The Hamamatsu R11410-21 photomultiplier tube is the photodetector of choice for the XENON1T dual-phase time projection chamber. The device has been optimized for a very low intrinsic radioactivity, a ...high quantum efficiency and a high sensitivity to single photon detection. A total of 248 tubes are currently operated in XENON1T, selected out of 321 tested units. In this article the procedures implemented to evaluate the large number of tubes prior to their installation in XENON1T are described. The parameter distributions for all tested tubes are shown, with an emphasis on those selected for XENON1T, of which the impact on the detector performance is discussed. All photomultipliers have been tested in a nitrogen atmosphere at cryogenic temperatures, with a subset of the tubes being tested in gaseous and liquid xenon, simulating their operating conditions in the dark matter detector. The performance and evaluation of the tubes in the different environments is reported and the criteria for rejection of PMTs are outlined and quantified.
Abstract
Background
The endocannabinoid (eCB) 2-arachidonoylglycerol (2-AG) is a known modulator of inflammation and few studies have addressed its influence on myeloid cells in the context of ...atherogenesis. However, the impact of 2-AG on endothelial cell function has not been studied before.
Methods
Endothelial repair was studied in two treatment groups of wildtype mice following electrical denudation of the common carotid artery at a length of 3000 μm. One group received the monoacylglycerol lipase (MAGL)-inhibitor JZL184 5 mg/kg i.p., which impairs 2-AG degradation and thus causes elevated 2-AG levels, the other group received vehicle. The residual endothelial gap at five days in either group was visualized by Evan's blue staining. In vitro, the effect of 2-AG on human coronary artery endothelial cell (HCAEC) viability was assessed by an XTT-based assay. Endothelial activation was studied by an adhesion assay of THP-1 monocytes to 2-AG-preconditioned HCAEC. HCAEC migration, ROS-production, expression of NADPH oxidases, and secretion of inflammatory cytokines were assessed by Boyden chamber, qPCR, and colorimetric assays.
Results
Treatment with JZL184 produced a significant increase in 2-AG levels and impaired reendothelialisation in wildtype mice following electrical injury of the common carotid artery. The residual denudation at 5 days yielded 2291±286 μm in JZL184-treated animals vs. 1505±223 μm in vehicle treated controls (n=18–19; p<0.05). In vitro, JZL184 significantly reduced viability of HCAEC at 24 hours (0.31±0.10 vs. 1.00±0.08; n=3; p<0.01). Finally, 2-AG promoted HCAEC activation resulting in a significant increase in THP-1 monocyte adhesion to HCAEC following pre-treatment of HCAEC with 2-AG (0.17±0.03 THP-1 cells per HCAEC vs. 0.07±0.01 THP-1 cells per HCAEC; n=3; p<0.05). Besides, HCAEC migration, ROS-production, expression of NADPH oxidases and secretion of inflammatory cytokines were unaffected by 2-AG.
Conclusion
Elevated 2-AG levels appear to hamper endothelial repair and to promote HCAEC activation and cell death. Our data suggest that besides its influence on myeloid cells, 2-AG is also adverse to endothelial integrity which might promote early atherosclerotic lesion formation. Thus, decreasing vascular 2-AG levels might represent a promising therapeutic strategy for the prevention of atherosclerosis and coronary heart disease.
Abstract
Background
The endocannabinoid 2-arachidonoylglycerol (2-AG) is an inflammatory mediator and ligand to the cannabinoid receptors CB1 and CB2, which are expressed on myeloid and endothelial ...cells. 2-AG has recently been described to promote atherogenesis in ApoE-deficient mice. While the CB2 receptor has previously been considered to solely exert anti-inflammatory and atheroprotective effects, newer data have raised the notion, that CB2 might exert atherogenic effects in the context of elevated 2-AG plasma levels. In the present study, we investigated the atherogenic mechanisms of 2-AG and the role of the CB2 receptor on myeloid and endothelial cells in atherogenesis using cell-specific knockout mouse models.
Methods
Two mouse models with atherogenic background and distinct cell-specific knockouts of the CB2 receptor on myeloid (ApoE−/−LysMcreCB2fl/fl) or endothelial cells (ApoE−/−Tie2creCB2fl/fl) were created. Mice were treated with JZL184, which inhibits 2-AG-degrading enzyme monoacylglycerol lipase, and thereby elevates 2-AG plasma levels, or with vehicle (DMSO), while being fed a high-fat diet for four weeks. Plaque volume and plaque composition were analyzed. In vitro, macrophages were treated with 2-AG and mRNA levels of adhesion molecules, scavenger receptors and chemokines, the production of reactive oxygen species (ROS) and the release of myeloperoxidase (MPO) were determined using qPCR, fluorometric assays and ELISA respectively.
Results
Elevated levels of 2-AG promote atherogenesis in ApoE-deficient mice (JZL184 vs. DMSO: 39.6±2.1% vs. 32.6±2.4%; n=14; p<0.05). The atherogenic effect of 2-AG is abrogated in mice lacking myeloid CB2 receptor expression (35.0±2.0% vs. 34.0±2.5%; n=14–16; p>0.05) but not in mice lacking endothelial CB2 receptor expression (37.1±3.1% vs. 20.9±2.6%; n=10–12; p<0.01). In vitro, 2-AG significantly increases transcription of adhesion molecule ICAM-1 (2.09±0.42 –fold; n=5–6; p<0.05), chemokine receptor CCR-1 (2.04±0.46 -fold; n=10–11; p<0.05) and scavenger receptor CD36 (8.02±1.89-fold; n=3; p<0.05) in 2-AG-treated macrophages. These effects are mitigated by pharmacological inhibition of CB2. Furthermore, 2-AG significantly increases myeloperoxidase (MPO) release in monocytes in a CB receptor-dependent fashion (451±23 pg/ml vs. 151±8.3 pg/ml; n=3–4; p<0.01) and promotes ROS production (2698±24 pdu vs. 1981±27 pdu; n=8; p<0.01).
Conclusion
Elevated 2-AG levels show an atherogenic effect in vivo which is dependent on the presence of the CB2 receptor on myeloid cells. Our in vitro data reveal 2-AG to promote pro-inflammatory signaling in macrophages and elucidate a previously unrecognized link between the endocannabinoid system and MPO in monocytes. In summary, cell-specific effects of the endocannabinoid system will have to be taken into account to facilitate its exploitation as an anti-atherosclerotic drug target.
Acknowledgement/Funding
This work was supported by the Bonfor program of the University of Bonn grant number O-109.0057 to JJ.
The Hamamatsu R11410-21 photomultiplier tube is the photodetector of choice for the XENON1T dual-phase time projection chamber. The device has been optimized for a very low intrinsic radioactivity, a ...high quantum efficiency and a high sensitivity to single photon detection. A total of 248 tubes are currently operated in XENON1T, selected out of 321 tested units. In this article the procedures implemented to evaluate the large number of tubes prior to their installation in XENON1T are described. The parameter distributions for all tested tubes are shown, with an emphasis on those selected for XENON1T, of which the impact on the detector performance is discussed. All photomultipliers have been tested in a nitrogen atmosphere at cryogenic temperatures, with a subset of the tubes being tested in gaseous and liquid xenon, simulating their operating conditions in the dark matter detector. The performance and evaluation of the tubes in the different environments is reported and the criteria for rejection of PMTs are outlined and quantified.
Beyond Civility Danisch, Robert; Keith, William
2020, Letnik:
23
eBook
From the pundits to the polls, nearly everyone seems to agree that US politics have rarely been more fractious, and calls for a return to “civil discourse” abound. Yet it is also true that the ...requirements of polite discourse effectively silence those who are not in power, gaming the system against the disenfranchised. What, then, should a democracy do?
This book makes a case for understanding civility in a different light. Examining the history of the concept and its basis in communication and political theory, William Keith and Robert Danisch present a clear, robust analysis of civil discourse. Distinguishing it from politeness, they claim that civil argument must be redirected from the goal of political comity to that of building and maintaining relationships of minimal respect in the public sphere. They also take into account how civility enables discrimination, indicating conditions under which uncivil resistance is called for. When viewed as a communication practice for uniting people with differences and making them more equal, civility is transformed from a preferable way of speaking into an essential component of democratic life.
Guarding against uncritical endorsement of civility as well as skepticism, Keith and Danisch show with rigor, nuance, and care that the practice of civil communication is both paradoxical and sorely needed. Beyond Civility is necessary reading for our times.
The type II transmembrane serine proteases TMPRSS2 and HAT activate influenza viruses and the SARS-coronavirus (TMPRSS2) in cell culture and may play an important role in viral spread and ...pathogenesis in the infected host. However, it is at present largely unclear to what extent these proteases are expressed in viral target cells in human tissues. Here, we show that both HAT and TMPRSS2 are coexpressed with 2,6-linked sialic acids, the major receptor determinant of human influenza viruses, throughout the human respiratory tract. Similarly, coexpression of ACE2, the SARS-coronavirus receptor, and TMPRSS2 was frequently found in the upper and lower aerodigestive tract, with the exception of the vocal folds, epiglottis and trachea. Finally, activation of influenza virus was conserved between human, avian and porcine TMPRSS2, suggesting that this protease might activate influenza virus in reservoir-, intermediate- and human hosts. In sum, our results show that TMPRSS2 and HAT are expressed by important influenza and SARS-coronavirus target cells and could thus support viral spread in the human host.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Chimeric antigen receptor (CAR) T cells brought substantial benefit to patients with B‐cell malignancies. Notwithstanding, CAR T‐cell manufacturing requires complex procedures impeding the broad ...supply chain. Here, we provide evidence that human CD19‐CAR T cells can be generated directly in vivo using the lentiviral vector CD8‐LV specifically targeting human CD8+ cells. Administration into mice xenografted with Raji lymphoma cells and human peripheral blood mononuclear cells led to CAR expression solely in CD8+ T cells and efficacious elimination of CD19+ B cells. Further, upon injection of CD8‐LV into mice transplanted with human CD34+ cells, induction of CAR T cells and CD19+ B‐cell depletion was observed in 7 out of 10 treated animals. Notably, three mice showed elevated levels of human cytokines in plasma. Tissue‐invading CAR T cells and complete elimination of the B‐lymphocyte‐rich zones in spleen were indicative of a cytokine release syndrome. Our data demonstrate the feasibility of in vivo reprogramming of human CD8+ CAR T cells active against CD19+ cells, yet with similar adverse effects currently notorious in the clinical practice.
Synopsis
T cells were reprogrammed with chimeric antigen receptors (CARs) in situ upon systemic injection of CD8‐targeted lentiviral vectors, thus providing proof‐of‐principle for converting CAR T cell therapy from an autologous, individualized to an off‐the‐shelf product.
In situ CAR T cell generation was evidenced in mice transplanted with human PBMC or CD34+ stem cells.
In situ generated CAR T cells expanded upon antigen recognition and eliminated CD19+ cells.
Some mice developed side‐effects resembling the cytokine release syndrome observed in patients treated with CAR T cells.
T cells were reprogrammed with chimeric antigen receptors (CARs) in situ upon systemic injection of CD8‐targeted lentiviral vectors, thus providing proof‐of‐principle for converting CAR T cell therapy from an autologous, individualized to an off‐the‐shelf product.
Human cytomegalovirus (HCMV) causes serious complications to immune compromised hosts. Dendritic cells (iDCgB) expressing granulocyte-macrophage colony-stimulating factor, interferon-alpha and ...HCMV-gB were developed to promote de novo antiviral adaptive responses. Mice reconstituted with a human immune system (HIS) were immunized with iDCgB and challenged with HCMV, resulting into 93% protection. Immunization stimulated the expansion of functional effector memory CD8+ and CD4+ T cells recognizing gB. Machine learning analyses confirmed bone marrow T/CD4+, liver B/IgA+ and spleen B/IgG+ cells as predictive biomarkers of immunization (≈87% accuracy). CD8+ and CD4+ T cell responses against gB were validated. Splenic gB-binding IgM-/IgG+ B cells were sorted and analyzed at a single cell level. iDCgB immunizations elicited human-like IgG responses with a broad usage of various IgG heavy chain V gene segments harboring variable levels of somatic hypermutation. From this search, two gB-binding human monoclonal IgGs were generated that neutralized HCMV infection in vitro. Passive immunization with these antibodies provided proof-of-concept evidence of protection against HCMV infection. This HIS/HCMV in vivo model system supported the validation of novel active and passive immune therapies for future clinical translation.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
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