as well as gene bodies.

Transcription of almost all mammalian genes occurs in stochastic bursts, however the fundamental control mechanisms that allow appropriate single-cell responses remain unresolved. Here we utilise single cell genomics data and stochastic models of transcription to perform global analysis of the toll-like receptor (TLR)-induced gene expression variability. Based on analysis of more than 2000 TLR-response genes across multiple experimental conditions we demonstrate that the single-cell, gene-by-gene expression variability can be empirically described by a linear function of the population mean. We show that response heterogeneity of individual genes can be characterised by the slope of the mean-variance line, which captures how cells respond to stimulus and provides insight into evolutionary differences between species. We further demonstrate that linear relationships theoretically determine the underlying transcriptional bursting kinetics, revealing different regulatory modes of TLR response heterogeneity. Stochastic modelling of temporal scRNA-seq count distributions demonstrates that increased response variability is associated with larger and more frequent transcriptional bursts, which emerge via increased complexity of transcriptional regulatory networks between genes and different species. Overall, we provide a methodology relying on inference of empirical mean-variance relationships from single cell data and new insights into control of innate immune response variability.

Autistic individuals and individuals with a range of other neurodevelopmental conditions (NDD) often present with lower levels of daily living skills (DLS) when compared to their neurotypical peers. Importantly, lower levels of DLS have been linked to a range of negative outcomes, including lower rates of post-secondary education, lower employment rates, and higher daily support needs across autism and NDD. However, there are currently no open-source informant-reported instruments for capturing key aspects of DLS. This study describes the development, refinement, and initial psychometric evaluation of a new, relatively brief (53-item). Daily Living Skills Scale (DLSS) in a sample of 1,361 children aged 2–17 years, Confirmatory Factor Analysis demonstrated an excellent fit of unidimensional model to the data (CFI = 0.953, TLI = 0.951, RMSEA = 0.073 [95% CI: 0.071–0.074]). The single-factor CFA model showed evidence of measurement invariance of factor loadings, thresholds, and residual variance (strict invariance) across sex, age, race, and ethnicity. Model reliability and internal consistency were excellent (ω = 0.98; α = 0.97). Conditional reliability estimates indicated very good reliability (= 0.80) for the total DLS scale from very low (θ = −4.2) to high (θ = +2.4) scores. Conceptually derived self-care, homecare, and community participation subscales also showed strong reliability and internal consistency. With further replication, the EFS has excellent potential for wide adoption across research and clinical contexts.

Lamins A/C are nuclear intermediate filament proteins that are involved in diverse cellular mechanical and biochemical functions. Here, we report that recognition of Lamins A/C by a commonly used antibody (JOL-2) that binds the Lamin A/C Ig-fold and other antibodies targeting similar epitopes is highly dependent on cell density, even though Lamin A/C protein levels do not change with cell density. The density-dependent Lamin A/C labeling was distinct from previously reported differential apico-basal labeling, which was independent of cell density. Comparison of the density-dependent labeling effects of antibodies recognizing different Lamin A/C epitopes suggests that the effect is caused by partial unfolding or masking of the C’E and/or EF loops of the Ig-fold in response to cell spreading. Seeding cells on micropatterned surfaces with different areas confirmed that increased cell spreading resulted in reduced Lamin A/C labeling with the JOL-2 antibody. Surprisingly, JOL-2 antibody labeling was insensitive to depolymerization of cytoskeletal filaments or disruption of the Linker of Nucleoskeleton and Cytoskeleton (LINC) complex. Although the density-dependent changes of the Ig-fold did not alter nuclear stiffness or nucleo-cytoskeletal force transmission, they may nonetheless modulate interaction with lamin binding partners and thereby affect cellular functions. Taken together, our results point to a previously unrecognized change in the Lamin A/C Ig-fold that affects recognition by the JOL-2 antibody. These findings are not only important for the interpretation of immunofluorescence data for Lamin A/C, but also raise the intriguing prospect that the conformational changes may play a role in Lamin A/C mediated cellular function.

Artykuł skupia się na kwestiach dotyczących mediów (szczególnie nowych, lecz nawiązuje także do tradycyjnych) w obliczu pandemii koronawirusa – zwłaszcza roli, jaką przyjęły, oraz nowych nawyków, które wykształciły się wśród ich odbiorców. Najogólniej rzecz biorąc, praca ma na celu ukazanie najważniejszych zagadnień związanych z mediami i komunikacją w tym specyficznym czasie – zarówno z perspektywy nadawców, jak i odbiorców. W pracy przedstawiono również refleksję etyczną nad mediami w kontekście pandemii COVID-19. Artykuł ma charakter przeglądowy i nakreśla wybrane problemy. Został opracowany na podstawie opublikowanych dotychczas raportów, analiz oraz opracowań naukowych w obszarze poruszanej tematyki.

Po II wojnie światowej, wznowiono „Wojskowy Przegląd Prawniczy” – czasopismo prawnicze znane i cenione przed 1939 r. Reaktywowany kwartalnik od 1945 r. był jednym z wielu narzędzi władzy komunistycznej do ideologicznej walki. Czasopismo propagowało nowe rewolucyjne podejście do teorii prawa i praktyki orzeczniczej. Treści prezentowane na łamach „WPP” były skierowane nie tylko dla prawników wojskowych, ale również dla dowódców wojska i przedstawicieli komunistycznego aparatu politycznego, wpływając na właściwy światopogląd czytelników zgodny z linią partii.

Morphological transitions are typically attributed to the actions of proteins and lipids. Largely overlooked in membrane shape regulation is the glycocalyx, a pericellular membrane coat that resides on all cells in the human body. Comprised of complex sugar polymers known as glycans as well as glycosylated lipids and proteins, the glycocalyx is ideally positioned to impart forces on the plasma membrane. Large, unstructured polysaccharides and glycoproteins in the glycocalyx can generate crowding pressures strong enough to induce membrane curvature. Stress may also originate from glycan chains that convey curvature preference on asymmetrically distributed lipids, which are exploited by binding factors and infectious agents to induce morphological changes. Through such forces, the glycocalyx can have profound effects on the biogenesis of functional cell surface structures as well as the secretion of extracellular vesicles. In this review, we discuss recent evidence and examples of these mechanisms in normal health and disease.

Objective Although theoretical conceptualizations of suicide hold that passive and active suicidal ideation are etiologically distinct, existing research observing this distinction is modest, with most prior studies focusing exclusively on active ideation. Understanding processes associated with passive ideation is clinically important insofar as passive ideation may precede active ideation, and thus serve as an earlier intervention target prior to potential onset of suicidal behavior. We aimed to evaluate intrapersonal and interpersonal vulnerability and resilience factors for passive ideation and differentiating passive from active ideation. Method Left-behind adolescents in rural China (n = 371) were assessed for passive and active ideation, depressive symptoms, rumination, grit, peer support, and peer victimization. Results Overall, 15.9% of the sample endorsed passive ideation without active ideation, and 17.8% endorsed active ideation. In multivariate analyses, rumination and grit differentiated left-behind children with passive ideation from those with no ideation. Depressive symptom severity predicted active ideation among adolescents with passive ideation. Conclusions The current findings suggest that rumination and grit may characterize passive ideation. Although passive and active ideation may differ modestly in vulnerability and resilience factors, depressive symptoms may be important to monitor among those with passive ideation and have not yet experience active ideation.

The intensive development of sport shooting and the significant increase in the number of people using sport and recreational shooting ranges also increases the risk of hearing damage to shooters, due to exposure to impulse noise from the shots of firearms. This article discusses the methodology for assessing shooters’ exposure to noise and identifies and extensively discusses possible ways to minimise this exposure. It then describes a research experiment carried out to measure and calculate the effectiveness of noise reduction by means of acoustic silencers when shooting with typical weapons in the most popular sporting and hunting calibres. The experiment also included firing tests with subsonic ammunition. On this basis, conclusions were drawn regarding the possibility of reducing impulse noise during shooting and the possibility of eliminating the risk of hearing damage.

The development of high-performance organic electrodes for potassium-ion batteries (KIBs) is attracting interest due to their sustainability and low costs. However, the electrolyte systems and moieties that generally proved to be successful in high-performance Li-ion batteries have found relatively little success in KIBs. Herein, two alkynyl-based covalent organic frameworks (COFs) containing 1,3,5-tris(arylethynyl)benzene (TAEB) and dehydrobenzoannulene (DBA) units are utilized as bulk anode materials for KIBs in a localized high-concentration electrolyte. TAEB-COF provides a high capacity value of 254.0 mAh g-1 at ∼100% efficiency after 300 cycles, and DBA-COF 3 provides a capacity of 76.3 mAh g-1 with 98.7% efficiency after 300 cycles. DFT calculations suggest that the alkynyl units of TAEB-COF facilitate the binding of K-ions through both enthalpic and geometric driving forces, leading to high reversible capacities.

Coronary artery disease (CAD) is associated with a prothrombotic tendency including increased factor (F) VIIa-antithrombin (FVIIa-AT) complexes, a measure of tissue factor (TF) exposure, and activated FXI (FXIa). We investigated whether increased FVIIa-AT complexes are associated with FXIa and active TF and if major adverse clinical outcomes are predicted by the complexes in CAD.In 120 CAD patients, we assessed FVIIa-AT complex concentrations and the presence of circulating FXIa and active TF. Levels of 8-iso-prostaglandin F2α (8-iso-PGF2α), interleukin-6, high-sensitivity C reactive protein, prothrombin fragment 1 + 2, and free Tissue Factor Pathway Inhibitor were determined. Myocardial infarction (MI), ischemic stroke, systemic thromboembolism (SE), and cardiovascular (CV) death were recorded separately and as a composite endpoint, during follow-up.FVIIa-AT complexes were positively associated with current smoking and multivessel CAD. Elevated FVIIa-AT complexes characterized patients with circulating FXIa and/or active TF in association with increased plasma isoprostanes but not with thrombin generation or inflammatory markers. During a median follow-up of 106 months (interquartile range 95-119), high baseline levels of FVIIa-AT complexes predicted ischemic stroke/SE (HR 4.61 [95% CI 1.48-18.42]) and a composite endpoint of MI, stroke/SE, and CV death (HR 7.47 [95% CI 2.81-19.87]).This study is the first to show that high FVIIa-AT complexes characterize advanced CAD patients with detectable FXIa and active TF, which is, in part, driven by oxidative stress. High FVIIa-AT complexes were associated with the risk of ischemic stroke/SE during long-term follow-up, highlighting the need for effective antithrombotic agents in CAD.

The cancer cell glycocalyx serves as a major line of defense against immune surveillance. However, how specific physical properties of the glycocalyx are regulated and contribute to immune evasion is not well understood. Here, we uncover how the surface density, glycosylation, and crosslinking of cancer-associated mucins contribute to the nanoscale material thickness of the glycocalyx, and further analyze the effect of the glycocalyx thickness on resistance to effector cell attack. Natural Killer (NK) cell-mediated cytotoxicity exhibits a near perfect inverse correlation with the glycocalyx thickness of target cells regardless of the specific glycan structures present, with changes in the glycocalyx thickness of approximately 10 nanometers significantly altering susceptibility to NK attack. Equipping the NK cell surface with a mucin-digesting enzyme can improve killing with a performance enhancement that can rival or exceed chimeric antigen receptors (CARs). Together, our results motivate the development of glycocalyx-editing immune cells for cancer immunotherapy.

There is a discrepancy between epicardial vessel patency and microcirculation perfusion in a third of patients treated with percutaneous coronary intervention (PCI) for ST-segment elevation myocardial infarction (STEMI). Optimization with aspiration thrombectomy (AT) may reduce distal embolization and microvascular obstruction. The effect of AT in the treatment of STEMI is debatable. The purpose of this study was to use cardiac magnetic resonance (CMR) to determine whether AT influences microvascular obstruction (MVO), infarct size and left ventricular (LV) remodelling in STEMI patients. Sixty STEMI patients with a thrombus-occluded coronary artery were randomized in a 2:1 fashion to receive PCI proceeded by AT (AT + PCI group), or PCI only. MVO, myocardial infarct size and LV remodelling were assessed by CMR during the index hospitalization and 6 months thereafter. The majority of patients had a large thrombus burden (TIMI thrombus grade 5 in over 70% of patients). PCI and AT were effective in all cases. There were no periprocedural strokes. CMR showed that the addition of AT to standard PCI was associated with lesser MVO when indexed to the infarct size and larger infarct size reduction. There were less patients with left ventricle remodelling in the AT + PCI vs. the PCI only group. To conclude, in STEMI patients with a high thrombus burden, AT added to PCI is effective in reducing infarct size, MVO and LV remodelling.

Bone metastasis is a leading cause of breast cancer-related deaths and often initiated by tumor cell dissemination to osteogenic niches. During new bone formation, osteoblasts first deposit osteoid, the collagen I-rich, unmineralized component of bone ECM, within which carbonated hydroxyapatite nanoparticles subsequently form. However, it remains elusive how bone matrix mineralization dictates tumor cell phenotype due in part to the lack of relevant model systems. Using biofunctional, collagen I-based bone matrix models with physiological, intrafibrillar mineralization, we show that mineralization inhibits proliferation, while inducing a stem-like phenotype in tumor cells. These changes were due to reduced mechanosignaling contradicting the conventional assumption that increased rigidity caused by mineralization stimulates metastatic progression. Our findings are translationally relevant as the presence of mineral reduced tumor growthin vivoand upregulated a gene signature that correlated with decreased patient mortality. Our results could help explain why decreased bone mineral density increases the risk for bone metastasis in patients and highlight that bone metastasis models should integrate organic and inorganic matrix components in a manner that mimics physiological mineralization.

Metastasis is the leading cause of breast cancer-related deaths and often driven by invasion and cancer-stem like cells (CSCs). Both the CSC phenotype and invasion have been associated with increased hyaluronic acid (HA) production. How these independent observations are connected, and which role metabolism plays in this process remains unclear due in part to the lack of convergent approaches that integrate engineered model systems, computational tools, and cancer biology. Using microfluidic invasion models, metabolomics, computational flux balance analysis (FBA), and bioinformatic analysis of patient data we investigated the functional links between the stem-like, invasive, and metabolic phenotype of breast cancer cells as a function of HA biosynthesis. Our results suggest that CSCs are more invasive than non-CSCs and that broad metabolic changes caused by overproduction of HA play a role in this process. Accordingly, overexpression of hyaluronic acid synthases (HAS) 2 or 3 induced a metabolic phenotype that promoted breast cancer cell stemness and invasion in vitro and upregulated a transcriptomic signature that was predictive of increased invasion and worse survival in patients. Collectively, this study suggests that HA overproduction leads to metabolic adaptations that help satisfy the energy demands necessary for 3D invasion of breast cancer stem cells further highlighting the importance of engineered model systems and multidisciplinary approaches in cancer research.

Pathogens have developed intricate strategies to overcome the host’s innate immune responses. In this paper we use live-cell microscopy with a single bacterium resolution to follow in real time interactions between the food-borne pathogen L. monocytogenes and host macrophages, a key event controlling the infection in vivo. We demonstrate that infection results in heterogeneous outcomes, with only a subset of bacteria able to establish a replicative invasion of macrophages. The fate of individual bacteria in the same host cell was independent from each other and non-cooperative, but a higher multiplicity of infection resulted in a reduced probability of replication. Using internalisation assays and conditional probabilities to mathematically describe the multi-stage invasion process, we demonstrate that the secreted Listeriolysin toxin (LLO) of the PrfA regulon regulates replication probability by compromising the ability to phagocytose bacteria. Using strains expressing fluorescent reporters to follow transcription of either the LLO-encoding hly or actA genes, we show that replicative bacteria exhibited higher PrfA regulon expression in comparison to those bacteria that did not replicate, however elevated PrfA expression per se was not sufficient to increase the probability of replication. Overall, this demonstrates a new role for the population-level, but not single cell PrfA-mediated cooperativity to regulate outcomes of host pathogen interactions.Key pointsL. monocytogenes invasion of innate immune macrophages results in heterogeneous infection outcomes at the single cell levelFate of individual bacteria in the same host cell is independent from each other and non-cooperativeBacterial populations coordinate host cell uptake via the rate of phagocytosis to reduce internalization at high MOIThe PrfA regulon system is necessary but not sufficient for L. monocytogenes replication, but population-level PrfA virulence regulates single cell outcome probability

Background: The beneficial metabolic effects of the PPARγ agonist rosiglitazone (Rosi) are associated with anti-inflammatory changes in adipose tissue macrophages (ATMs) . One study aim was to characterize the in vivo Rosi-induced phenotypic switch of ATMs using a variety of macrophage markers, including the lipid-associated macrophage markers CD9 and Trem2. As a second aim, since ATM-derived exosomes can exert insulin-sensitizing or desensitizing effects dependent on ATM cell context, we assessed the effects of ATM exosomes from Rosi-treated obese mice on in vitro insulin actions. Research Design and Method: ATMs were isolated from 4 months HFD/obese mice treated for the last month with Rosi (60 mg/kg HFD) compared to HFD or chow-fed controls. Among CD11b+F4/80+ or CD64+ macrophages, we identified pro-inflammatory (“M1-like” CD11c+ and/or CD9+) , anti-inflammatory (“M2-like” CD11c-CD206+) and lipid-associated (CD9+ and/or Trem2+) ATMs by flow cytometry. Bone marrow-derived macrophages (BMDMs) were treated in vitro with µM Rosi for 24h. 3T3-L1 adipocytes, L6 myotubes, and primary hepatocytes were cultured 24h with exosomes isolated from ATMs of Rosi treated obese mice or in vitro Rosi treated BMDMs and then assessed for insulin sensitivity. Results: Rosi treatment reduced adipose tissue inflammation, as seen by reduced numbers of pro-inflammatory (“M1-like”) ATMs and increased the Trem2 signature in “M2-like” ATMs. Importantly, Rosi directly induced Trem2 expression in vitro in BMDMs. Additionally, exosomes isolated from Rosi-treated obese mice or M1 BMDMs increased glucose uptake and insulin signalling (pAKT) in adipocytes and myotubes and reduced glucose production in hepatocytes compared to control exosomes. Conclusion: Our data indicate that Trem2 is a PPARγ target gene. Since exosomes from Rosi treated ATMs or BMDMs enhance cellular insulin action, this could provide a new explanation for the well-known insulin-sensitizing effects of PPARγ agonists. Disclosure T.V.Rohm: None. F.C.G.Reis: None. R.Isaac: None. M.Paszek: None. G.Bandyopadhyay: None. J.M.Olefsky: None. Funding Swiss National Science Foundation (SNF) Early Postdoc.Mobility grant (to T.R.) .

Aim: The inadequate system of triage in Emergency Departments leads to the situation in which the personnel is not always able to provide care on time. A regulation determining the conditions of performing triage in Emergency Departments (paragraph 1, section 2) was issued on 27 June 2019. In order to present the level of the fulfilment of rules from the regulation and to determine the level of organization in the Polish health care, the percentage of Emergency Departments which provided the required information on their websites, was examined. Material and methods: The study was conducted in April 2020. 236 websites of hospitals with Emergency Departments were analyzed. The criteria of data retrieval were as follows: 1. The presence of the information concerning the rules of redirecting patients to Primary Health Care/After Hours Medical Centers. 2. The presence of information concerning the time an Emergency Department doctor has to inform the patient about the admission or about refusal to admit a patient. The collected data were statistically analyzed. Each Emergency Department was given the following statuses: “yes”, “no”, or “incomplete”. Results: Information concerning the 1st criterion were available in case of 59 hospitals (25%), while no such information was on the website in Opole and West Pomeranian Provinces. Information concerning the 2nd criterion was placed by 51 hospitals. In 28 cases the information was described as ”complete” and in 23 as “incomplete”. Conclusions: The majority of hospitals failed to observe the obligation to provide the information included in the Regulation.

Immune cells fine tune their responses to infection and inflammatory cues. Here, using live-cell confocal microscopy and mathematical modelling, we investigate interferon induced JAK-STAT signalling in innate immune macrophages. We demonstrate that transient exposure to IFN-γ stimulation induces a long-term desensitisation of STAT1 signalling and gene expression responses, revealing a dose- and time-dependent regulatory feedback that controls JAK-STAT responses upon re-exposure to stimulus. We show that IFN-α/β1 elicit different level of desensitisation from IFN-γ, where cells refractory to IFN-α/β1 are sensitive to IFN-γ, but notvice versa. We experimentally demonstrate that the underlying feedback mechanism involves regulation of STAT1 phosphorylation but is independent of new mRNA synthesis and cognate receptor expression. A new feedback model of the protein tyrosine phosphatase activity recapitulates experimental data and demonstrates JAK-STAT network’s ability to decode relative changes of dose, timing, and type of temporal interferon stimulation. These findings reveal that STAT desensitisation renders cells with signalling memory of type I and II interferon stimulation, which in the future may improve administration of interferon therapy.

The aim of the article is to demonstrate the relationship between the work of Józef Hołard and literature that inspired his work. A valuable document explaining this issue is found in the unpublished text of the artist entitled Credo, written in connection with the conduct of proceedings at the university. Associated with Bielsko-Biała, Professor Józef Hołard died in January 2015, but his painting, drawing and design works have not been sufficiently explored. They contain many symbolic elements derived from the esoteric and occult sciences. However, they are used and understood very freely by the artist, who gives priority to artistic creation. For many years he drew inspiration from literature dealing with similar subjects, especially Foucault’s Pendulum by Umberto Eco. The series of drawings and paintings According to Umberto Eco is best known in Hołard’s oeuvre.

Precise pH measurements in the immediate environment of receptors is essential for elucidating the mechanisms through which local pH changes associated with diseased phenotypes manifest into aberrant receptor function. However, current pH sensors lack the ability to localize and target specific receptor molecules required to make these measurements. Herein we present the Litmus-body, our recombinant protein-based pH sensor, which through fusion to an anti-IgG nanobody is capable of piggybacking on IgG antibodies for molecular targeting to specific proteins on the cell surface. By normalizing a pH-dependent green fluorescent protein to a long Stokes shift red fluorophore or fluorescent protein, we readily report pH independent of sensor concentration using a single 488 nm excitation. Our Litmus-body showed excellent responsiveness in solution, with a greater than 50-fold change across the regime of physiological pH. The sensor was further validated for use on live cells and shown to be specific to the protein of interest. In complex with our Litmus-body, cetuximab therapeutic antibody retained its functionality in binding and inhibiting ligand interaction of its target epidermal growth factor receptor (EGFR), triggering receptor-mediated endocytosis that allowed tracking of local pH from the cell surface through the endocytic pathway.

The article presents crucial aspects related to examinations of records from cockpit voice recorders (CVR) and the possible effects of the application of digital correction of these type of records. It also presents the resulting limitations concerning phonoscopic recognition examinations of speakers and determining the content of their utterances. A method for the real reduction of the described limitations was suggested. Part II of the article describes research experiments conducted at Air Force Institute of Technology.