bluuebunny/crossref_metadata_2025 huggingface dataset:
Oncology Clinic-Based Hereditary Cancer Genetic Testing in a Population-Based Health Care System
Authors: Richardson Matthew , Min Hae Jung , Hong Quan , Compton Katie , Mung Sze Wing , Lohn Zoe , Nuk Jennifer , McCullum Mary , Portigal-Todd Cheryl , Karsan Aly , Regier Dean , Brotto Lori A. , Sun Sophie , Schrader Kasmintan A.
Published: February 2020

New streamlined models for genetic counseling and genetic testing have recently been developed in response to increasing demand for cancer genetic services. To improve access and decrease wait times, we implemented an oncology clinic-based genetic testing model for breast and ovarian cancer patients in a publicly funded population-based health care setting in British Columbia, Canada. This observational study evaluated the oncology clinic-based model as compared to a traditional one-on-one approach with a genetic counsellor using a multi-gene panel testing approach. The primary objectives were to evaluate wait times and patient reported outcome measures between the oncology clinic-based and traditional genetic counselling models. Secondary objectives were to describe oncologist and genetic counsellor acceptability and experience. Wait times from referral to return of genetic testing results were assessed for 400 patients with breast and/or ovarian cancer undergoing genetic testing for hereditary breast and ovarian cancer from June 2015 to August 2017. Patient wait times from referral to return of results were significantly shorter with the oncology clinic-based model as compared to the traditional model (403 vs. 191 days; p < 0.001). A subset of 148 patients (traditional n = 99; oncology clinic-based n = 49) completed study surveys to assess uncertainty, distress, and patient experience. Responses were similar between both models. Healthcare providers survey responses indicated they believed the oncology clinic-based model was acceptable and a positive experience. Oncology clinic-based genetic testing using a multi-gene panel approach and post-test counselling with a genetic counsellor significantly reduced wait times and is acceptable for patients and health care providers.
Comment on: Kadomoto, S. et al. “Tumor-Associated Macrophages Induce Migration of Renal Cell Carcinoma Cells via Activation of the CCL20-CCR6 Axis” Cancers 2020, 12, 89
Authors: Zins Karin , Abraham Dietmar
Published: February 2020

Macrophages form a major component of the leukocyte infiltrate in solid tumors and it has become increasingly clear that tumor-associated macrophages (TAMs) have tumor-promoting effects within the stroma [...]
Early Detection of Hyperprogressive Disease in Non-Small Cell Lung Cancer by Monitoring of Systemic T Cell Dynamics
Authors: Arasanz Hugo , Zuazo Miren , Bocanegra Ana , Gato María , Martínez-Aguillo Maite , Morilla Idoia , Fernández Gonzalo , Hernández Berta , López Paúl , Alberdi Nerea , Hernández Carlos , Chocarro Luisa , Teijeira Lucía , Vera Ruth , Kochan Grazyna , Escors David
Published: February 2020

Hyperprogressive disease (HPD) is an adverse outcome of immunotherapy consisting of an acceleration of tumor growth associated with prompt clinical deterioration. The definitions based on radiological evaluation present important technical limitations. No biomarkers have been identified yet. In this study, 70 metastatic NSCLC patients treated with anti-PD-1/PD-L1 immunotherapy after progression to platinum-based therapy were prospectively studied. Samples from peripheral blood were obtained before the first (baseline) and second cycles of treatment. Peripheral blood mononuclear cells (PBMCs) were isolated and differentiation stages of CD4 lymphocytes quantified by flow cytometry and correlated with HPD as identified with radiological criteria. A strong expansion of highly differentiated CD28− CD4 T lymphocytes (CD4 THD) between the first and second cycle of therapy was observed in HPD patients. After normalizing, the proportion of posttreatment/pretreatment CD4 THD was significantly higher in HPD when compared with the rest of patients (median 1.525 vs. 0.990; p = 0.0007), and also when stratifying by HPD, non-HPD progressors, and responders (1.525, 1.000 and 0.9700 respectively; p = 0.0025). A cut-off value of 1.3 identified HPD with 82% specificity and 70% sensitivity. An increase of CD28− CD4 T lymphocytes ≥ 1.3 (CD4 THD burst) was significantly associated with HPD (p = 0.008). The tumor growth ratio (TGR) was significantly higher in patients with expansion of CD4 THD burst compared to the rest of patients (median 2.67 vs. 0.86, p = 0.0049), and also when considering only progressors (median 2.67 vs. 1.03, p = 0.0126). A strong expansion of CD28− CD4 lymphocytes in peripheral blood within the first cycle of therapy is an early differential feature of HPD in NSCLC treated with immune-checkpoint inhibitors. The monitoring of T cell dynamics allows the early detection of this adverse outcome in clinical practice and complements radiological evaluation.
Inhibition of Wnt/β-Catenin Signaling in Neuroendocrine Tumors In Vitro: Antitumoral Effects
Authors: Jin Xi-Feng , Spöttl Gerald , Maurer Julian , Nölting Svenja , Auernhammer Christoph Josef
Published: February 2020

Background and aims: Inhibition of Wnt/β-catenin signaling by specific inhibitors is currently being investigated as an antitumoral strategy for various cancers. The role of Wnt/β-catenin signaling in neuroendocrine tumors still needs to be further investigated. Methods: This study investigated the antitumor activity of the porcupine (PORCN) inhibitor WNT974 and the β-catenin inhibitor PRI-724 in human neuroendocrine tumor (NET) cell lines BON1, QGP-1, and NCI-H727 in vitro. NET cells were treated with WNT974, PRI-724, or small interfering ribonucleic acids against β-catenin, and subsequent analyses included cell viability assays, flow cytometric cell cycle analysis, caspase3/7 assays and Western blot analysis. Results: Treatment of NET cells with WNT974 significantly reduced NET cell viability in a dose- and time-dependent manner by inducing NET cell cycle arrest at the G1 and G2/M phases without inducing apoptosis. WNT974 primarily blocked Wnt/β-catenin signaling by the dose- and time-dependent downregulation of low-density lipoprotein receptor-related protein 6 (LRP6) phosphorylation and non-phosphorylated β-catenin and total β-catenin, as well as the genes targeting the latter (c-Myc and cyclinD1). Furthermore, the WNT974-induced reduction of NET cell viability occurred through the inhibition of GSK-3-dependent or independent signaling (including pAKT/mTOR, pEGFR and pIGFR signaling). Similarly, treatment of NET cells with the β-catenin inhibitor PRI-724 caused significant growth inhibition, while the knockdown of β-catenin expression by siRNA reduced NET tumor cell viability of BON1 cells but not of NCI-H727 cells. Conclusions: The PORCN inhibitor WNT974 possesses antitumor properties in NET cell lines by inhibiting Wnt and related signaling. In addition, the β-catenin inhibitor PRI-724 possesses antitumor properties in NET cell lines. Future studies are needed to determine the role of Wnt/β-catenin signaling in NET as a potential therapeutic target.
WNT5A-Induced Activation of the Protein Kinase C Substrate MARCKS Is Required for Melanoma Cell Invasion
Authors: Mohapatra Purusottam , Yadav Vikas , Toftdahl Maren , Andersson Tommy
Published: February 2020

WNT5A is a well-known mediator of melanoma cell invasion and metastasis via its ability to activate protein kinase C (PKC), which is monitored by phosphorylation of the endogenous PKC substrate myristoylated alanine-rich c-kinase substrate (MARCKS). However, a possible direct contribution of MARCKS in WNT5A-mediated melanoma cell invasion has not been investigated. Analyses of melanoma patient databases suggested that similar to WNT5A expression, MARCKS expression appears to be associated with increased metastasis. A relationship between the two is suggested by the findings that recombinant WNT5A (rWNT5A) induces both increased expression and phosphorylation of MARCKS, whereas WNT5A silencing does the opposite. Moreover, WNT5A-induced invasion of melanoma cells was blocked by siRNA targeting MARCKS, indicating a crucial role of MARCKS expression and/or its phosphorylation. Next, we employed a peptide inhibitor of MARCKS phosphorylation that did not affect MARCKS expression and found that it abolished WNT5A-induced melanoma cell invasion. Similarly, rWNT5A induced the accumulation of phosphorylated MARCKS in membrane protrusions at the leading edge of melanoma cells. Our results demonstrate that WNT5A-induced phosphorylation of MARCKS is not only an indicator of PKC activity but also a crucial regulator of the metastatic behavior of melanoma and therefore an attractive future antimetastatic target in melanoma patients.
From Bench to Bedside and Beyond: Therapeutic Scenario in Acute Myeloid Leukemia
Authors: Gurnari Carmelo , Voso Maria Teresa , Maciejewski Jaroslaw P. , Visconte Valeria
Published: February 2020

Acute myeloid leukemia (AML) is a heterogeneous group of clonal disorders characterized by abnormal proliferation of undifferentiated myeloid progenitors, impaired hematopoiesis, and variable response to therapy. To date, only about 30% of adult patients with AML become long-term survivors and relapse and/or disease refractoriness are the major cause of treatment failure. Thus, this is an urgent unmet clinical need and new drugs are envisaged in order to ameliorate disease survival outcomes. Here, we review the latest therapeutic approaches (investigational and approved agents) for AML treatment. A specific focus will be given to molecularly targeted therapies for AML as a representation of possible agents for precision medicine. We will discuss experimental and preclinical data for FLT3, IDH1, BCL-2, Hedgehog pathway inhibitors, and epitherapy.
Viral miRNAs as Active Players and Participants in Tumorigenesis
Authors: Gallo Alessia , Miceli Vitale , Bulati Matteo , Iannolo Gioacchin , Contino Flavia , Conaldi Pier Giulio
Published: February 2020

The theory that viruses play a role in human cancers is now supported by scientific evidence. In fact, around 12% of human cancers, a leading cause of morbidity and mortality in some regions, are attributed to viral infections. However, the molecular mechanism remains complex to decipher. In recent decades, the uncovering of cellular miRNAs, with their invaluable potential as diagnostic and prognostic biomarkers, has increased the number of studies being conducted regarding human cancer diagnosis. Viruses develop clever mechanisms to succeed in the maintenance of the viral life cycle, and some viruses, especially herpesviruses, encode for miRNA, v-miRNAs. Through this viral miRNA, the viruses are able to manipulate cellular and viral gene expression, driving carcinogenesis and escaping the host innate or adaptive immune system. In this review, we have discussed the main viral miRNAs and virally influenced cellular pathways, and their capability to drive carcinogenesis.
Circulating Levels of PD-L1 in Mesothelioma Patients from the NIBIT-MESO-1 Study: Correlation with Survival
Authors: Chiarucci Carla , Cannito Sara , Daffinà Maria Grazia , Amato Giovanni , Giacobini Gianluca , Cutaia Ornella , Lofiego Maria Fortunata , Fazio Carolina , Giannarelli Diana , Danielli Riccardo , Di Giacomo Anna Maria , Coral Sandra , Calabrò Luana , Maio Michele , Covre Alessia
Published: February 2020

Targeting of the programmed cell death protein (PD)-1/programmed death-ligand 1 (PD-L1) axis has shown a significant clinical impact in several tumor types. Accordingly, our phase II NIBIT-MESO-1 study demonstrated an improved clinical efficacy in mesothelioma patients treated with the anti-PD-L1 durvalumab combined with the anti-cytotoxic T-lymphocyte antigen (CTLA)-4 tremelimumab, as compared to tremelimumab alone. Due to the promising therapeutic activity of immune check-point inhibitors (ICIs) in mesothelioma patients, the identification of biomarkers predictive of response to treatment is of crucial relevance. The prognostic role of soluble PD-L1 (sPD-L1) proposed in cancer patients prompted us to investigate this protein in sera from mesothelioma patients (n = 40) enrolled in the NIBIT-MESO-1 study. A significant (p < 0.001) increase in sPD-L1 levels was detected in patients after the first cycle and during therapy vs. baseline. A longer overall survival (OS) was observed in patients with sPD-L1 concentrations below (at baseline, d1C2, d1C5 (p < 0.01)) or FC values above (p < 0.05 at d1C2, d1C3, d1C5) their statistically calculated optimal cut-offs. On the basis of these initial results, the specific role of CTLA-4-, PD-L1-, or PD-1-targeting on sPD-L1 release was then investigated in sera from 81 additional ICI-treated solid cancer patients. Results showed a significant (p < 0.001) increase of sPD-L1 levels during therapy compared to baseline only in anti-PD-L1-treated patients, supporting the specific involvement of PD-L1 targeting in the release of its soluble form. Our findings suggest that sPD-L1 represents a predictive biomarker of clinical response to anti-PD-L1 cancer immunotherapy.
Patient-Derived Scaffolds of Colorectal Cancer Metastases as an Organotypic 3D Model of the Liver Metastatic Microenvironment
Authors: D’Angelo Edoardo , Natarajan Dipa , Sensi Francesca , Ajayi Omolola , Fassan Matteo , Mammano Enzo , Pilati Pierluigi , Pavan Piero , Bresolin Silvia , Preziosi Melissa , Miquel Rosa , Zen Yoh , Chokshi Shilpa , Menon Krishna , Heaton Nigel , Spolverato Gaya , Piccoli Martina , Williams Roger , Urbani Luca , Agostini Marco
Published: February 2020

The liver is the most common site for colorectal cancer (CRC) metastasis and there is an urgent need for new tissue culture models to study colorectal cancer liver metastasis (CRLM) as current models do not mimic the biological, biochemical, and structural characteristics of the metastatic microenvironment. Decellularization provides a novel approach for the study of the cancer extracellular matrix (ECM) as decellularized scaffolds retain tissue-specific features and biological properties. In the present study, we created a 3D model of CRC and matched CRLM using patient-derived decellularized ECM scaffolds seeded with the HT-29 CRC cell line. Here, we show an increased HT-29 cell proliferation and migration capability when cultured in cancer-derived scaffolds compared to same-patient healthy colon and liver tissues. HT-29 cells cultured in CRLM scaffolds also displayed an indication of epithelial-mesenchymal transition (EMT), with a loss of E-cadherin and increased Vimentin expression. EMT was confirmed by gene expression profiling, with the most represented biological processes in CRLM-seeded scaffolds involving demethylation, deacetylation, a cellular response to stress metabolic processes, and a response to the oxygen level and starvation. HT-29 cells cultured in cancer-specific 3D microenvironments showed a reduced response to treatment with 5-fluorouracil and 5-fluorouracil combined with Irinotecan when used at a standard IC50 (as determined in the 2D culture). Our 3D culture system with patient-derived tissue-specific decellularized ECM better recapitulates the metastatic microenvironment compared to conventional 2D culture conditions and represents a relevant approach for the study of CRLM progression and assessing the response to chemotherapy agents.
The IGF-II–Insulin Receptor Isoform-A Autocrine Signal in Cancer: Actionable Perspectives
Authors: Scalia Pierluigi , Giordano Antonio , Williams Stephen J.
Published: February 2020

Insulin receptor overexpression is a common event in human cancer. Its overexpression is associated with a relative increase in the expression of its isoform A (IRA), a shorter variant lacking 11 aa in the extracellular domain, conferring high affinity for the binding of IGF-II along with added intracellular signaling specificity for this ligand. Since IGF-II is secreted by the vast majority of malignant solid cancers, where it establishes autocrine stimuli, the co-expression of IGF-II and IRA in cancer provides specific advantages such as apoptosis escape, growth, and proliferation to those cancers bearing such a co-expression pattern. However, little is known about the exact role of this autocrine ligand–receptor system in sustaining cancer malignant features such as angiogenesis, invasion, and metastasis. The recent finding that the overexpression of angiogenic receptor kinase EphB4 along with VEGF-A is tightly dependent on the IGF-II/IRA autocrine system independently of IGFIR provided new perspectives for all malignant IGF2omas (those aggressive solid cancers secreting IGF-II). The present review provides an updated view of the IGF system in cancer, focusing on the biology of the autocrine IGF-II/IRA ligand–receptor axis and supporting its underscored role as a malignant-switch checkpoint target.
A Novel Reduced Graphene Oxide-Attapulgite (RGO-ATP) Supported Fe2O3 Catalyst for Heterogeneous Fenton-like Oxidation of Ciprofloxacin: Degradation Mechanism and Pathway
Authors: Zhang Ting , Qian Chunyuan , Guo Pengran , Gan Shuchai , Dong Lingyu , Bai Ge , Guo Qiyang
Published: February 2020

Ciprofloxacin, a third-generation fluoroquinolones (FQs) antibiotic, is observed to increasingly pollute the environment. In this study, a three-dimensional reduced graphene oxide-attapulgite-based catalyst Fe2O3/RGO-ATP was prepared and used to analyze the degradation of ciprofloxacin in a heterogeneous Fenton reaction. The heterogeneous catalyst Fe2O3/RGO-ATP was prepared by a one-step hydrothermal method, and the samples were characterized using BET(Brunauer-Emmett-Teller) surface area, Raman spectroscopy, X-ray diffraction (XRD), Fourier infrared (FTIR) spectroscopy, scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The effect of reaction time, temperature, pH, initial concentration, H2O2 dosage and reuse time on the degradation of ciprofloxacin by the catalyst Fe2O3/RGO-ATP was investigated. The optimum conditions of degradation of ciprofloxacin are observed to be 60 °C, pH 5, H2O2 concentration of 2.9724 mmol/L, and initial ciprofloxacin concentration of 50 mg/L. The catalyst could be reused several times with a decline in catalytic capacity. Fourier-transform ion cyclotron resonance mass spectrometer (FT) was also employed to study the degradation products of ciprofloxacin in the aqueous solution. The results show that the heterogeneous catalyst Fe2O3/RGO-ATP possessed an excellent ability for the catalytic degradation of ciprofloxacin. Direct hydroxyl oxidation is noted to be the main pathway of degradation of ciprofloxacin, and no defluorination reaction is observed during the degradation process.
Friedel–Crafts-Type Alkylation of Indoles in Water Using Amphiphilic Resin-Supported 1,10-Phenanthroline–Palladium Complex under Aerobic Conditions
Authors: Suzuka Toshimasa , Ooshiro Yuto , Ogihara Kazuhito
Published: February 2020

The palladium-catalyzed Friedel-Crafts-type alkylation of indoles in water has been achieved using amphiphilic polystyrene-poly(ethylene glycol) (PS-PEG) resin-supported phenanthroline-palladium complexes in water under aerobic conditions, affording the corresponding products with good-to-high yield. The polymeric catalyst was also found to promote the C3-alkylation reaction to give a thermodynamic alkylation product with high selectivity. The polymeric catalyst was recovered and reused several times without any loss of catalytic activity.
Beyond Tumor Suppression: Senescence in Cancer Stemness and Tumor Dormancy
Authors: Triana-Martínez Francisco , Loza María Isabel , Domínguez Eduardo
Published: February 2020

Here, we provide an overview of the importance of cellular fate in cancer as a group of diseases of abnormal cell growth. Tumor development and progression is a highly dynamic process, with several phases of evolution. The existing evidence about the origin and consequences of cancer cell fate specification (e.g., proliferation, senescence, stemness, dormancy, quiescence, and cell cycle re-entry) in the context of tumor formation and metastasis is discussed. The interplay between these dynamic tumor cell phenotypes, the microenvironment, and the immune system is also reviewed in relation to cancer. We focus on the role of senescence during cancer progression, with a special emphasis on its relationship with stemness and dormancy. Selective interventions on senescence and dormancy cell fates, including the specific targeting of cancer cell populations to prevent detrimental effects in aging and disease, are also reviewed. A new conceptual framework about the impact of synthetic lethal strategies by using senogenics and then senolytics is given, with the promise of future directions on innovative anticancer therapies.
Regulation of MT1-MMP Activity through Its Association with ERMs
Authors: Suárez Henar , López-Martín Soraya , Toribio Víctor , Zamai Moreno , Hernández-Riquer M. Victoria , Genís Laura , Arroyo Alicia G. , Yáñez-Mó María
Published: February 2020

Membrane-bound proteases play a key role in biology by degrading matrix proteins or shedding adhesion receptors. MT1-MMP metalloproteinase is critical during cancer invasion, angiogenesis, and development. MT1-MMP activity is strictly regulated by internalization, recycling, autoprocessing but also through its incorporation into tetraspanin-enriched microdomains (TEMs), into invadopodia, or by its secretion on extracellular vesicles (EVs). We identified a juxtamembrane positively charged cluster responsible for the interaction of MT1-MMP with ERM (ezrin/radixin/moesin) cytoskeletal connectors in breast carcinoma cells. Linkage to ERMs regulates MT1-MMP subcellular distribution and internalization, but not its incorporation into extracellular vesicles. MT1-MMP association to ERMs and insertion into TEMs are independent phenomena, so that mutation of the ERM-binding motif in the cytoplasmic region of MT1-MMP does not preclude its association with the tetraspanin CD151, but impairs the accumulation and coalescence of CD151/MT1-MMP complexes at actin-rich structures. Conversely, gene deletion of CD151 does not impact on MT1-MMP colocalization with ERM molecules. At the plasma membrane MT1-MMP autoprocessing is severely dependent on ERM association and seems to be the dominant regulator of the enzyme collagenolytic activity. This newly characterized MT1-MMP/ERM association can thus be of relevance for tumor cell invasion.
Curcumin Derivative GT863 Inhibits Amyloid-Beta Production via Inhibition of Protein N-Glycosylation
Authors: Urano Yasuomi , Takahachi Mina , Higashiura Ryo , Fujiwara Hitomi , Funamoto Satoru , Imai So , Futai Eugene , Okuda Michiaki , Sugimoto Hachiro , Noguchi Noriko
Published: February 2020

Amyloid-β (Aβ) peptides play a crucial role in the pathogenesis of Alzheimer’s disease (AD). Aβ production, aggregation, and clearance are thought to be important therapeutic targets for AD. Curcumin has been known to have an anti-amyloidogenic effect on AD. In the present study, we performed screening analysis using a curcumin derivative library with the aim of finding derivatives effective in suppressing Aβ production with improved bioavailability of curcumin using CHO cells that stably express human amyloid-β precursor protein and using human neuroblastoma SH-SY5Y cells. We found that the curcumin derivative GT863/PE859, which has been shown to have an inhibitory effect on Aβ and tau aggregation in vivo, was more effective than curcumin itself in reducing Aβ secretion. We further found that GT863 inhibited neither β- nor γ-secretase activity, but did suppress γ-secretase-mediated cleavage in a substrate-dependent manner. We further found that GT863 suppressed N-linked glycosylation, including that of the γ-secretase subunit nicastrin. We also found that mannosidase inhibitors that block the mannose trimming step of N-glycosylation suppressed Aβ production in a similar fashion, as was observed as a result of treatment with GT863. Collectively, these results suggest that GT863 downregulates N-glycosylation, resulting in suppression of Aβ production without affecting secretase activity.
A20 Promotes Ripoptosome Formation and TNF-Induced Apoptosis via cIAPs Regulation and NIK Stabilization in Keratinocytes
Authors: Feoktistova Maria , Makarov Roman , Brenji Sihem , Schneider Anne T. , Hooiveld Guido J. , Luedde Tom , Leverkus Martin , Yazdi Amir S. , Panayotova-Dimitrova Diana
Published: February 2020

The ubiquitin-editing protein A20 (TNFAIP3) is a known key player in the regulation of immune responses in many organs. Genome-wide associated studies (GWASs) have linked A20 with a number of inflammatory and autoimmune disorders, including psoriasis. Here, we identified a previously unrecognized role of A20 as a pro-apoptotic factor in TNF-induced cell death in keratinocytes. This function of A20 is mediated via the NF-κB-dependent alteration of cIAP1/2 expression. The changes in cIAP1/2 protein levels promote NIK stabilization and subsequent activation of noncanonical NF-κB signaling. Upregulation of TRAF1 expression triggered by the noncanonical NF-κB signaling further enhances the NIK stabilization in an autocrine manner. Finally, stabilized NIK promotes the formation of the ripoptosome and the execution of cell death. Thus, our data demonstrate that A20 controls the execution of TNF-induced cell death on multiple levels in keratinocytes. This signaling mechanism might have important implications for the development of new therapeutic strategies for the treatment of A20-associated skin diseases.
The miR-28-5p Targetome Discovery Identified SREBF2 as One of the Mediators of the miR-28-5p Tumor Suppressor Activity in Prostate Cancer Cells
Authors: Fazio Sofia , Berti Gabriele , Russo Francesco , Evangelista Monica , D’Aurizio Romina , Mercatanti Alberto , Pellegrini Marco , Rizzo Milena
Published: February 2020

miR-28-5p is downregulated in some tumor tissues in which it has been demonstrated to have tumor suppressor (TS) activity. Here, we demonstrate that miR-28-5p acts as a TS in prostate cancer (PCa) cells affecting cell proliferation/survival, as well as migration and invasion. Using the miRNA pull out assay and next generation sequencing, we collected the complete repertoire of miR-28-5p targets, obtaining a data set (miR-28-5p targetome) of 191 mRNAs. Filtering the targetome with TargetScan 7, PITA and RNA22, we found that 61% of the transcripts had miR-28-5p binding sites. To assign a functional value to the captured transcripts, we grouped the miR-28-5p targets into gene families with annotated function and showed that six transcripts belong to the transcription factor category. Among them we selected SREBF2, a gene with an important role in PCa. We validated miR-28-5p/SREBF2 interaction, demonstrating that SREBF2 inhibition affects almost all the tumor processes altered by miR-28-5p re-expression, suggesting that SREBF2 is an important mediator of miR-28-5p TS activity. Our findings support the identification of the targetome of cancer-related miRNAs as a tool to discover genes and pathways fundamental for tumor development, and potential new targets for anti-tumor therapy.
Impaired Nuclear Export of the Ribonucleoprotein Complex and Virus-Induced Cytotoxicity Combine to Restrict Propagation of the A/Duck/Malaysia/02/2001 (H9N2) Virus in Human Airway Cells
Authors: Kumar Sriram , Yeo Dawn , Harur Muralidharan Nisha , Lai Soak Kuan , Tong Cathlyn , Tan Boon Huan , Sugrue Richard J.
Published: February 2020

In humans, (A549) cells impaired H9N2 virus nuclear export of the ribonucleoprotein (RNP) complex contrasted with the early and efficient nuclear export of the H1N1/WSN and pH1N1 virus RNP complexes. Although nuclear export of the RNP complex occurred via the nuclear pore complex, H9N2 virus infection also induced modifications in the nuclear envelope and induced cell cytotoxicity. Reduced PA protein levels in H9N2 virus-infected A549 cells occurred, and this phenomenon was independent of virus infection. Silencing the H1N1/WSN PA protein expression leads to impaired nuclear export of RNP complexes, suggesting that the impaired nuclear export of the H9N2 virus RNP complex may be one of the consequences of reduced PA protein levels. Early and efficient export of the RNP complex occurred in H9N2 virus-infected avian (CEF) cells, although structural changes in the nuclear envelope also occurred. Collectively our data suggest that a combination of delayed nuclear export and virus-induced cell cytotoxicity restricts H9N2 virus transmission in A549 cells. However, the early and efficient export of the RNP complex mitigated the effects of virus-induced cytotoxicity on H9N2 virus transmission in CEF cells. Our findings highlight the multi-factorial nature of host-adaptation of the polymerase proteins of avian influenza viruses in non-avian cell environments.
A-Kinase Anchoring Proteins Diminish TGF-β1/Cigarette Smoke-Induced Epithelial-To-Mesenchymal Transition
Authors: Zuo Haoxiao , Trombetta-Lima Marina , Heijink Irene H. , van der Veen Christina H. T. J. , Hesse Laura , Faber Klaas Nico , Poppinga Wilfred J. , Maarsingh Harm , Nikolaev Viacheslav O. , Schmidt Martina
Published: February 2020

Epithelial-to-mesenchymal transition (EMT) plays a role in chronic obstructive pulmonary diseases (COPD). Cyclic adenosine monophosphate (cAMP) can inhibit transforming growth factor-β1 (TGF-β1) mediated EMT. Although compartmentalization via A-kinase anchoring proteins (AKAPs) is central to cAMP signaling, functional studies regarding their therapeutic value in the lung EMT process are lacking. The human bronchial epithelial cell line (BEAS-2B) and primary human airway epithelial (pHAE) cells were exposed to TGF-β1. Epithelial (E-cadherin, ZO-1) and mesenchymal markers (collagen Ӏ, α-SMA, fibronectin) were analyzed (mRNA, protein). ELISA measured TGF-β1 release. TGF-β1-sensitive AKAPs Ezrin, AKAP95 and Yotiao were silenced while using siRNA. Cell migration was analyzed by wound healing assay, xCELLigence, Incucyte. Prior to TGF-β1, dibutyryl-cAMP (dbcAMP), fenoterol, rolipram, cilostamide, and forskolin were used to elevate intracellular cAMP. TGF-β1 induced morphological changes, decreased E-cadherin, but increased collagen Ӏ and cell migration, a process that was reversed by the inhibitor of δ/epsilon casein kinase I, PF-670462. TGF-β1 altered (mRNA, protein) expression of Ezrin, AKAP95, and Yotiao. St-Ht31, the AKAP antagonist, decreased E-cadherin (mRNA, protein), but counteracted TGF-β1-induced collagen Ӏ upregulation. Cigarette smoke (CS) increased TGF-β1 release, activated TGF signaling, augmented cell migration, and reduced E-cadherin expression, a process that was blocked by TGF-β1 neutralizing antibody. The silencing of Ezrin, AKAP95, and Yotiao diminished TGF-β1-induced collagen Ӏ expression, as well as TGF-β1-induced cell migration. Fenoterol, rolipram, and cilostamide, in AKAP silenced cells, pointed to distinct cAMP compartments. We conclude that Ezrin, AKAP95, and Yotiao promote TGF-β1-mediated EMT, linked to a TGF-β1 release by CS. AKAP members might define the ability of fenoterol, rolipram, and cilostamide to modulate the EMT process, and they might represent potential relevant targets in the treatment of COPD.
Much More Than a Scaffold: Cytoskeletal Proteins in Neurological Disorders
Authors: Muñoz-Lasso Diana C. , Romá-Mateo Carlos , Pallardó Federico V. , Gonzalez-Cabo Pilar
Published: February 2020

Recent observations related to the structure of the cytoskeleton in neurons and novel cytoskeletal abnormalities involved in the pathophysiology of some neurological diseases are changing our view on the function of the cytoskeletal proteins in the nervous system. These efforts allow a better understanding of the molecular mechanisms underlying neurological diseases and allow us to see beyond our current knowledge for the development of new treatments. The neuronal cytoskeleton can be described as an organelle formed by the three-dimensional lattice of the three main families of filaments: actin filaments, microtubules, and neurofilaments. This organelle organizes well-defined structures within neurons (cell bodies and axons), which allow their proper development and function through life. Here, we will provide an overview of both the basic and novel concepts related to those cytoskeletal proteins, which are emerging as potential targets in the study of the pathophysiological mechanisms underlying neurological disorders.
Mechanotransduction in T Cell Development, Differentiation and Function
Authors: Rushdi Muaz , Li Kaitao , Yuan Zhou , Travaglino Stefano , Grakoui Arash , Zhu Cheng
Published: February 2020

Cells in the body are actively engaging with their environments that include both biochemical and biophysical aspects. The process by which cells convert mechanical stimuli from their environment to intracellular biochemical signals is known as mechanotransduction. Exemplifying the reliance on mechanotransduction for their development, differentiation and function are T cells, which are central to adaptive immune responses. T cell mechanoimmunology is an emerging field that studies how T cells sense, respond and adapt to the mechanical cues that they encounter throughout their life cycle. Here we review different stages of the T cell’s life cycle where existing studies have shown important effects of mechanical force or matrix stiffness on a T cell as sensed through its surface molecules, including modulating receptor–ligand interactions, inducing protein conformational changes, triggering signal transduction, amplifying antigen discrimination and ensuring directed targeted cell killing. We suggest that including mechanical considerations in the immunological studies of T cells would inform a more holistic understanding of their development, differentiation and function.
Blood Contamination in CSF and Its Impact on Quantitative Analysis of Alpha-Synuclein
Authors: Barkovits Katalin , Kruse Niels , Linden Andreas , Tönges Lars , Pfeiffer Kathy , Mollenhauer Brit , Marcus Katrin
Published: February 2020

Analysis of cerebrospinal fluid (CSF) is important for diagnosis of neurological diseases. Especially for neurodegenerative diseases, abnormal protein abundance in CSF is an important biomarker. However, the quality of CSF is a key factor for the analytic outcome. Any external contamination has tremendous impact on the analysis and the reliability of the results. In this study, we evaluated the effect of blood contamination in CSF with respect to protein biomarker identification. We compared three distinct measures: Combur10-Test® strips, a specific hemoglobin ELISA, and bottom-up mass spectrometry (MS)-based proteomics for the determination of the general blood contamination level. In parallel, we studied the impact of blood contamination on the detectability of alpha-synuclein (aSyn), a highly abundant protein in blood/erythrocytes and a potential biomarker for Parkinson’s disease. Comparable results were achieved, with all three approaches enabling detection of blood levels in CSF down to 0.001%. We found higher aSyn levels with increasing blood contamination, highlighting the difficulty of authentic quantification of this protein in CSF. Based on our results, we identified other markers for blood contamination beyond hemoglobin and defined a grading system for blood levels in CSF samples, including a lower limit of tolerable blood contamination for MS-based biomarker studies.
Impairment of Membrane Lipid Homeostasis by Bichalcone Analog TSWU-BR4 Attenuates Function of GRP78 in Regulation of the Oxidative Balance and Invasion of Cancer Cells
Authors: Lee Tsung-Lin , Wang Shyang-Guang , Chan Wen-Ling , Lee Ching-Hsiao , Wu Tian-Shung , Lin Meng-Liang , Chen Shih-Shun
Published: February 2020

The specialized cholesterol/sphingolipid-rich membrane domains termed lipid rafts are highly dynamic in the cancer cells, which rapidly assemble effector molecules to form a sorting platform essential for oncogenic signaling transduction in response to extra- or intracellular stimuli. Density-based membrane flotation, subcellular fractionation, cell surface biotinylation, and co-immunoprecipitation analyses of bichalcone analog ((E)-1-(4-Hydroxy-3-((4-(4-((E)-3-(pyridin-3-yl)acryloyl)phenyl)piperazin-1-yl)methyl)phenyl)-3-(pyridin-3-yl)prop-2-en-1-one (TSWU-BR4)-treated cancer cells showed dissociation between GRP78 and p85α conferring the recruitment of PTEN to lipid raft membranes associated with p85α. Ectopic expression of GRP78 could overcome induction of lipid raft membrane-associated p85α–unphosphorylated PTEN complex formation and suppression of GRP78−PI3K−Akt−GTP-Rac1-mediated and GRP78-regulated PERK−Nrf2 antioxidant pathway and cancer cell invasion by TSWU-BR4. Using specific inducer, inhibitor, or short hairpin RNA for ASM demonstrated that induction of the lipid raft membrane localization and activation of ASM by TSWU-BR4 is responsible for perturbing homeostasis of cholesterol and ceramide levels in the lipid raft and ER membranes, leading to alteration of GRP78 membrane trafficking and subsequently inducing p85α–unphosphorylated PTEN complex formation, causing disruption of GRP78−PI3K−Akt−GTP-Rac1-mediated signal and ER membrane-associated GRP78-regulated oxidative stress balance, thus inhibiting cancer cell invasion. The involvement of the enrichment of ceramide to lipid raft membranes in inhibition of NF-κB-mediated MMP-2 expression was confirmed through attenuation of NF-κB activation using C2-ceramide, NF-κB specific inhibitors, ectopic expression of NF-κB p65, MMP-2 promoter-driven luciferase, and NF-κB-dependent reporter genes. In conclusion, localization of ASM in the lipid raft membranes by TSWU-BR4 is a key event for initiating formation of ceramide-enriched lipid raft membrane platforms, which causes delocalization of GRP78 from the lipid raft and ER membranes to the cytosol and formation of p85α–unphosphorylated PTEN complexes to attenuate the GRP78-regulated oxidative stress balance and GRP78−p85α−Akt−GTP-Rac1−NF-κB−MMP-2-mediated cancer cell invasion.
CDKI-73 Is a Novel Pharmacological Inhibitor of Rab11 Cargo Delivery and Innate Immune Secretion
Authors: Sorvina Alexandra , Shandala Tetyana , Wang Shudong , Sharkey David J. , Parkinson-Lawrence Emma , Selemidis Stavros , Brooks Douglas A.
Published: February 2020

Innate immunity is critical for host defence against pathogen and environmental challenge and this involves the production and secretion of immune mediators, such as antimicrobial peptides and pro-inflammatory cytokines. However, when dysregulated, innate immunity can contribute to multifactorial diseases, including inflammatory rheumatic disorders, type 2 diabetes, cancer, neurodegenerative and cardiovascular diseases and even septic shock. During an innate immune response, antimicrobial peptides and cytokines are trafficked via Rab11 multivesicular endosomes, and then sorted into Rab11 vesicles for traffic to the plasma membrane and secretion. In this study, a cyclin-dependent kinase inhibitor CDKI-73 was used to determine its effect on the innate immune response, based on previously identified targets for this compound. Our results showed that CDKI-73 inhibited the delivery of Rab11 vesicles to the plasma membrane, resulting in the accumulation of large multivesicular Rab11 endosomes near the cell periphery. In addition to the effect on endosome delivery, CDKI-73 down-regulated the amount of innate immune cargo, including the antimicrobial peptide Drosomycin and pro-inflammatory cytokines interleukin-6 (IL-6) and tumour necrosis factor alpha (TNFα). We concluded that CDKI-73 has the potential to regulate the delivery and secretion of certain innate immune cargo, which could be used to control inflammation.
Obtaining Mn-Co Alloys in AISI 430 Steel from Lithium-Ion Battery Recycling: Application in SOFC Interconnectors
Authors: Gonçalves Sicele L. A. , Garcia Eric M. , Tarôco Hosane A. , Matencio Tulio
Published: February 2020

The recycling of exhausted lithium-ion batteries from mobile phones originate five solutions with different Co and Mn proportions that were used as electrolytic solutions to obtain Mn-Co spinel coatings on the surface of AISI430 stainless steel. The coatings are intended to contain chromium volatility in the working conditions of Solid Oxide Fuel Cells (SOFC) metallic interconnectors. Potentiostatic electrodeposition was the technique used to obtain Mn-Co coatings from low concentration electrolytes at pH = 3.0 and potential applied −1.3 V. Charge efficiency data were used for sample optimization. Three optimized samples were subjected to oxidation heat treatment at 800 °C for 300 h and then characterized by XRD, SEM and EDS. The results showed that the addition of manganese ions instead of cobalt ions in the electrolytic bath produces more stable and well-distributed deposits as the ratio of the two ions becomes equal in the electrolytic bath. Thin, homogeneous and stable spinel coatings (Mn, Co)3O4 2.8 μm and 3.9 μm thick were able to block chromium volatility when exposed to SOFC operating temperature.
Molecular Dynamics Simulations of Vacancy Generation and Migration near a Monocrystalline Silicon Surface during Energetic Cluster Ion Implantation
Authors: Liang Guoying , Zhong Haowen , Wang Yinong , Zhang Shijian , Xu Mofei , Kuang Shicheng , Ren Jianhui , Zhang Nan , Yan Sha , Yu Xiao , Remnev Gennady Efimovich , Le Xiaoyun
Published: February 2020

The process of ion implantation often involves vacancy generation and migration. The vacancy generation and migration near a monocrystalline silicon surface during three kinds of energetic Si35 cluster ion implantations were investigated by molecular dynamics simulations in the present work. The patterns of vacancy generation and migration, as well as the implantation-induced amorphous structure, were analyzed according to radial distribution function, Wigner–Seitz cell, and identify diamond structure analytical methods. A lot of vacancies rapidly generate and migrate in primary directions and form an amorphous structure in the first two picoseconds. The cluster with higher incident kinetic energy can induce the generation and migration of more vacancies and a deeper amorphous structure. Moreover, boundaries have a loading–unloading effect, where interstitial atoms load into the boundary, which then acts as a source, emitting interstitial atoms to the target and inducing the generation of vacancies again. These results provide more insight into doping silicon via ion implantation.
Crystal Strengths at Micro- and Nano-Scale Dimensions
Authors: Armstrong Ronald W. , Elban Wayne L.
Published: February 2020

Higher strength levels, achieved for dimensionally-smaller micro- and nano-scale materials or material components, such as MEMS devices, are an important enabler of a broad range of present-day engineering devices and structures. Beyond such applications, there is an important effort to understand the dislocation mechanics basis for obtaining such improved strength properties. Four particular examples related to these issues are described in the present report: (1) a compilation of nano-indentation hardness measurements made on silicon crystals spanning nano- to micro-scale testing; (2) stress–strain measurements made on iron and steel materials at micro- to nano-crystal (grain size) dimensions; (3) assessment of small dislocation pile-ups relating to Griffith-type fracture stress vs. crack-size calculations for cleavage fracturing of α-iron; and (4) description of thermally-dependent strain rate sensitivities for grain size strengthening and weakening for macro- to micro- to nano-polycrystalline copper and nickel materials.
Preface of the Special Issue “Pharmaceutical Crystals”
Authors: Yonemochi Etsuo , Uekusa Hidehiro
Published: February 2020

Dear Colleagues, [...]
Structure of Imidazolium-N-phthalolylglycinate Salt Hydrate: Combined Experimental and Quantum Chemical Calculations Studies
Authors: Usman Mohammad , Khan Rais Ahmad , Jaafar Mohammad H. , Alsalme Ali , Tabassum Sartaj
Published: February 2020

An organic supramolecular salt hydrate (imidazolium:N-phthalolylglycinate:H2O; IM+-NPG−-HYD) has been examined for its charge-transfer (CT) characteristics. Accordingly, IM+–NPG−–HYD has been characterized thoroughly using various spectroscopic techniques. Combined experimental and quantum chemical studies, along with wave function analysis, were performed to study the non-covalent interactions and their role in CT in the supramolecular salt hydrate. Notably, IM+–NPG−–HYD crystalizes in two configurations (A and B), both of which are held together via non-covalent interactions to result in a three-dimensional CT supramolecular assembly. The through-space CT occurs from NPG– (donor) to IM+ (acceptor), and this was mediated via non-covalent forces. We demonstrated the role of π–π stacking interactions (mixed-stacking donor-acceptor interactions) in the presence of charge-assisted hydrogen bonds in the regulation of CT properties in the self-assembly of the IM+–NPG−–HYD salt hydrate.
The Deep-Sea Genus Coronarctus (Tardigrada, Arthrotardigrada) in Brazil, South-Western Atlantic Ocean, with the Description of Three New Species
Authors: Gomes-Júnior Edivaldo , Santos Érika , da Rocha Clélia M.C. , Santos Paulo J. P. , Fontoura Paulo
Published: February 2020

Three new marine tardigrade species from the deep-sea genus Coronarctus are described from the South-Western Atlantic Ocean: Coronarctus dissimilis sp. nov., C. neptunus sp. nov., and C. yurupari sp. nov. These, and C. laubieri Renaud-Mornant, 1987, are the first records of deep-sea tardigrades from this marine region. Specimens of those species were collected from two localities of the Brazilian continental slope (Potiguar and Campos basins) at depths comprised between 150 and 3000 m. Specimens of the three new species have short cephalic appendages and heteromorphic claws, belonging to the, here designated, C. tenellus group of species. Each of the new species can be distinguished from all the other species of the group by their peculiar-shaped secondary clavae and claws. The most relevant morphological characters used for the taxonomy of the genus: shape of cephalic cirri, shape of secondary-clavae, size and number of accessory spines on claws, and shape of seminal receptacles, are discussed and an identification key to all ten known Coronarctus species is provided.
Residential Power Traces for Five Houses: The iHomeLab RAPT Dataset
Authors: Huber Patrick , Ott Melvin , Friedli Martin , Rumsch Andreas , Paice Andrew
Published: February 2020

Datasets with measurements of both solar electricity production and domestic electricity consumption separated into the major loads are interesting for research focussing on (i) local optimization of solar energy consumption and (ii) non-intrusive load monitoring. To this end, we publish the iHomeLab RAPT dataset consisting of electrical power traces from five houses in the greater Lucerne region in Switzerland spanning a period from 1.5 up to 3.5 years with a sampling frequency of five minutes. For each house, the electrical energy consumption of the aggregated household and specific appliances such as dishwasher, washing machine, tumble dryer, hot water boiler, or heating pump were metered. Additionally, the data includes electric production data from PV panels for all five houses, and battery power flow measurement data from two houses. Thermal metadata is also provided for the three houses with a heating pump.