bluuebunny/crossref_metadata_2025_split huggingface dataset:
An Investigation of the Effects of Isothermal Exposure on Microstructural Evolution and Oxidation in a Thermal Barrier Coating
Authors: Chang T. , Mercer C. , Walter M. E. , Soboyejo W. O. 
Published: November 2021

Abstract
This paper presents the results of a systematic investigation of the effects of isothermal exposure on the microstructural evolution and the isothermal oxidation behavior of a thermal barrier coating with a platinum aluminide bond coat. Sintering of the columnar partially stabilized zirconia is shown to occur during isothermal exposure at 1150°C for durations up to 200 hours. This is accompanied by roughening of the TGO layers, the formation of hemi-spheroidal defects, parabolic coarsening of the TGO and final pull-out of the zirconia layers. The implications of the results are discussed for the analysis and design of TBCs for aeroengine applications.
Measurements of Decomposed EPS Gases Pressure and Molten Metal-Polymeric Foam Interface Velocity During Lost Foam Casting Process: Part 1—Disconnect Casting
Authors: Bakhtiyarov Sayavur I. , Overfelt Ruel A. 
Published: November 2021

Abstract
Process parameters of lost foam casting were studied experimentally on a commercial scale for two casting techniques: conventional and vacuum assisted gravity castings. The paper presents the results of the foundry experiments focused on the time and spatial dependent aspects of mold filling of a commercial pattern (Disconnect). A small-scale commercial model was designed and fabricated to conduct the experiments to study the thermo- and hydrodynamics of the mold filling characteristics. A high sensitivity linear output capacitive sensor was used to record the rate of sand flow and compaction in flask while horizontal and/or vertical vibrations were applied. A computer aided thermometric technique was used to study the heat-related processes in the counter-gravity lost foam vacuum casting. A complete pattern of vacuum/pressure fluctuations is obtained during counter-gravity vacuum casting using bi-directional vacuum/pressure transducer. Tolerance analyses were performed for styrofoam patterns and their cast products produced by the conventional and vacuum assisted casting procedures.
Centennial to millennial variability of greenhouse climate across the mid-Cenomanian event
Authors: Ma Chao , Hinnov Linda A. , Eldrett James S. , Meyers Stephen R. , Bergman Steven C. , Minisini Daniel , Lutz Brendan 
Published: November 2021

Abstract
Centennial- to millennial-scale climate variations are often attributed to solar forcing or internal climate system variability, but recognition of such variations in the deep-time paleoclimate record is extremely rare. We present an exceptionally well-preserved, millimeter-scale laminated marlstone from a succession of precession-driven limestone-marlstone couplets deposited in the Western Interior Seaway (North America) immediately preceding and during the Cretaceous mid-Cenomanian event (ca. 96.5 Ma). Sedimentological, geochemical, and micropaleontological data indicate that individual pairs of light-dark laminae record alternations in the extent of water-column mixing and oxygenation. Principal component analysis of X-ray fluorescence element counts and a grayscale scan of a continuous thin section through the marlstone reveal variations with 80–100 yr, 200–230 yr, 350–500 yr, ∼1650 yr, and 4843 yr periodicities. A substantial fraction of the data indicates an anoxic bottom water variation with a pronounced 10,784 yr cycle. The centennial to millennial variations are reminiscent of those found in Holocene total solar irradiance variability, and the 10,784 yr anoxia cycle may be a manifestation of semi-precession-influenced Tethyan oxygen minimum zone waters entering the seaway.
Evaluating preservation bias in the continental growth record against the monazite archive
Authors: Mulder Jacob A. , Cawood Peter A. 
Published: November 2021

Abstract
Most recent models of continental growth are based on large global compilations of detrital zircon ages, which preserve a distinctly episodic record of crust formation over billion-year timescales. However, it remains unclear whether this uneven distribution of zircon ages reflects a true episodicity in the generation of continental crust through time or is an artifact of the selective preservation of crust isolated in the interior of collisional orogens. We address this issue by analyzing a new global compilation of monazite ages (n &gt;100,000), which is comparable in size, temporal resolution, and spatial distribution to the zircon continental growth record and unambiguously records collisional orogenesis. We demonstrate that the global monazite and zircon age distributions are strongly correlated throughout most of Earth history, implying a link between collisional orogenesis and the preserved record of continental growth. Our findings support the interpretation that the continental crust provides a preservational, rather than generational, archive of crustal growth.
21 Neutrophil extracellular traps (NETs) can be measured non-invasively with CPa9-HNE – biomarker potential across different solid tumor types in the immuno-oncology setting
Authors: Jensen Christina , Thorlacius-Ussing Jeppe , Mortensen Joachim , Karsdal Morten , Willumsen Nicholas 
Published: November 2021

BackgroundSerological predictive biomarkers to anti-PD-1 therapy across different solid tumor types represent an unmet medical need. High neutrophil activity is associated with poor response to immune checkpoint inhibitor therapy, amongst others due to the release of neutrophil extracellular traps (NETs) that have been shown to protect tumor cells against cytotoxic attacks. Human neutrophil elastase (HNE) and calprotectin are the major NETs constituents. We have previously shown that high levels of a serum calprotectin assay, which quantifies a specific HNE generated calprotectin fragment (CPa9-HNE), as a measure of neutrophil activity, associate with poor response to anti-PD-1 therapy in metastatic melanoma patients. The aim of this study was to investigate the performance of CPa9-HNE in serum from patients with other solid tumor types.MethodsHNE mediated degradation of calprotectin (CPa9-HNE) was measured in pretreatment serum from 223 patients with bladder cancer (n=20), breast cancer (n=20), colorectal cancer (n=20), head and neck cancer (n=20), kidney cancer (n=20), liver cancer (n=3), lung cancer (n=20), melanoma (n=20), ovarian cancer (n=20), pancreatic cancer (n=20), prostate cancer (n=20), and stomach cancer (n=20), and compared to age-matched healthy controls (n=33). Statistical differences were analyzed using the Kruskal-Wallis test adjusted for Dunn’s multiple comparisons test.ResultsCPa9-HNE serum levels were significantly elevated in patients with bladder cancer (p<0.0001), breast cancer (p<0.0001), colorectal cancer (p<0.0001), head and neck cancer (p<0.0001), kidney cancer (p<0.0001), lung cancer (p<0.0001), melanoma (p=0.0001), ovarian cancer (p<0.0001), pancreatic cancer (p<0.0001), prostate cancer (p=0.0009), and stomach cancer (p<0.0001) compared to healthy controls. In all solid tumor types, a large patient-to-patient variation was observed. CPa9-HNE was also significantly elevated in stage IV cancer patients compared to stage I cancer patients (p=0.044).ConclusionsThe novel serological biomarker CPa9-HNE measuring HNE mediated degradation of calprotectin is significantly elevated in patients with different solid tumor types suggesting that these patients have high neutrophil activity. As high neutrophil activity is associated with poor response to anti-PD-1 therapy, CPa9-HNE may be used as a biomarker for anti-PD-1 therapy across different solid tumor types.Ethics ApprovalThe serum samples in this study was obtained from the commercial vendor Proteogenex and BioIVT, and according to the vendors, sample collection was approved by an Institutional Review Board or Independent Ethical Committee and patients gave their informed consent (Protocol numbers PG-ONC 2003/1 and WIRB® Protocol #20161665). All investigations were carried out according to the Helsinki Declaration.
41 Assessment of the spatial distribution of CD4+ T cells subpopulations in the tumor microenvironment by Brightplex®, a sequential chromogenic multiplex assay
Authors: Collignon Aurelie , Trinh Alex , Olive Marion , Jaume Clémence , Chamourin Maïté , Sfeir Nour , Anselmo Dylan , Ramouz-Charpentier Raana , Culey Georgia , Haond Christophe , Galon Jerome , Fieschi-Meric Jacques 
Published: November 2021

BackgroundCancer immunotherapy reinvigorates tumor-specific T cell responses of CD8+ cytotoxic T lymphocytes that detect intracellular antigens that are presented by MHC class I molecules expressed by all tumor cell types. Because most tumors do not express MHC class II, the potential antitumor protective role of CD4 T cells, which bind MHC class II molecules on target cells, has been less studied. However, CD4+ T cells are also required for efficacious antitumor immunity; they are core components of adaptive immunity that differentiate into lineages responsible for effector activities. Both TH1 and TH2 cell types mediate antitumor immunity, although TH1 cells may be more potent due to the production of large amounts of IFN-γ, as well as chemokines that enhance the priming and expansion of CD8 cells. TH1 cells help in recruiting natural killer cells and type I macrophages to tumor sites, which can act in concert toward tumor eradication. The ability of TH2 cells to mobilize innate cells, may represent a general pathway for their impact on the host antitumor response. Tumor infiltrating TFH cells play a key role in immune cell recruitment to the tumor and in the formation of intratumoral follicular structures, which correlate with a positive prognosis. On the contrary, cells from the TH17 subset induces inflammatory responses resulting in a tumor-promoting environment. CD4+ Tregs which are critically important for the maintenance of self-tolerance, impede effective immunity against the tumor when they are present in the tumor microenvironment (TME).Therefore, beyond the detection of total CD4+ T cells within the TME, it is of critical importance to determine to which subpopulation each CD4+ T cell belongs to decipher their roles in tumor rejection.MethodsWe have developed a multiplex 7-plex panel of of antibodies against biomarkers to identify main types of CD4+ T cellsResultsOn a single FFPE tissue section, main types of CD4+ T cells: TH1, TH2, TFH, TH17 and Tregs are identified by a combination of antibodies against transcription factors and membrane proteins. Following images registration, complex cells phenotypes can be detected and quantified. Furthermore, digital pathology tools allow the evaluation of the spatial distribution of CD4+ T cells within the TME.ConclusionsThis new tool unravels the diversity of CD4+ T cells in TME and could help clinical researchers to design more effective immunotherapies in cancer treatment. Integrated into an Immunogram, this new Brightplex® Panel will also be critical to understand the immune contexture of tumors.
49 Highly multiplexed detection of critical immune checkpoints and immune cell subtypes in cancerous FFPE tissues using CODEX
Authors: Shang Olive , Gordon Judi , Nikulina Nadya , Mistry Sejal , Singh Jasmine , Zong Hailing , Yuan Jessica , Blackmer Trillium , Locke Darren , Braubach Oliver , Kennedy-Darling Julia , Miller Peter 
Published: November 2021

BackgroundThere is growing consensus that spatial biology is the key to unlocking the underlying mechanisms of cancer immunotherapy and to predicting patient outcomes. Indeed, a recent example using the Akoya Phenoptics technology revealed a unique phenotypic signature of CD8/Foxp3 positive cells embedded within the tumor microenvironment of patients that responded favorably to PD-1 checkpoint inhibition.1 In this case, the combination of both multiparameter and spatial readouts was required to correlate significantly with outcome. As the number of treatment options expands and knowledge regarding cell types that contribute to treatment mechanisms improves, so too do the number of markers required to analyze responses that enable discovery of new signatures.MethodsHere, we present data from the analysis of human FFPE cancer tissues using an expanded CODEX antibody catalog targeting a variety of immune, immune checkpoint and transcription factors. CODEX enables the highly multiplexed detection of more than 40 targets within the same tissue sample, with single cell resolution and without degradation of the sample.ResultsOur expanded target list enables detection of key macrophage populations, T and B cell subtypes, granulocytes, dendritic cells, natural killer cells, stromal, tumor and epithelial cells. Additionally, the activation state of these immune and tumor cell types can be measured through detection of key immune checkpoints, including PD-1 and PD-L1. Through the addition of these critical markers, both cells known to contribute to treatment outcome and new biomarker signatures can be identified.ConclusionsContinued expansion of spatial biology discovery capabilities will be critical to continuing to improve patient outcomes and to develop new treatment options of solid tumors using cancer immunotherapies.ReferenceBerry S, Taube J, et al. Analysis of multispectral imaging with the AstroPath platform informs efficacy of PD-1 blockade. Science. 2021; 372: 6547.
78 Novel RNA-seq platform improve patient outcome in clinical oncology and enable implementation of AI in the clinic
Authors: Nguyen Vy , Pedersen Gitte , Pedersen Morten 
Published: November 2021

BackgroundOnly 1 out of 4 cancer treatments prolongs life while expenditure for cancer treatment is greater than $100 billion/year. RNA-sequencing has allowed researchers to gain insight into the transcriptome of human cancers. However, RNA-sequencing remains widely unused in clinical oncology. We address this issue through the development and CLIA validation of OneRNA—an RNA-sequencing platform for cancer diagnostics and the design of new treatments. The development of OneRNA had to overcome the two main hurdles for implementation of RNA sequencing in the clinic: 1) clinical samples are typically embedded in FFPE which results in highly fragmented RNA making sequencing of these samples difficult. 2) how to interpret aberrant gene expression events and translate these results into clinical action. We demonstrate how OneRNA® would enable the design of sophisticated combinatorial clinical studies. An example is combining immune targeting agents such as checkpoint inhibitors with mRNA vaccines. OneRNA also supports the integration of gene expression algorithms because of its ability to interrogate the entire sample transcriptome. OneRNA® has been CLIA certified using FFPE, FF, blood, and saliva samples. Furthermore, the sample preparation method has demonstrated >95% concordance between FF and FFPE and 5–10X the sensitivity compared to Truseq.MethodsThis study aims to demonstrate the clinical utility of OneRNA in detecting aberrant gene expression events and connecting these to already approved drugs that targets these events to offer truly individualized treatment options.ResultsWe show that OneRNA has the ability to predict results for not only validated biomarkers used in standard of care such as ER, PR and HER2 in breast cancer, but also provide insight into biomarkers for response to already approved drugs independent of tissue type and with no standard test. Finally, we demonstrate the reproducibility of OneRNA in predicting IHC status in ER, PR and HER2.ConclusionsThese results demonstrate that OneRNA has applications in both cancer research, drug discovery and development, development of companion diagnostic algorithms and implementation of truly individualized treatment.
230 Preclinical efficacy of CLEC-1 antagonist as novel myeloid immune checkpoint therapy for oncology
Authors: Gauttier Vanessa , Drouin Marion , Pengam Sabrina , Saenz Javier , Evrard Bérangère , Neyton Stéphanie , Mary caroline , Teppaz Géraldine , Desselle Ariane , Thépénier Virginie , Wilhelm Emmanuelle , Poirier Nicolas , Chiffoleau Elise 
Published: November 2021

BackgroundC-type lectin receptors (CLRs) are powerful pattern recognition receptors shaping immune cell-mediated tissue damage by positively or negatively regulating myeloid cell functions and hence tumor elimination or evasion. We previously reported that the orphan CLR CLEC-1 expressed by dendritic cells (DCs) tempers T cell’s responses in vivo by limiting antigen cross-presentation by cDC1. Furthermore, we observed that CLEC-1 is highly expressed by myeloid cells purified from human tumor microenvironment, in particular tumor-associated macrophages.MethodsMacrophages were generated from monocytes of healthy volunteers for phagocytosis assays. MC38 and Hepa 1.6 murine tumor cells were implanted in Clec1a KO or KI mice for immunotherapeutic treatment evaluation.ResultsUsing newly developed anti-human CLEC-1 monoclonal antibodies (mAbs), we found that antagonist anti-CLEC-1 mAbs with the capacity to block CLEC-1/CLEC-1Ligand interaction, as opposed to non-antagonist CLEC-1 mAbs, increase the phagocytosis of CLEC-1Ligand-positive human tumor cells by human macrophages, in particular when opsonized by tumor-associated antigen mAbs (Rituximab, Cetuximab, Trastuzumab) or with anti-CD47 mAb (Magrolimab). In-vivo, CLEC-1 knock-out (KO) mice (n=19) display significant prolonged survival in monotherapy as compared to wild-type littermates (n=12) in an orthotopic hepatocellular carcinoma (HCC) model and anti-tumor memory responses was demonstrated by tumor rechallenge in cured mice. CLEC1 KO mice also illustrate significant eradication of MC38 colorectal tumors in combination with chemotherapy promoting CLEC-1Ligand expression by tumor cells (n=16 with Gemcitabine or n=11 with Cyclophosphamide). HCC tumor microenvironment analysis after 2 weeks of tumor implantation shows significantly higher number of CD8+ and memory CD8+ T cells with reduced PD1 expression in CLEC1 KO animals (n=16 versus n=12 for KO vs WT mice respectively). Finally, we recently generated human CLEC-1 knock-in mice expressing the extracellular human CLEC1 domain fused to the intracellular mouse CLEC1 tail and confirmed preclinical efficacy in vivo with anti-human CLEC1 antagonist mAb in monotherapy in the orthotopic HCC model.ConclusionsThese data illustrate that CLEC-1 inhibition represents a novel therapeutic target for immuno-oncology modifying T cell immune responses and tumor cell phagocytosis by macrophages.
279 ILT3 blockade reduces tumor blast cells in acute myeloid leukemia PBMC and inhibits tumor cell growth in a humanized AML tumor model
Authors: Byford Alan , Qu Yujie , Xu Haiyan , Robinson Gain , Singh Latika , Muise Eric , Pinheiro Elaine , Chen Mei , Moy Lily , Alves Stephen , Zhang-Hoover Jie 
Published: November 2021

BackgroundImmunoglobulin-like transcript 3 (ILT3), an immune-inhibitory receptor expressed on myeloid cells, is highly expressed in acute myeloid leukemia (AML) with monocytic differentiation (M4 and M5 subtypes) and clinically is negatively correlated with overall survival. We examined the effects of a highly selective and potent chimeric anti-ILT3 mAb (c52B8) on tumor cell survival/growth and human immune cell modulation in AML PBMC and AML tumor cell lines in pre-clinical in vitro and in vivo models.MethodsILT3 mRNA and cell surface expression levels were measured by RNAseq and FACS, respectively. PBMC from AML patients with different levels of ILT3 cell surface expression were treated with c52B8 in vitro.A systemic AML model was generated by IV inoculation of NSG mice with luciferase-transfected MV-4-11 human AML cells, with or without human PBMC engraftment. Tumor bearing mice were treated weekly with c52B8, modified to contain the human Fc of either the IgG1 or IgG4 subclass. MV-4-11 growth in vivo was measured by bioluminescence imaging (BLI).CyTOF was performed to phenotype AML PBMC in in vitro culture and bone marrow samples from the systemic MV-4-11 model after the treatment.THP-1 cells/human T cell co-cultures stimulated with anti-CD3/CD28 antibodies were treated with c52B8. Human T cell activity was assessed by levels of IFNγ production in the culture using MSD ELISA.ResultsIn ILT3 high AML patient PBMC, c52B8 treatment in vitro decreased the frequency of tumor blasts, modulated Tregs, and increased monocytic myeloid cells. In the MV-4-11 model, weekly treatment with c52B8 significantly reduced tumor burden in vivo, regardless of the IgG subclass. Importantly, tumor cell growth inhibition by c52B8 occurred in the absence of human PBMC. Additional post-mortem CyTOF analysis of bone marrow cells from MV-4-11/human PBMC inoculated mice confirmed a reduction in MV-4-11 cells, identified by CD3-CD19- human myeloid cells following c52B8 treatment. In THP-1/human T cell co-cultures, c52B8 treatment reversed THP-1 induced T cell suppression, as measured by enhanced IFNγ production.ConclusionsAnti-ILT3 mAb (c52B8) is efficacious in blocking progression of AML with monocytic differentiation by directly influencing tumor cell growth and enhancing human T cell activity in pre-clinical models. These studies support a potential role for this compound in humans.Ethics ApprovalAll animal work was reviewed and approved by Merck IACUC before experiments were conducted.
FGFR3 Destabilizes PD-L1 via NEDD4 to Control T-cell–Mediated Bladder Cancer Immune Surveillance
Authors: Jing Weiqiang , Wang Ganyu , Cui Zhiwei , Xiong Gaozhong , Jiang Xin , Li Yue , Li Wushan , Han Bo , Chen Shouzhen , Shi Benkang 
Published: November 2021

Abstract

Fibroblast growth factor receptor 3 (FGFR3) is frequently activated by mutation or overexpression, and it is a validated therapeutic target in urothelial carcinoma (UC) of the bladder. However, the role and detailed molecular mechanism of FGFR3 in the immune microenvironment of bladder cancer remain largely unknown. Here, we demonstrate that inhibition of FGFR3 in FGFR3-activated bladder cancer elevates PD-L1 protein levels by affecting its ubiquitination, thereby inhibiting the antitumor activity of CD8+ T cells. Tissue microarray analysis in human UC showed an inverse correlation between FGFR3 and PD-L1. Furthermore, NEDD4, an E3 ubiquitin ligase of the NEDD4 family of proteins, was phosphorylated by FGFR3 activation and served as a regulator of PD-L1 ubiquitination. Mechanistically, NEDD4 interacted with PD-L1 and catalyzed Lys48 (K48)-linked polyubiquitination of PD-L1. In mice bearing NEDD4 knockout bladder cancer, CD8+ T-cell infiltration and antitumor activity were significantly inhibited due to PD-L1 upregulation in bladder cancer cells. Furthermore, multiple FGFR3-activated tumor-bearing mouse models suggested that attenuated CD8+ T-cell–mediated antitumor efficacy following FGFR3-targeted therapy could be rescued by a combination with anti-PD-1 immunotherapy, which leads to effective tumor suppression. This study establishes a key molecular link between targeted therapy and immune surveillance and identifies NEDD4 as a crucial E3 ubiquitin ligase that targets PD-L1 for degradation in FGFR3-activated bladder cancer. These findings may potentially be exploited for combination therapies in UC of the bladder and possibly other malignancies with activated FGFR3.

Significance:
NEDD4 links two important molecules associated with targeted therapy and immune surveillance, providing mechanistic rationale and preclinical support for immuno-targeted combination therapy for FGFR3-activated bladder cancer.