bluuebunny/crossref_metadata_2025 huggingface dataset:
Preparation of Eggshell-Type Ru/Al2O3 Catalysts for Hydrogen Production Using Steam-Methane Reforming on PEMFC
Authors: Lee Jong-Heon , Jo Seongbin , Kim Tae-Young , Woo Jin-Hyeok , Lee Yeji , Kim Min-Seok , Park Hye-Ok , Lee Soo-Chool , Kim Jae-Chang
Published: August 2021

Ru-based eggshell-type catalysts, in which Ru is located at the outer region of the pellet, were prepared by the impregnation method, using spherically shaped γ-Al2O3 pellets for steam-methane reforming (SMR). Ru was only supported on the external region of the pellet because of the strong interaction between its precursor and the alumina pellet. The Ru precursor penetrated the inside of the pellet by adding nitric acid to the impregnation solution. The distribution and thickness of the Ru layer in the catalyst can be controlled using the HNO3/Ru molar ratio and contact time at the impregnation step. Among the catalysts, the graded eggshell-type catalyst showed the highest activity and long-term stability in the SMR reaction. In addition, in the daily startup and shutdown (DSS) operation, similar to the hydrogen production environment for domestic polymer electrolyte membrane fuel cells (PEMFC), the graded eggshell-type catalyst showed high activity and stability after multiple cycles. Based on the experimental studies, it was confirmed that Ru-based catalysts are suitable for steam-methane reforming for PEMFC.
Long-Lasting Photocatalytic and Antimicrobial Activity of Cotton Towels Modified with TiO2 and ZnO Nanoparticles
Authors: Gutarowska Beata , Matyjas-Zgondek Edyta , Kulpiński Piotr , Mroczyńska-Florczak Marta , Rutkowski Eugeniusz
Published: August 2021

This study aimed to evaluate the durability of the photocatalytic and antimicrobial activities of ZnO and TiO2 nanoparticles (NPs)-modified 100% cotton terry textiles. SEM-EDX confirmed the long-lasting durability of the washing materials, and TGA analysis revealed that ZnO and TiO2 NPs can be found on the terry fabric surface; however, the amount of NPs decreased 10 times after 15 washes and 1.6 times after the subsequent 15 washes. The efficiency of self-cleaning properties and antimicrobial activity against five microorganisms (Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 10536, Candida albicans ATCC 10231, Aspergillus niger ATCC 16404, and Bacillus subtilis NCAIM 01644) depended on UVA/B radiation intensity. The increase in UVA/B radiation intensity from 400 to 1400 µW/cm2 significantly increases the effectiveness of photocatalysis. Long-lasting self-cleaning properties characterised the tested fabric; however, stronger photocatalytic efficiency was observed in light with a greater intensity of UVA/B radiation. At the UVA/B radiation intensity of 1400 µW/cm2, a biocidal effect (R = 100%) against all tested microorganisms (E. coli, S. aureus. B. subtilis, C. albicans, and A. niger) was observed on the surface of materials. The lower UVA/B radiation intensity (400 µW/cm2) and 30 wash cycles reduce the antimicrobial activity of the material (R = 65.4–99.4%) for B. subtilis, C. albicans, and A. niger. The antimicrobial activity of washed materials modified with TiO2/ZnO nanoparticles can be increased by irradiation with a light bulb (1400 µW/cm2).
LuxAB-Based Microbial Cell Factories for the Sensing, Manufacturing and Transformation of Industrial Aldehydes
Authors: Bayer Thomas , Becker Aileen , Terholsen Henrik , Kim In Jung , Menyes Ina , Buchwald Saskia , Balke Kathleen , Santala Suvi , Almo Steven C. , Bornscheuer Uwe T.
Published: August 2021

The application of genetically encoded biosensors enables the detection of small molecules in living cells and has facilitated the characterization of enzymes, their directed evolution and the engineering of (natural) metabolic pathways. In this work, the LuxAB biosensor system from Photorhabdus luminescens was implemented in Escherichia coli to monitor the enzymatic production of aldehydes from primary alcohols and carboxylic acid substrates. A simple high-throughput assay utilized the bacterial luciferase—previously reported to only accept aliphatic long-chain aldehydes—to detect structurally diverse aldehydes, including aromatic and monoterpene aldehydes. LuxAB was used to screen the substrate scopes of three prokaryotic oxidoreductases: an alcohol dehydrogenase (Pseudomonas putida), a choline oxidase variant (Arthrobacter chlorophenolicus) and a carboxylic acid reductase (Mycobacterium marinum). Consequently, high-value aldehydes such as cinnamaldehyde, citral and citronellal could be produced in vivo in up to 80% yield. Furthermore, the dual role of LuxAB as sensor and monooxygenase, emitting bioluminescence through the oxidation of aldehydes to the corresponding carboxylates, promises implementation in artificial enzyme cascades for the synthesis of carboxylic acids. These findings advance the bio-based detection, preparation and transformation of industrially important aldehydes in living cells.
Dye Decoloring Peroxidase Structure, Catalytic Properties and Applications: Current Advancement and Futurity
Authors: Xu Lingxia , Sun Jianzhong , Qaria Majjid A. , Gao Lu , Zhu Daochen
Published: August 2021

Dye decoloring peroxidases (DyPs) were named after their high efficiency to decolorize and degrade a wide range of dyes. DyPs are a type of heme peroxidase and are quite different from known heme peroxidases in terms of amino acid sequences, protein structure, catalytic residues, and physical and chemical properties. DyPs oxidize polycyclic dyes and phenolic compounds. Thus they find high application potentials in dealing with environmental problems. The structure and catalytic characteristics of DyPs of different families from the amino acid sequence, protein structure, and enzymatic properties, and analyzes the high-efficiency degradation ability of some DyPs in dye and lignin degradation, which vary greatly among DyPs classes. In addition, application prospects of DyPs in biomedicine and other fields are also discussed briefly. At the same time, the research strategy based on genetic engineering and synthetic biology in improving the stability and catalytic activity of DyPs are summarized along with the important industrial applications of DyPs and associated challenges. Moreover, according to the current research findings, bringing DyPs to the industrial level may require improving the catalytic efficiency of DyP, increasing production, and enhancing alkali resistance and toxicity.
Synthetic and Post-Synthetic Strategies to Improve Photoluminescence Quantum Yields in Perovskite Quantum Dots
Authors: Lee ChaeHyun , Lee Soo Jeong , Shin YeJi , Woo Yeonsu , Han Sung-Hwan , Gualdrón-Reyes Andrés Fabián , Mora-Seró Iván , Yoon Seog Joon
Published: August 2021

Making high-quality raw materials is the key to open the versatile potential of next generation materials. All-inorganic CsPbX3 (X: Cl−, Br−, and/or I−) perovskite quantum dots (PQDs) have been applied in various optoelectronic devices, such as photocatalysis, hydrogen evolution, solar cells, and light-emitting diodes, due to their outstanding photophysical properties, such as high photoluminescence quantum yield (PLQY), absorption cross-section, efficient charge separation, and so on. Specifically, for further improvement of the PLQY of the PQDs, it is essential to diminish the non-radiative charge recombination processes. In this work, we approached two ways to control the non-radiative charge recombination processes through synthetic and post-synthetic processes. Firstly, we proposed how refinement of the conventional recrystallization process for PbI2 contributes to higher PLQY of the PQDs. Secondly, after halide exchange from CsPbI3 PQDs to CsPbBr3, through an in situ spectroelectrochemical setup, we monitored the positive correlation between bromide deposition of on the surface of the perovskite and photoluminescence improvement of the CsPbBr3 perovskite film through electrodeposition. These two strategies could provide a way to enhance the photophysical properties of the perovskites for application to various perovskite-based optoelectronic devices.
Catalytic Ozonation of Toluene over Acidic Surface Transformed Natural Zeolite: A Dual-Site Reaction Mechanism and Kinetic Approach
Authors: Alejandro-Martín Serguei , Valdés Héctor , Zaror Claudio A.
Published: August 2021

Volatile organic compounds (VOCs) are responsible for damage to health due to their carcinogenic effects. Catalytic ozonation using zeolite appears as a valuable process to eliminate VOCs from industrial emissions at room temperature. For full-scale application of this new abatement technology, an intrinsic reaction rate equation is needed for an effective process design and scale-up. Results obtained here provide a mechanistic approach during the initial stage of catalytic ozonation of toluene using an acidic surface transformed natural zeolite. In particular, the contribution of Lewis and Brønsted acid sites on the surface reaction mechanism and overall kinetic rate are identified through experimental data. The least-squares non-linear regression method allows the rate-determining step to be established, following a Langmuir–Hinshelwood surface reaction approximation. Experimental evidence suggest that ozone is adsorbed and decomposed at Lewis acid sites, forming active atomic oxygen that leads to the oxidation of adsorbed toluene at Brønsted acid sites.
What Guides Peripheral Immune Cells into the Central Nervous System?
Authors: Greiner Theresa , Kipp Markus
Published: August 2021

Multiple sclerosis (MS), an immune-mediated demyelinating disease of the central nervous system (CNS), initially presents with a relapsing-remitting disease course. During this early stage of the disease, leukocytes cross the blood–brain barrier to drive the formation of focal demyelinating plaques. Disease-modifying agents that modulate or suppress the peripheral immune system provide a therapeutic benefit during relapsing-remitting MS (RRMS). The majority of individuals with RRMS ultimately enter a secondary progressive disease stage with a progressive accumulation of neurologic deficits. The cellular and molecular basis for this transition is unclear and the role of inflammation during the secondary progressive disease stage is a subject of intense and controversial debate. In this review article, we discuss the following main hypothesis: during both disease stages, peripheral immune cells are triggered by CNS-intrinsic stimuli to invade the brain parenchyma. Furthermore, we outline the different neuroanatomical routes by which peripheral immune cells might migrate from the periphery into the CNS.
Diversity of Adult Neural Stem and Progenitor Cells in Physiology and Disease
Authors: Finkel Zachary , Esteban Fatima , Rodriguez Brianna , Fu Tianyue , Ai Xin , Cai Li
Published: August 2021

Adult neural stem and progenitor cells (NSPCs) contribute to learning, memory, maintenance of homeostasis, energy metabolism and many other essential processes. They are highly heterogeneous populations that require input from a regionally distinct microenvironment including a mix of neurons, oligodendrocytes, astrocytes, ependymal cells, NG2+ glia, vasculature, cerebrospinal fluid (CSF), and others. The diversity of NSPCs is present in all three major parts of the CNS, i.e., the brain, spinal cord, and retina. Intrinsic and extrinsic signals, e.g., neurotrophic and growth factors, master transcription factors, and mechanical properties of the extracellular matrix (ECM), collectively regulate activities and characteristics of NSPCs: quiescence/survival, proliferation, migration, differentiation, and integration. This review discusses the heterogeneous NSPC populations in the normal physiology and highlights their potentials and roles in injured/diseased states for regenerative medicine.
Smoking Cessation after a Cancer Diagnosis: A Cross-Sectional Analysis in the Setting of a Developing Country
Authors: Golčić Marin , Tomaš Ilijan , Stevanović Aleksandra , Golčić Goran , Dobrila-Dintinjana Renata , Erić Suzana , Šambić-Penc Mirela , Baretić Marinac Martina , Gović-Golčić Lidija , Majnarić Tea
Published: August 2021

Since smoking accounts for around 30% of all cancer deaths, public health campaigns often focus on smoking cessation as a means of primary prevention. However, smoking after cancer diagnosis is also associated with a higher symptom burden and lower survival rate. As data regarding smoking cessation vary dramatically between different populations, we aimed to analyze smoking prevalence in cancer patients, smoking cessation after cancer diagnosis, and the factors associated with smoking cessation in the setting of a developing country. We performed a cross-sectional survey on 695 patients in two clinical hospital centers. After cancer diagnosis, 15.6% of cancer patients stopped smoking. Male gender, younger age, and smoking-related cancer were the main factors associated with greater smoking cessation (p < 0.05). A total of 96% of breast cancer patients continued to smoke after cancer diagnosis and, compared to lung and colorectal cancer patients, exhibited a lower reduction in the number of cigarettes smoked (p = 0.023). An alarming rate of smoking prevalence was recorded in younger patients (45.6% at the time of cancer diagnosis) suggesting a future rise in smoking-related cancers and complications. These results should guide anti-smoking public health campaigns in transitional countries with a critical focus on younger and breast cancer patients.
Orbital Dynamics in the Restricted Three Body Problem: Overview of Recent Analytical Advances Obtained by Separating Rapid and Slow Subsystems in Non-Planar Configurations
Authors: Oks Eugene
Published: August 2021

Analytical solutions to a variety of simplified versions of the restricted three-body problem in celestial mechanics possess long running history that encompasses several centuries. Most of the successes were limited either to the planar configuration of the three bodies, to the motion around the Lagrange points, or to the Kozai–Lidov effect. We review some analytical advances obtained by separating rapid and slow subsystems as presented in recently published papers concerning the non-planar motion of the three bodies unrelated to the Lagrange points and to the Kozai–Lidov effect. Most (but not all) of the discussed advances correspond to the bound motion in the considered celestial systems.
Flipped Classroom in the Context of Higher Education: Learning, Satisfaction and Interaction
Authors: Sosa Díaz María José , Guerra Antequera Jorge , Cerezo Pizarro Mario
Published: August 2021

Educational governmental institutions have recommended implementing blended learning in higher education to respond to the “new educational normality” caused by the COVID-19 pandemic, although this is not a new challenge. Over the last few decades, higher education institutions have tried to incorporate the use of technological devices to university teaching, by redesigning and optimizing the learning experiences through a mixed teaching model. In this context, the Flipped Classroom (FC) model is one of the pedagogical models that is revolutionizing the scope of education. However, there is still not enough evidence of its advantages and disadvantages in the university stage. Therefore, it is important to analyze the impact of the FC on the learning, satisfaction and interaction of the different agents of the university community. Due to the idiosyncrasy of the present study, an exclusively qualitative and longitudinal methodology was selected; thus, 266 interviews based on open questions were conducted throughout the last five years. The results show that students value the FC model positively and corroborate its great potential from academic, competence, personal and social perspectives. Nevertheless, a small group of students are still very critical about the model and would prefer to keep using a traditional methodology, mainly due to the lack of habit using active methodologies and establishing the learning commitment required by such methodologies.
Review of Electric Vehicle Technologies, Charging Methods, Standards and Optimization Techniques
Authors: Arif Syed Muhammad , Lie Tek Tjing , Seet Boon Chong , Ayyadi Soumia , Jensen Kristian
Published: August 2021

This paper presents a state-of-the-art review of electric vehicle technology, charging methods, standards, and optimization techniques. The essential characteristics of Hybrid Electric Vehicle (HEV) and Electric Vehicle (EV) are first discussed. Recent research on EV charging methods such as Battery Swap Station (BSS), Wireless Power Transfer (WPT), and Conductive Charging (CC) are then presented. This is followed by a discussion of EV standards such as charging levels and their configurations. Next, some of the most used optimization techniques for the sizing and placement of EV charging stations are analyzed. Finally, based on the insights gained, several recommendations are put forward for future research.
FedResilience: A Federated Learning Application to Improve Resilience of Resource-Constrained Critical Infrastructures
Authors: Imteaj Ahmed , Khan Irfan , Khazaei Javad , Amini Mohammad Hadi
Published: August 2021

Critical infrastructures (e.g., energy and transportation systems) are essential lifelines for most modern sectors and have utmost significance in our daily lives. However, these important domains can fail to operate due to system failures or natural disasters. Though the major disturbances in such critical infrastructures are rare, the severity of such events calls for the development of effective resilience assessment strategies to mitigate relative losses. Traditional critical infrastructure resilience approaches consider that the available critical infrastructure agents are resource-sufficient and agree to exchange local data with the server and other agents. Such assumptions create two issues: (1) uncertainty in reaching convergence while applying learning strategies on resource-constrained critical infrastructure agents, and (2) a huge risk of privacy leakage. By understanding the pressing need to construct an effective resilience model for resource-constrained critical infrastructure, this paper aims at leveraging a distributed machine learning technique called Federated Learning (FL) to tackle an agent’s resource limitations effectively and at the same time keep the agent’s information private. Particularly, this paper is focused on predicting the probable outage and resource status of critical infrastructure agents without sharing any local data and carrying out the learning process even when most of the agents are incapable of accomplishing a given computational task. To that end, an FL algorithm is designed specifically for a resource-constrained critical infrastructure environment that could facilitate the training of each agent in a distributed fashion, restrict them from sharing their raw data with any other external entities (e.g., server, neighbor agents), choose proficient clients by analyzing their resources, and allow a partial amount of computation tasks to be performed by the resource-constrained agents. We considered a different number of agents with various stragglers and checked the performance of FedAvg and our proposed FedResilience algorithm with prediction tasks for a probable outage, as well as checking the agents’ resource-sharing scope. Our simulation results show that if the majority of the FL agents are stragglers and we drop them from the training process, then the agents learn very slowly and the overall model performance is negatively affected. We also demonstrate that the selection of proficient agents and allowing them to complete only parts of their tasks can significantly improve the knowledge of each agent by eliminating the straggler effects, and the global model convergence is accelerated.
Definition of Ecological Flow Using IHA and IARI as an Operative Procedure for Water Management
Authors: Greco Michele , Arbia Francesco , Giampietro Raffaele
Published: August 2021

It is widely recognized that the hydrological regime of natural flow plays a primary and crucial role in influencing the physical condition of habitats, which, in turn, determines the biotic composition and sustainability of aquatic ecosystems. The current hydro-ecological understanding states that all flow components might be considered as operational targets for water management, starting from base flows (including low flows) to high and flood regimes in terms of magnitude, frequency, duration, timing, and rate of change. Several codes have been developed and applied on different case studies in order to define common tools to be implemented for Eflow assessment. This work deals with the definition of an operative procedure for the evaluation of the Eflow monthly distribution to be adopted in a generic watercourse cross-section for sustainable surface water resource management and exploitation. The methodology proposes the application of the Indicators of Hydrologic Alteration methodology (IHA by TNC) coupled to the valuation of the Index of Hydrological Regime Alteration (IARI by ISPRA) as an operative tool to define the ecological flow in each monitoring cross-section to support sustainable water resource management and planning. The case study of the Agri River in Basilicata (Southern Italy) is presented. The analyses were carried out based on monthly discharge data derived by applying the HEC-Hydrological Modeling System at the basin scale using the daily rain data measurements obtained by the regional rainfall gauge stations and calibrated through the observed inlet water discharge registered at the Lago del Pertusillo reservoir station.
Homopolymer and ABC Triblock Copolymer Mixtures for Thermoresponsive Gel Formulations
Authors: Constantinou Anna P. , Provatakis Nikitas , Li Qian , Georgiou Theoni K.
Published: August 2021

Our group has recently invented a novel series of thermoresponsive ABC triblock terpolymers based on oligo(ethylene glycol) methyl ether methacrylate with average Mn 300 g mol−1 (OEGMA300, A unit), n-butyl methacrylate (BuMA, B unit) and di(ethylene glycol) methyl ether methacrylate (DEGMA, C unit) with excellent thermogelling properties. In this study, we investigate how the addition of OEGMA300x homopolymers of varying molar mass (MM) affects the gelation characteristics of the best performing ABC triblock terpolymer. Interestingly, the gelation is not disrupted by the addition of the homopolymers, with the gelation temperature (Tgel) remaining stable at around 30 °C, depending on the MM and content in OEGMA300x homopolymer. Moreover, stronger gels are formed when higher MM OEGMA300x homopolymers are added, presumably due to the homopolymer chains acting as bridges between the micelles formed by the triblock terpolymer, thus, favouring gelation. In summary, novel formulations based on mixtures of triblock copolymer and homopolymers are presented, which can provide a cost-effective alternative for use in biomedical applications, compared to the use of the triblock copolymer only.
“Our Antient Friends … Are Much Reduced”: Mary and James Wright, the Hopewell Friends Meeting, and Quaker Women in the Southern Backcountry, c. 1720–c. 1790
Authors: Knight Thomas Daniel
Published: August 2021

Although the existence of Quakers in Virginia is well known, the best recent surveys of Virginia history devote only passing attention to them, mostly in the context of expanding religious freedoms during the revolutionary era. Few discuss the Quakers themselves or the nature of Quaker settlements although notably, Warren Hofstra, Larry Gragg, and others have studied aspects of the Backcountry Quaker experience. Recent Quaker historiography has reinterpreted the origins of the Quaker faith and the role of key individuals in the movement, including the roles of Quaker women. Numerous studies address Quaker women collectively. Few, however, examine individual families or women of different generations within a single family, and Robynne Rogers Healey has argued for “more biographies of less well-known Quaker women”. This essay uses a four-generation genealogical case study of the Quaker Bowater-Wright family to analyze the development of the Quaker faith in the Virginia backcountry and the lower South and its spread into the Old Northwest. In the backcountry environment, with its geographically isolated settlements and widely dispersed population, early Quaker migrants found fertile ground for both their economic and religious activities. The way of life that developed there differed significantly from the hierarchical Anglican structure of the Tidewater region and the more vocal evangelical groups with their independent congregational structure in the southern backcountry. This article argues that Quaker women played a critical role in shaping Quaker migration and institutional growth in eighteenth and nineteenth century America. It also suggests that the Quaker institutional structure reinforced family connections by creating a close bond that united southern Quakers across a great geographical area.
The Danish SoL Project: Effects of a Multi-Component Community-Based Health Promotion Intervention on Prevention of Overweight among 3–8-Year-Old Children
Authors: Buch-Andersen Tine , Eriksson Frank , Bloch Paul , Glümer Charlotte , Mikkelsen Bent Egberg , Toft Ulla
Published: August 2021

The aim of the study was to determine the effects of a multi-component community-based health promotion intervention on body mass index (BMI) z-scores and waist circumference (WC) in three- to eight-year-old children. A quasi-experimental design was adopted to evaluate the effects of the SoL intervention involving three intervention and three control communities. The 19-month intervention was based on the supersetting approach and was designed to promote healthier eating and physical activity among children and their families. BMI z-scores and WC were measured at baseline and follow-up. At baseline, 238 (54%) and 214 (51%) of all eligible children were measured from intervention and control, respectively. The change over time in the BMI z-scores of children from the intervention group was significantly different from that of the control group (p = 0.001). BMI z-scores increased over time in the intervention group in contrast to the control group, whose BMI z-scores decreased (difference in change between groups 0.19 z-scores 95% CI 0.08, 0.30). No significant differences were observed for WC. The results showed no favourable effects of the intervention of Project SoL on BMI z-scores and WC in children. Further studies based on a larger sample size and a longer intervention duration are needed.
Teaching LGBT+ Health and Gender Education to Future Doctors: Implementation of Case-Based Teaching
Authors: Yang Hsing-Chen
Published: August 2021

Improving the education of medical students and physicians can address the disparities in LGBT+ (lesbian, gay, bisexual, transgender, and others) health care. This study explored how teachers used case-based teaching to teach medical students about gender and LGBT+ health care and discussed the implementation and effectiveness of case-based teaching from the perspective of the teachers and students. This study employed the case study method and collected data through semi-structured interviews. This study used two gender courses in clinical psychiatric education as case studies. Two teachers and 19 medical students were recruited as participants. The findings of this study were as follows: (1) effective cases links theory to clinical practice and competency learning; (2) experience sharing by LGBT+ is highly effective; (3) discussions promote the effectiveness of case-based teaching; and (4) the challenges of case-based teaching included time limitations, the multiplexity of the cases, and multilevel learning. This study also found that using narrative cases is a form of narrative pedagogy, which can help students to integrate medicine, gender, and LGBT+ competency education. A successful narrative case–based teaching strategy involves teachers integrating knowledge related to gender, guiding students through the cases to understand the importance of these cases, and reflecting on the medical profession to make improvements. However, teachers face challenges in this approach, such as changes in the school’s teaching culture and a lack of institutional support.
Risk Factors Associated with Nursing Home COVID-19 Outbreaks: A Retrospective Cohort Study
Authors: Cazzoletti Lucia , Zanolin Maria , Tocco Tussardi Ilaria , Alemayohu Mulubirhan , Zanetel Ernesto , Visentin Donatella , Fabbri Luca , Giordani Massimo , Ruscitti Giancarlo , Benetollo Pier , Tardivo Stefano , Torri Emanuele
Published: August 2021

Background: The coronavirus disease 2019 (COVID-19) pandemic had a devastating impact on nursing homes/long-term care facilities. This study examined the relationship between geography, size, design, organizational characteristics, and implementation of infection prevention and control (IPC) measures and the extent of COVID-19 outbreaks in nursing homes in the Autonomous Province of Trento (Italy) during the time frame of March-May 2020. Methods: The analysis included 57 nursing homes (5145 beds). The association between median cumulative incidence of COVID-19 cases among residents and characteristics of nursing homes was assessed by Mann–Whitney U test, Kruskal–Wallis test or Spearman rho. To evaluate the potential confounding of geographical area, a 2-level random intercept logistic model was fitted, with level 1 units (patients in nursing homes) nested into level 2 units (nursing homes), and “being a COVID-19 case” as the dependent variable. Results: Median cumulative incidence was not significantly associated with any of the variables, except for geographical region (p = 0.002). COVID-19 cases clustered in the part of the province bordering the Italian region most affected by the pandemic (Lombardy) (45.2% median cumulative incidence). Conclusions: Structural/organizational factors and standard IPC measures may not predict the epidemiology of COVID-19 outbreaks and be sufficient alone to protect nursing homes against them.
Antagonistic Interaction between Histone Deacetylase Inhibitor: Cambinol and Cisplatin—An Isobolographic Analysis in Breast Cancer In Vitro Models
Authors: Hałasa Marta , Łuszczki Jarogniew J. , Dmoszyńska-Graniczka Magdalena , Baran Marzena , Okoń Estera , Stepulak Andrzej , Wawruszak Anna
Published: August 2021

Breast cancer (BC) is the leading cause of death in women all over the world. Currently, combined chemotherapy with two or more agents is considered a promising anti-cancer tool to achieve better therapeutic response and to reduce therapy-related side effects. In our study, we demonstrated an antagonistic effect of cytostatic agent-cisplatin (CDDP) and histone deacetylase inhibitor: cambinol (CAM) for breast cancer cell lines with different phenotypes: estrogen receptor positive (MCF7, T47D) and triple negative (MDA-MB-231, MDA-MB-468). The type of pharmacological interaction was assessed by an isobolographic analysis. Our results showed that both agents used separately induced cell apoptosis; however, applying them in combination ameliorated antiproliferative effect for all BC cell lines indicating antagonistic interaction. Cell cycle analysis showed that CAM abolished cell cycle arrest in S phase, which was induced by CDDP. Additionally, CAM increased cell proliferation compared to CDDP used alone. Our data indicate that CAM and CDDP used in combination produce antagonistic interaction, which could inhibit anti-cancer treatment efficacy, showing importance of preclinical testing.
The Impact of Early Life Exposure to Cannabis: The Role of the Endocannabinoid System
Authors: Martínez-Peña Annia A. , Perono Genevieve A. , Gritis Sarah Alexis , Sharma Reeti , Selvakumar Shamini , Walker O’Llenecia S. , Gurm Harmeet , Holloway Alison C. , Raha Sandeep
Published: August 2021

Cannabis use during pregnancy has continued to rise, particularly in developed countries, as a result of the trend towards legalization and lack of consistent, evidence-based knowledge on the matter. While there is conflicting data regarding whether cannabis use during pregnancy leads to adverse outcomes such as stillbirth, preterm birth, low birthweight, or increased admission to neonatal intensive care units, investigations into long-term effects on the offspring’s health are limited. Historically, studies have focused on the neurobehavioral effects of prenatal cannabis exposure on the offspring. The effects of cannabis on other physiological aspects of the developing fetus have received less attention. Importantly, our knowledge about cannabinoid signaling in the placenta is also limited. The endocannabinoid system (ECS) is present at early stages of development and represents a potential target for exogenous cannabinoids in utero. The ECS is expressed in a broad range of tissues and influences a spectrum of cellular functions. The aim of this review is to explore the current evidence surrounding the effects of prenatal exposure to cannabinoids and the role of the ECS in the placenta and the developing fetus.
Small Molecule Receptor Binding Inhibitors with In Vivo Efficacy against Botulinum Neurotoxin Serotypes A and E
Authors: Ben David Alon , Barnea Ada , Diamant Eran , Dor Eyal , Schwartz Arieh , Torgeman Amram , Zichel Ran
Published: August 2021

Botulinum neurotoxins (BoNTs) are the most poisonous substances in nature. Currently, the only therapy for botulism is antitoxin. This therapy suffers from several limitations and hence new therapeutic strategies are desired. One of the limitations in discovering BoNT inhibitors is the absence of an in vitro assay that correlates with toxin neutralization in vivo. In this work, a high-throughput screening assay for receptor-binding inhibitors against BoNT/A was developed. The assay is composed of two chimeric proteins: a receptor-simulating protein, consisting of the fourth luminal loop of synaptic vesicle protein 2C fused to glutathione-S-transferase, and a toxin-simulating protein, consisting of the receptor-binding domain of BoNT/A fused to beta-galactosidase. The assay was applied to screen the LOPAC1280 compound library. Seven selected compounds were evaluated in mice exposed to a lethal dose of BoNT/A. The compound aurintricarboxylic acid (ATA) conferred 92% protection, whereas significant delayed time to death (p < 0.005) was observed for three additional compounds. Remarkably, ATA was also fully protective in mice challenged with a lethal dose of BoNT/E, which also uses the SV2 receptor. This study demonstrates that receptor-binding inhibitors have the potential to serve as next generation therapeutics for botulism, and therefore the assay developed may facilitate discovery of new anti-BoNT countermeasures.
Platelet-Derived Extracellular Vesicles for Regenerative Medicine
Authors: Antich-Rosselló Miquel , Forteza-Genestra Maria Antònia , Monjo Marta , Ramis Joana M.
Published: August 2021

Extracellular vesicles (EVs) present a great potential for the development of new treatments in the biomedical field. To be used as therapeutics, many different sources have been used for EVs obtention, while only a few studies have addressed the use of platelet-derived EVs (pEVs). In fact, pEVs have been shown to intervene in different healing responses, thus some studies have evaluated their regenerative capability in wound healing or hemorrhagic shock. Even more, pEVs have proven to induce cellular differentiation, enhancing musculoskeletal or neural regeneration. However, the obtention and characterization of pEVs is widely heterogeneous and differs from the recommendations of the International Society for Extracellular Vesicles. Therefore, in this review, we aim to present the main advances in the therapeutical use of pEVs in the regenerative medicine field while highlighting the isolation and characterization steps followed. The main goal of this review is to portray the studies performed in order to enhance the translation of the pEVs research into feasible therapeutical applications.
Identification of a Common Epitope in Nucleocapsid Proteins of Euro-America Orthotospoviruses and Its Application for Tagging Proteins
Authors: Cheng Hao-Wen , Tsai Wei-Ting , Hsieh Yi-Ying , Chen Kuan-Chun , Yeh Shyi-Dong
Published: August 2021

The NSs protein and the nucleocapsid protein (NP) of orthotospoviruses are the major targets for serological detection and diagnosis. A common epitope of KFTMHNQIF in the NSs proteins of Asia orthotospoviruses has been applied as an epitope tag (nss-tag) for monitoring recombinant proteins. In this study, a monoclonal antibody TNP MAb against the tomato spotted wilt virus (TSWV) NP that reacts with TSWV-serogroup members of Euro-America orthotospoviruses was produced. By truncation and deletion analyses of TSWV NP, the common epitope of KGKEYA was identified and designated as the np sequence. The np sequence was successfully utilized as an epitope tag (np-tag) to monitor various proteins, including the green fluorescence protein, the coat protein of the zucchini yellow mosaic virus, and the dust mite chimeric allergen Dp25, in a bacterial expression system. The np-tag was also applied to investigate the protein–protein interaction in immunoprecipitation. In addition, when the np-tag and the nss-tag were simultaneously attached at different termini of the expressed recombinant proteins, they reacted with the corresponding MAbs with high sensitivity. Here, we demonstrated that the np sequence and TNP MAb can be effectively applied for tagging and detecting proteins and can be coupled with the nss-tag to form a novel epitope-tagging system for investigating protein–protein interactions.
Transcranial Magnetic Stimulation Improves Muscle Involvement in Experimental Autoimmune Encephalomyelitis
Authors: Peña-Toledo Maria Angeles , Luque Evelio , Ruz-Caracuel Ignacio , Agüera Eduardo , Jimena Ignacio , Peña-Amaro Jose , Tunez Isaac
Published: August 2021

Skeletal muscle is affected in experimental autoimmune encephalomyelitis (EAE), which is a model of multiple sclerosis that produces changes including muscle atrophy; histological features of neurogenic involvement, and increased oxidative stress. In this study, we aimed to evaluate the therapeutic effects of transcranial magnetic stimulation (TMS) on the involvement of rat skeletal muscle and to compare them with those produced by natalizumab (NTZ). EAE was induced by injecting myelin oligodendrocyte glycoprotein (MOG) into Dark Agouti rats. Both treatments, NTZ and TMS, were implemented from day 15 to day 35. Clinical severity was studied, and after sacrifice, the soleus and extensor digitorum longus muscles were extracted for subsequent histological and biochemical analysis. The treatment with TMS and NTZ had a beneficial effect on muscle involvement in the EAE model. There was a clinical improvement in functional motor deficits, atrophy was attenuated, neurogenic muscle lesions were reduced, and the level of oxidative stress biomarkers was lower in both treatment groups. Compared to NTZ, the best response was obtained with TMS for all the parameters analyzed. The myoprotective effect of TMS was higher than that of NTZ. Thus, the use of TMS may be an effective strategy to reduce muscle involvement in multiple sclerosis.
Cathepsin L, a Target of Hypoxia-Inducible Factor-1-α, Is Involved in Melanosome Degradation in Melanocytes
Authors: Kim Ji Young , Lee Eun Jung , Ahn Yuri , Park Sujin , Bae Yu Jeong , Kim Tae Gyun , Oh Sang Ho
Published: August 2021

Hypoxic conditions induce the activation of hypoxia-inducible factor-1α (HIF-1α) to restore the supply of oxygen to tissues and cells. Activated HIF-1α translocates into the nucleus and binds to hypoxia response elements to promote the transcription of target genes. Cathepsin L (CTSL) is a lysosomal protease that degrades cellular proteins via the endolysosomal pathway. In this study, we attempted to determine if CTSL is a hypoxia responsive target gene of HIF-1α, and decipher its role in melanocytes in association with the autophagic pathway. The results of our luciferase reporter assay showed that the expression of CTSL is transcriptionally activated through the binding of HIF1-α at its promoter. Under autophagy-inducing starvation conditions, HIF-1α and CTSL expression is highly upregulated in melan-a cells. The mature form of CTSL is closely involved in melanosome degradation through lysosomal activity upon autophagosome–lysosome fusion. The inhibition of conversion of pro-CTSL to mature CTSL leads to the accumulation of gp100 and tyrosinase in addition to microtubule-associated protein 1 light chain 3 (LC3) II, due to decreased lysosomal activity in the autophagic pathway. In conclusion, we have identified that CTSL, a novel target of HIF-1α, participates in melanosome degradation in melanocytes through lysosomal activity during autophagosome–lysosome fusion.
Epigenetic Regulation of Cardiomyocyte Differentiation from Embryonic and Induced Pluripotent Stem Cells
Authors: Kim Yong-Jin , Tamadon Amin , Kim Yoon-Young , Kang Byeong-Cheol , Ku Seung-Yup
Published: August 2021

With the intent to achieve the best modalities for myocardial cell therapy, different cell types are being evaluated as potent sources for differentiation into cardiomyocytes. Embryonic stem cells and induced pluripotent stem cells have great potential for future progress in the treatment of myocardial diseases. We reviewed aspects of epigenetic mechanisms that play a role in the differentiation of these cells into cardiomyocytes. Cardiomyocytes proliferate during fetal life, and after birth, they undergo permanent terminal differentiation. Upregulation of cardiac-specific genes in adults induces hypertrophy due to terminal differentiation. The repression or expression of these genes is controlled by chromatin structural and epigenetic changes. However, few studies have reviewed and analyzed the epigenetic aspects of the differentiation of embryonic stem cells and induced pluripotent stem cells into cardiac lineage cells. In this review, we focus on the current knowledge of epigenetic regulation of cardiomyocyte proliferation and differentiation from embryonic and induced pluripotent stem cells through histone modification and microRNAs, the maintenance of pluripotency, and its alteration during cardiac lineage differentiation.
Molecular Chaperones and miRNAs in Epilepsy: Pathogenic Implications and Therapeutic Prospects
Authors: Zummo Leila , Vitale Alessandra Maria , Caruso Bavisotto Celeste , De Curtis Marco , Garbelli Rita , Giallonardo Anna Teresa , Di Bonaventura Carlo , Fanella Martina , Conway de Macario Everly , Cappello Francesco , Macario Alberto J. L. , Marino Gammazza Antonella
Published: August 2021

Epilepsy is a pathologic condition with high prevalence and devastating consequences for the patient and its entourage. Means for accurate diagnosis of type, patient monitoring for predicting seizures and follow up, and efficacious treatment are desperately needed. To improve this adverse outcome, miRNAs and the chaperone system (CS) are promising targets to understand pathogenic mechanisms and for developing theranostics applications. miRNAs implicated in conditions known or suspected to favor seizures such as neuroinflammation, to promote epileptic tolerance and neuronal survival, to regulate seizures, and others showing variations in expression levels related to seizures are promising candidates as useful biomarkers for diagnosis and patient monitoring, and as targets for developing novel therapies. Components of the CS are also promising as biomarkers and as therapeutic targets, since they participate in epileptogenic pathways and in cytoprotective mechanisms in various epileptogenic brain areas, even if what they do and how is not yet clear. The data in this review should help in the identification of molecular targets among the discussed miRNAs and CS components for research aiming at understanding epileptogenic mechanisms and, subsequently, develop means for predicting/preventing seizures and treating the disease.
Geraniin Inhibits the Entry of SARS-CoV-2 by Blocking the Interaction between Spike Protein RBD and Human ACE2 Receptor
Authors: Kim Young Soo , Chung Hwan-Suck , Noh Sang Gyun , Lee Bonggi , Chung Hae Young , Choi Jang-Gi
Published: August 2021

The coronavirus disease 2019 (COVID-19) pandemic is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Despite the development of vaccines, the emergence of SARS-CoV-2 variants and the absence of effective therapeutics demand the continual investigation of COVID-19. Natural products containing active ingredients may be good therapeutic candidates. Here, we investigated the effectiveness of geraniin, the main ingredient in medical plants Elaeocarpus sylvestris var. ellipticus and Nephelium lappaceum, for treating COVID-19. The SARS-CoV-2 spike protein binds to the human angiotensin-converting enzyme 2 (hACE2) receptor to initiate virus entry into cells; viral entry may be an important target of COVID-19 therapeutics. Geraniin was found to effectively block the binding between the SARS-CoV-2 spike protein and hACE2 receptor in competitive enzyme-linked immunosorbent assay, suggesting that geraniin might inhibit the entry of SARS-CoV-2 into human epithelial cells. Geraniin also demonstrated a high affinity to both proteins despite a relatively lower equilibrium dissociation constant (KD) for the spike protein (0.63 μM) than hACE2 receptor (1.12 μM), according to biolayer interferometry-based analysis. In silico analysis indicated geraniin’s interaction with the residues functionally important in the binding between the two proteins. Thus, geraniin is a promising therapeutic agent for COVID-19 by blocking SARS-CoV-2’s entry into human cells.
Ortho Isomeric Mn(III) N-Alkyl- and Alkoxyalkylpyridylporphyrins—Enhancers of Hyaluronan Degradation Induced by Ascorbate and Cupric Ions
Authors: Valachová Katarína , Rapta Peter , Moura Nuno M. M. , Batinic-Haberle Ines , Šoltés Ladislav
Published: August 2021

High levels of hyaluronic acid (HA) in tumors correlate with poor outcomes with several types of cancers due to HA-driven support of adhesion, migration and proliferation of cells. In this study we explored how to enhance the degradation of HA into low-molecular fragments, which cannot prevent the immune system to fight tumor proliferation and metastases. The physiological solution of HA was exposed to oxidative degradation by ascorbate and cupric ions in the presence of either one of three ortho isomeric Mn(III) substituted N-alkyl- and alkoxyalkylpyridylporphyrins or para isomeric Mn(III) N-methylpyridyl analog, commonly known as mimics of superoxide dismutase. The changes in hyaluronan degradation kinetics by four Mn(III) porphyrins were monitored by measuring the alteration in the dynamic viscosity of the HA solution. The ortho compounds MnTE-2-PyP5+ (BMX-010, AEOL10113), MnTnBuOE-2-PyP5+ (BMX-001) and MnTnHex-2-PyP5+ are able to redox cycle with ascorbate whereby producing H2O2 which is subsequently coupled with Cu(I) to produce the •OH radical essential for HA degradation. Conversely, with the para analog, MnTM-4-PyP5+, no catalysis of HA degradation was demonstrated, due to its inertness towards redox cycling with ascorbate. The impact of different Mn(III)-porphyrins on the HA decay was further clarified by electron paramagnetic resonance spectrometry. The ability to catalyze the degradation of HA in a biological milieu, in the presence of cupric ions and ascorbate under the conditions of high tumor oxidative stress provides further insight into the anticancer potential of redox-active ortho isomeric Mn(III) porphyrins.