18 research outputs found
Chemokines are secreted by monocytes following OK-432 (lyophilized Streptococcus pyogenes) stimulation
Background: OK-432, penicillin-killed Streptococcus pyogenes, is used in treating lymphangiomas and carcinomas. We have studied in vitro the role of mononuclear phagocytes (MNPs), including purified monocytes (MOs), in the immune response to OK-432. MIP-1α/β and MCP-1 secretions were assessed in whole blood (WB), peripheral blood mononuclear cells (PBMCs) and purified MOs, after in vitro stimulation with OK-432 with or without adherence for 24 hours. Results: OK-432 stimulated MNPs to secrete MCP-1 and MIP-1α/β in healthy individuals and in head and neck squamous cell carcinoma (HNSCC) patients, except for OK-432 stimulation of WB giving a minimal MIP-1α/β response. Upon culture on low-attachment wells, a spontaneous chemokine secretion was observed, with an unchanged secretion following OK-432 stimulation. Inhibition of Syk kinase and/or PI-3 kinase did not significantly change the chemokine response to OK-432, except for MIP-1α production being increased upon Syk inhibitor addition and an increased MCP-1 response upon addition of both inhibitors. Adhesion may possibly involve β1 and/ or β3 integrins, not β2, whereas β1–3 integrins may act as co-stimulatory receptors for OK-432. Based on direct blockage of CD36 or CD18 by antibodies, MCP-1 production may be mediated by CD18 while MIP-1β and MCP-1 production may occur upon binding to CD36. Conclusion: Adherent human MOs produce MCP-1 and MIP-1α/β upon stimulation with OK-432. CD36 modulates MIP-1β and MCP-1 response. Thus, to some extent OK-432 acts as a substance whereby only MOs adhered to surfaces secrete MCP-1 and MIP-1α/β, in part explaining why OK- 432 is suited as a biological response modifying drug.publishedVersio
Clathrin- and Dynamin-Independent Endocytosis of FGFR3 – Implications for Signalling
Endocytosis of tyrosine kinase receptors can influence both the duration and the specificity of the signal emitted. We have investigated the mechanisms of internalization of fibroblast growth factor receptor 3 (FGFR3) and compared it to that of FGFR1 which is internalized predominantly through clathrin-mediated endocytosis. Interestingly, we observed that FGFR3 was internalized at a slower rate than FGFR1 indicating that it may use a different endocytic mechanism than FGFR1. Indeed, after depletion of cells for clathrin, internalization of FGFR3 was only partly inhibited while endocytosis of FGFR1 was almost completely abolished. Similarly, expression of dominant negative mutants of dynamin resulted in partial inhibition of the endocytosis of FGFR3 whereas internalization of FGFR1 was blocked. Interfering with proposed regulators of clathrin-independent endocytosis such as Arf6, flotillin 1 and 2 and Cdc42 did not affect the endocytosis of FGFR1 or FGFR3. Furthermore, depletion of clathrin decreased the degradation of FGFR1 resulting in sustained signalling. In the case of FGFR3, both the degradation and the signalling were only slightly affected by clathrin depletion. The data indicate that clathrin-mediated endocytosis is required for efficient internalization and downregulation of FGFR1 while FGFR3, however, is internalized by both clathrin-dependent and clathrin-independent mechanisms
The replication crisis has led to positive structural, procedural, and community changes
The emergence of large-scale replication projects yielding successful rates substantially lower than expected caused the behavioural, cognitive, and social sciences to experience a so-called ‘replication crisis’. In this Perspective, we reframe this ‘crisis’ through the lens of a credibility revolution, focusing on positive structural, procedural and community-driven changes. Second, we outline a path to expand ongoing advances and improvements. The credibility revolution has been an impetus to several substantive changes which will have a positive, long-term impact on our research environment
Recommended from our members
The replication crisis has led to positive structural, procedural, and community changes
In response to large-scale replication projects yielding successful replication rates substantially lower than expected, the behavioural, cognitive and social sciences have found themselves amidst a ‘replication crisis’. In this narrative review, we reframe this ’crisis’ through the lens of a credibility revolution, focusing on positive structural, procedural and community-driven changes. Second, we outline a path to expand ongoing advances and improvements. The credibility revolution has been an impetus to several substantive changes which will have a positive, long-term impact upon our research environment
Meditation and Gender Affect the Attentional Blink: An ERP and Bayesian Learning Analysis
Nyere modeller anser hjernen som en Bayesiansk probabilistisk inferensmaskin som bruker fri-energiprinsipper til å oppdatere internaliserte modeller om omgivelsene. Dette impliserer at top-down læringsfunksjoner i hjernen blir forårsaket av interne prediksjoner om utfallet av en situasjon, og at læring er et produkt av oppdaterte modeller basert på prediksjonsfeil. Dette gav opphav til Bayesianske læringsmodeller som ‘Hierarchical Gaussian Filter’. Disse er i stand til å mer effektivt modellere læring på individnivå, og predikere adferd når de sammenlignes med eldre modeller som Rescorla Wagner. Vi modellerte oppnådd læring i en EEG Attentional Blink (AB) oppgave for deltakerne uten tidligere meditasjonstrening (N = 32). Vi sammenlignet grad av læring mellom kjønn for to meditasjonstyper: «Open Monitoring Meditation» (OMM) og «Focused Attention Meditation» (FAM). Vi antok at responser til det andre AB-målet, i innen- versus utenfor-AB svar ‘Target 2’, ville generere prediksjonsfeil som fører til en implisitt læringseffekt. Vi fant at kvinner i FAM-gruppen viste høyere ω nivåer for ‘volatility’-estimater (tredje nivå), noe som antydet at de oppfattet en høyere endringsrate i miljøet. Deltakere i OMM-gruppen scoret høyere på T2-treffsikkerhet, og denne effekten var hovedsakelig forårsaket av kvinnelige deltagere. EEG-data støtter denne konklusjonen, og viser at kvinner i OMM-gruppen viste høyere ERP-amplituder av P300-komponenten for utenfor-AB-gjennomføringer. Dette indikerer at kvinner blir raskt påvirket av ‘mindfulness’-meditasjon under selektive oppmerksomhetsoppgaver, og at denne effekten kan forklares gjennom antagelser om hvor stabile omgivelsene er
Chemokines are secreted by monocytes following OK-432 (lyophilized Streptococcus pyogenes) stimulation
Background: OK-432, penicillin-killed Streptococcus pyogenes, is used in treating lymphangiomas and carcinomas. We have studied in vitro the role of mononuclear phagocytes (MNPs), including purified monocytes (MOs), in the immune response to OK-432. MIP-1α/β and MCP-1 secretions were assessed in whole blood (WB), peripheral blood mononuclear cells (PBMCs) and purified MOs, after in vitro stimulation with OK-432 with or without adherence for 24 hours. Results: OK-432 stimulated MNPs to secrete MCP-1 and MIP-1α/β in healthy individuals and in head and neck squamous cell carcinoma (HNSCC) patients, except for OK-432 stimulation of WB giving a minimal MIP-1α/β response. Upon culture on low-attachment wells, a spontaneous chemokine secretion was observed, with an unchanged secretion following OK-432 stimulation. Inhibition of Syk kinase and/or PI-3 kinase did not significantly change the chemokine response to OK-432, except for MIP-1α production being increased upon Syk inhibitor addition and an increased MCP-1 response upon addition of both inhibitors. Adhesion may possibly involve β1 and/ or β3 integrins, not β2, whereas β1–3 integrins may act as co-stimulatory receptors for OK-432. Based on direct blockage of CD36 or CD18 by antibodies, MCP-1 production may be mediated by CD18 while MIP-1β and MCP-1 production may occur upon binding to CD36. Conclusion: Adherent human MOs produce MCP-1 and MIP-1α/β upon stimulation with OK-432. CD36 modulates MIP-1β and MCP-1 response. Thus, to some extent OK-432 acts as a substance whereby only MOs adhered to surfaces secrete MCP-1 and MIP-1α/β, in part explaining why OK- 432 is suited as a biological response modifying drug
Vesicle transmembrane potential is required for translocation to the cytosol of externally added FGF-1
Externally added fibroblast growth factor-1 (FGF-1) is capable of crossing cellular membranes to reach the cytosol and the nucleus in a number of cell types. We have monitored the translocation of the growth factor by two methods: phosphorylation of FGF-1, and prenylation of an FGF-1 mutant that contains a C-terminal prenylation signal. Inhibition of endosomal acidification by ammonium chloride or monensin did not block the translocation of FGF-1, whereas bafilomycin A1, a specific inhibitor of vacuolar proton pumps, blocked translocation completely. A combination of ionophores expected to dissipate the vesicular membrane potential (valinomycin plus monensin) also fully inhibited the translocation. The inhibition of translocation by bafilomycin A1 was overcome in the presence of monensin or nigericin, while ouabain blocked translocation under these conditions. The data indicate that translocation of FGF-1 to cytosol occurs from the lumen of intracellular vesicles possessing vacuolar proton pumps, and that a vesicular membrane potential is required. Apparently, activation of vesicular Na(+)/K(+)-ATPase by monensin or nigericin generates a membrane potential that can support translocation when the proton pump is blocked
Translocation of FGF-1 and FGF-2 across Vesicular Membranes Occurs during G(1)-Phase by a Common Mechanism
The entry of exogenous fibroblast growth factor 2 (FGF-2) to the cytosolic/nuclear compartment was studied and compared with the translocation mechanism used by FGF-1. To differentiate between external and endogenous growth factor, we used FGF-2 modified to contain a farnesylation signal, a CaaX-box. Because farnesylation occurs only in the cytosol and nucleoplasm, farnesylation of exogenous FGF-2-CaaX was taken as evidence that the growth factor had translocated across cellular membranes. We found that FGF-2 translocation occurred in endothelial cells and fibroblasts, which express FGF receptors, and that the efficiency of translocation was increased in the presence of heparin. Concomitantly with translocation, the 18-kDa FGF-2 was N-terminally cleaved to yield a 16-kDa form. Translocation of FGF-2 required PI3-kinase activity but not transport through the Golgi apparatus. Inhibition of endosomal acidification did not prevent translocation, whereas dissipation of the vesicular membrane potential completely blocked it. The data indicate that translocation occurs from intracellular vesicles containing proton pumps and that an electrical potential across the vesicle membrane is required. Translocation of both FGF-1 and FGF-2 occurred during most of G(1) but decreased shortly before the G(1)→S transition. A common mechanism for FGF-1 and FGF-2 translocation into cells is postulated