Basement membrane product, endostatin, as a link between inflammation, coagulation and vascular permeability in COVID-19 and non-COVID-19 acute respiratory distress syndrome

Background: Immune cell recruitment, endothelial cell barrier disruption, and platelet activation are hallmarks of lung injuries caused by COVID-19 or other insults which can result in acute respiratory distress syndrome (ARDS). Basement membrane (BM) disruption is commonly observed in ARDS, however, the role of newly generated bioactive BM fragments is mostly unknown. Here, we investigate the role of endostatin, a fragment of the BM protein collagen XVIIIα1, on ARDS associated cellular functions such as neutrophil recruitment, endothelial cell barrier integrity, and platelet aggregation in vitro. Methods: In our study we analyzed endostatin in plasma and post-mortem lung specimens of patients with COVID-19 and non-COVID-19 ARDS. Functionally, we investigated the effect of endostatin on neutrophil activation and migration, platelet aggregation, and endothelial barrier function in vitro. Additionally, we performed correlation analysis for endostatin and other critical plasma parameters. Results: We observed increased plasma levels of endostatin in our COVID-19 and non-COVID-19 ARDS cohort. Immunohistochemical staining of ARDS lung sections depicted BM disruption, alongside immunoreactivity for endostatin in proximity to immune cells, endothelial cells, and fibrinous clots. Functionally, endostatin enhanced the activity of neutrophils, and platelets, and the thrombin-induced microvascular barrier disruption. Finally, we showed a positive correlation of endostatin with soluble disease markers VE-Cadherin, c-reactive protein (CRP), fibrinogen, and interleukin (IL)-6 in our COVID-19 cohort. Conclusion: The cumulative effects of endostatin on propagating neutrophil chemotaxis, platelet aggregation, and endothelial cell barrier disruption may suggest endostatin as a link between those cellular events in ARDS pathology

Basement membrane product, endostatin, as a link between inflammation, coagulation and vascular permeability in COVID-19 and non-COVID-19 acute respiratory distress syndrome

BackgroundImmune cell recruitment, endothelial cell barrier disruption, and platelet activation are hallmarks of lung injuries caused by COVID-19 or other insults which can result in acute respiratory distress syndrome (ARDS). Basement membrane (BM) disruption is commonly observed in ARDS, however, the role of newly generated bioactive BM fragments is mostly unknown. Here, we investigate the role of endostatin, a fragment of the BM protein collagen XVIIIα1, on ARDS associated cellular functions such as neutrophil recruitment, endothelial cell barrier integrity, and platelet aggregation in vitro.MethodsIn our study we analyzed endostatin in plasma and post-mortem lung specimens of patients with COVID-19 and non-COVID-19 ARDS. Functionally, we investigated the effect of endostatin on neutrophil activation and migration, platelet aggregation, and endothelial barrier function in vitro. Additionally, we performed correlation analysis for endostatin and other critical plasma parameters.ResultsWe observed increased plasma levels of endostatin in our COVID-19 and non-COVID-19 ARDS cohort. Immunohistochemical staining of ARDS lung sections depicted BM disruption, alongside immunoreactivity for endostatin in proximity to immune cells, endothelial cells, and fibrinous clots. Functionally, endostatin enhanced the activity of neutrophils, and platelets, and the thrombin-induced microvascular barrier disruption. Finally, we showed a positive correlation of endostatin with soluble disease markers VE-Cadherin, c-reactive protein (CRP), fibrinogen, and interleukin (IL)-6 in our COVID-19 cohort.ConclusionThe cumulative effects of endostatin on propagating neutrophil chemotaxis, platelet aggregation, and endothelial cell barrier disruption may suggest endostatin as a link between those cellular events in ARDS pathology

Role of connexin 43 in TGF-β₁-induced fibroblast to myofibroblast transition in bronchial fibroblasts derived from asthmatic patients

Obserwowana, w trakcie rozwoju astmy, przebudowa (remodeling) dróg oddechowych jest bezpośrednio związana z różnicowaniem fibroblastów w miofibroblasty (FMT). Ważnym mediatorem tego procesu jest TGF-β₁, który jako aktywator ścieżki sygnalizacyjnej białek Smad, stymuluje ekspresję genów wielu białek profibrotycznych m.in. α-SMA. Molekularne mechanizmy leżące u podstaw astmy nadal nie są dobrze znane. Celem badań niniejszej pracy, było ustalenie roli koneksyny 43 (Cx43) w FMT ludzkich fibroblastów oskrzelowych, pochodzących od osób ze zdiagnozowaną astmą (HBF AS). Z wykorzystaniem technik immunocytofluorescenscencyjnych przeprowadzono analizy intensywności FMT i translokacji p-Smad2 do jądra komórkowego, w fibroblastach stymulowanych TGF-β₁ w obecności inhibitora komunikacji międzykomórkowej przez złącza szczelinowe (GJIC) lub po wyciszeniu ekspresji Cx43. Analiza obejmowała również pomiar poziomu ekspresji α-SMA metodą Western Blot oraz α-SMA i Cx43 przy pomocy cytometrii przepływowo-obrazowej.Uzyskane wyniki wskazują na istnienie dodatniej korelacji między poziomem ekspresji Cx43 i α-SMA w HBF AS. Wykazano również, że inhibitor GJIC, obniża intensywność FMT i ekspresję α-SMA, jednocześnie nie wpływając na rozmiar kontaktów zogniskowanych i organizację cytoszkieletu aktynowego i nie hamując istotnie translokacji p-Smad2 do jądra. Wyciszenie w HBF AS ekspresji Cx43 obniża intensywność FMT i hamuje translokację p-Smad2 do jądra. Otrzymane wyniki pozwalają po raz pierwszy stwierdzić, że Cx43 jest ważnym białkiem regulującym intensywność FMT w fibroblastach oskrzelowych astmatyków, poprzez wpływ na przekaz sygnału od receptora TGF-β do jądra komórkowego (z udziałem białek Smad). Uzyskane rezultaty mogą przyczynić się do lepszego poznania molekularnych mechanizmów leżących u podłoża zmian chorobowych towarzyszących astmie.The fibroblast to myofibroblast transition (FMT) is directly related to airway remodeling observed during the development of asthma. An important mediator of this process is TGF-β₁, which stimulates gene expression of many profibrotic proteins like α-SMA, through the activation of the Smad signaling pathway. Molecular mechanisms underlying asthma onset and progression are still not well known.The aim of this work was to investigate the role of connexin 43 (Cx43) in the process of FMT in human bronchial fibroblasts derived from asthmatic patients (HBF AS). Fibroblasts with silenced Cx43 expression or in the presence of inhibitor of gap junctional intercellular communication (GJIC) were simulated with TGF-β₁. Subsequently, the intensity of FMT and phospho-Smad2 accumulation in the nucleus was evaluated. Expression levels of α-SMA were assessed by Western Blotting, and α-SMA and Cx43 levels were determined with the use of Imaging Flow Cytometry. The obtained results showed a positive correlation between the expression levels of Cx43 and α-SMA in HBF AS. It was also found that GJIC inhibitor reduced FMT activity and lowerd α-SMA expression. However, GJIC inhibitor did not effect either actin cytoskeleton organization or focal contacts size, and did not significantly inhibit phospho-Smad2 nucleus accumulation. HBFs with silenced Cx43 expression showed substantialy limited FMT intensity and reduction of translocation of phospho-Smad2 to the nucleus. Results presented in this paper demonstrate a novel mechanism for Cx43 in control of FMT intensity in asthmatic bronchial fibroblasts by positive regulation of TGF-β signaling mediated by Smad proteins. Thesis findings may contribute to a better understanding of the molecular mechanisms underlying the pathological changes associated with onset and development of asthma

Students of University of Warsaw in the era of the COVID-19 pandemic – legal criminological and social aspects.

Przedmiotem artykułu jest ocena  możliwego wpływu pandemii COVID-19 na wybrane sfery życia studentów Uniwersytetu Warszawskiego – ich zdrowie, edukację (która przeniosła się do formuły zdalnej), stosunek studentów do wprowadzanych na poziomie uniwersyteckim, jak i krajowym obostrzeń oraz ryzyko pokrzywdzenia przestępstwem i/lub przemocą.  Ocenę tę przeprowadzono w oparciu o wyniki badania ankietowanego, które zostało przeprowadzone wśród studentów UW w 2021 roku. W artykule przeanalizowano także rozwiązania normatywne wprowadzane przez władze UW w okresie pandemii COVID-19, z uwagi na fakt, że  ich przyjęcie w istotnym stopniu zmodyfikowało model życia i kształcenia społeczności studenckiej.   The subject of the article is to assess the possible impact of the COVID-19 pandemic on selected spheres of life of students of the University of Warsaw - their health, education (which has moved to the remote formula), students’ attitude to the restrictions introduced at the university and national level, and the risk of becoming a victim of crime and / or violence. This assessment was based on the results of a survey conducted among students of the University of Warsaw in 2021. The article also analyzes the normative solutions introduced by the University of Warsaw authorities during the COVID-19 pandemic, due to the fact that their adoption significantly modified the model of life and education of the student community

Impact of Pneumococcal Vaccination on Nasopharyngeal Carriage of Streptococcus pneumoniae and Microbiota Profiles in Preschool Children in South East Poland

In 2017, Poland introduced the 10-valent pneumococcal conjugate vaccine (PCV) into its national immunization schedule. This prospective study was conducted between March and June 2020 to determine the impact of vaccination on prevalence of the nasopharyngeal carriage of S. pneumoniae in 176 healthy children and to determine how conjugate vaccines indirectly affect colonization of nasopharyngeal microbiota. Pneumococcal isolates were analyzed by serotyping and antimicrobial resistance tests. Nasopharyngeal microbiota were detected and identified using the culture method and real-time PCR amplification primers and hydrolysis-probe detection with the 16S rRNA gene as the target. In the vaccinated group of children, colonization was in 24.2% of children, compared to 21.4% in the unvaccinated group. Serotypes 23A and 23B constituted 41.5% of the isolates. Serotypes belonging to PCV10 and PCV13 constituted 4.9% and 17.1% of the isolates, respectively. S. pneumoniae isolates were resistant to penicillin (34.1%), erythromycin (31.7%), and co-trimoxazole (26.8%). Microbial DNA qPCR array correlated to increased amounts of Streptococcus mitis and S. sanguinis in vaccinated children, with reduced amounts of C. pseudodiphtericum, S. aureus, and M. catarrhalis. Introduction of PCV for routine infant immunization was associated with significant reductions in nasopharyngeal carriage of PCV serotypes and resistant strains amongst vaccine serotypes, yet carriage of non-PCV serotypes increased modestly, particularly serotype 23B

BMP7 aberrantly induced in the psoriatic epidermis instructs inflammation-associated Langerhans cells.

BACKGROUND Epidermal hyperplasia represents a morphologic hallmark of psoriatic skin lesions. Langerhans cells (LCs) in the psoriatic epidermis engage with keratinocytes (KCs) in tight physical interactions; moreover, they induce T cell-mediated immune responses critical to psoriasis. OBJECTIVE Epidermal factors in psoriasis pathogenesis remain poorly understood. METHODS We phenotypically characterized BMP7-LCs vs. TGF-β1-LCs and analyzed their functional properties using flow cytometry, cell kinetic studies, co-culture with CD4 T-cells and cytokine measurements. Furthermore, immunohistology of healthy and psoriatic skin was performed. Additionally, in vivo experiments with Junf/fJunBf/fK5creER mice were carried out to assess the role of BMP signaling in psoriatic skin inflammation. RESULTS Here we identified a KC-derived signal, i.e. bone morphogenetic protein (BMP) signaling, to promote epidermal changes in psoriasis. Whereas BMP7 is strictly confined to the basal KC layer in the healthy skin, it is expressed at high levels throughout the lesional psoriatic epidermis. BMP7 instructs precursor cells to differentiate into LCs that phenotypically resemble psoriatic LCs. These BMP7-LCs exhibit proliferative activity and increased sensitivity to bacterial stimulation. Moreover, aberrant high BMP signaling in the lesional epidermis is mediated by a KC intrinsic mechanism, as suggested from murine data and clinical outcome after topical anti-psoriatic treatment in human patients. CONCLUSION Our data indicate that available TGF-β family members within the lesional psoriatic epidermis preferentially signal through the canonical BMP signaling cascade to instruct inflammatory-type LCs and to promote psoriatic epidermal changes. Targeting BMP signaling might allow to therapeutically interfere with cutaneous psoriatic manifestations

Psoriatic skin inflammation is promoted by c‐Jun/AP‐1‐dependent CCL2 and IL‐23 expression in dendritic cells

Abstract Toll‐like receptor (TLR) stimulation induces innate immune responses involved in many inflammatory disorders including psoriasis. Although activation of the AP‐1 transcription factor complex is common in TLR signaling, the specific involvement and induced targets remain poorly understood. Here, we investigated the role of c‐Jun/AP‐1 protein in skin inflammation following TLR7 activation using human psoriatic skin, dendritic cells (DC), and genetically engineered mouse models. We show that c‐Jun regulates CCL2 production in DCs leading to impaired recruitment of plasmacytoid DCs to inflamed skin after treatment with the TLR7/8 agonist Imiquimod. Furthermore, deletion of c‐Jun in DCs or chemical blockade of JNK/c‐Jun signaling ameliorates psoriasis‐like skin inflammation by reducing IL‐23 production in DCs. Importantly, the control of IL‐23 and CCL2 by c‐Jun is most pronounced in murine type‐2 DCs. CCL2 and IL‐23 expression co‐localize with c‐Jun in type‐2/inflammatory DCs in human psoriatic skin and JNK‐AP‐1 inhibition reduces the expression of these targets in TLR7/8‐stimulated human DCs. Therefore, c‐Jun/AP‐1 is a central driver of TLR7‐induced immune responses by DCs and JNK/c‐Jun a potential therapeutic target in psoriasis