Perfringolysin O-induced plasma membrane pores trigger actomyosin remodeling and endoplasmic reticulum redistribution

Clostridium perfringens produces an arsenal of toxins that act together to cause severe infections in humans and livestock animals. Perfringolysin O (PFO) is a cholesterol-dependent pore-forming toxin encoded in the chromosome of virtually all C. perfringens strains and acts in synergy with other toxins to determine the outcome of the infection. However, its individual contribution to the disease is poorly understood. Here, we intoxicated human epithelial and endothelial cells with purified PFO to evaluate the host cytoskeletal responses to PFO-induced damage. We found that, at sub-lytic concentrations, PFO induces a profound reorganization of the actomyosin cytoskeleton culminating into the assembly of well-defined cortical actomyosin structures at sites of plasma membrane (PM) remodeling. The assembly of such structures occurs concomitantly with the loss of the PM integrity and requires pore-formation, calcium influx, and myosin II activity. The recovery from the PM damage occurs simultaneously with the disassembly of cortical structures. PFO also targets the endoplasmic reticulum (ER) by inducing its disruption and vacuolation. ER-enriched vacuoles were detected at the cell cortex within the PFO-induced actomyosin structures. These cellular events suggest the targeting of the endothelium integrity at early stages of C. perfringens infection, in which secreted PFO is at sub-lytic concentrations.This work was financed by FEDER—Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020—Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020, and by Portuguese funds through FCT - Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior in the framework of the project POCI-01-0145-FEDER-030863 (PTDC/BIA-CEL/30863/2017) and Norte-01-0145-FEDER-000012—Structured program on bioengineered therapies for infectious diseases and tissue regeneration, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (FEDER). Publication Fees were supported by ICBAS, University of Porto. CB and FSM were supported by FCT fellowships (SFRH/BD/112217/2015, SFRH/BPD/94458/2013, respectively). CB was a Fulbright and FLAD fellow. SS received support from FCT Investigator program (COMPETE, POPH, and FCT)

On the origin and pathway of the saline inflow to the Nordic Seas: insights from models

The behaviours of three high-resolution ocean circulation models of the North Atlantic, differing chiefly in their description of the vertical coordinate, are investigated in order to elucidate the routes and mechanisms by which saline water masses of southern origin provide inflows to the Nordic Seas. An existing hypothesis is that Mediterranean Overflow Water (MOW) is carried polewards in an eastern boundary undercurrent, and provides a deep source for these inflows. This study, however, provides an alternative view that the inflows are derived from shallow sources, and are comprised of water masses of western origin, carried by branches of the North Atlantic Current (NAC), and also more saline Eastern North Atlantic Water (ENAW), transported northwards from the Bay of Biscay region via a ‘Shelf Edge Current’ (SEC) flowing around the continental margins. In two of the models, the MOW flows northwards, but reaches only as far as the Porcupine Bank (53°N). In third model, the MOW also invades the Rockall Trough (extending to 60°N). However, none of the models allows the MOW to flow northwards into the Nordic Seas. Instead, they all support the hypothesis of there being shallow pathways, and that the saline inflows to the Nordic Seas result from NAC-derived and ENAW water masses, which meet and partially mix in the Rockall Trough. Volume and salinity transports into the southern Rockall Trough via the SEC are, in the various models, between 25 and 100% of those imported by the NAC, and are also a similarly significant proportion (20–75%) of the transports into the Nordic Seas. Moreover, the highest salinities are carried northwards by the SEC (these being between 0.13 and 0.19 psu more saline at the southern entrance to the Trough than those in the NAC-derived waters). This reveals for the first time the importance of the SEC in carrying saline water masses through the RockallTrough and into the Nordic Seas. Furthermore, the high salinities found on density surfaces appropriate to the MOW in the Nordic Seas are shown to result from the wintertime mixing of the saline near-surface waters advected northwards by the SEC/NAC system. Throughout, we have attempted to demonstrate the extent to which the models agree or disagree with interpretations derived from observations, so that the study also contributes to an ongoing community effort to assess the realism of our current generation of ocean models

Autoantibodies to aS1-Casein Are Induced by Breast-Feeding

Background: The generation of antibodies is impaired in newborns due to an immature immune system and reduced exposure to pathogens due to maternally derived antibodies and placental functions. During nursing, the immune system of newborns is challenged with multiple milk-derived proteins. Amongst them, caseins are the main constituent. In particular, human aS1-casein (CSN1S1) was recently shown to possess immunomodulatory properties. We were thus interested to determine if auto-antibodies to CSN1S1 are induced by breast-feeding and may be sustained into adulthood. Methods: 62 sera of healthy adult individuals who were (n = 37) or were not (n = 25) breast-fed against human CSN1S1 were investigated by a new SD (surface display)-ELISA. For cross-checking, these sera were tested for anti Epstein-Barr virus (EBV) antibodies by a commercial ELISA. Results: IgG-antibodies were predominantly detected in individuals who had been nursed. At a cut-off value of 0.4, the SDELISA identified individuals with a history of having been breast-fed with a sensitivity of 80% and a specificity of 92%. Under these conditions, 35 out of 37 sera from healthy donors, who where breast-fed, reacted positively but only 5 sera of the 25 donors who were not breast-fed. The duration of breast-feeding was of no consequence to the antibody reaction as some healthy donors were only short term breast-fed (5 days minimum until 6 weeks maximum), but exhibited significant serum reaction against human CSN1S1 nonetheless. Conclusion: We postulate that human CSN1S1 is an autoantigen. The antigenicity is orally determined, caused by breastfeeding, and sustained into adulthood

Worms:Pernicious parasites or allies against allergies?

Host-parasite interactio

VEGF induces signalling and angiogenesis by directing VEGFR2 internalisation via macropinocytosis

Endocytosis plays critical role in receptor signalling. VEGFR2 and its ligand VEGFA are fundamental in neovascularization. Yet, our understanding of the role of endocytosis in VEGFR2 signalling remains limited. Despite the existence of diverse internalisation routes, the only known endocytic pathway of VEGFR2 is the clathrin-mediated. Here, we show that this pathway is the predominant internalisation route of VEGFR2 only in the absence of ligand. Intriguingly, VEGF introduces a novel internalisation itinerary for VEGFR2, the pathway of macropinocytosis, which becomes the prevalent endocytic route of the receptor in the presence of ligand, while the route of clathrin becomes minor. Macropinocytic internalisation of VEGFR2, which mechanistically is mediated via the small GTPase CDC42, takes place via macropinosomes generated at ruffling areas of the membrane. Interestingly, macropinocytosis plays critical role in VEGF-induced signalling, endothelial cell functions in vitro and angiogenesis in vivo, while clathrin-mediated endocytosis is not essential for VEGF signalling. These findings expand our knowledge on the endocytic pathways of VEGFR2 and suggest that VEGF-driven internalisation of VEGFR2 via macropinocytosis is essential for endothelial cell signalling and angiogenesis

Інженерна комп’ютерна графіка

Розглянуто відомості про систему тривимірного моделювання КОМПАС-3D при виконанні практичних завдань, побудову тривимірних моделей деталей і складальних одиниць машин та обладнання, будівельних споруд, а також про випуск асоціативних креслеників, розробку специфікацій, експлікацій, інших текстових документів

Prolonged high-dose intravenous magnesium therapy for severe tetanus in the intensive care unit: a case series

<p>Abstract</p> <p>Introduction</p> <p>Tetanus rarely occurs in developed countries, but it can result in fatal complications including respiratory failure due to generalized muscle spasms. Magnesium infusion has been used to treat spasticity in tetanus, and its effectiveness is supported by several case reports and a recent randomized controlled trial.</p> <p>Case presentations</p> <p>Three Caucasian Greek men aged 30, 50 and 77 years old were diagnosed with tetanus and admitted to a general 12-bed intensive care unit in 2006 and 2007 for respiratory failure due to generalized spasticity. Intensive care unit treatment included antibiotics, hydration, enteral nutrition, early tracheostomy and mechanical ventilation. Intravenous magnesium therapy controlled spasticity without the need for additional muscle relaxants. Their medications were continued for up to 26 days, and adjusted as needed to control spasticity. Plasma magnesium levels, which were measured twice a day, remained in the 3 to 4.5 mmol/L range. We did not observe hemodynamic instability, arrhythmias or other complications related to magnesium therapy in these patients. All patients improved, came off mechanical ventilation, and were discharged from the intensive care unit in a stable condition.</p> <p>Conclusion</p> <p>In comparison with previous reports, our case series contributes the following meaningful additional information: intravenous magnesium therapy was used on patients already requiring mechanical ventilation and remained effective for up to 26 days (significantly longer than in previous reports) without significant toxicity in two patients. The overall outcome was good in all our patients. However, the optimal dose, optimal duration and maximum safe duration of intravenous magnesium therapy are unknown. Therefore, until more data on the safety and efficacy of magnesium therapy are available, its use should be limited to carefully selected tetanus cases.</p

International consensus recommendations for management of new onset refractory status epilepticus including febrile infection-related epilepsy syndrome: Statements and supporting evidence

Objective: This study was undertaken to develop consensus-based recommendations for the management of adult and pediatric patients with new onset refractory status epilepticus (NORSE)/febrile infection-related epilepsy syndrome (FIRES) based on best evidence and experience. Methods: The Delphi methodology was followed. A facilitator group of nine experts was established, who defined the scope, users, and suggestions for recommendations. Following a review of the current literature, recommendation statements concerning diagnosis, treatment, and research directions were generated, which were then rated on a scale of 1 (strongly disagree) to 9 (strongly agree) by a panel of 48 experts in the field. Consensus that a statement was appropriate was reached if the median score was ≥7 and inappropriate if the median score was ≤3. The analysis of evidence was mapped to the results of each statement included in the Delphi survey. Results: Overall, 85 recommendation statements achieved consensus. The recommendations are divided into five sections: (1) disease characteristics; (2) diagnostic testing and sampling; (3) acute treatment; (4) treatment in the postacute phase; and (5) research, registries, and future directions in NORSE/FIRES. The detailed results and discussion of all 85 statements are outlined herein. A corresponding summary of findings and practical flowsheets are presented in a companion article. Significance: This detailed analysis offers insight into the supporting evidence and the current gaps in the literature that are associated with expert consensus statements related to NORSE/FIRES. The recommendations generated by this consensus can be used as a guide for the diagnosis, evaluation, and management of patients with NORSE/FIRES, and for planning of future research