Extracellular ATP released by osteoblasts is a key local inhibitor of bone mineralisation

Previous studies have shown that exogenous ATP (>1µM) prevents bone formation in vitro by blocking mineralisation of the collagenous matrix. This effect is thought to be mediated via both P2 receptor-dependent pathways and a receptor-independent mechanism (hydrolysis of ATP to produce the mineralisation inhibitor pyrophosphate, PPi). Osteoblasts are also known to release ATP constitutively. To determine whether this endogenous ATP might exert significant biological effects, bone-forming primary rat osteoblasts were cultured with 0.5-2.5U/ml apyrase (which sequentially hydrolyses ATP to ADP to AMP + 2Pi). Addition of 0.5U/ml apyrase to osteoblast culture medium degraded extracellular ATP to <1% of control levels within 2 minutes; continuous exposure to apyrase maintained this inhibition for up to 14 days. Apyrase treatment for the first 72 hours of culture caused small decreases (≤25%) in osteoblast number, suggesting a role for endogenous ATP in stimulating cell proliferation. Continuous apyrase treatment for 14 days (≥0.5U/ml) increased mineralisation of bone nodules by up to 3-fold. Increases in bone mineralisation were also seen when osteoblasts were cultured with the ATP release inhibitors, NEM and brefeldin A, as well as with P2X1 and P2X7 receptor antagonists. Apyrase decreased alkaline phosphatase (TNAP) activity by up to 60%, whilst increasing the activity of the PPi-generating ecto-nucleotide pyrophosphatase/phosphodiesterases (NPPs) up to 2.7-fold. Both collagen production and adipocyte formation were unaffected. These data suggest that nucleotides released by osteoblasts in bone could act locally, via multiple mechanisms, to limit mineralisation

Identification of plasma lipid biomarkers for prostate cancer by lipidomics and bioinformatics

Background: Lipids have critical functions in cellular energy storage, structure and signaling. Many individual lipid molecules have been associated with the evolution of prostate cancer; however, none of them has been approved to be used as a biomarker. The aim of this study is to identify lipid molecules from hundreds plasma apparent lipid species as biomarkers for diagnosis of prostate cancer. Methodology/Principal Findings: Using lipidomics, lipid profiling of 390 individual apparent lipid species was performed on 141 plasma samples from 105 patients with prostate cancer and 36 male controls. High throughput data generated from lipidomics were analyzed using bioinformatic and statistical methods. From 390 apparent lipid species, 35 species were demonstrated to have potential in differentiation of prostate cancer. Within the 35 species, 12 were identified as individual plasma lipid biomarkers for diagnosis of prostate cancer with a sensitivity above 80%, specificity above 50% and accuracy above 80%. Using top 15 of 35 potential biomarkers together increased predictive power dramatically in diagnosis of prostate cancer with a sensitivity of 93.6%, specificity of 90.1% and accuracy of 97.3%. Principal component analysis (PCA) and hierarchical clustering analysis (HCA) demonstrated that patient and control populations were visually separated by identified lipid biomarkers. RandomForest and 10-fold cross validation analyses demonstrated that the identified lipid biomarkers were able to predict unknown populations accurately, and this was not influenced by patient's age and race. Three out of 13 lipid classes, phosphatidylethanolamine (PE), ether-linked phosphatidylethanolamine (ePE) and ether-linked phosphatidylcholine (ePC) could be considered as biomarkers in diagnosis of prostate cancer. Conclusions/Significance: Using lipidomics and bioinformatic and statistical methods, we have identified a few out of hundreds plasma apparent lipid molecular species as biomarkers for diagnosis of prostate cancer with a high sensitivity, specificity and accuracy

Generation of peptide-specific cytotoxic T cells and presence of regulatory T cells during vaccination with hTERT (class I and II) peptide-pulsed DCs

Optimal techniques for DC generation for immunotherapy in cancer are yet to be established. Study aims were to evaluate: (i) DC activation/maturation milieu (TNF-α +/- IFN-α) and its effects on CD8+ hTERT-specific T cell responses to class I epitopes (p540 or p865), (ii) CD8+ hTERT-specific T cell responses elicited by vaccination with class I alone or both class I and II epitope (p766 and p672)-pulsed DCs, prepared without IFN-α, (iii) association between circulating T regulatory cells (Tregs) and clinical responses

Spondin-2 (SPON2), a More Prostate-Cancer-Specific Diagnostic Biomarker

BACKGROUND: Prostate-specific antigen (PSA) screening, although common, has recently been called into question. To find prostate cancer (PCa) diagnostic biomarkers that can make up for the defects of PSA, we compared the secretomes of several benign and PCa cell lines, selected candidate molecules, and then confirmed their clinical value. METHODOLOGY/PRINCIPAL FINDINGS: We first identified extracellular proteins by two-dimensional gel electrophoresis (2-DE) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) identification. We then validated the secreted proteins on a cellular level, and finally determined whether they could be used as PCa diagnostic biomarkers using prostate tissue and serum specimens of Chinese volunteers by immunohistostaining and sandwich ELISA. We obtained credible extracellular protein 2-DE graphs of prostate cell lines. The 5 spots that showed superior repeatability were selected for LC-MS/MS analysis, which identified seven candidate molecules. One of the candidate molecules, spondin-2 (SPON2), was only expressed in the conditioned media (CM) of androgen receptor (AR) positive PCa cell lines. Using tissue microarray by immunohistostaining, we found SPON2 to be over-expressed in PCa. SPON2 staining was more intense in Gleason score sum 7-8 and in PCa patients with metastasis. By receiver operator characteristic (ROC) curve analysis, we found that the serum SPON2 level was elevated in PCa patients, showing sensitivity and specificity suitable for diagnostic use. We also found that SPON2 could be used to identify PCa patients with serum PSA levels no higher than 10 ng/ml from healthy elderly men. CONCLUSION/SIGNIFICANCE: SPON2 is a new serum and histological diagnostic biomarker for PCa. It can avoid some of the problems of PSA testing and was here found to offer relatively high sensitivity and specificity relative to PSA

Central Role of Pyrophosphate in Acellular Cementum Formation

Background: Inorganic pyrophosphate (PPi) is a physiologic inhibitor of hydroxyapatite mineral precipitation involved in regulating mineralized tissue development and pathologic calcification. Local levels of PPi are controlled by antagonistic functions of factors that decrease PPi and promote mineralization (tissue-nonspecific alkaline phosphatase, Alpl/TNAP), and those that increase local PPi and restrict mineralization (progressive ankylosis protein, ANK; ectonucleotide pyrophosphatase phosphodiesterase-1, NPP1). The cementum enveloping the tooth root is essential for tooth function by providing attachment to the surrounding bone via the nonmineralized periodontal ligament. At present, the developmental regulation of cementum remains poorly understood, hampering efforts for regeneration. To elucidate the role of PPi in cementum formation, we analyzed root development in knock-out ((-/-)) mice featuring PPi dysregulation. Results: Excess PPi in the Alpl(-/-) mouse inhibited cementum formation, causing root detachment consistent with premature tooth loss in the human condition hypophosphatasia, though cementoblast phenotype was unperturbed. Deficient PPi in both Ank and Enpp1(-/-) mice significantly increased cementum apposition and overall thickness more than 12-fold vs. controls, while dentin and cellular cementum were unaltered. Though PPi regulators are widely expressed, cementoblasts selectively expressed greater ANK and NPP1 along the root surface, and dramatically increased ANK or NPP1 in models of reduced PPi output, in compensatory fashion. In vitro mechanistic studies confirmed that under low PPi mineralizing conditions, cementoblasts increased Ank (5-fold) and Enpp1 (20-fold), while increasing PPi inhibited mineralization and associated increases in Ank and Enpp1 mRNA. Conclusions: Results from these studies demonstrate a novel developmental regulation of acellular cementum, wherein cementoblasts tune cementogenesis by modulating local levels of PPi, directing and regulating mineral apposition. These findings underscore developmental differences in acellular versus cellular cementum, and suggest new approaches for cementum regeneration

ATLAS searches for additional scalars and exotic Higgs boson decays with the LHC Run 2 dataset

This report reviews the published results of searches for possible additional scalar particles and exotic decays of the Higgs boson performed by the ATLAS Collaboration using up to 140 fb−1 of 13 TeV proton–proton collision data collected during Run 2 of the Large Hadron Collider. Key results are examined, and observed excesses, while never statistically compelling, are noted. Constraints are placed on parameters of several models which extend the Standard Model, for example by adding one or more singlet or doublet fields, or offering exotic Higgs boson decay channels. Summaries of new searches as well as extensions of previous searches are discussed. These new results have a wider reach or attain stronger exclusion limits. New experimental techniques that were developed for these searches are highlighted. Search channels which have not yet been examined are also listed, as these provide insight into possible future areas of exploration

Precise test of lepton flavour universality in \varvec{W}-boson decays into muons and electrons in \varvec{pp} collisions at \varvec{\sqrt{s}}=13\,\text {T}\text {e}\hspace{-1.00006pt}\text {V} with the ATLAS detector

Abstract: The ratio of branching ratios of the W boson to muons and electrons, $$R^{\,\mu /e}_W={{\mathcal {B}}(W\rightarrow \mu u )}$$ R W μ / e = B ( W → μ ν ) /$${{\mathcal {B}}(W\rightarrow e u )}$$ B ( W → e ν ) , has been measured using $$140\,\text{ fb}^{-1}\,$$ 140 fb - 1 of pp collision data at $$\sqrt{s}=13$$ s = 13 &nbsp;$$\text {T}\text {e}\hspace{-1.00006pt}\text {V}$$ Te V collected with the ATLAS detector at the LHC, probing the universality of lepton couplings. The ratio is obtained from measurements of the $$t\bar{t}$$ t t ¯ production cross-section in the ee, $$e\mu$$ e μ and $$\mu \mu$$ μ μ dilepton final states. To reduce systematic uncertainties, it is normalised by the square root of the corresponding ratio $$R^{\,\mu \mu /ee}_Z$$ R Z μ μ / e e for the Z boson measured in inclusive $$Z\rightarrow ee$$ Z → e e and $$Z\rightarrow \mu \mu$$ Z → μ μ events. By using the precise value of $$R^{\,\mu \mu /ee}_Z$$ R Z μ μ / e e determined from $$e^+e^-$$ e + e - colliders, the ratio $$R^{\,\mu /e}_W$$ R W μ / e is determined to be $$\begin{aligned} R^{\,\mu /e}_W&amp;= 0.9995\pm 0.0022\,\mathrm {(stat)}\,\pm 0.0036\,\mathrm {(syst)}\\ &amp;\quad \pm 0.0014\,\mathrm {(ext)} . \end{aligned}$$ R W μ / e = 0.9995 ± 0.0022 ( stat ) ± 0.0036 ( syst ) ± 0.0014 ( ext ) . The three uncertainties correspond to data statistics, experimental systematics and the external measurement of $$R^{\,\mu \mu /ee}_Z$$ R Z μ μ / e e , giving a total uncertainty of 0.0045, and confirming the Standard Model assumption of lepton flavour universality in W-boson decays at the 0.5% level

Search for diphoton resonances in the 66 to 110 GeV mass range using pp collisions at s = 13 TeV with the ATLAS detector

Abstract : A search is performed for light, spin-0 bosons decaying into two photons in the 66 to 110 GeV mass range, using 140 fb −1 of proton-proton collisions at $$\sqrt{s}$$ s = 13 TeV produced by the Large Hadron Collider and collected by the ATLAS detector. Multivariate analysis techniques are used to define event categories that improve the sensitivity to new resonances beyond the Standard Model. A model-independent search for a generic spin-0 particle and a model-dependent search for an additional low-mass Higgs boson are performed in the diphoton invariant mass spectrum. No significant excess is observed in either search. Mass-dependent upper limits at the 95% confidence level are set in the model-independent scenario on the fiducial cross-section times branching ratio into two photons in the range of 8 fb to 53 fb. Similarly, in the model-dependent scenario upper limits are set on the total cross-section times branching ratio into two photons as a function of the Higgs boson mass in the range of 19 fb to 102 fb

Differential cross-section measurements of Higgs boson production in the H → τ+τ− decay channel in pp collisions at s = 13 TeV with the ATLAS detector

Abstract : Differential measurements of Higgs boson production in the τ-lepton-pair decay channel are presented in the gluon fusion, vector-boson fusion (VBF), VH and $$t\overline{t}H$$ t t ¯ H associated production modes, with particular focus on the VBF production mode. The data used to perform the measurements correspond to 140 fb −1 of proton-proton collisions collected by the ATLAS experiment at the LHC. Two methods are used to perform the measurements: the Simplified Template Cross-Section (STXS) approach and an Unfolded Fiducial Differential measurement considering only the VBF phase space. For the STXS measurement, events are categorized by their production mode and kinematic properties such as the Higgs boson’s transverse momentum ( $${p}_{\textrm{T}}^{\textrm{H}}$$ p T H ), the number of jets produced in association with the Higgs boson, or the invariant mass of the two leading jets (m jj ). For the VBF production mode, the ratio of the measured cross-section to the Standard Model prediction for m jj &gt; 1.5 TeV and $${p}_{\textrm{T}}^{\textrm{H}}$$ p T H &gt; 200 GeV ( $${p}_{\textrm{T}}^{\textrm{H}}$$ p T H &lt; 200 GeV) is $${1.29}_{-0.34}^{+0.39}$$ 1.29 − 0.34 + 0.39 ( $${0.12}_{-0.33}^{+0.34}$$ 0.12 − 0.33 + 0.34 ). This is the first VBF measurement for the higher- $${p}_{\textrm{T}}^{\textrm{H}}$$ p T H criteria, and the most precise for the lower- $${p}_{\textrm{T}}^{\textrm{H}}$$ p T H criteria. The fiducial cross-section measurements, which only consider the kinematic properties of the event, are performed as functions of variables characterizing the VBF topology, such as the signed ∆ϕ jj between the two leading jets. The measurements have a precision of 30%–50% and agree well with the Standard Model predictions. These results are interpreted in the SMEFT framework, and place the strongest constraints to date on the CP-odd Wilson coefficient $${c}_{H\overset{\sim }{W}}$$ c H W ~

Search for a resonance decaying into a scalar particle and a Higgs boson in the final state with two bottom quarks and two photons in proton–proton collisions at s = 13 TeV with the ATLAS detector

A search for the resonant production of a heavy scalar X decaying into a Higgs boson and a new lighter scalar S, through the process X → S(→bb¯)H(→γγ), where the two photons are consistent with the Higgs boson decay, is performed. The search is conducted using an integrated luminosity of 140 fb−1 of proton-proton collision data at a centre-of-mass energy of 13 TeV recorded with the ATLAS detector at the Large Hadron Collider. The search is performed over the mass range 170 ≤ mX ≤ 1000 GeV and 15 ≤ mS ≤ 500 GeV. Parameterised neural networks are used to enhance the signal purity and to achieve continuous sensitivity in a domain of the (mX, mS) plane. No significant excess above the expected background is found and 95% CL upper limits are set on the cross section times branching ratio, ranging from 39 fb to 0.09 fb. The largest deviation from the background-only expectation occurs for (mX, mS) = (575, 200) GeV with a local (global) significance of 3.5 (2.0) standard deviations