bluuebunny/crossref_metadata_2025 huggingface dataset:
Investigating the Relationship between the Built Environment and Relative Risk of COVID-19 in Hong Kong
Authors: Huang Jianwei , Kwan Mei-Po , Kan Zihan , Wong Man , Kwok Coco , Yu Xinyu
Published: October 2020

Understanding the relationship between the built environment and the risk of COVID-19 transmission is essential to respond to the pandemic. This study explores the relationship between the built environment and COVID-19 risk using the confirmed cases data collected in Hong Kong. Using the information on the residential buildings and places visited for each case from the dataset, we assess the risk of COVID-19 and explore their geographic patterns at the level of Tertiary Planning Unit (TPU) based on incidence rate (R1) and venue density (R2). We then investigate the associations between several built-environment variables (e.g., nodal accessibility and green space density) and COVID-19 risk using global Poisson regression (GPR) and geographically weighted Poisson regression (GWPR) models. The results indicate that COVID-19 risk tends to be concentrated in particular areas of Hong Kong. Using the incidence rate as an indicator to assess COVID-19 risk may underestimate the risk of COVID-19 transmission in some suburban areas. The GPR and GWPR models suggest a close and spatially heterogeneous relationship between the selected built-environment variables and the risk of COVID-19 transmission. The study provides useful insights that support policymakers in responding to the COVID-19 pandemic and future epidemics.
Aging-Related Phenotypic Conversion of Medullary Microglia Enhances Intraoral Incisional Pain Sensitivity
Authors: Ikutame Daisuke , Urata Kentaro , Oto Tatsuki , Fujiwara Shintaro , Iinuma Toshimitsu , Shibuta Ikuko , Hayashi Yoshinori , Hitomi Suzuro , Iwata Koichi , Shinoda Masamichi
Published: October 2020

Activated microglia involved in the development of orofacial pain hypersensitivity have two major polarization states. The aim of this study was to assess the involvement of the aging-related phenotypic conversion of medullary microglia in the enhancement of intraoral pain sensitivity using senescence-accelerated mice (SAM)-prone/8 (SAMP8) and SAM-resistant/1 (SAMR1) mice. Mechanical head-withdrawal threshold (MHWT) was measured for 21 days post palatal mucosal incision. The number of CD11c-immunoreactive (IR) cells [affective microglia (M1)] and CD163-IR cells [protective microglia (M2)], and tumor-necrosis-factor-α (TNF-α)-IR M1 and interleukin (IL)-10-IR M2 were analyzed via immunohistochemistry on days 3 and 11 following incision. The decrease in MHWT observed following incision was enhanced in SAMP8 mice. M1 levels and the number of TNF-α-IR M1 were increased on day 3 in SAMP8 mice compared with those in SAMR1 mice. On day 11, M1 and M2 activation was observed in both groups, whereas IL-10-IR M2 levels were attenuated in SAMP8 mice, and the number of TNF-α-IR M1 cells increased, compared to those in SAMR1 mice. These results suggest that the mechanical allodynia observed following intraoral injury is potentiated and sustained in SAMP8 mice due to enhancement of TNF-α signaling, M1 activation, and an attenuation of M2 activation accompanying IL-10 release.
Crystal Structure of the Kinase Domain of MerTK in Complex with AZD7762 Provides Clues for Structure-Based Drug Development
Authors: Park Tae Hyun , Bae Seung-Hyun , Bong Seoung Min , Ryu Seong Eon , Jang Hyonchol , Lee Byung Il
Published: October 2020

Aberrant tyrosine-protein kinase Mer (MerTK) expression triggers prosurvival signaling and contributes to cell survival, invasive motility, and chemoresistance in many kinds of cancers. In addition, recent reports suggested that MerTK could be a primary target for abnormal platelet aggregation. Consequently, MerTK inhibitors may promote cancer cell death, sensitize cells to chemotherapy, and act as new antiplatelet agents. We screened an inhouse chemical library to discover novel small-molecule MerTK inhibitors, and identified AZD7762, which is known as a checkpoint-kinase (Chk) inhibitor. The inhibition of MerTK by AZD7762 was validated using an in vitro homogeneous time-resolved fluorescence (HTRF) assay and through monitoring the decrease in phosphorylated MerTK in two lung cancer cell lines. We also determined the crystal structure of the MerTK:AZD7762 complex and revealed the binding mode of AZD7762 to MerTK. Structural information from the MerTK:AZD7762 complex and its comparison with other MerTK:inhibitor structures gave us new insights for optimizing the development of inhibitors targeting MerTK.
Versatile Roles of the Receptor-Like Kinase Feronia in Plant Growth, Development and Host-Pathogen Interaction
Authors: Ji Dongchao , Chen Tong , Zhang Zhanquan , Li Boqiang , Tian Shiping
Published: October 2020

As a member of the Catharanthus roseus receptor-like kinase 1-like (CrRLK1L) protein kinase subfamily, FERONIA (FER) has emerged as a versatile player regulating multifaceted functions in growth and development, as well as responses to environmental factors and pathogens. With the concerted efforts of researchers, the molecular mechanism underlying FER-dependent signaling has been gradually elucidated. A number of cellular processes regulated by FER-ligand interactions have been extensively reported, implying cell type-specific mechanisms for FER. Here, we provide a review on the roles of FER in male-female gametophyte recognition, cell elongation, hormonal signaling, stress responses, responses to fungi and bacteria, and present a brief outlook for future efforts.
Selective Formation, Reactivity, Redox and Magnetic Properties of MnIII and FeIII Dinuclear Complexes with Shortened Salen-Type Schiff Base Ligands
Authors: Rigamonti Luca , Zardi Paolo , Carlino Stefano , Demartin Francesco , Castellano Carlo , Pigani Laura , Ponti Alessandro , Ferretti Anna Maria , Pasini Alessandro
Published: October 2020

The reactivity of the shortened salen-type ligands H3salmp, H2salmen and H2sal(p-X)ben with variable para-substituent on the central aromatic ring (X = tBu, Me, H, F, Cl, CF3, NO2) towards the trivalent metal ions manganese(III) and iron(III) is presented. The selective formation of the dinuclear complexes [M2(μ-salmp)2], M = Mn (1a), Fe (2a), [M2(μ-salmen)2(μ-OR)2)], R = Et, Me, H and M = Mn (3a–c) or Fe (4a–c), and (M2(μ-sal[p-X]ben)2(μ-OMe)2), X = tBu, Me, H, F, Cl, CF3, NO2 and M = Mn (5a–g) or Fe (6a–g), could be identified by reaction of the Schiff bases with metal salts and the base NEt3, and their characterization through elemental analysis, infrared spectroscopy, mass spectrometry and single-crystal X-ray diffraction of 2a·2AcOEt, 2a·2CH3CN and 3c·2DMF was performed. In the case of iron(III) and H3salmp, when using NaOH as a base instead of NEt3, the dinuclear complexes [Fe2(μ-salmp)(μ-OR)(salim)2], R = Me, H (2b,c) could be isolated and spectroscopically characterized, including the crystal structure of 2b·1.5H2O, which showed that rupture of one salmp3− to two coordinated salim− ligands and release of one salH molecule occurred. The same hydrolytic tendency could be identified with sal(p-X)ben ligands in the case of iron(III) also by using NEt3 or upon standing in solution, while manganese(III) did not promote such a C–N bond breakage. Cyclic voltammetry studies were performed for 3b, 4b, 5a and 6a, revealing that the iron(III) complexes can be irreversibly reduced to the mixed-valence FeIIFeIII and FeII2 dinuclear species, while the manganese(III) derivatives can be reversibly oxidized to either the mixed-valence MnIIIMnIV or to the MnIV2 dinuclear species. The super-exchange interaction between the metal centers, mediated by the bridging ligands, resulted in being antiferromagnetic (AFM) for the selected dinuclear compounds 3b, 4b, 5a, 5e,5f, 6a and 6e. The coupling constants J (–2JŜ1·Ŝ2 formalism) had values around −13 cm−1 for manganese(III) compounds, among the largest AFM coupling constants reported so far for dinuclear MnIII2 derivatives, while values between −3 and −10 cm−1 were obtained for iron(III) compounds.
Diameters and Fluorescence Calibration for Extracellular Vesicle Analyses by Flow Cytometry
Authors: Simeone Pasquale , Celia Christian , Bologna Giuseppina , Ercolino Eva , Pierdomenico Laura , Cilurzo Felisa , Grande Rossella , Diomede Francesca , Vespa Simone , Canonico Barbara , Guescini Michele , Stocchi Vilberto , Lotti Lavinia Vittoria , Guagnano Maria Teresa , Stellin Luisa , Papa Stefano , Trubiani Oriana , Marchisio Marco , Miscia Sebastiano , Lanuti Paola
Published: October 2020

Extracellular vesicles (EVs) play a crucial role in the intercellular crosstalk. Mesenchymal stem cell-derived EVs (MSC-EVs), displaying promising therapeutic roles, contribute to the strong rationale for developing EVs as an alternative therapeutic option. EV analysis still represents one of the major issues to be solved in order to translate the use of MSC-EV detection in clinical settings. Even if flow cytometry (FC) has been largely applied for EV studies, the lack of consensus on protocols for FC detection of EVs generated controversy. Standard FC procedures, based on scatter measurements, only allows the detection of the “tip of the iceberg” of all EVs. We applied an alternative FC approach based on the use of a trigger threshold on a fluorescence channel. The EV numbers obtained by the application of the fluorescence triggering resulted significantly higher in respect to them obtained from the same samples acquired by placing the threshold on the side scatter (SSC) channel. The analysis of EV concentrations carried out by three different standardized flow cytometers allowed us to achieve a high level of reproducibility (CV < 20%). By applying the here-reported method highly reproducible results in terms of EV analysis and concentration measurements were obtained.
The Molecular Mechanisms of Iron Metabolism and Its Role in Cardiac Dysfunction and Cardioprotection
Authors: Ravingerová Tanya , Kindernay Lucia , Barteková Monika , Ferko Miroslav , Adameová Adriana , Zohdi Vladislava , Bernátová Iveta , Ferenczyová Kristina , Lazou Antigone
Published: October 2020

Iron is an essential mineral participating in different functions of the organism under physiological conditions. Numerous biological processes, such as oxygen and lipid metabolism, protein production, cellular respiration, and DNA synthesis, require the presence of iron, and mitochondria play an important role in the processes of iron metabolism. In addition to its physiological role, iron may be also involved in the adaptive processes of myocardial “conditioning”. On the other hand, disorders of iron metabolism are involved in the pathological mechanisms of the most common human diseases and include a wide range of them, such as type 2 diabetes, obesity, and non-alcoholic fatty liver disease, and accelerate the development of atherosclerosis. Furthermore, iron also exerts potentially deleterious effects that may be manifested under conditions of ischemia/reperfusion (I/R) injury, myocardial infarction, heart failure, coronary artery angioplasty, or heart transplantation, due to its involvement in reactive oxygen species (ROS) production. Moreover, iron has been recently described to participate in the mechanisms of iron-dependent cell death defined as “ferroptosis”. Ferroptosis is a form of regulated cell death that is distinct from apoptosis, necroptosis, and other types of cell death. Ferroptosis has been shown to be associated with I/R injury and several other cardiac diseases as a significant form of cell death in cardiomyocytes. In this review, we will discuss the role of iron in cardiovascular diseases, especially in myocardial I/R injury, and protective mechanisms stimulated by different forms of “conditioning” with a special emphasis on the novel targets for cardioprotection.
Production of MCM-41 Nanoparticles with Control of Particle Size and Structural Properties: Optimizing Operational Conditions during Scale-Up
Authors: Castillo Rafael R. , de la Torre Lorena , García-Ochoa Félix , Ladero Miguel , Vallet-Regí María
Published: October 2020

The synthesis of Mobil Composition of Matter 41 (MCM-41) mesoporous silica nanoparticles (MSNs) of controlled sizes and porous structure has been performed at laboratory and pilot plant scales. Firstly, the effects of the main operating conditions (TEOS –Tetraethyl ortosilicate– addition rate, nanoparticle maturation time, temperature, and CTAB –Cetrimonium bromide– concentration) on the synthesis at laboratory scale (1 L round-bottom flask) were studied via a Taguchi experimental design. Subsequently, a profound one-by-one study of operating conditions was permitted to upscale the process without significant particle enlargement and pore deformation. To achieve this, the temperature was set to 60 °C and the CTAB to TEOS molar ratio to 8. The final runs were performed at pilot plant scale (5 L cylindrical reactor with temperature and stirring speed control) to analyze stirring speed, type of impeller, TEOS addition rate, and nanoparticle maturation time effects, confirming results at laboratory scale. Despite slight variations on the morphology of the nanoparticles, this methodology provided MSNs with adequate sizes and porosities for biomedical applications, regardless of the reactor/scale. The process was shown to be robust and reproducible using mild synthesis conditions (2 mL⋅min−1 TEOS addition rate, 400 rpm stirred by a Rushton turbine, 60 min maturation time, 60 °C, 2 g⋅L−1 CTAB, molar ratio TEOS/CTAB = 8), providing ca. 13 g of prismatic short mesoporous 100–200 nm nanorods with non-connected 3 nm parallel mesopores.
Clonal Hematopoiesis, Cardiovascular Diseases and Hematopoietic Stem Cells
Authors: Kandarakov Oleg , Belyavsky Alexander
Published: October 2020

Cardiovascular diseases and cancer, the leading causes of morbidity and mortality in the elderly, share some common mechanisms, in particular inflammation, contributing to their progression and pathogenesis. However, somatic mutagenesis, a driving force in cancer development, has not been generally considered as an important factor in cardiovascular disease pathology. Recent studies demonstrated that during normal aging, somatic mutagenesis occurs in blood cells, often resulting in expansion of mutant clones that dominate hematopoiesis at advanced age. This clonal hematopoiesis is primarily associated with mutations in certain leukemia-related driver genes and, being by itself relatively benign, not only increases the risks of subsequent malignant hematopoietic transformation, but, unexpectedly, has a significant impact on progression of atherosclerosis and cardiovascular diseases. In this review, we discuss the phenomenon of clonal hematopoiesis, the most important genes involved in it, its impact on cardiovascular diseases, and relevant aspects of hematopoietic stem cell biology.
SARS-CoV-2/Renin–Angiotensin System: Deciphering the Clues for a Couple with Potentially Harmful Effects on Skeletal Muscle
Authors: Gonzalez Andrea , Orozco-Aguilar Josué , Achiardi Oscar , Simon Felipe , Cabello-Verrugio Claudio
Published: October 2020

Severe acute respiratory syndrome coronavirus (SARS-CoV-2) has produced significant health emergencies worldwide, resulting in the declaration by the World Health Organization of the coronavirus disease 2019 (COVID-19) pandemic. Acute respiratory syndrome seems to be the most common manifestation of COVID-19. A high proportion of patients require intensive care unit admission and mechanical ventilation (MV) to survive. It has been well established that angiotensin-converting enzyme type 2 (ACE2) is the primary cellular receptor for SARS-CoV-2. ACE2 belongs to the renin–angiotensin system (RAS), composed of several peptides, such as angiotensin II (Ang II) and angiotensin (1-7) (Ang-(1-7)). Both peptides regulate muscle mass and function. It has been described that SARS-CoV-2 infection, by direct and indirect mechanisms, affects a broad range of organ systems. In the skeletal muscle, through unbalanced RAS activity, SARS-CoV-2 could induce severe consequences such as loss of muscle mass, strength, and physical function, which will delay and interfere with the recovery process of patients with COVID-19. This article discusses the relationship between RAS, SARS-CoV-2, skeletal muscle, and the potentially harmful consequences for skeletal muscle in patients currently infected with and recovering from COVID-19.
The HGF/Met/NF-κB Pathway Regulates RANKL Expression in Osteoblasts and Bone Marrow Stromal Cells
Authors: Tsubaki Masanobu , Seki Shiori , Takeda Tomoya , Chihara Akiko , Arai Yuuko , Morii Yuusuke , Imano Motohiro , Satou Takao , Shimomura Kazunori , Nishida Shozo
Published: October 2020

Multiple myeloma (MM)-induced bone disease occurs through hyperactivation of osteoclasts by several factors secreted by MM cells. MM cell-secreted factors induce osteoclast differentiation and activation via direct and indirect actions including enhanced expression of receptor activator of nuclear factor κB ligand (RANKL) in osteoblasts and bone marrow stromal cells (BMSCs). Hepatocyte growth factor (HGF) is elevated in MM patients and is associated with MM-induced bone disease, although the mechanism by which HGF promotes bone disease remains unclear. In the present study, we demonstrated that HGF induces RANKL expression in osteoblasts and BMSCs, and investigated the mechanism of induction. We found that HGF and MM cell supernatants induced RANKL expression in ST2 cells, MC3T3-E1 cells, and mouse BMSCs. In addition, HGF increased phosphorylation of Met and nuclear factor κB (NF-κB) in ST2 cells, MC3T3-E1 cells, or mouse BMSCs. Moreover, Met and NF-κB inhibitors suppressed HGF-induced RANKL expression in ST2 cells, MC3T3-E1 cells, and mouse BMSCs. These results indicated that HGF promotes RANKL expression in osteoblasts and BMSCs via the Met/NF-κB signaling pathway, and Met and NF-κB inhibitors suppressed HGF-induced RANKL expression. Our findings suggest that Met and NF-κB inhibitors are potentially useful in mitigating MM-induced bone disease in patients expressing high levels of HGF.
Molecular Drivers of Platelet Activation: Unraveling Novel Targets for Anti-Thrombotic and Anti-Thrombo-Inflammatory Therapy
Authors: Chatterjee Madhumita , Ehrenberg Agnes , Toska Laura Mara , Metz Lisa Maria , Klier Meike , Krueger Irena , Reusswig Friedrich , Elvers Margitta
Published: October 2020

Cardiovascular diseases (CVDs) are the leading cause of death globally—partly a consequence of increased population size and ageing—and are major contributors to reduced quality of life. Platelets play a major role in hemostasis and thrombosis. While platelet activation and aggregation are essential for hemostasis at sites of vascular injury, uncontrolled platelet activation leads to pathological thrombus formation and provokes thrombosis leading to myocardial infarction or stroke. Platelet activation and thrombus formation is a multistage process with different signaling pathways involved to trigger platelet shape change, integrin activation, stable platelet adhesion, aggregation, and degranulation. Apart from thrombotic events, thrombo-inflammation contributes to organ damage and dysfunction in CVDs and is mediated by platelets and inflammatory cells. Therefore, in the past, many efforts have been made to investigate specific signaling pathways in platelets to identify innovative and promising approaches for novel antithrombotic and anti-thrombo-inflammatory strategies that do not interfere with hemostasis. In this review, we focus on some of the most recent data reported on different platelet receptors, including GPIb-vWF interactions, GPVI activation, platelet chemokine receptors, regulation of integrin signaling, and channel homeostasis of NMDAR and PANX1.
Mapping Powdery Mildew (Blumeria graminis f. sp. tritici) Resistance in Wild and Cultivated Tetraploid Wheats
Authors: Simeone Rosanna , Piarulli Luciana , Nigro Domenica , Signorile Massimo Antonio , Blanco Emanuela , Mangini Giacomo , Blanco Antonio
Published: October 2020

Wheat is the most widely grown crop and represents the staple food for one third of the world’s population. Wheat is attacked by a large variety of pathogens and the use of resistant cultivars is an effective and environmentally safe strategy for controlling diseases and eliminating the use of fungicides. In this study, a collection of wild and cultivated tetraploid wheats (Triticum turgidum) were evaluated for seedling resistance (SR) and adult plant resistance (APR) to powdery mildew (Blumeria graminis) and genotyped with a 90K single nucleotide polymorphism (SNP) array to identify new sources of resistance genes. The genome-wide association mapping detected 18 quantitative trait loci (QTL) for APR and 8 QTL for SR, four of which were identical or at least closely linked to four QTL for APR. Thirteen candidate genes, containing nucleotide binding sites and leucine-rich repeats, were localized in the confidence intervals of the QTL-tagging SNPs. The marker IWB6155, associated to QPm.mgb-1AS, was located within the gene TRITD1Av1G004560 coding for a disease resistance protein. While most of the identified QTL were described previously, five QTL for APR (QPm.mgb-1AS, QPm.mgb-2BS, QPm.mgb-3BL.1, QPm.mgb-4BL, QPm.mgb-7BS.1) and three QTL for SR (QPm.mgb-3BL.3, QPm.mgb-5AL.2, QPm.mgb-7BS.2) were mapped on chromosome regions where no resistance gene was reported before. The novel QTL/genes can contribute to enriching the resistance sources available to breeders.
Hevein-Like Antimicrobial Peptides Wamps: Structure–Function Relationship in Antifungal Activity and Sensitization of Plant Pathogenic Fungi to Tebuconazole by WAMP-2-Derived Peptides
Authors: Odintsova Tatyana , Shcherbakova Larisa , Slezina Marina , Pasechnik Tatyana , Kartabaeva Bakhyt , Istomina Ekaterina , Dzhavakhiya Vitaly
Published: October 2020

Hevein-like antimicrobial peptides (AMPs) comprise a family of plant AMPs with antifungal activity, which harbor a chitin-binding site involved in interactions with chitin of fungal cell walls. However, the mode of action of hevein-like AMPs remains poorly understood. This work reports the structure–function relationship in WAMPs—hevein-like AMPs found in wheat (Triticum kiharae Dorof. et Migush.) and later in other Poaceae species. The effect of WAMP homologues differing at position 34 and the antifungal activity of peptide fragments derived from the central, N- and C-terminal regions of one of the WAMPs, namely WAMP-2, on spore germination of different plant pathogenic fungi were studied. Additionally, the ability of WAMP-2-derived peptides to potentiate the fungicidal effect of tebuconazole, one of the triazole fungicides, towards five cereal-damaging fungi was explored in vitro by co-application of WAMP-2 fragments with Folicur® EC 250 (25% tebuconazole). The antifungal activity of WAMP homologues and WAMP-2-derived peptides varied depending on the fungus, suggesting multiple modes of action for WAMPs against diverse pathogens. Folicur® combined with the WAMP-2 fragments inhibited the spore germination at a much greater level than the fungicide alone, and the type of interactions was either synergistic or additive, depending on the target fungus and concentration combinations of the compounds. The combinations, which resulted in synergism and drastically enhanced the sensitivity to tebuconazole, were revealed for all five fungi by a checkerboard assay. The ability to synergistically interact with a fungicide and exacerbate the sensitivity of plant pathogenic fungi to a commercial antifungal agent is a novel and previously uninvestigated property of hevein-like AMPs.
Discovery of Tricyclic Pyranochromenone as Novel Bruton’s Tyrosine Kinase Inhibitors with In Vivo Antirheumatic Activity
Authors: Cho Hyewon , Lee Eun , Kwon Hye Ah , Seul Lee , Jeon Hui-Jeon , Yu Ji Hoon , Ryu Jae-Ha , Jeon Raok
Published: October 2020

Bruton’s tyrosine kinase (BTK) is an attractive target for treating patients with B cell malignancies and autoimmune diseases. Many BTK inhibitors have been identified; however, like other kinase inhibitors, they lack diversity in their core structures. Therefore, it is important to secure a novel scaffold that occupies the adenine-binding site of BTK. We screened an in-house library of natural products and their analogs via a biochemical assay to identify a novel scaffold for targeting BTK. A pyranochromenone scaffold, derived from a natural active component decursin, was found to be effective at targeting BTK and was selected for further optimization. A series of pyranochromenone analogs was synthesized through the modification of pyranochromenone at the C7 position. Pyranochromenone compounds with an electrophilic warhead exhibited promising BTK inhibitory activity, with IC50 values in the range of 0.5–0.9 µM. A docking study of the representative compound 8 provided a reasonable explanation for compound activity. Compound 8 demonstrated good selectivity over other associated kinases and decreased the production of proinflammatory cytokines in THP cells. Moreover, compound 8 presented significant in vivo efficacy in a murine model of collagen-induced arthritis.
Optimizing the Profile of [99mTc]Tc–NT(7–13) Tracers in Pancreatic Cancer Models by Means of Protease Inhibitors
Authors: Kanellopoulos Panagiotis , Nock Berthold A. , Krenning Eric P. , Maina Theodosia
Published: October 2020

Background: The overexpression of neurotensin subtype 1 receptors (NTS1Rs) in human tumors may be elegantly exploited for directing neurotensin (NT)-based radionuclide carriers specifically to cancer sites for theranostic purposes. We have recently shown that [99mTc]Tc–DT1 ([99mTc]Tc–[N4–Gly7]NT(7–13)) and [99mTc]Tc–DT5 ([99mTc]Tc–[N4–βAla7,Dab9]NT(7–13)) show notably improved uptake in human colon adenocarcinoma WiDr xenografts in mice treated with neprilysin (NEP) inhibitors and/or angiotensin-converting enzyme (ACE) inhibitors compared with untreated controls. Aiming toward translation of this promising approach in NTS1R-positive pancreatic ductal adenocarcinoma (PDAC) patients, we now report on the impact of registered NEP/ACE inhibitors on the performance of [99mTc]Tc–DT1 and [99mTc]Tc–DT5 in pancreatic cancer models. Methods: The cellular uptake of [99mTc]Tc–DT1 and [99mTc]Tc–DT5 was tested in a panel of pancreatic cell lines, and their stability was assessed in mice treated or not treated with Entresto, lisinopril, or their combinations. Biodistribution was conducted in severe combined immunodeficiency (SCID) mice bearing pancreatic AsPC-1 xenografts. Results: The Entresto + lisinopril combination maximized the metabolic stability of the fast-internalizing [99mTc]Tc–DT1 in mice, resulting in notably enhanced tumor uptake (7.05 ± 0.80% injected activity (IA)/g vs. 1.25 ± 0.80% IA/g in non-treated controls at 4 h post-injection; p < 0.0001). Conclusions: This study has shown the feasibility of optimizing the uptake of [99mTc]Tc–DT1 in pancreatic cancer models with the aid of clinically established NEP/ACE inhibitors, in favor of clinical translation prospects.
Determination and Dissection of DNA-Binding Specificity for the Thermus thermophilus HB8 Transcriptional Regulator TTHB099
Authors: Moncja Kristi , Van Dyke Michael W.
Published: October 2020

Transcription factors (TFs) have been extensively researched in certain well-studied organisms, but far less so in others. Following the whole-genome sequencing of a new organism, TFs are typically identified through their homology with related proteins in other organisms. However, recent findings demonstrate that structurally similar TFs from distantly related bacteria are not usually evolutionary orthologs. Here we explore TTHB099, a cAMP receptor protein (CRP)-family TF from the extremophile Thermus thermophilus HB8. Using the in vitro iterative selection method Restriction Endonuclease Protection, Selection and Amplification (REPSA), we identified the preferred DNA-binding motif for TTHB099, 5′–TGT(A/g)NBSYRSVN(T/c)ACA–3′, and mapped potential binding sites and regulated genes within the T. thermophilus HB8 genome. Comparisons with expression profile data in TTHB099-deficient and wild type strains suggested that, unlike E. coli CRP (CRPEc), TTHB099 does not have a simple regulatory mechanism. However, we hypothesize that TTHB099 can be a dual-regulator similar to CRPEc.
Efficacy and Safety of Human Serum Albumin–Cisplatin Complex in U87MG Xenograft Mouse Models
Authors: Park Cho Rong , Kim Hyo Young , Song Myung Geun , Lee Yun-Sang , Youn Hyewon , Chung June-Key , Cheon Gi Jeong , Kang Keon Wook
Published: October 2020

Cisplatin (cis-diamminedichloroplatinum (II), CDDP) is a chemotherapeutic drug widely used against many solid tumors. A pharmacokinetics study found that CDDP can bind to human serum albumin (HSA), which is the most abundant plasma protein in serum. HSA has the advantage of being a nanocarrier and can accumulate in tumors by passive targeting and active targeting mediated by the secreted protein acidic and rich in cysteine (SPARC). In this study, we investigated the possibility of using a CDDP–HSA complex (HSA–CDDP) as a SPARC-mediated therapeutic agent. To investigate the HSA-dependent therapeutic effect of HSA–CDDP, we used two types of U87MG glioma cells that express SPARC differently. HSA–CDDP was highly taken up in SPARC expressing cells and this uptake was enhanced with exogenous SPARC treatment in cells with low expression of SPARC. The cytotoxicity of HSA–CDDP was also higher in SPARC-expressing cells. In the tumor model, HSA–CDDP showed a similar tumor growth and survival rate to CDDP only in SPARC-expressing tumor models. The biosafety test indicated that HSA–CDDP was less nephrotoxic than CDDP, based on blood markers and histopathology examination. Our findings show that HSA–CDDP has the potential to be a novel therapeutic agent for SPARC-expressing tumors, enhancing the tumor targeting effect by HSA and reducing the nephrotoxicity of CDDP.
White Adipose Tissue Expansion in Multiple Symmetric Lipomatosis Is Associated with Upregulation of CK2, AKT and ERK1/2
Authors: Sanna Marta , Borgo Christian , Compagnin Chiara , Favaretto Francesca , Vindigni Vincenzo , Trento Mariangela , Bettini Silvia , Comin Alessandra , Belligoli Anna , Rugge Massimo , Bassetto Franco , Donella-Deana Arianna , Vettor Roberto , Busetto Luca , Milan Gabriella
Published: October 2020

Multiple symmetric lipomatosis (MSL) is a rare disorder characterized by overgrowing lipomatous tissue (LT) in the subcutaneous adipose tissue (SAT). What LT is and how it expands are not completely understood; previous data suggested that it could derive from brown AT precursors. In six MSL type I patients, we compared LT morphology by histological and immunohistochemistry (IHC) analysis, gene expression, by qPCR, kinase activity, by Western Blot and in vitro assay to paired-control SAT using AT from patients with pheochromocytoma as a human browning reference. In the stromal vascular fraction (SVF), we quantified adipose stem cells (ASCs) by flow cytometry, the proliferation rate, white and beige adipogenic potential and clonogenicity and adipogenicity by a limiting dilution assay. LT displayed white AT morphology and expression pattern and did not show increased levels of the brown-specific marker UCP1. In LT, we evidenced AKT, CK2 and ERK1/2 hyperactivation. LT-SVF contained increased ASCs, proliferated faster, sprouted clones and differentiated into adipocytes better than the control, displaying enhanced white adipogenic potential but not increased browning compared to SAT. In conclusion, LT is a white AT depot expanding by hyperplasia through increased stemness and enhanced white adipogenesis upregulating AKT, CK2 and ERK1/2, which could represent new targets to counteract MSL.
The Grapevine Calmodulin-Like Protein Gene CML21 Is Regulated by Alternative Splicing and Involved in Abiotic Stress Response
Authors: Aleynova Olga A. , Kiselev Konstantin V. , Ogneva Zlata V. , Dubrovina Alexandra S.
Published: October 2020

Calmodulin-like proteins (CMLs) represent a large family of plant calcium sensor proteins involved in the regulation of plant responses to environmental cues and developmental processes. In the present work, we identified four alternatively spliced mRNA forms of the grapevine CML21 gene that encoded proteins with distinct N-terminal regions. We studied the transcript abundance of CML21v1, CML21v2, CML21v3, and CML21v4 in wild-growing grapevine Vitis amurensis Rupr. in response to desiccation, heat, cold, high salinity, and high mannitol stress using quantitative real-time RT-PCR. The levels of all four splice variants of VaCML21 were highly induced in response to cold stress. In addition, VaCML21v1 and VaCML21v2 forms were highly modulated by all other abiotic stress treatments. Constitutive expression of VaCML21v2 and VaCML21v4 improved biomass accumulation of V. amurensis callus cell cultures under prolonged low temperature stress. Heterologous expression of the grapevine CML21v2 and VaCML21v4 splice variants in Arabidopsis improved survival rates of the transgenic plants after freezing. The VaCML21v2 overexpression enhanced activation of the cold stress-responsive marker genes AtDREB1A and AtDREB2A, while VaCML21v4 overexpression—AtCOR47, AtRD29A, AtRD29B, and AtKIN1 genes after freezing stress in the transgenic Arabidopsis. The results indicate that the grapevine CML21 gene acts as a positive regulator in the plant response to cold stress. The detected variety of CML21 transcripts and their distinct transcriptional responses suggested that this expansion of mRNA variants could contribute to the diversity of grapevine adaptive reactions.
Skeletal Muscle Recovery from Disuse Atrophy: Protein Turnover Signaling and Strategies for Accelerating Muscle Regrowth
Authors: Mirzoev Timur M.
Published: October 2020

Skeletal muscle fibers have a unique capacity to adjust their metabolism and phenotype in response to alternations in mechanical loading. Indeed, chronic mechanical loading leads to an increase in skeletal muscle mass, while prolonged mechanical unloading results in a significant decrease in muscle mass (muscle atrophy). The maintenance of skeletal muscle mass is dependent on the balance between rates of muscle protein synthesis and breakdown. While molecular mechanisms regulating protein synthesis during mechanical unloading have been relatively well studied, signaling events implicated in protein turnover during skeletal muscle recovery from unloading are poorly defined. A better understanding of the molecular events that underpin muscle mass recovery following disuse-induced atrophy is of significant importance for both clinical and space medicine. This review focuses on the molecular mechanisms that may be involved in the activation of protein synthesis and subsequent restoration of muscle mass after a period of mechanical unloading. In addition, the efficiency of strategies proposed to improve muscle protein gain during recovery is also discussed.
Enzymatic Dissociation Induces Transcriptional and Proteotype Bias in Brain Cell Populations
Authors: Mattei Daniele , Ivanov Andranik , van Oostrum Marc , Pantelyushin Stanislav , Richetto Juliet , Mueller Flavia , Beffinger Michal , Schellhammer Linda , vom Berg Johannes , Wollscheid Bernd , Beule Dieter , Paolicelli Rosa Chiara , Meyer Urs
Published: October 2020

Different cell isolation techniques exist for transcriptomic and proteotype profiling of brain cells. Here, we provide a systematic investigation of the influence of different cell isolation protocols on transcriptional and proteotype profiles in mouse brain tissue by taking into account single-cell transcriptomics of brain cells, proteotypes of microglia and astrocytes, and flow cytometric analysis of microglia. We show that standard enzymatic digestion of brain tissue at 37 °C induces profound and consistent alterations in the transcriptome and proteotype of neuronal and glial cells, as compared to an optimized mechanical dissociation protocol at 4 °C. These findings emphasize the risk of introducing technical biases and biological artifacts when implementing enzymatic digestion-based isolation methods for brain cell analyses.
Document Recommendations and Feedback Collection Analysis within the Slovenian Open-Access Infrastructure
Authors: Borovič Mladen , Ferme Marko , Brezovnik Janez , Majninger Sandi , Kac Klemen , Ojsteršek Milan
Published: October 2020

This paper presents a hybrid document recommender system intended for use in digital libraries and institutional repositories that are part of the Slovenian Open Access Infrastructure. The recommender system provides recommendations of similar documents across different digital libraries and institutional repositories with the aim to connect researchers and improve collaboration efforts. The hybrid recommender system makes use of document processing techniques, document metadata, and the similarity ranking function BM25 to provide content-based recommendations as a primary method. It also uses collaborative-filtering methods as a secondary method in a cascade hybrid recommendation technique. We also provide a real-world data feedback collection analysis for our hybrid recommender system on an academic digital repository in order to be able to identify suitable time-frames for direct feedback collection during the year.
Does SEO Matter for Startups? Identifying Insights from UGC Twitter Communities
Authors: Saura José Ramón , Reyes-Menendez Ana , Van Nostrand Chris
Published: October 2020

In the present study, we analyzed User Generated Content (UGC) to measure the importance of Search Engine Optimization (SEO) for startups. For this purpose, we used several clustering algorithms to identify user communities on Twitter. The dataset contained a total of 67,126 tweets. A three-step UGC analysis process was applied to the data. First, a Latent Dirichlet allocation (LDA) was developed to divide the UGC-sample into topics. Next, a sentiment analysis (SA) with machine-learning was applied to divide the sample of topics into negative, positive, and neutral feelings. Finally, a textual analysis (TA) process with data mining techniques was used to extract indicators related to the SEO technique optimization in startups. The results helped us identify UGC communities in Twitter about SEO for startups and the main optimization indicators according to the feelings expressed in tweets. Our results also demonstrated that Black Hack SEO is not the most relevant strategy of positioning of digital marketing for startups and that, although this strategy is used by the startups, it is predominantly negatively perceived by SEO UGC communities.
Colonization Patterns, Phenology and Seasonal Abundance of the Nearctic Leafhopper Erasmoneura vulnerata (Fitch), a New Pest in European Vineyards
Authors: Duso Carlo , Zanettin Giulia , Gherardo Pamela , Pasqualotto Giulia , Raniero Damiano , Rossetto Filippo , Tirello Paola , Pozzebon Alberto
Published: October 2020

The Nearctic leafhopper Erasmoneura vulnerata (Fitch), detected in Europe for the first time (north-eastern Italy) in 2004, has remained a minor pest of grapevine for more than 10 years. The first outbreaks of E. vulnerata were reported in 2016 in commercial vineyards located in north-eastern Italy. High population densities and severe leaf symptoms (i.e., leaf discoloration and fall) were observed in late summer despite the application of insecticides. Investigations were carried out from 2017 to 2019 in 10 vineyards located in Veneto region (Vicenza and Verona provinces) to shed light on the seasonal abundance of E. vulnerata on different Vitis vinifera cultivars. Pest phenology was studied in six vineyards where the impact of insecticides was minimal. Erasmoneura vulnerata completed three generations in each of the growing seasons. Vineyard colonization by overwintered adults showed a clear edge effect, suggesting the influence of overwintering sites (e.g., rural buildings and hedgerows) in vineyard margins. The impact of natural enemies on pest populations appeared to be limited and mostly related to egg parasitoids. Organic vineyards were more heavily infested by E. vulnerata compared to conventional vineyards, likely due to the minimal efficacy of natural insecticides typically used in the former farms.
Evaluation of Low-Cost Sensors for Weather and Carbon Dioxide Monitoring in Internet of Things Context
Authors: Araújo Tiago , Silva Lígia , Moreira Adriano
Published: October 2020

In a context of increased environmental awareness, the Internet of Things has allowed individuals or entities to build their own connected devices to share data about the environment. These data are often obtained from widely available low-cost sensors. Some companies are also selling low-cost sensing kits for in-house or outdoor use. The work described in this paper evaluated, in the short term, the performance of a set of low-cost sensors for temperature, relative humidity, atmospheric pressure and carbon dioxide, commonly used in these platforms. The research challenge addressed with this work was assessing how trustable the raw data obtained from these sensors are. The experiments made use of 18 climatic sensors from six different models, and they were evaluated in a controlled climatic chamber that reproduced controlled situations for temperature and humidity. Four CO2 sensors from two different models were analysed through exposure to different gas concentrations in an indoor environment. Our results revealed temperature sensors with a very high positive coefficient of determination (r2 ≥ 0.99), as well as the presence of bias and almost zero random error; the humidity sensors demonstrated a very high positive correlation (r2 ≥ 0.98), significant bias and small-yet-relevant random error; the atmospheric pressure sensors presented good reproducibility, but further studies are required to evaluate their accuracy and precision. For carbon dioxide, the non-dispersive infra-red sensors demonstrated very satisfactory results (r2 ≥ 0.97, with a minimum root mean squared error (RMSE) value of 26 ppm); the metal oxide sensors, despite their moderate results (minimum RMSE equal to 40 ppm and r2 of 0.8–0.96), presented hysteresis, environmental dependence and even positioning interference. The results suggest that most of the evaluated low-cost sensors can provide a good sense of reality at a very good cost–benefit ratio in certain situations.
The German Hemophilia Registry: Growing with Its Tasks
Authors: Duda Heike , Hesse Janina , Haschberger Birgit , Hilger Anneliese , Keipert Christine
Published: October 2020

Hemophilia is a rare heredity bleeding disorder that requires treatment for life. While few therapeutic options were available in the past, multiple recent breakthroughs have fundamentally altered and diversified hemophilia therapy, with even more new therapeutic options forthcoming. These changes are mirrored by significant regulatory and legal changes, which have redefined the role of hemophilia registries in the European Union (EU). This dual paradigm shift poses new regulatory, scientific but also structural requirements for hemophilia registries. The aim of this manuscript is to enumerate these significant challenges and to demonstrate their incorporation into the redesign of the German Hemophilia Registry (Deutsches Hämophilieregister, dhr). To identify the spectrum of hemophilia therapies and the degree of regulatory changes, a horizon screening was performed. Consequently, a core dataset for the dhr was defined by harmonization with regulatory guidelines as well as other hemophilia registries and by heeding the needs of different stakeholders (patients, clinicians, regulators, and scientists). Based on this information, a new registry structure was established, which is optimized for capturing data on new and established hemophilia therapies in a changing therapeutic and regulatory landscape
Impact of COVID-19 Quarantine on Advanced Hemorrhoidal Disease and the Role of Telemedicine in Patient Management
Authors: Campennì Paola , Marra Angelo A. , Ferri Lorenzo , Orefice Raffaele , Parello Angelo , Litta Francesco , De Simone Veronica , Goglia Marta , Ratto Carlo
Published: October 2020

The aims of this population study were to assess the lockdown impact on patients waiting for hemorrhoidal surgery, and the role of telemedicine in patient management. All patients on our waiting list for hemorrhoidal surgery were considered. Eligible patients were contacted by phone. Rørvik score was evaluated and compared to the baseline score. Univariate and multivariate analyses were performed. A numeric rating scale was used to estimate patient satisfaction regarding telemedicine. One-hundred and ten patients were found to be eligible. Finally, 103 patients were included in the study of whom 16 (15.3%) were already considered for priority surgery. Patients waiting for a longer time showed significantly worse scores (p < 0.001). After telemedicine evaluation the priority waiting list increased by 43.8% (plus 23 patients). Weight loss of at least 3 kg and physical activity were found to be protective factors (p = 0.02 and p = 0.002 respectively). A high grade of satisfaction (almost 80%) towards telemedicine was registered. COVID-19-related delays are linked to a deterioration of hemorrhoidal symptoms and patients’ well-being. Weight and lifestyle changes were deemed key factors in determining disease severity. Telemedicine was a valuable tool to evaluate and re-evaluate patients waiting for hemorrhoidal surgery and was considered satisfactory by patients.
Revision Arthroscopic Bankart Repair: A Systematic Review of Clinical Outcomes
Authors: Yon Chang-Jin , Cho Chul-Hyun , Kim Du-Han
Published: October 2020

Although the frequency of arthroscopic revision surgery is increasing in patients with recurrent dislocation after a primary shoulder stabilization, the literature describing arthroscopic revision Bankart repair has been limited. Preferred reporting items for systematic meta-analyses guidelines were followed by utilizing PubMed, EMBASE, Scopus, and Cochrane Library databases. Keywords included shoulder dislocation, anterior shoulder instability, revision surgery, stabilization, and arthroscopic Bankart repair. Quality assessments were performed with criteria from the methodological index for nonrandomized studies (MINORS). A total of 14 articles were included in this analysis. The mean MINORS score was 12.43. A total of 339 shoulders (337 patients) were included (281 males and 56 females). The mean follow-up period was 36.7 months. Primary surgeries were as follows: arthroscopic procedures (n = 172, 50.7%), open procedure (n = 87, 25.7%), and unknown (n = 80, 23.6%). The mean rate of recurrent instability after revision arthroscopic Bankart repair was 15.3% (n = 52), and an additional re-revision procedure was needed in 6.5% of cases (n = 22). Overall, there were 18.0% (n = 61) of complications reported. This systematic review suggests that arthroscopic revision Bankart repair can lead to an improvement in functional outcomes and reasonable patient satisfaction with proper patient selection.