Analysis of the B cell repertoire in the systemic and mucosal tissues of rainbow trout (Oncorhynchus mykiss)

University of Technology, Sydney. Faculty of Science.Acquired immune responses against hapten-carrier systems have been used in mammals and teleosts in vivo as an effective means of identifying and characterizing T and B cell activity. Fish immunized by intraperitoneal (ip) injection of the hapten-carrier system FITC-KLH develop strong serum antibody responses to both the hapten (FITC) and carrier protein (KLH; Jones et al., 1999). In contrast, fish immunized with FITC-KLH by peranal (pa) intubation results in a failure to develop a detectable anti-KLH response in the serum, yet they still develop a significant anti-FITC response. To investigate the nature of the antibody secreting cell (ASC) response in the systemic (head kidney and spleen) and mucosal tissues (hindgut and gills) of trout, ELISA-based serum and cellular assays were utilized to detect serum and tissue-specific antibody responses to FITC and KLH. Given the distinction in serum responsiveness to FITC- KLH through the ip and pa routes, an identification of where the B cell tissue-specific responses were occurring would identify if there is a restricted B cell population in the mucosal tissues of rainbow trout. In the primary response to immunization with FITC-KLH, systemically challenged fish presented ASC activity in systemic tissues, while mucosally challenged fish presented ASC activity in a mixture of both systemic and mucosal tissues. In the secondary response to FITC-KLH, in fish immunized via a combination of systemic and mucosal routes there was a utilization of both systemic and mucosal tissues in the response. It is possible that immunization with FITC-KLH through the mucosal or systemic routes may cause presentation of FITC-KLH in the systemic tissues, resulting in trafficking of ASC or antigen presenting cells (APC) between the mucosal and systemic tissues. To examine the relationships within and between the B cell populations of the lymphoid tissues, the rearranged Vh gene repertoire was examined in the tissues with ASC activity. Identification of preferential or restricted use of the different Vh genes may provide further insight into the restricted serum response to KLH in mucosally challenged fish, and whether restricted populations of B cells exist within the mucosal tissues. A qRT-PCR assay was developed and optimized using non-immunized trout to analyze the use of Vh gene families VH-I to Vh-XI in rearranged Ig genes in the lymphoid tissues of trout. All Vh gene families were amplified across the tissues tested. In the primary response to immunization with FITC-KLH, families with the highest fold changes in gene expression for both systemic and mucosal tissues were Vh-VI, Vh-VIII, Vh-IX, Vh-X and Vh-XI. In the secondary response, families Vh-I, Vh-II, Vh-V, Vh-IX, Vh-X and Vh-XI had the highest fold changes in gene expression. Gaps in the repertoire were also apparent within the primary ASC response to FITC-KLH, and were mainly associated with families Vh-I, Vh-II, VH-III, VH-IV, VH-V and VH-VII. In the secondary ASC response study to FITC-KLH, systemic tissue contained few repertoire gaps, however in mucosal lymphoid tissues there was evidence of repertoire gaps for families VH-I, Vh-II, VH-III, Vh-IV, Vh-V, Vh-VI, Vh-VII and Vh-VIII. A similar pattern of expression for the Vh gene families could suggest B cells migrate to the different tissues from a common source, perhaps from the head kidney given its role as a primary lymphoid tissue. Alternatively, families present in high abundance may have a larger number of germline members, or are highly expressed due to an unknown antigenic challenge. This study is one of the first to identify the extent of the usage of the Vh gene families in these tissues

Estimation of Diameter of Quadrupled Hamstring Graft for ACL Reconstruction using Pre-operative MRI Measurement as a Predictive Tool

Introduction: The diameter of the quadrupled Hamstring graft plays a significant role in the incidence of graft failures for ACL reconstruction. The ability to predict the graft size pre-operatively can prepare the surgeon for alternatives in the event of an inadequate graft diameter. Materials and methods: We retrospectively measured the diameter of the Semitendinosus tendon (ST) on the MRI in all patients who underwent arthroscopic ACL reconstruction using quadrupled Semitendinosus as their graft. We also estimated any correlation between various anthropometric data with pre-operative MRI based Cross Sectional Area (CSA) of the Hamstring tendon and final graft diameter in the South Asian population. The patients were included from Jan 2018 - Dec 2020. Results: The minimum CSA of ST to predict an eventual graft diameter of 7.5mm was 10.7mm2. The MRI based cross-sectional area measurement showed moderate correlation with the intra-operative graft diameter obtained. (r=0.62, p<0.001). The intra-class correlation coefficient between the radiologist and the surgeon was 0.82, 95% CI (0.57, 0.92) and a p-value <0.001. Conclusion: Pre-operative MRI can be a useful tool to predict the graft diameter. This coupled with the anthropometric data of the patient can be used as an adjunct to estimate the probable graft diameter. Thus, the surgeon can be better prepared for the surgery and can seek alternate graft options if the graft size is deemed inadequate pre-operatively

Nano-Architecture of nitrogen-doped graphene films synthesized from a solid CN source

New synthesis routes to tailor graphene properties by controlling the concentration and chemical configuration of dopants show great promise. Herein we report the direct reproducible synthesis of 2-3% nitrogen-doped ‘few-layer’ graphene from a solid state nitrogen carbide a-C:N source synthesized by femtosecond pulsed laser ablation. Analytical investigations, including synchrotron facilities, made it possible to identify the configuration and chemistry of the nitrogen-doped graphene films. Auger mapping successfully quantified the 2D distribution of the number of graphene layers over the surface, and hence offers a new original way to probe the architecture of graphene sheets. The films mainly consist in a Bernal ABA stacking three-layer architecture, with a layer number distribution ranging from 2 to 6. Nitrogen doping affects the charge carrier distribution but has no significant effects on the number of lattice defects or disorders, compared to undoped graphene synthetized in similar conditions. Pyridinic, quaternary and pyrrolic nitrogen are the dominant chemical configurations, pyridinic N being preponderant at the scale of the film architecture. This work opens highly promising perspectives for the development of self-organized nitrogen-doped graphene materials, as synthetized from solid carbon nitride, with various functionalities, and for the characterization of 2D materials using a significant new methodology

Beta-HPV E6 Contributes To Skin Cancer by Hindering DNA Repair

<div><p>Recent work has explored a putative role for the E6 protein from some β-human papillomavirus genus (β-HPVs) in the development of non-melanoma skin cancers, specifically β-HPV 5 and 8 E6. Because these viruses are not required for tumor maintenance, they are hypothesized to act as co-factors that enhance the mutagenic capacity of UV-exposure by disrupting the repair of the resulting DNA damage. Supporting this proposal, we have previously demonstrated that UV damage signaling is hindered by β-HPV 5 and 8 E6 resulting in an increase in both thymine dimers and UV-induced double strand breaks (DSBs). Here we show that β-HPV 5 and 8 E6 further disrupt the repair of these DSBs and provide a mechanism for this attenuation. By binding and destabilizing a histone acetyltransferase, p300, β-HPV 5 and 8 E6 reduce the enrichment of the transcription factor at the promoter of two genes critical to the homology dependent repair of DSBs (BRCA1 and BRCA2). The resulting diminished BRCA1/2 transcription not only leads to lower protein levels but also curtails the ability of these proteins to form repair foci at DSBs. Using a GFP-based reporter, we confirm that this reduced foci formation leads to significantly diminished homology dependent repair of DSBs. By deleting the p300 binding domain of β-HPV 8 E6, we demonstrate that the loss of robust repair is dependent on viral-mediated degradation of p300 and confirm this observation using a combination of p300 mutants that are β-HPV 8 E6 destabilization resistant and p300 knock-out cells. In conclusion, this work establishes an expanded ability of β-HPV 5 and 8 E6 to attenuate UV damage repair, thus adding further support to the hypothesis that β-HPV infections play a role in skin cancer development by increasing the oncogenic potential of UV exposure.</p></div

Combination of searches for heavy spin-1 resonances using 139 fb−1 of proton-proton collision data at s = 13 TeV with the ATLAS detector

A combination of searches for new heavy spin-1 resonances decaying into different pairings of W, Z, or Higgs bosons, as well as directly into leptons or quarks, is presented. The data sample used corresponds to 139 fb−1 of proton-proton collisions at = 13 TeV collected during 2015–2018 with the ATLAS detector at the CERN Large Hadron Collider. Analyses selecting quark pairs (qq, bb, , and tb) or third-generation leptons (τν and ττ) are included in this kind of combination for the first time. A simplified model predicting a spin-1 heavy vector-boson triplet is used. Cross-section limits are set at the 95% confidence level and are compared with predictions for the benchmark model. These limits are also expressed in terms of constraints on couplings of the heavy vector-boson triplet to quarks, leptons, and the Higgs boson. The complementarity of the various analyses increases the sensitivity to new physics, and the resulting constraints are stronger than those from any individual analysis considered. The data exclude a heavy vector-boson triplet with mass below 5.8 TeV in a weakly coupled scenario, below 4.4 TeV in a strongly coupled scenario, and up to 1.5 TeV in the case of production via vector-boson fusion

Measurement of the Higgs boson production via vector boson fusion and its decay into bottom quarks in proton-proton collisions at s \sqrt{s} = 13 TeV

A preprint version of the article is available at arXiv:2308.01253v2 [hep-ex], https://arxiv.org/abs/2308.01253v2 . Comments: Replaced with the published version. Added the journal reference and the DOI. All the figures and tables can be found at https://cms-results.web.cern.ch/cms-results/public-results/publications/HIG-22-009 (CMS Public Pages). Report number: CMS-HIG-22-009, CERN-EP-2023-110.A measurement of the Higgs boson (H) production via vector boson fusion (VBF) and its decay into a bottom quark-antiquark pair (bb¯) is presented using proton-proton collision data recorded by the CMS experiment at s√ = 13 TeV and corresponding to an integrated luminosity of 90.8 fb−1. Treating the gluon-gluon fusion process as a background and constraining its rate to the value expected in the standard model (SM) within uncertainties, the signal strength of the VBF process, defined as the ratio of the observed signal rate to that predicted by the SM, is measured to be μqqHHbb¯ = 1.01 +0.55−0.46. The VBF signal is observed with a significance of 2.4 standard deviations relative to the background prediction, while the expected significance is 2.7 standard deviations. Considering inclusive Higgs boson production and decay into bottom quarks, the signal strength is measured to be μincl.Hbb¯ = 0.99 +0.48−0.41, corresponding to an observed (expected) significance of 2.6 (2.9) standard deviations.SCOAP3, STFC; Marie-Curie program and the European Research Council and Horizon 2020 Gran