bluuebunny/crossref_metadata_2025 huggingface dataset:
Volatile Organic Compounds Monitored Online at Three Photochemical Assessment Monitoring Stations in the Pearl River Delta (PRD) Region during Summer 2016: Sources and Emission Areas
Authors: Zhang Tao , Xiao Shaoxuan , Wang Xinming , Zhang Yanli , Pei Chenglei , Chen Duohong , Jiang Ming , Liao Tong
Published: March 2021

Volatile organic compounds (VOCs) were monitored online at three photochemical assessment monitoring stations (MDS, WQS and HGS) in the Pearl River Delta region during the summer of 2016. Measured levels of VOCs at the MDS, WQS and HGS sites were 34.78, 8.54 and 8.47 ppbv, respectively, with aromatics and alkenes as major ozone precursors and aromatics as major precursors to secondary organic aerosol (SOA). The positive matrix factorization (PMF) model revealed that VOCs at the sites mainly came from vehicle exhaust, petrochemical industry, and solvent use. Vehicle exhaust and industrial processes losses contributed most to ozone formation potentials (OFP) of VOCs, while industrial processes losses contributed most to SOA formation potentials of VOCs. Potential source contribution function (PSCF) analysis revealed a north-south distribution for source regions of aromatics occurring at MDS with emission sources in Guangzhou mainly centered in the Guangzhou central districts, and source regions of aromatics at WQS showed an east-west distribution across Huizhou, Dongguan and east of Guangzhou, while that at HGS showed a south-north distribution across Guangzhou, Foshan, Zhaoqing and Yangjiang. This study demonstrates that multi-point high time resolution data can help resolve emission sources and locate emission areas of important ozone and SOA precursors.
Air Pollutant Analysis and AQI Prediction Based on GRA and Improved SOA-SVR by Considering COVID-19
Authors: Xu Ting , Yan Huichao , Bai Yanping
Published: March 2021

Since COVID-19 pneumonia broke out, the Chinese government has taken a series of measures to control the spread of the epidemic, which has made the air quality of Taiyuan in February 2020 significantly better than during the same period in previous years. In this paper, the Gray Relational Analysis (GRA) method was first applied to evaluate and analyze the influence of six major pollutants on air quality. Then, the improved seagull optimization algorithm (ISOA) was proposed and combined with Support Vector Regression (SVR) to establish a hybrid predicted model ISOA-SVR. Finally, the proposed ISOA-SVR was utilized to predict air quality index (AQI). The experimental results on two kinds of different data showed that the proposed ISOA-SVR had the better generalization ability and robustness compared with other predicted models. Further, the proposed ISOA-SVR is suitable for the prediction of AQI.
Mesenchymal–Epithelial Transition in Fibroblasts of Human Normal Lungs and Interstitial Lung Diseases
Authors: Becerril Carina , Montaño Martha , Cisneros José , Mendoza-Milla Criselda , Pardo Annie , Ortiz-Quintero Blanca , Selman Moisés , Ramos Carlos
Published: March 2021

. In passages above ten and growing very actively, we observed that some human lung fibroblasts cultured under standard conditions were transformed into a lineage of epithelial-like cells (ELC). To systematically evaluate the possible mesenchymal–epithelial transition (MET) occurrence, fibroblasts were obtained from normal lungs and also from lungs affected by idiopathic interstitial diseases. When an unusual epithelial-like phenotypic change was observed, cultured cells were characterized by confocal immunofluorescence microscopy, immunoblotting, immunocytochemistry, cytofluorometry, gelatin zymography, RT-qPCR, and hybridization in a whole-transcript human microarray. Additionally, microvesicles fraction (MVs) from ELC and fibroblasts were used to induce MET, while the microRNAs (miRNAs) contained in the MVs were identified. Pattern-gene expression of the original fibroblasts and the derived ELC revealed profound changes, upregulating characteristic epithelial-cell genes and downregulating mesenchymal genes, with a marked increase of E-cadherin, cytokeratin, and ZO-1, and the loss of expression of α-SMA, collagen type I, and Thy-1 cell surface antigen (CD90). Fibroblasts, exposed to culture media or MVs from the ELC, acquired ELC phenotype. The miRNAs in MVs shown six expressed exclusively in fibroblasts, and three only in ELC; moreover, twelve miRNAs were differentially expressed between fibroblasts and ELC, all of them but one was overexpressed in fibroblasts. These findings suggest that the MET-like process can occur in human lung fibroblasts, either from normal or diseased lungs. However, the biological implication is unclear.
Angiogenesis Is Differentially Modulated by Platelet-Derived Products
Authors: Berndt Sarah , Carpentier Gilles , Turzi Antoine , Borlat Frédéric , Cuendet Muriel , Modarressi Ali
Published: March 2021

Platelet-derived preparations are being used in clinic for their role in tissue repair and regenerative processes. The release of platelet-derived products such as autologous growth factors, cytokines and chemokines can trigger therapeutic angiogenesis. In this in vitro study, we evaluated and compared the ability of three platelet-derived preparations: platelet-rich-plasma (PRP), PRP-hyaluronic acid (PRP-HA) and platelet lysates (PL) at various concentrations (5–40%) to modulate human umbilical vein endothelial cells (HUVEC) biological effects on metabolism, viability, senescence, angiogenic factors secretion and angiogenic capacities in 2D (endothelial tube formation assay or EFTA) and in 3D (fibrin bead assay or FBA). HUVEC exocytosis was stimulated with PRP and PRP-HA. Cell viability was strongly increased by PRP and PRP-HA but mildly by PL. The three preparations inhibit HUVEC tube formation on Matrigel, while PRP enhanced the complexity of the network. In the fibrin bead assay (FBA), PRP and PRP-HA stimulated all steps of the angiogenic process resulting in massive sprouting of a branched microvessel network, while PL showed a weaker angiogenic response. Secretome profiling revealed modulation of 26 human angiogenic proteins upon treatment with the platelet derived preparations. These in vitro experiments suggest that PRP and PRP-HA are effective biological therapeutic tools when sustained therapeutic angiogenesis is needed.
Blood Serum Cytokines in Patients with Subacute Spinal Cord Injury: A Pilot Study to Search for Biomarkers of Injury Severity
Authors: Ogurcov Sergei , Shulman Iliya , Garanina Ekaterina , Sabirov Davran , Baichurina Irina , Kuznetcov Maxim , Masgutova Galina , Kostennikov Alexander , Rizvanov Albert , James Victoria , Mukhamedshina Yana
Published: March 2021

Background. Despite considerable interest in the search for a spinal cord injury (SCI) therapy, there is a critical need to develop a panel of diagnostic biomarkers to determine injury severity. In this regard, there is a requirement for continuing research into the fundamental processes of neuroinflammatory and autoimmune reactions in SCI, identifying changes in the expression of cytokines. Methods. In this pilot study, an extended multiplex analysis of the cytokine profiles in the serum of patients at 2 weeks post-SCI (n = 28) was carried out, together with an additional assessment of neuron-specific enolase (NSE) and vascular endothelial growth factor (VEGF) levels by enzyme-linked immunosorbent assay. A total of 16 uninjured subjects were enrolled as controls. Results. The data obtained showed a large elevation of IFNγ (>52 fold), CCL27 (>13 fold), and CCL26 (>8 fold) 2 weeks after SCI. The levels of cytokines CXCL5, CCL11, CXCL11, IL10, TNFα, and MIF were different between patients with baseline American Spinal Injury Association Impairment Scale (AIS) grades of A or B, whilst IL2 (>2 fold) and MIP-3a (>6 fold) were significantly expressed in the cervical and thoracic regions. There was a trend towards increasing levels of NSE. However, the difference in NSE was lost when the patient set was segregated based on AIS group. Conclusions. Our pilot research demonstrates that serum concentrations of cytokines can be used as an affordable and rapid detection tool to accurately stratify SCI severity in patients.
Endoscopic Bilateral Optic Nerve Decompression for Treatment of Idiopathic Intracranial Hypertension
Authors: Göksu Ethem , Bozkurt Baran , İlhan Deniz , Özak Ahmet , Çırak Musa , Yağmurlu Kaan
Published: March 2021

Objective: To evaluate the results of bilateral endoscopic optic nerve decompression (EOND) with the opening nerve sheath (ONS) technique in patients with idiopathic intracranial hypertension (IIH). Methods: Between the years of 2017 and 2019, we retrospectively evaluated nine IIH patients with progressive visual impairment despite medical treatment and who were treated with the EOND and ONS techniques. We also demonstrated our surgical technique recipe on postmortem human heads in a stepwise manner. Results: There were 9 patients (7 females and 2 males) between the ages of 21 and 72 included in this study, and the mean age was 40.8. All patients had an impairment in visual acuity and/or their visual field, with signs of papilledema and/or optic atrophy. The patients were followed up with for 9–48 months. Improvements in visual acuity were observed in 7 out of 9 patients (78%). Visual field defects improved in 5 out of 8 patients (62.5%). Papilledema was resolved in all patients (100%). Headaches improved in all symptomatic patients (100%). No intraoperative or postoperative complications were observed. Conclusions: EOND is a safe and effective surgical procedure in selected patients with IIH. Bilateral wide bony decompression and nerve fenestration can also be an additional benefit for headache relief. Further clinical series and long-term follow-up are needed for more precise results.
Protective Effects of PGC-1α Activators on Ischemic Stroke in a Rat Model of Photochemically Induced Thrombosis
Authors: Shakova Fatima M. , Kirova Yuliya I. , Silachev Denis N. , Romanova Galina A. , Morozov Sergey G.
Published: March 2021

The pharmacological induction and activation of peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α), a key regulator of ischemic brain tolerance, is a promising direction in neuroprotective therapy. Pharmacological agents with known abilities to modulate cerebral PGC-1α are scarce. This study focused on the potential PGC-1α-modulating activity of Mexidol (2-ethyl-6-methyl-3-hydroxypyridine succinate) and Semax (ACTH(4–7) analog) in a rat model of photochemical-induced thrombosis (PT) in the prefrontal cortex. Mexidol (100 mg/kg) was administered intraperitoneally, and Semax (25 μg/kg) was administered intranasally, for 7 days each. The expression of PGC-1α and PGC-1α-dependent protein markers of mitochondriogenesis, angiogenesis, and synaptogenesis was measured in the penumbra via immunoblotting at Days 1, 3, 7, and 21 after PT. The nuclear content of PGC-1α was measured immunohistochemically. The suppression of PGC-1α expression was observed in the penumbra from 24 h to 21 days following PT and reflected decreases in both the number of neurons and PGC-1α expression in individual neurons. Administration of Mexidol or Semax was associated with preservation of the neuron number and neuronal expression of PGC-1α, stimulation of the nuclear translocation of PGC-1α, and increased contents of protein markers for PGC-1α activation. This study opens new prospects for the pharmacological modulation of PGC-1α in the ischemic brain.
Preconditioning Stimulus Intensity Alters Paired-Pulse TMS Evoked Potentials
Authors: Rawji Vishal , Kaczmarczyk Isabella , Rocchi Lorenzo , Fong Po-Yu , Rothwell John C. , Sharma Nikhil
Published: March 2021

Motor cortex (M1) paired-pulse TMS (ppTMS) probes excitatory and inhibitory intracortical dynamics by measurement of motor-evoked potentials (MEPs). However, MEPs reflect cortical and spinal excitabilities and therefore cannot isolate cortical function. Concurrent TMS-EEG has the ability to measure cortical function, while limiting peripheral confounds; TMS stimulates M1, whilst EEG acts as the readout: the TMS-evoked potential (TEP). Whilst varying preconditioning stimulus intensity influences intracortical inhibition measured by MEPs, the effects on TEPs is undefined. TMS was delivered to the left M1 using single-pulse and three, ppTMS paradigms, each using a different preconditioning stimulus: 70%, 80% or 90% of resting motor threshold. Corticospinal inhibition was present in all three ppTMS conditions. ppTMS TEP peaks were reduced predominantly under the ppTMS 70 protocol but less so for ppTMS 80 and not at all for ppTMS 90. There was a significant negative correlation between MEPs and N45 TEP peak for ppTMS 70 reaching statistical trends for ppTMS 80 and 90. Whilst ppTMS MEPs show inhibition across a range of preconditioning stimulus intensities, ppTMS TEPs do not. TEPs after M1 ppTMS vary as a function of preconditioning stimulus intensity: smaller preconditioning stimulus intensities result in better discriminability between conditioned and unconditioned TEPs. We recommend that preconditioning stimulus intensity should be minimized when using ppTMS to probe intracortical inhibition.
Design of Fear and Anxiety of COVID-19 Assessment Tool in Spanish Adult Population
Authors: Gómez-Salgado Juan , Allande-Cussó Regina , Domínguez-Salas Sara , García-Iglesias Juan Jesús , Coronado-Vázquez Valle , Ruiz-Frutos Carlos
Published: March 2021

The aim of this study was to develop a specific scale to measure anxiety and fear levels in the general Spanish population. For this, a transcultural adaptation to Spanish of the fear of coronavirus disease 2019 (COVID-19) scale, in its original version of 10 items, was carried out. Then, the Anxiety and Fear of COVID-19 Assessment Scale (AMICO, for its acronym in Spanish) was designed by translating the tool and Delphi technique into three rounds. Ten experts participated voluntarily, and inter-observer match rates and the reliability study of the designed scale were calculated. A pilot study was carried out with the final version of the scale for the validity and reliability study. The instrument did not raise problems in semantic and cultural terms during the first and second rounds of the translation process, with an overall weighted Kappa value of 0.9. In the third round, eight new items were designed and consensual, obtaining a weighted overall value of 0.89. The pilot study sample was made up of 445 subjects, of which 60.3% were women with a mean age of 46.2 years. The final version consisted of 16 items, 2 factors, and a Cronbach’s alpha value of 0.92. The AMICO scale was developed to assess the level of anxiety and fear of COVID-19 and proved to be valid and reliable for its use in the adult Spanish population.
Brain Plasticity Mechanisms Underlying Motor Control Reorganization: Pilot Longitudinal Study on Post-Stroke Subjects
Authors: Gandolla Marta , Niero Lorenzo , Molteni Franco , Guanziroli Elenora , Ward Nick S. , Pedrocchi Alessandra
Published: March 2021

Functional Electrical Stimulation (FES) has demonstrated to improve walking ability and to induce the carryover effect, long-lasting persisting improvement. Functional magnetic resonance imaging has been used to investigate effective connectivity differences and longitudinal changes in a group of chronic stroke patients that attended a FES-based rehabilitation program for foot-drop correction, distinguishing between carryover effect responders and non-responders, and in comparison with a healthy control group. Bayesian hierarchical procedures were employed, involving nonlinear models at within-subject level—dynamic causal models—and linear models at between-subjects level. Selected regions of interest were primary sensorimotor cortices (M1, S1), supplementary motor area (SMA), and angular gyrus. Our results suggest the following: (i) The ability to correctly plan the movement and integrate proprioception information might be the features to update the motor control loop, towards the carryover effect, as indicated by the reduced sensitivity to proprioception input to S1 of FES non-responders; (ii) FES-related neural plasticity supports the active inference account for motor control, as indicated by the modulation of SMA and M1 connections to S1 area; (iii) SMA has a dual role of higher order motor processing unit responsible for complex movements, and a superintendence role in suppressing standard motor plans as external conditions changes.
Modified Lime Binders for Restoration Work
Authors: Loganina Valentina , Sergeeva Kristina , Fediuk Roman , Klyuev Sergey , Vatin Nikolai , Vasilev Yuriy
Published: March 2021

Lime mixes are the primary material for restoration work in historic buildings. The research object of this study is modifying lime binders with specially synthesized calcium silicate hydrates (CSHs). This study aimed to improve lime render mixes’ weather resistance. The following factors were considered: the density of the liquid glass, the amount of the precipitating additive, the rate of introduction of the precipitating additive, the drying mode of the precipitate, and the storage time of the precipitate. The research methods were X-ray diffraction analysis, differential thermal analysis, Fourier transform infrared spectrometry, and optical and electron microscopy. It was revealed that lime compositions with CSH have a higher strength gain rate than the control compositions. A mathematical model of the kinetics of hardening a lime composite based on a binder filled with CSH was obtained. The regularities of the change in the lime composite’s strength depending on the filler grinding’s fineness, its content, and the amount of mixing water have been established. It was revealed that the introduction of CSH into the lime composition increases the weather resistance of facade lime mixtures by reducing the porosity and increasing the volume of closed pores of the composite.
Pancreatic Cancers with High Grade Tumor Budding Exhibit Hallmarks of Diminished Anti-Tumor Immunity
Authors: Sadozai Hassan , Acharjee Animesh , Gruber Thomas , Gloor Beat , Karamitopoulou Eva
Published: March 2021

Tumor budding is associated with epithelial-mesenchymal transition and diminished survival in a number of cancer types including pancreatic ductal adenocarcinoma (PDAC). In this study, we dissect the immune landscapes of patients with high grade versus low grade tumor budding to determine the features associated with immune escape and disease progression in pancreatic cancer. We performed immunohistochemistry-based quantification of tumor-infiltrating leukocytes and tumor bud assessment in a cohort of n = 111 PDAC patients in a tissue microarray (TMA) format. Patients were divided based on the ITBCC categories of tumor budding as Low Grade (LG: categories 1 and 2) and High Grade (HG: category 3). Tumor budding numbers and tumor budding grade demonstrated a significant association with diminished overall survival (OS). HG cases exhibit notably reduced densities of stromal (S) and intratumoral (IT) T cells. HG cases also display lower M1 macrophages (S) and increased M2 macrophages (IT). These findings were validated using gene expression data from TCGA. A published tumor budding gene signature demonstrated a significant association with diminished survival in PDAC patients in TCGA. Immune-related gene expression revealed an immunosuppressive TME in PDAC cases with high expression of the budding signature. Our findings highlight a number of immune features that permit an improved understanding of disease progression and EMT in pancreatic cancer.
From Liver Fat to Cancer: Perils of the Western Diet
Authors: Kim Ju Youn , He Feng , Karin Michael
Published: March 2021

Hepatocellular carcinoma (HCC), the most common type of primary liver cancer provides the prototypical example of an obesity-related cancer. The obesity epidemic gave rise to an enormous increase in the incidence of non-alcoholic fatty liver disease (NAFLD), a condition that affects one third of American adults. In about 20% of these individuals, simple liver steatosis (hepatosteatosis) progresses to non-alcoholic steatohepatitis (NASH) characterized by chronic liver injury, inflammation, and fibrosis. In addition to liver failure, NASH greatly increases the risk of HCC. Here we discuss the metabolic processes that control the progression from NAFLD to NASH and from NASH to HCC, with a special emphasis on the role of free-non-esterified cholesterol in the process.
Response to Immune Checkpoint Inhibitor Therapy in Patients with Unresectable Recurrent Malignant Pleural Mesothelioma Shown by FDG-PET and CT
Authors: Kitajima Kazuhiro , Maruyama Mitsunari , Yokoyama Hiroyuki , Minami Toshiyuki , Yokoi Takashi , Nakamura Akifumi , Hashimoto Masaki , Kondo Nobuyuki , Kuribayashi Kozo , Kijima Takashi , Hasegawa Seiki , Yamakado Koichiro
Published: March 2021

Background: To compare three FDG-PET criteria (EORTC, PERCIST, imPERCIST) with CT criteria (combined modified RECIST and RECIST 1.1) for response evaluation and prognosis prediction in patients with recurrent MPM treated with ICI monotherapy. Methods: Thirty MPM patients underwent FDG-PET/CT and contrast-enhanced CT at the baseline and during nivolumab therapy (median 10 cycles). Therapeutic response was evaluated according to EORTC, PERCIST, imPERCIST, and CT criteria. PFS and OS were examined using log-rank and Cox methods. Results: CMR/PMR/SMD/PMD numbered 5/3/4/18 for EORTC, 5/1/7/17 for PERCIST, and 5/3/9/13 for imPERCIST. With CT, CR/PR/SD/PD numbered 0/6/10/14. There was high concordance between EORTC and PERCIST (κ = 0.911), and PERCIST and imPERCIST (κ = 0.826), while that between EORTC and imPERCIST (κ = 0.746) was substantial, and between CT and the three PET criteria moderate (κ = 0.516–0.544). After median 14.9 months, 26 patients showed progression and nine died. According to both PET and CT findings, patients with no progression (CMR/PMR/SMD or CR/PR/SD) showed significantly longer PFS and somewhat longer OS than PMD and PD patients (EORTC p = 0.0004 and p = 0.055, respectively; PERCIST p = 0.0003 and p = 0.052; imPERCIST p < 0.0001 and p = 0.089; CT criteria p = 0.0015 and p = 0.056). Conclusions: Both FDG-PET and CT criteria are accurate for response evaluation of ICI therapy and prediction of MPM prognosis. In comparison with CT, all three FDG-PET/CT criteria judged a greater percentage of patients (16.7%) as CMR, while two (EORTC, PERCIST) judged a greater percentage (10–13.3%) as PMD. For predicting PFS, the three FDG-PET criteria were superior to the CT criteria, and imPERCIST demonstrated the highest rate of accurate prediction.
Impact of Carbon Ion Radiotherapy on Inoperable Bone Sarcoma
Authors: Shiba Shintaro , Okamoto Masahiko , Kiyohara Hiroki , Okazaki Shohei , Kaminuma Takuya , Shibuya Kei , Kohama Isaku , Saito Kenichi , Yanagawa Takashi , Chikuda Hirotaka , Nakano Takashi , Ohno Tatsuya
Published: March 2021

Management of patients with bone sarcoma who are unsuitable for surgery is challenging. We aimed to analyze the clinical outcomes among such patients who were treated with carbon ion radiotherapy (C-ion RT). We reviewed the medical records of the patients treated with C-ion RT between April 2011 and February 2019 and analyzed the data of 53 patients. Toxicities were classified using the National Cancer Institute’s Common Terminology Criteria for Adverse Events (Version 4.0). The median follow-up duration for all patients was 36.9 months. Histologically, 32 patients had chordoma, 9 had chondrosarcoma, 8 had osteosarcoma, 3 had undifferentiated pleomorphic sarcoma, and 1 had sclerosing epithelioid fibrosarcoma. The estimated 3-year overall survival (OS), local control (LC), and progression-free survival (PFS) rates were 79.7%, 88.6%, and 68.9%, respectively. No patients developed grade 3 or higher acute toxicities. Three patients developed both grade 3 radiation dermatitis and osteomyelitis, one developed both grade 3 radiation dermatitis and soft tissue infection, and one developed rectum-sacrum-cutaneous fistula. C-ion RT showed favorable clinical outcomes in terms of OS, LC, and PFS and low rates of toxicity in bone sarcoma patients. These results suggest a potential role for C-ion RT in the management of this population.
Cancer-Associated Fibroblasts as Players in Cancer Development and Progression and Their Role in Targeted Radionuclide Imaging and Therapy
Authors: Koustoulidou Sofia , Hoorens Mark W. H. , Dalm Simone U. , Mahajan Shweta , Debets Reno , Seimbille Yann , de Jong Marion
Published: March 2021

Cancer Associated Fibroblasts (CAFs) form a major component of the tumour microenvironment, they have a complex origin and execute diverse functions in tumour development and progression. As such, CAFs constitute an attractive target for novel therapeutic interventions that will aid both diagnosis and treatment of various cancers. There are, however, a few limitations in reaching successful translation of CAF targeted interventions from bench to bedside. Several approaches targeting CAFs have been investigated so far and a few CAF-targeting tracers have successfully been developed and applied. This includes tracers targeting Fibroblast Activation Protein (FAP) on CAFs. A number of FAP-targeting tracers have shown great promise in the clinic. In this review, we summarize our current knowledge of the functional heterogeneity and biology of CAFs in cancer. Moreover, we highlight the latest developments towards theranostic applications that will help tumour characterization, radioligand therapy and staging in cancers with a distinct CAF population.
Bleeding by Bruton Tyrosine Kinase-Inhibitors: Dependency on Drug Type and Disease
Authors: von Hundelshausen Philipp , Siess Wolfgang
Published: March 2021

Bruton tyrosine kinase (Btk) is expressed in B-lymphocytes, myeloid cells and platelets, and Btk-inhibitors (BTKi) are used to treat patients with B-cell malignancies, developed against autoimmune diseases, have been proposed as novel antithrombotic drugs, and been tested in patients with severe COVID-19. However, mild bleeding is frequent in patients with B-cell malignancies treated with the irreversible BTKi ibrutinib and the recently approved 2nd generation BTKi acalabrutinib, zanubrutinib and tirabrutinib, and also in volunteers receiving in a phase-1 study the novel irreversible BTKi BI-705564. In contrast, no bleeding has been reported in clinical trials of other BTKi. These include the brain-penetrant irreversible tolebrutinib and evobrutinib (against multiple sclerosis), the irreversible branebrutinib, the reversible BMS-986142 and fenebrutinib (targeting rheumatoid arthritis and lupus erythematodes), and the reversible covalent rilzabrutinib (against pemphigus and immune thrombocytopenia). Remibrutinib, a novel highly selective covalent BTKi, is currently in clinical studies of autoimmune dermatological disorders. This review describes twelve BTKi approved or in clinical trials. By focusing on their pharmacological properties, targeted disease, bleeding side effects and actions on platelets it attempts to clarify the mechanisms underlying bleeding. Specific platelet function tests in blood might help to estimate the probability of bleeding of newly developed BTKi.
Evaluation of Fast Molecular Detection of Lymph Node Metastases in Prostate Cancer Patients Using One-Step Nucleic Acid Amplification (OSNA)
Authors: Engels Svenja , Brautmeier Lutz , Reinhardt Lena , Wasylow Clara , Hasselmann Friederike , Henke Rolf P. , Wawroschek Friedhelm , Winter Alexander
Published: March 2021

Background: In clinical routine, only fractions of lymph nodes (LNs) are examined histopathologically, often resulting in missed (micro-)metastases and incorrect staging of prostate cancer (PCa). One-step nucleic acid amplification (OSNA) analyzes the entire LN by detecting cytokeratin 19 (CK19) mRNA as a surrogate for LN metastases requiring less effort than conventional biomolecular techniques. We aimed to evaluate performance of OSNA in detecting sentinel LN (SLN) metastases in PCa. Methods: SLNs (n = 534) of 64 intermediate- or high-risk PCa patients undergoing radical prostatectomy with extended and sentinel-guided lymphadenectomy were cut into slices and alternatingly assigned to OSNA and histopathology (hematoxylin-eosin staining, CK19, and CK AE1/AE3 immunohistochemistry). Sensitivity and specificity of OSNA and concordance and measure of agreement (Cohen’s kappa (κ)) between OSNA and histopathology were assessed. Results: Histopathology revealed metastases in 76 SLNs. Sensitivity and specificity of OSNA were 84.2% and 96.1%, respectively. Discordant results were recorded for 30 of 534 SLNs, revealing high concordance (94.4%). Twenty-four discordant cases were classified as micrometastases, indicating a possible allocation bias. In 18 cases, positive results were conferred only by OSNA resulting in seven LN-positive patients who were missed by histopathology. Overall, the level of agreement was high (κ = 0.78). Conclusions: OSNA provided a diagnosis that was as least as accurate as detailed histological examination and might improve LN staging in PCa.
Structural Basis for the Functional Changes by EGFR Exon 20 Insertion Mutations
Authors: Tamirat Mahlet Z. , Kurppa Kari J. , Elenius Klaus , Johnson Mark S.
Published: March 2021

Activating somatic mutations of the epidermal growth factor receptor (EGFR) are frequently implicated in non-small cell lung cancer (NSCLC). While L858R and exon 19 deletion mutations are most prevalent, exon 20 insertions are often observed in NSCLC. Here, we investigated the structural implications of two common EGFR exon 20 insertions in NSCLC, V769insASV and D770insNPG. The active and inactive conformations of wild-type, D770insNPG and V769insASV EGFRs were probed with molecular dynamics simulations to identify local and global alterations that the mutations exert on the EGFR kinase domain, highlighting mechanisms for increased enzymatic activity. In the active conformation, the mutations increase interactions that stabilize the αC helix that is essential for EGFR activity. Moreover, the key Lys745–Glu762 salt bridge was more conserved in the insertion mutations. The mutants also preserved the state of the structurally critical aspartate–phenylalanine–glycine (DFG)-motif and regulatory spine (R-spine), which were altered in wild-type EGFR. The insertions altered the structure near the ATP-binding pocket, e.g., the P-loop, which may be a factor for the clinically observed tyrosine kinase inhibitor (TKI) insensitivity by the insertion mutants. The inactive state simulations also showed that the insertions disrupt the Ala767–Arg776 interaction that is key for maintaining the “αC-out” inactive conformation, which could consequently fuel the transition from the inactive towards the active EGFR state.
Adapter Chimeric Antigen Receptor (AdCAR)-Engineered NK-92 Cells for the Multiplex Targeting of Bone Metastases
Authors: Grote Stefan , Traub Frank , Mittelstaet Joerg , Seitz Christian , Kaiser Andrew , Handgretinger Rupert , Schleicher Sabine
Published: March 2021

Background: Since metastatic spreading of solid tumor cells often leads to a fatal outcome for most cancer patients, new approaches for patient-individualized, targeted immunotherapy are urgently needed. Methods: Here, we established cell lines from four bone metastases of different tumor entities. We assessed AdCAR NK-92-mediated cytotoxicity in vitro in standard cytotoxicity assays as well as 3D spheroid models Results: AdCAR-engineered NK-92 cells successfully demonstrated distinct and specific cytotoxic potential targeting different tumor antigens expressed on cell lines established from bone metastases of mammary, renal cell and colorectal carcinoma as well as melanomas. In that process AdCAR NK-92 cells produced a multitude of NK effector molecules as well as pro inflammatory cytokines. Furthermore, AdCAR NK-92 showed increased cytotoxicity in 3D spheroid models which can recapitulate in vivo architecture, thereby bridging the gap between in vitro and in vivo models. Conclusions: AdCAR NK-92 cells may provide an interesting and promising “off-the-shelf” cellular product for the targeted therapy of cancers metastasizing to the bone, while utilization of clinically approved, therapeutic antibodies, as exchangeable adapter molecules can facilitate quick clinical translation.
Poisoning Effects of Alkali and Alkaline Earth Metal Doping on Selective Catalytic Reduction of NO with NH3 over the Nb-Ce/Zr-PILC Catalysts
Authors: Li Chenxi , Cheng Jin , Ye Qing , Meng Fanwei , Wang Xinpeng , Dai Hongxing
Published: March 2021

The poisoning effects of alkali metals (K and Na) and alkaline earth metals (Ca and Mg) on catalytic performance of the 2Nb4Ce/Zr-PILC catalyst for the selective catalytic reduction of NOx with NH3 (NH3-SCR) were investigated, and physicochemical properties of the catalysts were characterized by means of the X-ray diffraction XRD (XRD), Brunner−Emmet−Teller (BET), hydrogen temperature-programmed reduction (H2-TPR), X-ray Photoelectron Spectroscopy (XPS), ammonia temperature-programmed desorption (NH3-TPD), and in situ diffuse reflectance infrared Fourier transform spectroscopy (in situ DRIFTS) techniques. Doping of M (M = K, Na, Ca, and Mg) deactivated the 2Nb4Ce/Zr-PILC catalyst according to the sequence of 0.8 K > 0.8 Na > 0.8 Ca > 0.8 Mg (M/Ce molar ratio = 0.8). The characterization results showed that the decreases in redox ability, NH3 adsorption, Ce3+/Ce4+ atomic ratio, and amount of the chemisorbed oxygen (Oβ) were the important factors influencing catalytic activities of the alkali metal-and alkaline earth metal-doped samples. Consequently, compared with the Mg- and Ca-doped samples, doping of K caused the 2Nb4Ce/Zr-PILC sample to possess the lowest redox ability, NH3 adsorption, and amount of the Oβ species, which resulted in an obvious deactivation effect.
Mayenite-Based Electride C12A7e−: A Reactivity and Stability Study
Authors: Weber Sebastian , Schäfer Sebastian , Saccoccio Mattia , Ortner Nils , Bertmer Marko , Seidel Karsten , Berendts Stefan , Lerch Martin , Gläser Roger , Kohlmann Holger , Schunk Stephan A.
Published: March 2021

Ru supported on mayenite electride, [Ca24Al28O64]4+(e−)4 a calcium aluminum oxide denoted as C12A7e−, are described in the literature as highly active catalysts for ammonia synthesis, especially under conditions of low absolute pressure. In this study, we investigated the application of recently reported plasma arc melting synthesized C12A7e− (aluminum solid reductant) as supports of Ru/C12A7e− catalysts in ammonia synthesis up to pressures of 7.6 MPa. Together with the plasma-arc-melting-based catalyst support, we investigated a similar plasma-synthesized C12A7e− (graphite solid reductant) and a vacuum-sintering-based C12A7e−. Complementary to the catalytic tests, we applied 2H solid-state NMR spectroscopy, DRUVVis-spectroscopy, thermal analysis and PXRD to study and characterize the reactivity of different plasma-synthesized and vacuum-sintered C12A7e− towards H2/D2 and H2O. The catalysts showed an immediate deactivation at pressures > 1 MPa, which can be explained by irreversible hydride formation at higher pressures, as revealed by reactivity tests of C12A7e− towards H2/D2. The direct formation of C12A7:D from C12A7e− is proven. It can be concluded that the application of Ru/C12A7e− catalysts at the industrial scale has limited prospects due to irreversible hydride formation at relevant pressures > 1 MPa. Furthermore, we report an in-depth study relating to structural changes in the material in the presence of H2O.
Cellular Immunotherapy Targeting Cancer Stem Cells: Preclinical Evidence and Clinical Perspective
Authors: Donini Chiara , Rotolo Ramona , Proment Alessia , Aglietta Massimo , Sangiolo Dario , Leuci Valeria
Published: March 2021

The term “cancer stem cells” (CSCs) commonly refers to a subset of tumor cells endowed with stemness features, potentially involved in chemo-resistance and disease relapses. CSCs may present peculiar immunogenic features influencing their homeostasis within the tumor microenvironment. The susceptibility of CSCs to recognition and targeting by the immune system is a relevant issue and matter of investigation, especially considering the multiple emerging immunotherapy strategies. Adoptive cellular immunotherapies, especially those strategies encompassing the genetic redirection with chimeric antigen receptors (CAR), hold relevant promise in several tumor settings and might in theory provide opportunities for selective elimination of CSC subsets. Initial dedicated preclinical studies are supporting the potential targeting of CSCs by cellular immunotherapies, indirect evidence from clinical studies may be derived and new studies are ongoing. Here we review the main issues related to the putative immunogenicity of CSCs, focusing on and highlighting the existing evidence and opportunities for cellular immunotherapy approaches with T and non-T antitumor lymphocytes.
Extracellular Vesicles Analysis in the COVID-19 Era: Insights on Serum Inactivation Protocols towards Downstream Isolation and Analysis
Authors: Frigerio Roberto , Musicò Angelo , Brucale Marco , Ridolfi Andrea , Galbiati Silvia , Vago Riccardo , Bergamaschi Greta , Ferretti Anna Maria , Chiari Marcella , Valle Francesco , Gori Alessandro , Cretich Marina
Published: March 2021

Since the outbreak of the COVID-19 crisis, the handling of biological samples from confirmed or suspected SARS-CoV-2-positive individuals demanded the use of inactivation protocols to ensure laboratory operators’ safety. While not standardized, these practices can be roughly divided into two categories, namely heat inactivation and solvent-detergent treatments. These routine procedures should also apply to samples intended for Extracellular Vesicles (EVs) analysis. Assessing the impact of virus-inactivating pre-treatments is therefore of pivotal importance, given the well-known variability introduced by different pre-analytical steps on downstream EVs isolation and analysis. Arguably, shared guidelines on inactivation protocols tailored to best address EVs-specific requirements will be needed among the analytical community, yet deep investigations in this direction have not yet been reported. We here provide insights into SARS-CoV-2 inactivation practices to be adopted prior to serum EVs analysis by comparing solvent/detergent treatment vs. heat inactivation. Our analysis entails the evaluation of EVs recovery and purity along with biochemical, biophysical and biomolecular profiling by means of a set of complementary analytical techniques: Nanoparticle Tracking Analysis, Western Blotting, Atomic Force Microscopy, miRNA content (digital droplet PCR) and tetraspanin assessment by microarrays. Our data suggest an increase in ultracentrifugation (UC) recovery following heat treatment; however, it is accompanied by a marked enrichment in EVs-associated contaminants. On the other hand, solvent/detergent treatment is promising for small EVs (<150 nm range), yet a depletion of larger vesicular entities was detected. This work represents a first step towards the identification of optimal serum inactivation protocols targeted to EVs analysis.
Combined Inactivation of Pocket Proteins and APC/CCdh1 by Cdk4/6 Controls Recovery from DNA Damage in G1 Phase
Authors: Shaltiel Indra A. , Llopis Alba , Aprelia Melinda , Klompmaker Rob , Menegakis Apostolos , Krenning Lenno , Medema René H.
Published: March 2021

Most Cyclin-dependent kinases (Cdks) are redundant for normal cell division. Here we tested whether these redundancies are maintained during cell cycle recovery after a DNA damage-induced arrest in G1. Using non-transformed RPE-1 cells, we find that while Cdk4 and Cdk6 act redundantly during normal S-phase entry, they both become essential for S-phase entry after DNA damage in G1. We show that this is due to a greater overall dependency for Cdk4/6 activity, rather than to independent functions of either kinase. In addition, we show that inactivation of pocket proteins is sufficient to overcome the inhibitory effects of complete Cdk4/6 inhibition in otherwise unperturbed cells, but that this cannot revert the effects of Cdk4/6 inhibition in DNA damaged cultures. Indeed, we could confirm that, in addition to inactivation of pocket proteins, Cdh1-dependent anaphase-promoting complex/cyclosome (APC/CCdh1) activity needs to be inhibited to promote S-phase entry in damaged cultures. Collectively, our data indicate that DNA damage in G1 creates a unique situation where high levels of Cdk4/6 activity are required to inactivate pocket proteins and APC/CCdh1 to promote the transition from G1 to S phase.
Radiologic Assessment of Osteosarcoma Lung Metastases: State of the Art and Recent Advances
Authors: Chiesa Anna Maria , Spinnato Paolo , Miceli Marco , Facchini Giancarlo
Published: March 2021

The lung is the most frequent site of osteosarcoma (OS) metastases, which are a critical point in defining a patient’s prognosis. Chest computed tomography (CT) represents the gold standard for the detection of lung metastases even if its sensitivity widely ranges in the literature since lung localizations are often atypical. ESMO guidelines represent one of the major references for the follow-up program of OS patients. The development of new reconstruction techniques, such as the iterative method and the deep learning-based image reconstruction (DLIR), has led to a significant reduction of the radiation dose with the low-dose CT. The improvement of these techniques has great importance considering the young-onset of the disease and the strict chest surveillance during follow-up programs. The use of 18F-fluorodeoxyglucose (FDG) positron emission tomography (PET)/CT is still controversial, while volume doubling time (VDT) and computer-aided diagnosis (CAD) systems are recent diagnostic tools that could support radiologists for lung nodules evaluation. Their use, well-established for other malignancies, needs to be further evaluated, focusing on OS patients.
Downregulation of Mcl-1 by Panobinostat Potentiates Proton Beam Therapy in Hepatocellular Carcinoma Cells
Authors: Choi Changhoon , Lee Ga Haeng , Son Arang , Yoo Gyu Sang , Yu Jeong Il , Park Hee Chul
Published: March 2021

Epigenetic modulation by histone deacetylase (HDAC) inhibitors is an attractive anti-cancer strategy for diverse hematological and solid cancers. Herein, we explored the relative effectiveness of the pan-HDAC inhibitor panobinostat in combination with proton over X-ray irradiation in HCC cells. Clonogenic survival assays revealed that radiosensitization of Huh7 and Hep3B cells by panobinostat was more evident when combined with protons than X-rays. Panobinostat increased G2/M arrest and production of intracellular reactive oxygen species, which was further enhanced by proton irradiation. Immunofluorescence staining of γH2AX showed that panobinostat enhanced proton-induced DNA damage. Panobinostat dose-dependently decreased expression of an anti-apoptotic protein, Mcl-1, concomitant with increasing acetylation of histone H4. The combination of panobinostat with proton irradiation enhanced apoptotic cell death to a greater extent than that with X-ray irradiation. Depletion of Mcl-1 by RNA interference enhanced proton-induced apoptosis and proton radiosensitization, suggesting a potential role of Mcl-1 in determining proton sensitivity. Together, our findings suggest that panobinostat may be a promising combination agent for proton beam therapy in HCC treatment.
Treatment with Cyclic AMP Activators Reduces Glioblastoma Growth and Invasion as Assessed by Two-Photon Microscopy
Authors: Wartchow Krista Minéia , Schmid Benjamin , Tripal Philipp , Stadlbauer Andreas , Buchfelder Michael , Gonçalves Carlos-Alberto , Kleindienst Andrea
Published: March 2021

(1) Background: Despite progress in surgery and radio-chemotherapy of glioblastoma (GB), the prognosis remains very poor. GB cells exhibit a preference for hypoxia to maintain their tumor-forming capacity. Enhancing oxidative phosphorylation—known as the anti-Warburg effect—with cyclic AMP activators has been demonstrated to drive GB cells from proliferation to differentiation thereby reducing tumor growth in a cell culture approach. Here we re-evaluate this treatment in a more clinically relevant model. (2) Methods: The effect of treatment with dibutyryl cyclic AMP (dbcAMP, 1 mM) and the cAMP activator forskolin (50µM) was assessed in a GB cell line (U87GFP+, 104 cells) co-cultured with mouse organotypic brain slices providing architecture and biochemical properties of normal brain tissue. Cell viability was determined by propidium-iodide, and gross metabolic effects were excluded in the extracellular medium. Tumor growth was quantified in terms of area, volume, and invasion at the start of culture, 48 h, 7 days, and 14 days after treatment. (3) Results: The tumor area was significantly reduced following dbcAMP or forskolin treatment (F2,249 = 5.968, p = 0.0029). 3D volumetric quantification utilizing two-photon fluorescence microscopy revealed that the treated tumors maintained a spheric shape while the untreated controls exhibited the GB typical invasive growth pattern. (4) Conclusions: Our data demonstrate that treatment with a cAMP analog/activator reduces GB growth and invasion.
Rap1a Overlaps the AGE/RAGE Signaling Cascade to Alter Expression of α-SMA, p-NF-κB, and p-PKC-ζ in Cardiac Fibroblasts Isolated from Type 2 Diabetic Mice
Authors: Burr Stephanie D. , Stewart James A.
Published: March 2021

Cardiovascular disease, specifically heart failure, is a common complication for individuals with type 2 diabetes mellitus. Heart failure can arise with stiffening of the left ventricle, which can be caused by “active” cardiac fibroblasts (i.e., myofibroblasts) remodeling the extracellular matrix (ECM). Differentiation of fibroblasts to myofibroblasts has been demonstrated to be an outcome of AGE/RAGE signaling. Hyperglycemia causes advanced glycated end products (AGEs) to accumulate within the body, and this process is greatly accelerated under chronic diabetic conditions. AGEs can bind and activate their receptor (RAGE) to trigger multiple downstream outcomes, such as altering ECM remodeling, inflammation, and oxidative stress. Previously, our lab has identified a small GTPase, Rap1a, that possibly overlaps the AGE/RAGE signaling cascade to affect the downstream outcomes. Rap1a acts as a molecular switch connecting extracellular signals to intracellular responses. Therefore, we hypothesized that Rap1a crosses the AGE/RAGE cascade to alter the expression of AGE/RAGE associated signaling proteins in cardiac fibroblasts in type 2 diabetic mice. To delineate this cascade, we used genetically different cardiac fibroblasts from non-diabetic, diabetic, non-diabetic RAGE knockout, diabetic RAGE knockout, and Rap1a knockout mice and treated them with pharmacological modifiers (exogenous AGEs, EPAC, Rap1a siRNA, and pseudosubstrate PKC-ζ). We examined changes in expression of proteins implicated as markers for myofibroblasts (α-SMA) and inflammation/oxidative stress (NF-κB and SOD-1). In addition, oxidative stress was also assessed by measuring hydrogen peroxide concentration. Our results indicated that Rap1a connects to the AGE/RAGE cascade to promote and maintain α-SMA expression in cardiac fibroblasts. Moreover, Rap1a, in conjunction with activation of the AGE/RAGE cascade, increased NF-κB expression as well as hydrogen peroxide concentration, indicating a possible oxidative stress response. Additionally, knocking down Rap1a expression resulted in an increase in SOD-1 expression suggesting that Rap1a can affect oxidative stress markers independently of the AGE/RAGE signaling cascade. These results demonstrated that Rap1a contributes to the myofibroblast population within the heart via AGE/RAGE signaling as well as promotes possible oxidative stress. This study offers a new potential therapeutic target that could possibly reduce the risk for developing diabetic cardiovascular complications attributed to AGE/RAGE signaling.