bluuebunny/crossref_metadata_2025 huggingface dataset:
Investigating the Self-Reported Health Status of Domestic and Overseas Chinese Populations during the COVID-19 Pandemic
Authors: Mao Zhuxin , Chen Bohao , Wang Wei , Kind Paul , Wang Pei
Published: March 2021

To control the spread of COVID-19, governments in different countries and regions implemented various types of lockdown and outdoor restrictions. The research aimed to describe and compare the health status of Chinese people both domestically and abroad in this global health crisis. An online questionnaire survey was distributed to Chinese mainland citizens living in Hubei (the lockdown province), outside Hubei, and those living abroad in 2020. A total of 1000 respondents were recruited and reported worse health status compared with Chinese population norms. People living in Hubei reported worse health status than those living outside Hubei but revealed better health status than overseas respondents. It was clear that the pandemic as well as strict lockdown and outdoor restriction policies affected Chinese people’s health. It is important for the Chinese government to be aware of the negative impact of such strict policies and to take measures to reduce the panic of society when implementing similar policies in the future. It also implies that governments in other countries should promote social support for those who live far from home and actively call for support for non-discriminatory attitudes toward ethnic minorities.
Identifying Cognitive Mechanism Underlying Situation Awareness of Pilots’ Unsafe Behaviors Using Quantitative Modeling
Authors: Jiang Shaoqi , Chen Weijiong , Kang Yutao , Liu Jiahao , Kuang Wanglai
Published: March 2021

Situation awareness (SA) of pilots’ unsafe behavior can ensure safety onboard. Thus, the cognitive mechanism that controls the SA leading to unsafe behavior must be articulated. This study employs the SA model and theory of planned behavior (TPB) to articulate a quantitative model of ship safe piloting. Firstly, the hierarchical classification framework of unsafe behaviors was constructed as an analytical foundation for rational and unconscious behaviors in sight of cognitive processes, and then the measurement elements of the cognitive mechanisms for behaviors were identified. Subsequently, based on the structural model, a hypothetical model of the cognitive path for unsafe behaviors was proposed by using the extended TPB, where there are four independent variables (i.e., attitude (ATD), subjective norm (SN), and perceived behavioral control (PBC)), one mediating variables (i.e., SA) and two dependent variables (i.e., behavioral intention (BI) and unsafe behaviors (BE)). Finally, this hypothetical model was analyzed with the data resources from extended TPB questionnaire of 295 pilots. Analysis results show that relationships of causation and mediation in the cognitive mechanism are in line with the behavior pattern and SA have a pronounced mediating effect and a strong relevance to the causal chain of extended TPB framework. This study integrated the SA three-level model to understand the motivation–cognition–action–feedback (MCAF) mechanism of pilots’ unsafe behaviors under cognitive mode of information processing through structural model. It would make a valuable contribution to the assessment and intervention of safety behaviors, and provide a basic framework for monitoring the situation awareness of pilot by man-machine interactive measurement technology in the future.
Possible Roles of Permafrost Melting, Atmospheric Transport, and Solar Irradiance in the Development of Major Coronavirus and Influenza Pandemics
Authors: Hofmeister Anne M. , Seckler James M. , Criss Genevieve M.
Published: March 2021

Major pandemics involving respiratory viruses develop semi-regularly and require a large flux of novel viruses, yet their origination is equivocal. This paper explores how natural processes could give rise to this puzzling combination of characteristics. Our model is based on available data regarding the emergence of historic influenzas, early COVID-19 cases and spreading, the microbiome of permafrost, long-distance airborne transport of viruses reaching stratospheric levels, ultraviolet immunosuppression, sunlight variations, weather patterns, Arctic thawing, and global warming. Atmospheric conveyance is supported by hemispheric distribution disparities, ties of COVID-19 cases to air pollution particulate concentrations, and contemporaneous animal infections. The following sequence is proposed: (1) virus emergence after hot Arctic summers, predominantly near solar irradiance maxima or involving wildfires, indicates release of large amounts of ancient viruses during extensive permafrost melting, which are then incorporated in autumn polar air circulation, where cold storage and little sunlight permit survival. (2) Pandemics onset in winter to spring at rather few locations: from climate data on Wuhan, emergence occurs where the North Polar Jet stream hovers while intersecting warmer, moist air, producing rain which deposits particulates with the viral harvest on a vulnerable human population. (3) Spring and summer increases in COVID-19 cases link to high solar irradiance, implicating ultraviolet immune suppression as one means of amplification. (4) Viruses multiplied by infected humans at close range being incorporated in atmospheric circulation explains rapid global spread, periodic case surges (waves), and multi-year durations. Pollution and wind geography affect uptake and re-distribution. Our model can be tested, e.g., against permafrost stored in laboratories as well as Artic air samples, and suggests mitigating actions.
Association of Infrastructure and Route Environment Factors with Cycling Injury Risk at Intersection and Non-Intersection Locations: A Case-Crossover Study of Britain
Authors: Aldred Rachel , Kapousizis Georgios , Goodman Anna
Published: March 2021

Objective: This paper examines infrastructural and route environment correlates of cycling injury risk in Britain for commuters riding in the morning peak. Methods: The study uses a case-crossover design which controls for exposure. Control sites from modelled cyclist routes (matched on intersection status) were compared with sites where cyclists were injured. Conditional logistic regression for matched case–control groups was used to compare characteristics of control and injury sites. Results: High streets (defined by clustering of retail premises) raised injury odds by 32%. Main (Class A or primary) roads were riskier than other road types, with injury odds twice that for residential roads. Wider roads, and those with lower gradients increased injury odds. Guard railing raised injury odds by 18%, and petrol stations or car parks by 43%. Bus lanes raised injury odds by 84%. As in other studies, there was a ‘safety in numbers’ effect from more cyclists. Contrary to other analysis, including two recent studies in London, we did not find a protective effect from cycle infrastructure and the presence of painted cycle lanes raised injury odds by 54%. At intersections, both standard and mini roundabouts were associated with injury odds several times higher than other intersections. Presence of traffic signals, with or without an Advanced Stop Line (‘bike box’), had no impact on injury odds. For a cyclist on a main road, intersections with minor roads were riskier than intersections with other main roads. Conclusions: Typical cycling environments in Britain put cyclists at risk, and infrastructure must be improved, particularly on busy main roads, high streets, and bus routes.
Understanding Public Health Adaptation to Climate Change: An Explorative Study on the Development of Adaptation Strategies Relating to the Oak Processionary Moth in The Netherlands
Authors: Buist Yvette , Bekker Marleen , Vaandrager Lenneke , Koelen Maria
Published: March 2021

Understanding of public health adaptation (PHA) to climate change and implementation is limited. This study therefore focuses on one specific PHA issue: adaptation to the oak processionary moth (OPM). The aim is to examine the development of OPM adaptation in order to offer a problem description of the complexities involved in OPM adaptation. In this explorative case study, we investigate adaptation strategies based on semi-structured interviews with 26 actors involved in OPM adaptation in The Netherlands. The results indicate that the context of OPM adaptation is relatively complex, given the involvement of many interdependent actors. OPM adaptation was developed with limited knowledge and strategies were based on ad hoc approaches in which there was ambiguity about tasks and expertise. In addition, different actors have different perceptions and values concerning health, sustainability, risks and responsibilities influencing decision-making processes, while also posing a challenge to collaboration and the development of a coordinated approach. The generation of knowledge and its translation into practical strategies calls for interdisciplinary cooperation in knowledge development. PHA adaptation involves more than technical and organisational solutions alone. It also entails the development of a shared problem perception and solution space in which citizens are also engaged.
Evaluating the Impact of the Healthy Kids Community Challenge (HKCC) on Physical Activity of Older Youth
Authors: Leatherdale Scott T. , Burns Kathleen E. , Qian Wei , Faulkner Guy , Carson Valerie
Published: March 2021

(1) Background: The Healthy Kids Community Challenge (HKCC) was a community-based obesity prevention intervention funded by the Government of Ontario (Canada). (2) Methods: A quasi-experimental design was used to examine the impact of the HKCC on physical activity (PA) outcomes using both repeat cross-sectional (T1 2014–2015, n = 31,548; T2 2015–2016, n = 31,457; and T3 2016–2017, n = 30,454) and longitudinal data (n = 3906) from the COMPASS study. Grade 9–12 students in HKCC communities were placed into one of three intervention groups [T2 data collection post-HKCC finishing (IG1), T2 data collection during HKCC (IG2), and T2 data collection pre-HKCC starting (IG3)], Ontario students in non-HKCC communities were Control Group 1 (CG1) and Alberta students were Control Group 2 (CG2). (3) Results: Repeat cross-sectional results show over time the HKCC had no significant impact on PA outcomes in any of the intervention groups. Longitudinal results show a significant decrease in time spent in moderate-to-vigorous PA (IG2: −3.15 min/day) between T1 and T3 in IG2. (4) Conclusions: These results suggest the HKCC did not have an impact on improving PA outcomes among older youth in HKCC communities. Moving forward, there is a need to provide effective and sustainable interventions to promote PA among older youth.
Attachment Security and Suicide Ideation and Behaviour: The Mediating Role of Reflective Functioning
Authors: Green Jessica , Berry Katherine , Danquah Adam , Pratt Daniel
Published: March 2021

Background: To understand why attachment difficulties predispose individuals to suicidal thinking (suicide ideation) and behaviour, a leading cause of death, we need to explore the role of pertinent psychological mechanisms. Attachment processes are closely linked to the development of mentalisation capabilities, or reflective functioning; the ability to understand and interpret self and other behaviour as an expression of mental states. Interventions designed to improve mentalisation have been associated with a reduction in suicidal behaviour, yet reflective functioning has not been directly investigated in relation to suicidal ideation and behaviour. Aim: We aim to further verify the link between adult attachment security and suicidal ideation and examine whether deficits in reflective functioning mediate this relationship. Methods: Sixty-seven participants who experienced suicidal ideation within the past 12 months completed self-report measures of adult attachment, current suicidal ideation, reflective functioning, depressive symptomology and hopelessness. Partial correlations, mediation analyses and group comparisons were conducted to explore relationships between these factors. Results: Findings did not support a mediational role for reflective functioning in the relationship between attachment security and suicidal ideation. A direct relationship was established between attachment avoidance and suicidal ideation, after controlling for age, gender and depressive symptoms. However, participants with a history of attempted suicide were higher in anxious attachment compared to participants with no such history. Conclusions: This study shows that the attachment dimensions of attachment anxiety and avoidance may play differential roles in increasing risk for suicidal attempts versus ideation. This has important implications for tailoring interventions. Those aimed at reducing suicide attempts should focus on reducing attachment anxiety by helping people develop skills in emotional regulation. Interventions aimed at reducing suicidal ideation should focus on reducing attachment avoidance by helping people develop closer relationships with significant others. However, longitudinal and experimental designs are required to verify causality.
Successful Aging Perception in Middle-Aged Korean Men: A Q Methodology Approach
Authors: Seong Mi Hyeon , Shin Eunyoung , Sok Sohyune
Published: March 2021

The purpose of this study is to identify the types of perception of successful aging in middle-aged men and to analyze and describe the characteristics of each type of successful aging perception of middle-aged men in South Korea. This study uses an exploratory study design, applying the Q methodology, which is a subjective research method. The participants were 25 middle-aged men (40 to 60 years old) living in C, Y, and B cities, which were P-samples that were judged to best reveal the successful aging of middle-aged men. In this study, principal component analysis of the PQ method program was used. The five perception types of successful aging among middle-aged men are Type 1 for the “leisure type”, Type 2 for the “mature type”, Type 3 for the “health-oriented type”, Type 4 for the “patriarchal type”, and Type 5 for the “family-centered type”. The mature type had the highest variance among the five types, and the leisure type was the type that showed the second-highest variance. In nursing practice, nurses need to pay attention to the successful aging perceptions of middle-aged Korean men for their successful aging in the future.
Data Mining in Healthcare: Applying Strategic Intelligence Techniques to Depict 25 Years of Research Development
Authors: Kolling Maikel Luis , Furstenau Leonardo B. , Sott Michele Kremer , Rabaioli Bruna , Ulmi Pedro Henrique , Bragazzi Nicola Luigi , Tedesco Leonel Pablo Carvalho
Published: March 2021

In order to identify the strategic topics and the thematic evolution structure of data mining applied to healthcare, in this paper, a bibliometric performance and network analysis (BPNA) was conducted. For this purpose, 6138 articles were sourced from the Web of Science covering the period from 1995 to July 2020 and the SciMAT software was used. Our results present a strategic diagram composed of 19 themes, of which the 8 motor themes (‘NEURAL-NETWORKS’, ‘CANCER’, ‘ELETRONIC-HEALTH-RECORDS’, ‘DIABETES-MELLITUS’, ‘ALZHEIMER’S-DISEASE’, ‘BREAST-CANCER’, ‘DEPRESSION’, and ‘RANDOM-FOREST’) are depicted in a thematic network. An in-depth analysis was carried out in order to find hidden patterns and to provide a general perspective of the field. The thematic network structure is arranged thusly that its subjects are organized into two different areas, (i) practices and techniques related to data mining in healthcare, and (ii) health concepts and disease supported by data mining, embodying, respectively, the hotspots related to the data mining and medical scopes, hence demonstrating the field’s evolution over time. Such results make it possible to form the basis for future research and facilitate decision-making by researchers and practitioners, institutions, and governments interested in data mining in healthcare.
Cognitive Differences in the Older Adults Living in the General Community: Gender and Mental Occupational State Study
Authors: Calatayud Estela , Salavera Carlos , Gómez-Soria Isabel
Published: March 2021

Older adults are particularly vulnerable to cognitive impairment with age, and gender differences are remarkable. However, there is very little evidence to identify both baseline cognitive and occupational gender differences prior to older adults’ retirement to design more efficient personalized cognitive interventions. This descriptive observational study examined gender differences in initial cognitive performance in 367 older adults with subjective memory complaints from a primary healthcare center in Zaragoza (Spain). To evaluate initial cognitive performance, the Spanish version of the Mini-Mental State Examination (MEC-35) and the set test were used to measure verbal fluency. Sociodemographic and clinical characteristics were evaluated, and cognitive and occupational differences were analyzed per gender. Men had higher educational and occupational levels, were older and more of them were married (p < 0.001) than women. Regarding cardiovascular risk factors, diabetes and cerebrovascular accidents were more frequent in women, while hypercholesterolemia and obesity were more frequent in men (p < 0.001). High blood pressure was more frequent in women, but not significantly so (p = 0.639). Global cognition was higher in men (p < 0.001) for attention, calculation, and language (p < 0.001). Verbal fluency was higher in women, but the difference was not statistically significant (p = 0.105). These results could be generalized to other health centers in the province and other Spanish autonomous communities as their sociodemographic variables are similar. Individualized interventions that adapt to gender, cognitive and initial occupational performance should be developed and adapted to elderly populations living in the general community to maintain their cognitive capacity and prevent their cognitive impairment and the social health costs this would imply.
Impact of Innovation City Projects on National Balanced Development in South Korea: Identifying Regional Network and Centrality
Authors: Ahn Jane , Seo Ducksu , Kwon Youngsang
Published: March 2021

Innovation City projects, aimed at balanced national development in South Korea, have relocated public institutions from the Seoul metropolitan area to provinces, decentralizing population and economic functions, over the past decade. This study measured changes in regional centrality (the central and local location or hierarchy of objects in a network) at the 14 cities where Innovation City projects were constructed. Commuter Origin-Destination data were analyzed using Rstudio. In the case of connectivity centrality, 13 out of 14 regions saw a rise in centrality values; among them, Busan, Daegu, and Ulsan belong to large cities. This suggests that the impact of Innovation City projects on established metropolitan areas may not be very significant. Five of the 14 projects increased the value of eigenvector centrality, while 10 increased the centrality ranking. This means that the absolute traffic volume of Innovation Cities across the country had increased, while the centrality of areas around these cities declined, suggesting that Innovation Cities should pursue co-prosperity with surrounding areas. In this way, Innovation Cities can have a positive impact on surrounding areas, and positive externalities of relocation projects are maximized. However, such development effects are confined to Innovation City areas, negatively influencing balanced regional development.
Consideration of Uncertainty Information in Accessibility Analyses for an Effective Use of Urban Infrastructures
Authors: Schiewe Jochen , Knura Martin
Published: March 2021

Accessibility analyses are an essential step in the evaluation and planning of urban infrastructures such as transport or pipeline networks. However, these studies generally produce sharply defined lines (called isovarones) or areas (called isovarone areas) that represent the same or similar accessibility. Uncertainties in the input data are usually not taken into account. The aim of this contribution is, therefore, to set up a structured framework that describes the integration of uncertainty information for accessibility analyses. This framework takes uncertainties in the input data, in the processing step, in the target variables, and in the final visualization into account. Particular attention is paid, on the one hand, to the impact of the uncertainties in the target values, as these are key factors for reasoning and decision making. On the other hand, the visualization component is emphasized by applying a dichotomous classification of uncertainty visualization methods. This framework leads to a large set of possible combinations of uncertainty categories. Five selected examples that have been generated with a new software tool and that cover important combinations are presented and discussed.
Evaluating Natural Ecological Land Change in Function-Oriented Planning Regions Using the National Land Use Survey Data from 2009 to 2018 in China
Authors: Zhang Zhijie , Zhang Yuanjie , Yu Xiao , Lei Liping , Chen Yuqi , Guo Xudong
Published: March 2021

The natural ecological lands, such as forest land, grassland, wetland, etc., constitute the most important factor for maintaining and preserving the earth’s ecosystem, which must be well concerned in the regional function-oriented planning for the sustainability of human economic development. We analyzed and evaluated the change of natural ecological land in the function-oriented planning regions where we applied the major function-oriented zones introduced as a new concept in China. Using the land-use data from 2009 to 2018 that were produced by the National Land Use Survey, we re-classified natural ecological land types into the forest, grassland, wetland, and bare land, and then addressed the changes of natural ecological land types from 2009 to 2018 in the major function-oriented zones. As a result, the area of natural ecological lands generally tended to decrease from 2009 to 2018, while the decreasing trend of natural ecological land areas was controlled after 2015 with the implementation of governmental policies for environment protection and eco-logical projects. Especially, the decrease of forest land area significantly tended to be zero in 2018 in optimal development zones. The decreased areas of natural ecological lands were mostly converted from artificial land from 2008 to 2019. On the other side, the forest lands mostly changed from cropland and grassland in key development zones, agricultural production zones, and key ecological function zones, due to the fact that grassland conversed in afforestation during this period. The evaluation of natural ecological land changes, which could be implemented by using the annual updates of national land-use data in China, is significant to support the government’s spatial regulation design, to reshape the planned regions, and make policies for environmental restoration and protection management.
Epigallocatechin-3-Gallate-Loaded Liposomes Favor Anti-Inflammation of Microglia Cells and Promote Neuroprotection
Authors: Cheng Chun-Yuan , Barro Lassina , Tsai Shang-Ting , Feng Tai-Wei , Wu Xiao-Yu , Chao Che-Wei , Yu Ruei-Siang , Chin Ting-Yu , Hsieh Ming Fa
Published: March 2021

Microglia-mediated neuroinflammation is recognized to mainly contribute to the progression of neurodegenerative diseases. Epigallocatechin-3-gallate (EGCG), known as a natural antioxidant in green tea, can inhibit microglia-mediated inflammation and protect neurons but has disadvantages such as high instability and low bioavailability. We developed an EGCG liposomal formulation to improve its bioavailability and evaluated the neuroprotective activity in in vitro and in vivo neuroinflammation models. EGCG-loaded liposomes have been prepared from phosphatidylcholine (PC) or phosphatidylserine (PS) coated with or without vitamin E (VE) by hydration and membrane extrusion method. The anti-inflammatory effect has been evaluated against lipopolysaccharide (LPS)-induced BV-2 microglial cells activation and the inflammation in the substantia nigra of Sprague Dawley rats. In the cellular inflammation model, murine BV-2 microglial cells changed their morphology from normal spheroid to activated spindle shape after 24 h of induction of LPS. In the in vitro free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, EGCG scavenged 80% of DPPH within 3 min. EGCG-loaded liposomes could be phagocytized by BV-2 cells after 1 h of cell culture from cell uptake experiments. EGCG-loaded liposomes improved the production of BV-2 microglia-derived nitric oxide and TNF-α following LPS. In the in vivo Parkinsonian syndrome rat model, simultaneous intra-nigral injection of EGCG-loaded liposomes attenuated LPS-induced pro-inflammatory cytokines and restored motor impairment. We demonstrated that EGCG-loaded liposomes exert a neuroprotective effect by modulating microglia activation. EGCG extracted from green tea and loaded liposomes could be a valuable candidate for disease-modifying therapy for Parkinson’s disease (PD).
Metabolic Fingerprinting of Murine L929 Fibroblasts as a Cell-Based Tumour Suppressor Model System for Methionine Restriction
Authors: Schmitz Werner , Koderer Corinna , El-Mesery Mohamed , Gubik Sebastian , Sampers Rene , Straub Anton , Kübler Alexander Christian , Seher Axel
Published: March 2021

Since Otto Warburg reported in 1924 that cancer cells address their increased energy requirement through a massive intake of glucose, the cellular energy level has offered a therapeutic anticancer strategy. Methionine restriction (MetR) is one of the most effective approaches for inducing low-energy metabolism (LEM) due to the central position in metabolism of this amino acid. However, no simple in vitro system for the rapid analysis of MetR is currently available, and this study establishes the murine cell line L929 as such a model system. L929 cells react rapidly and efficiently to MetR, and the analysis of more than 150 different metabolites belonging to different classes (amino acids, urea and tricarboxylic acid cycle (TCA) cycles, carbohydrates, etc.) by liquid chromatography/mass spectrometry (LC/MS) defines a metabolic fingerprint and enables the identification of specific metabolites representing normal or MetR conditions. The system facilitates the rapid and efficient testing of potential cancer therapeutic metabolic targets. To date, MS studies of MetR have been performed using organisms and yeast, and the current LC/MS analysis of the intra- and extracellular metabolites in the murine cell line L929 over a period of 5 days thus provides new insights into the effects of MetR at the cellular metabolic level.
A High-Throughput Screening System Based on Fluorescence-Activated Cell Sorting for the Directed Evolution of Chitinase A
Authors: Menghiu Gheorghita , Ostafe Vasile , Prodanović Radivoje , Fischer Rainer , Ostafe Raluca
Published: March 2021

Chitinases catalyze the degradation of chitin, a polymer of N-acetylglucosamine found in crustacean shells, insect cuticles, and fungal cell walls. There is great interest in the development of improved chitinases to address the environmental burden of chitin waste from the food processing industry as well as the potential medical, agricultural, and industrial uses of partially deacetylated chitin (chitosan) and its products (chito-oligosaccharides). The depolymerization of chitin can be achieved using chemical and physical treatments, but an enzymatic process would be more environmentally friendly and more sustainable. However, chitinases are slow-acting enzymes, limiting their biotechnological exploitation, although this can be overcome by molecular evolution approaches to enhance the features required for specific applications. The two main goals of this study were the development of a high-throughput screening system for chitinase activity (which could be extrapolated to other hydrolytic enzymes), and the deployment of this new method to select improved chitinase variants. We therefore cloned and expressed the Bacillus licheniformis DSM8785 chitinase A (chiA) gene in Escherichia coli BL21 (DE3) cells and generated a mutant library by error-prone PCR. We then developed a screening method based on fluorescence-activated cell sorting (FACS) using the model substrate 4-methylumbelliferyl β-d-N,N′,N″-triacetyl chitotrioside to identify improved enzymes. We prevented cross-talk between emulsion compartments caused by the hydrophobicity of 4-methylumbelliferone, the fluorescent product of the enzymatic reaction, by incorporating cyclodextrins into the aqueous phases. We also addressed the toxicity of long-term chiA expression in E. coli by limiting the reaction time. We identified 12 mutants containing 2–8 mutations per gene resulting in up to twofold higher activity than wild-type ChiA.
Targeting mTOR-CCL20 Signaling May Improve Response to Docetaxel in Head and Neck Squamous Cell Carcinoma
Authors: Chou Ming-Huei , Chuang Hui-Ching , Lin Yu-Tsai , Tsai Ming-Hsien , Kao Ying-Hsien , Lin I-Chun , Huang Tai-Lin , Fang Fu-Min , Chien Chih-Yen
Published: March 2021

Patients with advanced head and neck squamous cell carcinoma (HNSCC) usually show a dismal prognosis. It is this worthwhile to develop new, effective therapeutic regimens for these patients, such as molecular targeted therapy, which is promising as an alternative or combination treatment for HNSCC. The mammalian target of rapamycin (mTOR) pathway, which plays an important role in the carcinogenesis of HNSCC, is the most frequently activated, and is thus worthy of further investigation. In this study, two human HNSCC cell lines, FaDu and SAS, were evaluated for cell growth with trypan blue staining and tumor growth using an orthotopic xenograft model. The immunohistochemical expression of mTOR in the subcutaneous xenograft model and the inhibitory effects of docetaxel on the growth and state of activation of the PI3K/mTOR pathway were also evaluated and examined by colony formation and Western blot, respectively. Cell proliferation and migration were measured by water-soluble tetrazolium salt (WST-1) and OrisTM cell migration assay, respectively. Furthermore, the effects of rapamycin and BEZ235, a phosphatidylinositol 3-kinases (PI3K) and mTOR inhibitor in combination with docetaxel or CCL20 were evaluated in the FaDu and SAS cells. The results showed that the expression of mTOR was significantly higher in the SAS and FaDu xenograft models than in the control. Docetaxel treatment significantly suppressed HNSCC cell proliferation and migration in vitro via the PI3K/mTOR/CCL-20 signaling pathway. Additionally, when administered in a dose-dependent fashion, mTOR inhibitors inhibited the growth and migration of the HNSCC cells. This combination was synergistic with docetaxel, resulting in almost complete cell growth and migration arrest. In conclusion, docetaxel significantly inhibited HNSCC cell proliferation and migration in vitro via the PI3K/mTOR/CCL-20 signaling pathway. The synergistic and additive activity of mTOR inhibitors combined with docetaxel shows potential as a new treatment strategy for HNSCC.
β-Hydroxy-β-Methylbutyrate (HMB) Supplementation Prevents Bone Loss during Pregnancy—Novel Evidence from a Spiny Mouse (Acomys cahirinus) Model
Authors: Tomaszewska Ewa , Donaldson Janine , Kosiński Jakub , Dobrowolski Piotr , Tomczyk-Warunek Agnieszka , Hułas-Stasiak Monika , Lamorski Krzysztof , Laskowska-Woźniak Dorota , Muszyński Siemowit , Blicharski Rudolf , Blicharski Tomasz
Published: March 2021

The aim of this study was to determine the effects of ß-hydroxy-ß-methylbutyrate (HMB) supplementation during pregnancy on postpartum bone tissue quality by assessing changes in trabecular and compact bone as well as in hyaline and epiphyseal cartilage. The experiment was carried out on adult 6-month-old female spiny mice (Acomys cahirinus) divided into three groups: pregnant control (PregCont), pregnant HMB-treated (supplemented with 0.02 g/kg b.w of HMB during the second trimester of pregnancy, PregHMB), and non-pregnant females (NonPreg). Cross-sectional area and cortical index of the femoral mid-shaft, stiffness, and Young modulus were significantly greater in the PregHMB group. Whole-bone mineral density was similar in all groups, and HMB supplementation increased trabecular number. Growth plate cartilage was the thinnest, while the articular cartilage was the thickest in the PregHMB group. HMB supplementation increased the content of proteoglycans in the articular cartilage and the percentage of immature collagen content in metaphyseal trabeculae and compact bone. In summary, dietary HMB supplementation during the second trimester of pregnancy intensifies bone metabolic processes and prevents bone loss during pregnancy.
Predialysis and Dialysis Therapies Differently Affect Nitric Oxide Synthetic Pathway in Red Blood Cells from Uremic Patients: Focus on Peritoneal Dialysis
Authors: Palmerini Carola , Piscitani Luca , Bologna Giuseppina , Riganti Chiara , Lanuti Paola , Mandatori Domitilla , Di Liberato Lorenzo , Di Fulvio Giorgia , Sirolli Vittorio , Renda Giulia , Pipino Caterina , Marchisio Marco , Bonomini Mario , Pandolfi Assunta , Di Pietro Natalia
Published: March 2021

Red blood cells (RBCs) have been found to synthesize and release both nitric oxide (NO) and cyclic guanosine monophosphate (cGMP), contributing to systemic NO bioavailability. These RBC functions resulted impaired in chronic kidney disease (CKD). This study aimed to evaluate whether predialysis (conservative therapy, CT) and dialysis (peritoneal dialysis, PD; hemodialysis, HD) therapies used during CKD progression may differently affect NO-synthetic pathway in RBCs. Our data demonstrated that compared to PD, although endothelial-NO-synthase activation was similarly increased, HD and CT were associated to cGMP RBCs accumulation, caused by reduced activity of cGMP membrane transporter (MRP4). In parallel, plasma cGMP levels were increased by both CT and HD and they significantly decreased after hemodialysis, suggesting that this might be caused by reduced cGMP renal clearance. As conceivable, compared to healthy subjects, plasma nitrite levels were significantly reduced by HD and CT but not in patients on PD. Additionally, the increased carotid intima-media thickness (IMT) values did not reach the significance exclusively in patients on PD. Therefore, our results show that PD might better preserve the synthetic NO-pathway in CKD-erythrocytes. Whether this translates into a reduced development of uremic vascular complications requires further investigation.
Vascular Endothelial Integrity Affects the Severity of Enterovirus-Mediated Cardiomyopathy
Authors: Park Jin-Ho , Shin Ha-Hyeon , Rhyu Hyun-Seung , Kim So-Hee , Jeon Eun-Seok , Lim Byung-Kwan
Published: March 2021

Coxsackievirus and adenovirus receptor (CAR) is present in epithelial and vascular endothelial cell junctions. We have previously shown a hemorrhagic phenotype in germ-line CAR knock-out mouse embryos; we have also found that CAR interacts with ZO-1 and β-catenin. However, the role of CAR in vascular endothelial junction permeability has not been proven. To understand the roles of CAR in the vascular endothelial junctions, we generated endothelium-specific CAR knockout (CAR-eKO) mice. In the absence of CAR, the endothelial cell layer showed an increase in transmembrane electrical resistance (TER, Ω) and coxsackievirus permeability. Evans blue dye and 70 kDa dextran-FITC were delivered by tail vein injection. We observed increased vascular permeability in the hearts of adult CAR-eKO mice compare with wild-type (WT) mice. There was a marked increase in monocyte and macrophage penetration into the peritoneal cavity caused by thioglycolate-induced peritonitis. We found that CAR ablation in endothelial cells was not significantly increased coxsackievirus B3 (CVB3) induced myocarditis in murine model. However, tissue virus titers were significantly higher in CAR-eKO mice compared with WT. Moreover, CVB3 was detected in the brain of CAR-eKO mice. Endothelial CAR deletion affects the expression of major endothelial junction proteins, such as cadherin and platelet endothelial cell adhesion molecule-1 (PECAM-1) in the cultured endothelial cells as well as liver vessel. We suggest that CAR expression is required for normal vascular permeability and endothelial tight junction homeostasis. Furthermore, CVB3 organ penetration and myocarditis severities were dependent on the endothelial CAR level.
Anti-Cancer Effects and Tumor Marker Role of Glutathione S-Transferase Mu 5 in Human Bladder Cancer
Authors: Jou Yeong-Chin , Wang Shou-Chieh , Dia Yuan-Chang , Wang Shou-Tsung , Yu Min-Hua , Yang Hsin-Yi , Chen Lei-Chin , Shen Cheng-Huang , Liu Yi-Wen
Published: March 2021

Our previous study demonstrated that the glutathione S-transferase Mu 5 (GSTM5) gene is highly CpG-methylated in bladder cancer cells and that demethylation by 5-aza-dC activates GSTM5 gene expression. The aim of the present study was to investigate the role of GSTM5 in bladder cancer. The levels of GSTM5 gene expression and DNA methylation were analyzed in patients with bladder cancer, and functional studies of GSTM5 were conducted using GSTM5 overexpression in cultured bladder cancer cells. Clinical analysis revealed that the GSTM5 mRNA expression was lower in bladder cancer tissues than in normal tissues and that the level of GSTM5 DNA methylation was higher in bladder cancer tissues than in normal urine pellets. Overexpression of GSTM5 decreased cell proliferation, migration and colony formation capacity. Glutathione (GSH) assay results indicated that cellular GSH concentration was decreased by GSTM5 expression and that GSH supplementation reversed the decrease in proliferation and migration of cells overexpressing GSTM5. By contrast, a GSH synthesis inhibitor significantly decreased 5637 cell GSH levels, survival and migration. Furthermore, GSTM5 overexpression inhibited the adhesion of cells to the extracellular matrix protein fibronectin. To elucidate the effect of GSTM5 on anticancer drugs used to treat bladder cancer, cellular viability was compared between cells with or without GSTM5 overexpression. GSTM5-overexpressed cells showed no significant change in the cytotoxicity of cisplatin or mitomycin C in 5637, RT4 and BFTC 905 cells. Though a degree of resistance to doxorubicin was noted in 5637 cells overexpressing GSTM5, no such resistance was observed in RT4 and BFTC 905 cells. In summary, GSTM5 plays a tumor suppressor role in bladder cancer cells without significantly affecting chemoresistance to cisplatin and mitomycin C, and the cellular GSH levels highlight a key mechanism underlying the cancer inhibition effect of GSTM5. These findings suggest that low gene expression and high DNA methylation levels of GSTM5 may act as tumor markers for bladder cancer.
The Role of Z-disc Proteins in Myopathy and Cardiomyopathy
Authors: Wadmore Kirsty , Azad Amar J. , Gehmlich Katja
Published: March 2021

The Z-disc acts as a protein-rich structure to tether thin filament in the contractile units, the sarcomeres, of striated muscle cells. Proteins found in the Z-disc are integral for maintaining the architecture of the sarcomere. They also enable it to function as a (bio-mechanical) signalling hub. Numerous proteins interact in the Z-disc to facilitate force transduction and intracellular signalling in both cardiac and skeletal muscle. This review will focus on six key Z-disc proteins: α-actinin 2, filamin C, myopalladin, myotilin, telethonin and Z-disc alternatively spliced PDZ-motif (ZASP), which have all been linked to myopathies and cardiomyopathies. We will summarise pathogenic variants identified in the six genes coding for these proteins and look at their involvement in myopathy and cardiomyopathy. Listing the Minor Allele Frequency (MAF) of these variants in the Genome Aggregation Database (GnomAD) version 3.1 will help to critically re-evaluate pathogenicity based on variant frequency in normal population cohorts.
Therapeutic Role of Tocilizumab in SARS-CoV-2-Induced Cytokine Storm: Rationale and Current Evidence
Authors: Pelaia Corrado , Calabrese Cecilia , Garofalo Eugenio , Bruni Andrea , Vatrella Alessandro , Pelaia Girolamo
Published: March 2021

Among patients suffering from coronavirus disease 2019 (COVID-19) syndrome, one of the worst possible scenarios is represented by the critical lung damage caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)-induced cytokine storm, responsible for a potentially very dangerous hyperinflammatory condition. Within such a context, interleukin-6 (IL-6) plays a key pathogenic role, thus being a suitable therapeutic target. Indeed, the IL-6-receptor antagonist tocilizumab, already approved for treatment of refractory rheumatoid arthritis, is often used to treat patients with severe COVID-19 symptoms and lung involvement. Therefore, the aim of this review article is to focus on the rationale of tocilizumab utilization in the SARS-CoV-2-triggered cytokine storm, as well as to discuss current evidence and future perspectives, especially with regard to ongoing trials referring to the evaluation of tocilizumab’s therapeutic effects in patients with life-threatening SARS-CoV-2 infection.
SARS-CoV-2 Entry Related Viral and Host Genetic Variations: Implications on COVID-19 Severity, Immune Escape, and Infectivity
Authors: Huang Szu-Wei , Wang Sheng-Fan
Published: March 2021

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has evolved to display particular patterns of genetic diversity in the genome across geographical regions. These variations in the virus and genetic variation in human populations can determine virus transmissibility and coronavirus disease 2019 (COVID-19) severity. Genetic variations and immune differences in human populations could be the driving forces in viral evolution. Recently emerged SARS-CoV-2 variants show several mutations at the receptor binding domain in the spike (S) glycoprotein and contribute to immune escape and enhanced binding with angiotensin 1-converting enzyme 2 (ACE2). Since ACE2 and transmembrane protease serine 2 (TMPRSS2) play important roles in SARS-CoV-2 entry into the cell, genetic variation in these host entry-related proteins may be a driving force for positive selection in the SARS-CoV-2 S glycoprotein. Dendritic or liver/lymph cell-specific intercellular adhesion molecule (ICAM)-3-grabbing non-integrin is also known to play vital roles in several pathogens. Genetic variations of these host proteins may affect the susceptibility to SARS-CoV-2. This review summarizes the latest research to describe the impacts of genetic variation in the viral S glycoprotein and critical host proteins and aims to provide better insights for understanding transmission and pathogenesis and more broadly for developing vaccine/antiviral drugs and precision medicine strategies, especially for high risk populations with genetic risk variants.
Therapeutic Potential of AAV1-Rheb(S16H) Transduction against Neurodegenerative Diseases
Authors: Nam Youngpyo , Moon Gyeong Joon , Kim Sang Ryong
Published: March 2021

Neurotrophic factors (NTFs) are essential for cell growth, survival, synaptic plasticity, and maintenance of specific neuronal population in the central nervous system. Multiple studies have demonstrated that alterations in the levels and activities of NTFs are related to the pathology and symptoms of neurodegenerative disorders, such as Parkinson’s disease (PD), Alzheimer’s disease (AD), and Huntington’s disease. Hence, the key molecule that can regulate the expression of NTFs is an important target for gene therapy coupling adeno-associated virus vector (AAV) gene. We have previously reported that the Ras homolog protein enriched in brain (Rheb)–mammalian target of rapamycin complex 1 (mTORC1) axis plays a vital role in preventing neuronal death in the brain of AD and PD patients. AAV transduction using a constitutively active form of Rheb exerts a neuroprotective effect through the upregulation of NTFs, thereby promoting the neurotrophic interaction between astrocytes and neurons in AD conditions. These findings suggest the role of Rheb as an important regulator of the regulatory system of NTFs to treat neurodegenerative diseases. In this review, we present an overview of the role of Rheb in neurodegenerative diseases and summarize the therapeutic potential of AAV serotype 1 (AAV1)-Rheb(S16H) transduction in the treatment of neurodegenerative disorders, focusing on diseases, such as AD and PD.
The Role of TRPA1 in Skin Physiology and Pathology
Authors: Maglie Roberto , Souza Monteiro de Araujo Daniel , Antiga Emiliano , Geppetti Pierangelo , Nassini Romina , De Logu Francesco
Published: March 2021

The transient receptor potential ankyrin 1 (TRPA1), a member of the TRP superfamily of channels, acts as ‘polymodal cellular sensor’ on primary sensory neurons where it mediates the peripheral and central processing of pain, itch, and thermal sensation. However, the TRPA1 expression extends far beyond the sensory nerves. In recent years, much attention has been paid to its expression and function in non-neuronal cell types including skin cells, such as keratinocytes, melanocytes, mast cells, dendritic cells, and endothelial cells. TRPA1 seems critically involved in a series of physiological skin functions, including formation and maintenance of physico-chemical skin barriers, skin cells, and tissue growth and differentiation. TRPA1 appears to be implicated in mechanistic processes in various immunological inflammatory diseases and cancers of the skin, such as atopic and allergic contact dermatitis, psoriasis, bullous pemphigoid, cutaneous T-cell lymphoma, and melanoma. Here, we report recent findings on the implication of TRPA1 in skin physiology and pathophysiology. The potential use of TRPA1 antagonists in the treatment of inflammatory and immunological skin disorders will be also addressed.
The ATXN2 Orthologs CID3 and CID4, Act Redundantly to In-Fluence Developmental Pathways throughout the Life Cycle of Arabidopsis thaliana
Authors: López-Juárez Zaira M. , Aguilar-Henonin Laura , Guzmán Plinio
Published: March 2021

RNA-binding proteins (RBPs) are key elements involved in post-transcriptional regulation. Ataxin-2 (ATXN2) is an evolutionarily conserved RBP protein, whose function has been studied in several model organisms, from Saccharomyces cerevisiae to the Homo sapiens. ATXN2 interacts with poly(A) binding proteins (PABP) and binds to specific sequences at the 3′UTR of target mRNAs to stabilize them. CTC-Interacting Domain3 (CID3) and CID4 are two ATXN2 orthologs present in plant genomes whose function is unknown. In the present study, phenotypical and transcriptome profiling were used to examine the role of CID3 and CID4 in Arabidopsis thaliana. We found that they act redundantly to influence pathways throughout the life cycle. cid3cid4 double mutant showed a delay in flowering time and a reduced rosette size. Transcriptome profiling revealed that key factors that promote floral transition and floral meristem identity were downregulated in cid3cid4 whereas the flowering repressor FLOWERING LOCUS C (FLC) was upregulated. Expression of key factors in the photoperiodic regulation of flowering and circadian clock pathways, were also altered in cid3cid4, as well as the expression of several transcription factors and miRNAs encoding genes involved in leaf growth dynamics. These findings reveal that ATXN2 orthologs may have a role in developmental pathways throughout the life cycle of plants.