268 research outputs found

    Complement C7 and clusterin form a complex in circulation

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    IntroductionThe complement system is part of innate immunity and is comprised of an intricate network of proteins that are vital for host defense and host homeostasis. A distinct mechanism by which complement defends against invading pathogens is through the membrane attack complex (MAC), a lytic structure that forms on target surfaces. The MAC is made up of several complement components, and one indispensable component of the MAC is C7. The role of C7 in MAC assembly is well documented, however, inherent characteristics of C7 are yet to be investigated.MethodsTo shed light on the molecular characteristics of C7, we examined the properties of serum-purified C7 acquired using polyclonal and novel monoclonal antibodies. The properties of serum‚ÄĎpurified C7 were investigated through a series of proteolytic analyses, encompassing Western blot and mass spectrometry. The nature of C7 protein-protein interactions were further examined by a novel enzyme-linked immunosorbent assay (ELISA), as well as size‚ÄĎexclusion chromatography. ResultsProtein analyses showcased an association between C7 and clusterin, an inhibitory complement regulator. The distinct association between C7 and clusterin was also demonstrated in serum-purified clusterin. Further assessment revealed that a complex between C7 and clusterin (C7-CLU) was detected. The C7-CLU complex was also identified in healthy serum and plasma donors, highlighting the presence of the complex in circulation. DiscussionClusterin is known to dissociate the MAC structure by binding to polymerized C9, nevertheless, here we show clusterin binding to the native form of a terminal complement protein in vivo. The presented data reveal that C7 exhibits characteristics beyond that of MAC assembly, instigating further investigation of the effector role that the C7-CLU complex plays in the complement cascade

    DataSheet_1_Complement C7 and clusterin form a complex in circulation.docx

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    IntroductionThe complement system is part of innate immunity and is comprised of an intricate network of proteins that are vital for host defense and host homeostasis. A distinct mechanism by which complement defends against invading pathogens is through the membrane attack complex (MAC), a lytic structure that forms on target surfaces. The MAC is made up of several complement components, and one indispensable component of the MAC is C7. The role of C7 in MAC assembly is well documented, however, inherent characteristics of C7 are yet to be investigated.MethodsTo shed light on the molecular characteristics of C7, we examined the properties of serum-purified C7 acquired using polyclonal and novel monoclonal antibodies. The properties of serum‚ÄĎpurified C7 were investigated through a series of proteolytic analyses, encompassing Western blot and mass spectrometry. The nature of C7 protein-protein interactions were further examined by a novel enzyme-linked immunosorbent assay (ELISA), as well as size‚ÄĎexclusion chromatography. ResultsProtein analyses showcased an association between C7 and clusterin, an inhibitory complement regulator. The distinct association between C7 and clusterin was also demonstrated in serum-purified clusterin. Further assessment revealed that a complex between C7 and clusterin (C7-CLU) was detected. The C7-CLU complex was also identified in healthy serum and plasma donors, highlighting the presence of the complex in circulation. DiscussionClusterin is known to dissociate the MAC structure by binding to polymerized C9, nevertheless, here we show clusterin binding to the native form of a terminal complement protein in vivo. The presented data reveal that C7 exhibits characteristics beyond that of MAC assembly, instigating further investigation of the effector role that the C7-CLU complex plays in the complement cascade. </p

    Measurement of the <math><mrow><mmultiscripts><mi>Se</mi><mprescripts/><none/><mn>77</mn></mmultiscripts><mo>(</mo><mi>n</mi><mo>,</mo><mi>ő≥</mi><mo>)</mo></mrow></math> cross section¬†up to 200 keV at the n_TOF facility at CERN

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    International audienceThe Se77(n,ő≥) reaction is of importance for Se77 abundance during the slow neutron capture process in massive stars. We have performed a new measurement of the Se77 radiative neutron capture cross section¬†at the Neutron Time-of-Flight facility at CERN. Resonance capture kernels were derived up to 51 keV and cross sections¬†up to 200 keV. Maxwellian-averaged cross sections¬†were calculated for stellar temperatures between kT=5keV and kT=100keV, with uncertainties between 4.2% and 5.7%. Our results lead to substantial decreases of 14% and 19% in Se77 abundances produced through the slow neutron capture process in selected stellar models of 15M‚äô and 2M‚äô, respectively, compared to using previous recommendation of the cross section

    Evolution of CRISPR-associated endonucleases as inferred from resurrected proteins

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    Clustered regularly interspaced short palindromic repeats (CRISPR)-associated Cas9 is an effector protein that targets invading DNA and plays a major role in the prokaryotic adaptive immune system. Although Streptococcus pyogenes CRISPR‚ÄďCas9 has been widely studied and repurposed for applications including genome editing, its origin and evolution are poorly understood. Here, we investigate the evolution of Cas9 from resurrected ancient nucleases (anCas) in extinct firmicutes species that last lived 2.6‚ÄČbillion years before the present. We demonstrate that these ancient forms were much more flexible in their guide RNA and protospacer-adjacent motif requirements compared with modern-day Cas9 enzymes. Furthermore, anCas portrays a gradual palaeoenzymatic adaptation from nickase to double-strand break activity, exhibits high levels of activity with both single-stranded DNA and single-stranded RNA targets and is capable of editing activity in human cells. Prediction and characterization of anCas with a resurrected protein approach uncovers an evolutionary trajectory leading to functionally flexible ancient enzymes.This work has been supported by grant nos. PID2019-109087RB-I00 (to R.P.-J.) and RTI2018-101223-B-I00 and PID2021-127644OB-I00 (to L.M.) from the Spanish Ministry of Science and Innovation. This project has received funding from the European Union‚Äôs Horizon 2020 research and innovation programme under grant agreement no. 964764 (to R.P.-J.). The content presented in this document represents the views of the authors, and the European Commission has no liability in respect to the content. We acknowledge financial support from the Spanish Foundation for the Promotion of Research of Amyotrophic Lateral Sclerosis. A.F. acknowledges Spanish Center for Biomedical Network Research on Rare Diseases (CIBERE) intramural funds (no. ER19P5AC756/2021). F.J.M.M. acknowledges research support by Conselleria d‚ÄôEducaci√≥, Investigaci√≥, Cultura i Esport from Generalitat Valenciana, research project nos. PROMETEO/2017/129 and PROMETEO/2021/057. M.M. acknowledges funding from CIBERER (grant no. ER19P5AC728/2021). The work has received funding from the Regional Government of Madrid (grant no. B2017/BMD3721 to M.A.M.-P.) and from Instituto de Salud Carlos III, cofounded with the European Regional Development Fund ‚ÄėA way to make Europe‚Äô within the National Plans for Scientific and Technical Research and Innovation 2017‚Äď2020 and 2021‚Äď2024 (nos. PI17/1659, PI20/0429 and IMP/00009; to M.A.M.-P. B.P.K. was supported by an MGH ECOR Howard M. Goodman Award and NIH P01 HL142494

    miniBELEN: A modular neutron counter for (, ) reactions

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    miniBELEN is a modular and transportable neutron moderated counter with a nearly flat neutron detection efficiency up to 10 MeV. Modularity implies that the moderator can be reassembled in different ways in order to obtain different types of response. The detector has been developed in the context of the Measurement of Alpha Neutron Yields (MANY) collaboration, which is a scientific effort aiming to carry out measurements of (, ) production yields, reaction cross-sections and neutron energy spectra. In this work we present and discuss several configurations of the miniBELEN detector. The experimental validation of the efficiency calculations using 252Cf sources and the measurement of the 27Al(, ) 30P reaction is also presented

    Overview of the dissemination of n_TOF experimental data and resonance parameters

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    The n_TOF neutron time-of-flight facility at CERN is used for nuclear data measurements. The n_TOF Collaboration works closely with the Nuclear Reaction Data Centres (NRDC) network to disseminate the experimental data through the international EXFOR library. In addition, the Collaboration helps integrate the results in the evaluated library projects. The present contribution describes the dissemination status of n_TOF results, their impact on evaluated libraries and ongoing efforts to provide n_TOF resonance parameters in ENDF-6 format for further use by evaluation projects

    Measurement of the neutron-induced fission cross section of <math><mmultiscripts><mi>Th</mi><mprescripts/><none/><mn>230</mn></mmultiscripts></math> at the CERN n_TOF facility

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    International audienceThe neutron-induced fission cross section of Th230 has been measured at the neutron time-of-flight facility n_TOF located at CERN. The experiment was performed at the experimental area EAR-1 with a neutron flight path of 185 m, using Micromegas detectors for the detection of the fission fragments. The Th230(n,f) cross section was determined relative to the U235(n,f) one, covering the energy range from the fission threshold up to 400 MeV. The results from the present work are compared with existing cross-section datasets and the observed discrepancies are discussed and analyzed. Finally, using the code empire 3.2.3 a theoretical study, based on the statistical model, was performed leading to a satisfactory reproduction of the experimental results with the proper tuning of the respective parameters, while for incident neutron energy beyond 200 MeV the fission of Th230 was described by Monte Carlo simulations

    Maresin 1 activates brown adipose tissue and promotes browning of white adipose tissue in mice

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    Objective: Maresin 1 (MaR1) is a docosahexaenoic acid-derived proresolving lipid mediator with insulin-sensitizing and anti-steatosis properties. Here, we aim to unravel MaR1 actions on brown adipose tissue (BAT) activation and white adipose tissue (WAT) browning. Methods: MaR1 actions were tested in cultured murine brown adipocytes and in human mesenchymal stem cells (hMSC)-derived adipocytes. In vivo effects of MaR1 were tested in diet-induced obese (DIO) mice and lean WT and Il6 knockout (Il6 / ) mice. Results: In cultured differentiated murine brown adipocytes, MaR1 reduces the expression of inflammatory genes, while stimulates glucose uptake, fatty acid utilization and oxygen consumption rate, along with the upregulation of mitochondrial mass and genes involved in mitochondrial biogenesis and function and the thermogenic program. In Leucine Rich Repeat Containing G Protein-Coupled Receptor 6 (LGR6)-depleted brown adipocytes using siRNA, the stimulatory effect of MaR1 on thermogenic genes was abrogated. In DIO mice, MaR1 promotes BAT remodeling, characterized by higher expression of genes encoding for master regulators of mitochondrial biogenesis and function and iBAT thermogenic activation, together with increased M2 macrophage markers. In addition, MaR1-treated DIO mice exhibit a better response to cold-induced BAT activation. Moreover, MaR1 induces a beige adipocyte signature in inguinal WAT of DIO mice and in hMSC-derived adipocytes. MaR1 potentiates Il6 expression in brown adipocytes and BAT of cold exposed lean WT mice. Interestingly, the thermogenic properties of MaR1 were abrogated in Il6 / mice. Conclusions: These data reveal MaR1 as a novel agent that promotes BAT activation and WAT browning by regulating thermogenic program in adipocytes and M2 polarization of macrophages. Moreover, our data suggest that LGR6 receptor is mediating MaR1 actions on brown adipocytes, and that IL-6 is required for the thermogenic effects of MaR1

    Neutron-induced fission cross sections of <math><mmultiscripts><mi>Th</mi><mprescripts/><none/><mn>232</mn></mmultiscripts></math> and <math><mmultiscripts><mi mathvariant="normal">U</mi><mprescripts/><none/><mn>233</mn></mmultiscripts></math> up to 1 GeV using parallel plate avalanche counters at the CERN n_TOF facility

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    International audienceThe neutron-induced fission cross sections¬†of Th232 and U233 were measured relative to U235 in a wide neutron energy range up to 1 GeV (and from fission threshold in the case of Th232, and from 0.7¬†eV in case of U233), using the white-spectrum neutron source at the CERN Neutron Time-of-Flight (n_TOF) facility. Parallel plate avalanche counters (PPACs) were used, installed at the Experimental Area 1 (EAR1), which is located at 185¬†m from the neutron spallation target. The anisotropic emission of fission fragments were taken into account in the detection efficiency by using, in the case of U233, previous results available in EXFOR, whereas in the case of Th232 these data were obtained from our measurement, using PPACs and targets tilted 45‚ąė with respect to the neutron beam direction. Finally, the obtained results are compared with past measurements and major evaluated nuclear data libraries. Calculations using the high-energy reaction models INCL++ and ABLA07 were performed and some of their parameters were modified to reproduce the experimental results. At high energies, where no other neutron data exist, our results are compared with experimental data on proton-induced fission. Moreover, the dependence of the fission cross section¬†at 1 GeV with the fissility parameter of the target nucleus is studied by combining those (p,f) data with our (n,f) data on Th232 and U233 and on other isotopes studied earlier at n_TOF using the same experimental setup

    First measurement of the 94Nb(n,ő≥) cross section at the CERN n_TOF facility

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    One of the crucial ingredients for the improvement of stellar models is the accurate knowledge of neutron capture cross-sections for the different isotopes involved in the s-,r- and i- processes. These measurements can shed light on existing discrepancies between observed and predicted isotopic abundances and help to constrain the physical conditions where these reactions take place along different stages of stellar evolution. In the particular case of the radioactive 94Nb, the 94Nb(n,ő≥) cross-section could play a role in the determination of the s-process production of 94Mo in AGB stars, which presently cannot be reproduced by state-of-the-art stellar models. There are no previous 94Nb(n,ő≥) experimental data for the resolved and unresolved resonance regions mainly due to the difficulties in producing highquality samples and also due to limitations in conventional detection systems commonly used in time-of-flight experiments. Motivated by this situation, a first measurement of the 94Nb(n,ő≥) reaction was carried out at CERN n_TOF, thereby exploiting the high luminosity of the EAR2 area in combination with a new detection system of small-volume C6D6-detectors and a high quality 94Nb-sample. The latter was based on hyper-pure 93Nb material activated at the high-flux reactor of ILL-Grenoble. An innovative ring-configuration detection system in close geometry around the capture sample allowed us to significantly enhance the signal-to-background ratio. This set-up was supplemented with two conventional C6D6-detectors and a highresolution LaCl3(Ce)-detector, which will be employed for addressing reliably systematic effects and uncertainties. At the current status of the data analysis, 18 resonance in 94Nb+n have been observed for the first time in the neutron energy range from thermal up to 10 keV.Title in Web of Science: First measurement of the Nb-94(n,gamma) cross section at the CERN n_TOF facility</p
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