bluuebunny/crossref_metadata_2025 huggingface dataset:
Relationship between Type and Intensity of Sports Activities and the Prevalence of Overweight in Serbian School Children
Authors: Malićević Sead , Mazić Sanja , Cataldi Stefania , Fischetti Francesco , Greco Gianpiero
Published: July 2022

This study aimed to explore the relationship between different forms and intensities of sports activities and the prevalence of overweight in primary school children. Using International Obesity Task Force BMI criteria, we have identified body weight status in a group of 2893 children aged 9–15 years participating in 27 different sports and within a group of 4987 non-active children of the same age. We have compared the prevalence of overweight and obesity between these groups, as well as between genders and different forms and intensities of sports, within the group of active children. We have found lower prevalence of overweight (Χ2 = 41.689, DF = 1, p < 0.001), as well as prevalence of obesity (Χ2 = 175.184, DF = 1, p < 0.001) in physically active children compared with their non-active counterparts, as expected. Overweight (including obesity) had the highest prevalence in mixed sports of the Classification of Sports of the European Association of Preventive Cardiology, in boys (p = 0.003), as well as in girls (p = 0.043). A lower prevalence of overweight was noted in boys (p = 0.001), as well as in girls (p = 0.025) with more than 7 years of training. Regarding the number of hours of training per week, a lower prevalence of overweight was noted only in girls with 4 and more hours of training per week (p = 0.025). Concerning intensity, we have found a significant drop in the prevalence of overweight in children with sports activities whose intensity is more than 60 MET-hours per week. We conclude that a significant relationship between sports and the prevalence of overweight is found in children with more than 60 MET-hours of sports activities per week, as well as among children involved in mixed sports, and after more than 7 years spent in regular sports training.
Exploring Scientific Discourse on Marine Litter in Europe: Review of Sources, Causes and Solutions
Authors: Havas Vilma , Løkke Søren , Kørnøv Lone
Published: July 2022

Marine litter is a transboundary environmental issue that affects all the world’s oceans. Marine litter research is a young discipline but one that has exploded during the last five years. However, the increased knowledge of sources and underlying causes to marine litter, as well as knowledge regarding solutions, lack systematic review and synthesis. This study reviews the scientific discourses around plastic marine litter in Europe, and more specifically, in Norway and Denmark, and explores emerging discourse coalitions. Four main thematic storylines on the source-cause-solution causal relationship, as well as two emerging storylines within marine litter research, are found. This study concludes that in order to secure sustainability of solutions and to avoid risk transformation and greenwashing, more interdisciplinary research, including life cycle assessment and investigations of scientific and societal discourses, is needed.
Does Food Insecurity in Early Life Make People More Depressed?—Evidence from CHARLS
Authors: Liu Yanrong , Zhao Xuecun
Published: July 2022

Based on the China Health and Retirement Longitudinal Study (CHARLS) data, this paper estimates the long-term association between food insecurity and later adult health and health behaviors with the Probit model. The results show that food insecurity in early life significantly increases adults’ depression likelihood (measured by the CES-D scale). The food insecurity experience is also negatively and significantly associated with individual self-rated health status, memory, sleep quality, and life satisfaction. The negative association between food insecurity and cognitive ability and sleep hours is larger for females.
The Culture-Oriented Urban Regeneration: Place Narrative in the Case of the Inner City of Haiyan (Zhejiang, China)
Authors: Diao Jiahui , Lu Shaoming
Published: July 2022

Recent decades have seen an increasing interest in the urban regeneration of inner-city areas in China. As urban areas take shape based on cultural aspects as much as on physical aspects, this paper employs the theory of place narrative to explore the strategies involved in culture-oriented urban regeneration. Initially, this paper sets out the basic theory of place narrative, which is relevant not only for the way in which a particular regenerated urban area is presented to the general public, but also for the way in which the hidden cultural information of the city translates into concrete urban developments. It also examines the urban regeneration of Haiyan (Zhejiang), an ongoing case, by applying methods and concepts of narrative theory. The main idea of urban regeneration is illustrated, based on the use of ‘culture’ as the revitalizing mechanism. Firstly, the method of place narrative is used to sort out and interpret the cultural information, which is commonly attached to the cultural resources in the context of urban space. Secondly, based on the idea of narrative structures, cultural resources can be integrated into the urban system connotationally and physically as well. Thirdly, cultural events are organized in the urban space to promote the experience of the place. The greatest contribution of this research is to provide a new approach, i.e., place narrative, to culture-oriented urban regeneration and for its sustainable development as well.
Agricultural Structures Management Based on Nonpoint Source Pollution Control in Typical Fuel Ethanol Raw Material Planting Area
Authors: Cui Guannan , Bai Xinyu , Wang Pengfei , Wang Haitao , Wang Shiyu , Dong Liming
Published: July 2022

Increasing the promotion and application of biofuel ethanol has been a national strategy in China, which in turn has affected changes in the raw material planting structure. This study analyzed the effects of agricultural land-use changes on water quality in a typical maize fuel ethanol raw material planting area. The results revealed that an increase in cultivated land and construction land would also increase the load of TN (total nitrogen) and TP (total phosphorus), while an expansion in forest land would reduce the load. As for crop structures, maize might have a remarkable positive effect on TN and TP, while rice and soybean performed in no significant manner. Furthermore, scenarios under the carbon neutralization policy and water pollution control were carried out to forecast the nonpoint source pollutants based on the quantitative relations coefficients. It was proven that maize planting was not suitable for vigorous fuel ethanol development. Reducing maize area in the Hulan River Basin was beneficial to reducing nonpoint source pollution. However, the area of maize should not be less than 187 km2, otherwise, the food security of the population in the basin would be threatened. Under the change in fuel ethanol policy, this study could provide scientific support for local agriculture land-use management in realizing the carbon neutralization vision and set a good example for the development of the fuel ethanol industry in other maize planting countries.
Service Quality of Bus Performance in Asia: A Systematic Literature Review and Conceptual Framework
Authors: Bakar Muhammad Fadhlullah Abu , Norhisham Shuhairy , Katman Herda Yati , Fai Chow Ming , Azlan Nor Najwa Irina Mohd , Samsudin Nur Sarah Shaziah
Published: July 2022

Bus services have played a significant role in public transportation, especially in urban areas throughout the years. Since bus services compete greatly with other types of public transportation, such as e-hailing services and private vehicles, they have recently attracted scholars to conduct many relevant studies. However, research in assessing public transport networks in urban areas by researchers by using systematic literature review is lacking. There are definitive gaps between quality standards set by the transportation regulatory authority and what bus operators can provide based on their obligations. Therefore, the present study had concentrated on the service quality of bus services in Asia by using the systematic literature review of articles. This study was based on previous studies, specifically on service quality of performance. Several previous studies were selected by using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRIMSA) approach. SCOPUS and Science Direct were chosen as the main journal database. Through this approach, 41 articles were selected for further analysis. This study was focused on three primary themes: study approach, stakeholders, and service quality attributes. An advanced analysis on these primary themes was used to formulate another 18 sub-themes. All themes and sub-themes which reflected the significant impacts of service quality towards bus services were discussed in detail. This study addressed several qualities of bus services and bus performance in regard to the improvement of urban transportation polices. Several recommendations that can provide necessary knowledge and information for future research were also presented.
A Study of the Influencing Factors on the Carbon Emission Trading Price in China Based on the Improved Gray Relational Analysis Model
Authors: Song Xiaohua , Zhang Wen , Ge Zeqi , Huang Siqi , Huang Yamin , Xiong Sijia
Published: July 2022

Carbon emission trading market construction is an important policy tool to promote the realization of China’s “double carbon” goal. However, problems still exist, such as the lack of market trading vitality, the large difference in carbon trading prices between the eight pilot markets and the instability of the prices. In order to explore the key influencing factors on carbon trading prices, 15 factors were selected to study in detail according to the policy, green industry, economy and environment. Taking China’s eight pilot carbon trading markets as research subjects, we explored the correlation degree of each factor by using the improved gray relational analysis model (GRAM) from the two dimensions of space and time. The research results show that from the space dimension, the industrial development level, development degree of low-carbon industries, air pollution degree and green technology maturity are the main factors that affect the carbon trading price in the eight pilot areas. Meanwhile, from the time dimension, the correlation degree between various factors and carbon trading price both showed a downward trend as a whole, and the fluctuation of the correlation degree of individual factors was different from the overall trend. In conclusion, we can put forward recommendations on the pricing mechanism of the carbon trading market after this comprehensive study.
Spatial-Heterogeneity Analysis of the Heavy Metals Cd and Pb in Road Dust in the Main Urban Area of Harbin
Authors: Cheng Zhiying , Xu Siruo , Na Xueying , Zhang Xujia , Ma Dalong , Zhang Peng
Published: July 2022

To provide a scientific basis for pollution prevention and control of the urban environment, the current status of heavy-metal pollution in road dust in Harbin was studied. In total, 63 road dust samples in the main urban area of Harbin were collected, and the contents of Cd and Pb, two representative heavy metals, were detected. Using the Moran Index coupled with semi-variable function and geostatistical methods, the spatial correlation, variation structure, and distribution pattern were analyzed, and the estimated probability of the heavy metals exceeding the safety standard was determined. The study showed that: The mean concentrations of Cd and Pb were higher than the background values in Heilongjiang province, and both exhibited moderate variability, while the coefficient of variation of Cd was larger than that of Pb; Cd was weakly correlated in space and randomly distributed, Pb was moderately correlated in space and exhibited good spatial structure, and both were spatially aggregated. The optimal model for fitting the variance function showed that Cd was a spherical model, and Pb was an exponential model. The variation of Cd was mainly influenced by human factors, and the variation of Pb was influenced by both structural and random factors. The optimized interpolation results of the variance function had high accuracy, and the spatial distribution of Cd was elliptical, whereas the distribution of Pb was stripe-shaped, Cd was mainly influenced by traffic factors, such as industrial enterprise distribution and road painting, while Pb was influenced by natural factors such as river sediment or the study area belonging to a geologically high background area, in addition to the above factors. The estimated probabilities indicate a higher potential risk of Cd in the northeastern part of the study area.
Digital Economy, Industry Heterogeneity, and Service Industry Resource Allocation
Authors: Qian Wei , Liu Huan , Pan Fanghui
Published: July 2022

Based on the traditional framework of resource mismatch theory analysis and existing literature studies, this paper constructs a model of resource mismatch efficiency loss including the digitalization factor of the service industry, measures the resource mismatch of China’s service industry and its sub-sectors, and empirically analyzes the impact of digital economy development on resource mismatch of service industry using panel data of each sub-sector of China’s service industry from 2001 to 2020. The findings reveal that: (1) Resource mismatch exists in China’s service industry, and the degree of mismatch of capital is more serious than that of labor. (2) Traditional service industries with low digitalization have serious efficiency losses, while emerging service industries with high digitalization have almost no efficiency losses. (3) The increase in the development of the digital economy can significantly improve the resource mismatch in the service industry; appropriate government intervention can improve the capital mismatch but not the labor mismatch; the increase in the proportion of state-owned enterprises is conducive to improving the labor mismatch but not the capital mismatch. Meanwhile, the results of the industry heterogeneity test show that the increase in the digital economy can improve the resource mismatch of both emerging and traditional service industries, but the improvement is more obvious for emerging service industries. Therefore, in the context of the development of the digital economy, we make the following suggestions. The government intervenes appropriately in the capital market, develops emerging service industries, and formulates different digital transformation policies for different industries. Relevant enterprises increase their efforts in technology research and development, and actively explore the direction of digital transformation of service industries. The government and enterprises work together to promote the improvement of China’s economic development level.
Spatial Planning and Systems Thinking Tools for Climate Risk Reduction: A Case Study of the Andaman Coast, Thailand
Authors: McMillan Joanna M. , Birkmann Joern , Tangwanichagapong Siwaporn , Jamshed Ali
Published: July 2022

The impact of climate change and related hazards such as floods, heatwaves, and sea level rise on human lives, cities, and their hinterlands depends not only on the nature of the hazard, but also on urban development, adaptation, and other socioeconomic processes that determine vulnerability and exposure. Spatial planning can reduce climate risk not just by influencing the exposure, but also by addressing social vulnerability. This requires that relevant information is available to planners and that plans are implemented and coordinated between sectors. This article is based on a research project in Thailand, particularly on the results of multi-sectoral workshops in the case study region of the Andaman Coast in southern Thailand, and draws upon climate risk, spatial planning, and systems thinking discourses. The article formulates recommendations for planning in the context of Thailand that are relevant for other rapidly growing and urbanizing regions. Among other conclusions, it suggests that systems thinking approaches and cross-sectoral strategies are ways to grasp the interdependencies between and within climate risk and spatial development challenges.
Understanding the Human Dimensions of Recycling and Source Separation Practices at the Household Level: An Evidence in Perak, Malaysia
Authors: Yu Pei Lin , Ab Ghafar Norafida , Adam Mastura , Goh Hong Ching
Published: July 2022

Recycling and source separation (R&SS) are believed to have been the first attempt to minimise waste. This research adopted mixed methods that followed sequential quantitative then qualitative data collection, combining questionnaire surveys from 100 households, semi-structured interviews, and participatory observations to study the human dimension of waste generation and management. Scoring Assessment (with modified Bloom’s Cut Off point) indicated that households had moderate knowledge and positive attitudes yet poor behaviour, and these three components indicated no linear associations, tested using Pearson’s Correlation Coefficient. However, age group, marital status, educational level and living duration showed statistical significance with households’ participation in source separation through Chi-Square Test. Meanwhile, observation data showed that waste management mechanisms and environment had inefficiently supported households’ participation in R&SS practices (external factors: poor accessibility to services, lack of tangible incentives, and absence of restriction in consumption). Elicited data indicated that a satisfactory level of intentions, knowledge, and willingness, together with good habit and quality persuasion (internal factors), were required to drive good behaviour. Subsequently, a series of recommendations were formulated to promote gradual yet solid transformation of the waste management system, tapping on existing initiatives by considering additional parameters upon the gap in households’ knowledge, attitude, and behaviour.
Antibiotic Resistance Genes Associated with Marine Surface Sediments: A Baseline from the Shores of Kuwait
Authors: Habibi Nazima , Uddin Saif , Lyons Brett , Al-Sarawi Hanan A. , Behbehani Montaha , Shajan Anisha , Razzack Nasreem Abdul , Zakir Farhana , Alam Faiz
Published: July 2022

Marine sediments are a sink for antibiotic resistance genes (ARGs) and antibiotic-resistant microbes (ARMs). Wastewater discharge into the aquatic environment is the dominant pathway for pharmaceuticals reaching aquatic organisms. Hence, the characterization of ARGs is a priority research area. This baseline study reports the presence of ARGs in 12 coastal sediment samples covering the urban coastline of Kuwait through whole-genome metagenomic sequencing. The presence of 402 antibiotic resistance genes (ARGs) were recorded in these samples; the most prevalent were patA, adeF, ErmE, ErmF, TaeA, tetX, mphD, bcrC, srmB, mtrD, baeS, Erm30, vanTE, VIM-7, AcrF, ANT4-1a, tet33, adeB, efmA, and rpsL, which showed resistance against 34 drug classes. Maximum resistance was detected against the beta-lactams (cephalosporins and penam), and 46% of genes originated from the phylum Proteobacteria. Low abundances of ESKAPEE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumonia, Acinetobacter baumanii, Pseudomonas aeruginosa, Enterobacter sps., and Escherichia coli) were also recorded. Approximately 42% of ARGs exhibited multiple drug resistance. All the ARGs exhibited spatial variations. The major mode of action was antibiotic efflux, followed by antibiotic inactivation, antibiotic target alteration, antibiotic target protection, and antibiotic target replacement. Our findings supported the occurrence of ARGs in coastal marine sediments and the possibility of their dissemination to surrounding ecosystems.
Use of Hydrochar Produced by Hydrothermal Carbonization of Lignocellulosic Biomass for Thermal Power Plants in Chile: A Techno-Economic and Environmental Study
Authors: Carrasco Samuel , Pino-Cortés Ernesto , Barra-Marín Andrés , Fierro-Gallegos Alejandro , León Marcelo
Published: July 2022

Hydrothermal carbonization makes it possible to transform lignocellulosic biomass into hydrochar, a carbon-rich material that can be used as fuel. Hydrochar has less calorific value than standard coal but generates less ashes during combustion. This study is a pre-feasibility analysis carried out to evaluate technically-economically and environmentally the use of hydrochar as fuel or co-fuel in thermal power plants in Chile. Until now there are no reports about it. The proposal of this work was to study the replacement of coal with a fuel that uses the same equipment and processes in power generation but with less air emission and with an economically profitable change. The results suggest that a plant with a supply of 104 t/h of bituminous coal could be replaced between 18 and 37 t/h of hydrochar, with a reduction of 8 and 27% in NOx and SO2 emissions, a reduction in 7 to 24% in ashes and a marginal increase in CO2 emission. The proposed use of hydrochar was economically profitable, with internal rates of return of up to 40% and with cash flows that reached USD 22 million.
A New Clustering Method to Generate Training Samples for Supervised Monitoring of Long-Term Water Surface Dynamics Using Landsat Data through Google Earth Engine
Authors: Taheri Dehkordi Alireza , Valadan Zoej Mohammad Javad , Ghasemi Hani , Ghaderpour Ebrahim , Hassan Quazi K.
Published: July 2022

Water resources are vital to the survival of living organisms and contribute substantially to the development of various sectors. Climatic diversity, topographic conditions, and uneven distribution of surface water flows have made reservoirs one of the primary water supply resources in Iran. This study used Landsat 5, 7, and 8 data in Google Earth Engine (GEE) for supervised monitoring of surface water dynamics in the reservoir of eight Iranian dams (Karkheh, Karun-1, Karun-3, Karun-4, Dez, UpperGotvand, Zayanderud, and Golpayegan). A novel automated method was proposed for providing training samples based on an iterative K-means refinement procedure. The proposed method used the Function of the Mask (Fmask) initial water map to generate final training samples. Then, Support Vector Machines (SVM) and Random Forest (RF) models were trained with the generated samples and used for water mapping. Results demonstrated the satisfactory performance of the trained RF model with the samples of the proposed refinement procedure (with overall accuracies of 95.13%) in comparison to the trained RF with direct samples of Fmask initial water map (with overall accuracies of 78.91%), indicating the proposed approach’s success in producing training samples. The performance of three feature sets was also evaluated. Tasseled-Cap (TC) achieved higher overall accuracies than Spectral Indices (SI) and Principal Component Transformation of Image Bands (PCA). However, simultaneous use of all features (TC, SI, and PCA) boosted classification overall accuracy. Moreover, long-term surface water changes showed a downward trend in five study sites. Comparing the latest year’s water surface area (2021) with the maximum long-term extent showed that all study sites experienced a significant reduction (16–62%). Analysis of climate factors’ impacts also revealed that precipitation (0.51 ≤ R2 ≤ 0.79) was more correlated than the temperature (0.22 ≤ R2 ≤ 0.39) with water surface area changes.
Negative Poisson’s Ratio Re-Entrant Base Modeling and Vibration Isolation Performance Analysis
Authors: Pan Kun , Zhang Wei , Ding Jieyu
Published: July 2022

Negative Poisson’s ratio materials are increasingly used in the design of vibration isolation bases due to their unique tensile properties. In this paper, based on the expansion feature of the negative Poisson’s ratio re-entrant structure, the influence of the size of the re-entrant structure within a single structure was analyzed, and a honeycomb base was designed with a negative Poisson’s ratio re-entrant structure. A new modeling method for the honeycomb base is proposed. In the modeling process, the honeycomb base was analyzed according to its symmetry using the Lagrange equation for base modeling and the finite element consistent mass matrix was introduced to simplify the calculation. The vibration isolation performance of the honeycomb base was evaluated by vibration level difference. COMSOL software was used to simulate and analyze the cellular base in order to verify the correctness of the results obtained from numerical modeling. In conclusion, the honeycomb base had a vibration isolation effect on external excitation in the vertical direction of the base. Furthermore, the vibration isolation performance of the base was greatly related to the wall thickness and Poisson’s ratio of the re-entrant structure.
Machine Learning-Based Investigation on the Impact of Chinese Venture Capital Institutions’ Performance: Evaluation Factors of Venture Enterprises to Venture Capital Institutions
Authors: Hu Jitao , Hu Longying , Hu Mingzhu , He Qiuzhi
Published: July 2022

Studying the influencing factors of venture capital fund investment performance is crucial for the decision making of venture capital institutions. This paper explored the influencing factors of venture capital institutions from the perspective of startups, aiming to elucidate the mechanisms of these factors on the performance of venture capital funds and to propose a novel and effective predictive model of investment performance. Linear regression and one-way ANOVA were used to analyze the influence of each variable on investment performance, and the weight proportion of each influencing factor was obtained under the linear model. Two machine learning models, including the random forest algorithm and extreme learning machine algorithm, are established, and the particle swarm algorithm and machine learning algorithm were combined to optimize the random parameters in the two models. Compare the reliability and accuracy of machine learning models and multivariate linear regression models. The analysis results indicate that the PSO-ELM hybrid model has a better predictive performance than other prediction models. A convenient machine learning algorithm provided in this paper can quickly and effectively predict the investment performance of various investment portfolios and provide investors with decision-making assistance.
The Role of Nitrogen Fertilization on the Occurrence of Regulated, Modified and Emerging Mycotoxins and Fungal Metabolites in Maize Kernels
Authors: Scarpino Valentina , Sulyok Michael , Krska Rudolf , Reyneri Amedeo , Blandino Massimo
Published: July 2022

The European Food Safety Authority is currently evaluating the risks related to the presence of emerging mycotoxins in food and feeds. The aim of this study was to investigate the role of soil fertility, resulting from different nitrogen fertilization rates, on the contamination of regulated mycotoxins and emerging fungal metabolites in maize grains. The trial was carried out in the 2012–2013 growing seasons as part of a long-term (20-year) experimental platform area in North-West Italy, where five different N rates, ranging from 0 to 400 kg N ha−1, were applied to maize each year. Maize samples were analyzed by means of a dilute-and-shoot multi-mycotoxin LC-MS/MS method, and more than 25 of the most abundant mycotoxins and fungal metabolites were detected. Contamination by fumonisins and other fungal metabolites produced by Fusarium spp. of the section Liseola was observed to have increased in soils that showed a poor fertility status. On the other hand, an overload of nitrogen fertilization was generally associated with higher deoxynivalenol and zearalenone contamination in maize kernels, as well as a higher risk of other fungal metabolites produced by Fusarium spp. sections Discolor and Roseum. A balanced application of N fertilizer, in accordance with maize uptake, generally appears to be the best solution to guarantee an overall lower contamination by regulated mycotoxins and emerging fungal metabolites.
Staphylococcus aureus Alpha-Toxin in Deep Tracheal Aspirates—Preliminary Evidence for Its Presence in the Lungs of Sepsis Patients
Authors: Ziesemer Sabine , Kuhn Sven-Olaf , Hahnenkamp Anke , Gerber Manuela , Lutjanov Elvira , Gruendling Matthias , Hildebrandt Jan-Peter
Published: July 2022

The pore forming alpha-toxin (hemolysin A, Hla) of Staphylococcus aureus (S. aureus) is a major virulence factor with relevance for the pathogenicity of this bacterium, which is involved in many cases of pneumonia and sepsis in humans. Until now, the presence of Hla in the body fluids of potentially infected humans could only be shown indirectly, e.g., by the presence of antibodies against Hla in serum samples or by hemolysis testing on blood agar plates of bacterial culture supernatants of the clinical isolates. In addition, nothing was known about the concentrations of Hla actually reached in the body fluids of the infected hosts. Western blot analyses on 36 samples of deep tracheal aspirates (DTA) isolated from 22 hospitalized sepsis patients using primary antibodies against different epitopes of the Hla molecule resulted in the identification of six samples from five patients containing monomeric Hla (approx. 33 kDa). Two of these samples showed also signals at the molecular mass of heptameric Hla (232 kDa). Semiquantitative analyses of the samples revealed that the concentrations of monomeric Hla ranged from 16 to 3200 ng/mL. This is, to our knowledge, the first study directly showing the presence of S. aureus Hla in samples of airway surface liquid in human patients.
Chikungunya Virus Asian Lineage Infection in the Amazon Region Is Maintained by Asiatic and Caribbean-Introduced Variants
Authors: de Oliveira Ribeiro Geovani , Gill Danielle Elise , do Socorro Foro Ramos Endrya , Villanova Fabiola , Soares D’Athaide Ribeiro Edcelha , Monteiro Fred Julio Costa , Morais Vanessa S. , Rego Marlisson Octavio da S. , Araújo Emerson Luiz Lima , Pandey Ramendra Pati , Raj V. Samuel , Deng Xutao , Delwart Eric , da Costa Antonio Charlys , Leal Élcio
Published: July 2022

The simultaneous transmission of two lineages of the chikungunya virus (CHIKV) was discovered after the pathogen’s initial arrival in Brazil. In Oiapoque (Amapá state, north Brazil), the Asian lineage (CHIKV-Asian) was discovered, while in Bahia state, the East-Central-South-African lineage (CHIKV-ECSA) was discovered (northeast Brazil). Since then, the CHIKV-Asian lineage has been restricted to the Amazon region (mostly in the state of Amapá), whereas the ECSA lineage has expanded across the country. Despite the fact that the Asian lineage was already present in the Amazon region, the ECSA lineage brought from the northeast caused a large outbreak in the Amazonian state of Roraima (north Brazil) in 2017. Here, CHIKV spread in the Amazon region was studied by a Zika–Dengue–Chikungunya PCR assay in 824 serum samples collected between 2013 and 2016 from individuals with symptoms of viral infection in the Amapá state. We found 11 samples positive for CHIKV-Asian, and, from these samples, we were able to retrieve 10 full-length viral genomes. A comprehensive phylogenetic study revealed that nine CHIKV sequences came from a local transmission cluster related to Caribbean strains, whereas one sequence was related to sequences from the Philippines. These findings imply that CHIKV spread in different ways in Roraima and Amapá, despite the fact that both states had similar climatic circumstances and mosquito vector frequencies.
Genetic Analysis of HPIV3 That Emerged during the SARS-CoV-2 Pandemic in Gwangju, South Korea
Authors: Lee Hongsu , Kim Sun-Hee , Cho Sun-Ju , Lee Yeong-Un , Lee Kwangho , Lee Yong-Pyo , Seo Jinjong , Chung Yoon-Seok
Published: July 2022

Community mitigation measures taken owing to the COVID-19 pandemic have caused a decrease in the number of respiratory viruses, including the human parainfluenza virus type 3 (HPIV3), and a delay in their occurrence. HPIV3 was rarely detected as a consequence of monitoring respiratory viral pathogens in Gwangju, Korea, in 2020; however, it resurfaced as a delayed outbreak and peaked in September–October 2021. To understand the genetic characteristics of the reemerging virus, antigenic gene sequences and evolutionary analyses of the hemagglutinin-neuraminidase (HN) and fusion (F) genes were performed for 129 HPIV3 pathogens prevalent in Gwangju from 2018 to 2021. Unlike the prevalence of various HPIV3 strains in 2018-2019, the prevalence of HPIV3 by strains with reduced diversity was confirmed in 2021. It could be inferred that this decrease in genetic diversity was due to the restriction of inflow from other regions at home and abroad following the community mitigation measures and the spread within the region. The HPIV3 that emerged in 2021 consisted of HN coding regions that were 100% consistent with the sequence identified in Saitama, Japan, in 2018, and F coding regions exhibiting 99.6% homology to a sequence identified in India in 2017, among the ranks reported to the National Center for Biotechnology Information. The emergence of a new lineage in a community can lead to a mass outbreak by collapsing the collective immunity of the existing acquired area; therefore, continuous monitoring is necessary.
Human ACE2 Polymorphisms from Different Human Populations Modulate SARS-CoV-2 Infection
Authors: Hu Pan , Bauer Vanessa L. , Sawyer Sara L. , Diaz-Griffero Felipe
Published: July 2022

The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has resulted in over 6 million deaths worldwide. The high variability in COVID-19 symptoms remains one of the most interesting mysteries of the pandemic. Genetic and environmental factors are likely to be key determinants of COVID-19 symptomatology. Here, we explored ACE2 as a genetic determinant for SARS-CoV-2 infection and COVID-19 symptomatology. Each human genome encodes two alleles of ACE2, which encodes the cell entry receptor for SARS-CoV-2. Here, we determined whether naturally occurring human ACE2 (hACE2) polymorphisms in the human population affect SARS-CoV-2 infection and the severity of COVID-19 symptoms. ACE2 variants S19P, I21V, E23K, K26R, K31R, N33I, H34R, E35K, and T92I showed increased virus infection compared to wild-type ACE2; thus, these variants could increase the risk for COVID-19. In contrast, variants D38V, Y83H, I468V, and N638S showed reduced infection, indicating a potential protective effect. hACE2 variants K26R and T92I increased infection by three-fold without changing the levels of ACE2 on the surface of the cells, suggesting that these variants may increase the risk of severe COVID-19. On the contrary, hACE2 variants D38V and Y83H decreased SARS-CoV-2 infection by four- and ten-fold, respectively, without changing surface expression, suggesting that these variants may protect against severe COVID-19. Remarkably, all protective hACE2 Polymorphisms were found almost exclusively in Asian populations, which may provide a partial explanation for the low COVID-19 mortality rates in Asian countries. Thus, hACE2 polymorphisms may modulate susceptibility to SARS-CoV-2 in the host and partially account for the differences in severity of COVID-19 among different ethnic groups.
Combination of E- and NS1-Derived DNA Vaccines: The Immune Response and Protection Elicited in Mice against DENV2
Authors: Pinto Paolla Beatriz A. , Barros Tamiris A. C. , Lima Lauro M. , Pacheco Agatha R. , Assis Maysa L. , Pereira Bernardo A. S. , Gonçalves Antônio J. S. , Azevedo Adriana S. , Neves-Ferreira Ana Gisele C. , Costa Simone M. , Alves Ada M. B.
Published: July 2022

The occurrence of dengue disease has increased radically in recent decades. Previously, we constructed the pE1D2 and pcTPANS1 DNA vaccines encoding the DENV2 envelope (E) and non-structural 1 (NS1) proteins, respectively. To decrease the number of plasmids in a tetravalent candidate vaccine, we constructed a bicistronic plasmid, pNS1/E/D2, encoding these two proteins simultaneously. We evaluated the protective immunity induced in mice vaccinated with the pNS1/E/D2 candidate and compared to the responses elicited by immunization with the former vaccines isolated or in combination. We transfected BHK-21 cells with the different plasmids and detected recombinant proteins by immunofluorescence and mass spectrometry assays to confirm antigen expression. BALB/c mice were inoculated with the DNA vaccines followed by a lethal DENV2 challenge. ELISA, PRNT50, and IFN-gamma ELISPOT assays were performed for the investigation of the humoral and cellular responses. We observed the concomitant expression of NS1 and E proteins in pNS1/E/D2-transfected cells. All E-based vaccines induced anti-E and neutralizing antibodies. However, anti-NS1 antibodies were only observed after immunization with the pcTPANS1 administered alone or combined with pE1D2. In contrast, splenocytes from pNS1/E/D2- or pcTPANS1 + pE1D2-vaccinated animals responded to NS1- and E-derived synthetic peptides. All the DNA vaccines conferred protection against DENV2.
Design of a Multiepitope Vaccine against Chicken Anemia Virus Disease
Authors: Fatoba Abiodun Joseph , Adeleke Victoria T. , Maharaj Leah , Okpeku Moses , Adeniyi Adebayo A. , Adeleke Matthew A.
Published: July 2022

Chicken anemia virus (CAV) causes severe clinical and sub-clinical infection in poultry globally and thus leads to economic losses. The drawbacks of the commercially available vaccines against CAV disease signal the need for a novel, safe, and effective vaccine design. In this study, a multiepitope vaccine (MEV) consisting of T-cell and B-cell epitopes from CAV viral proteins (VP1 and VP2) was computationally constructed with the help of linkers and adjuvant. The 3D model of the MEV construct was refined and validated by different online bioinformatics tools. Molecular docking showed stable interaction of the MEV construct with TLR3, and this was confirmed by Molecular Dynamics Simulation. Codon optimization and in silico cloning of the vaccine in pET-28a (+) vector also showed its potential expression in the E. coli K12 system. The immune simulation also indicated the ability of this vaccine to induce an effective immune response against this virus. Although the vaccine in this study was computationally constructed and still requires further in vivo study to confirm its effectiveness, this study marks a very important step towards designing a potential vaccine against CAV disease.
Differential Dynamics of Humoral and Cell-Mediated Immunity with Three Doses of BNT162b2 SARS-CoV-2 Vaccine in Healthcare Workers in Japan: A Prospective Cohort Study
Authors: Yamashita Keita , Suzuki Akira , Takebayashi Shiori , Toguchi Akihiro , Ogitani Kenya , Niizeki Noriyasu , Nagura Osanori , Furuhashi Kazuki , Iwaizumi Moriya , Maekawa Masato
Published: July 2022

Vaccines against SARS-CoV-2 with good efficacy are now available worldwide. However, gained immunity diminishes over time. Here, we investigate the course of both humoral and cell-mediated immunity in response to three doses of the Pfizer mRNA BNT162b2 SARS-CoV-2 vaccine in healthcare workers in Japan. SARS-CoV-2 anti-receptor-binding domain (RBD) antibodies (total Ig, IgG), neutralizing antibodies (NAb), and ELISpot were measured in serum and whole blood samples collected after each vaccine dose. ELISpot numbers were higher than the cutoff values in most participants at all times. It was suggested that the difference in behavior between humoral immunity and cell-mediated immunity with age is complementary. Anti-RBD total Ig, IgG, and NAb indicated a high correlation at each time point after vaccine doses. Total Ig was retained long-term after the second dose and increased significantly faster by the booster dose than IgG. Nab levels of all subjects were ≤20% six months after the second dose, and the correlation coefficient was greatly reduced. These are due to the avidity of each antibody and differences among commercial kits, which may affect the evaluation of immunokinetics in previous COVID-19 studies. Therefore, it is necessary to harmonize reagents categorized by the same characteristics.
Cross-Species Transmission of Rabbit Hepatitis E Virus to Pigs and Evaluation of the Protection of a Virus-like Particle Vaccine against Rabbit Hepatitis E Virus Infection in Pigs
Authors: Han Sang-Hoon , Ahn Hee-Seop , Go Hyeon-Jeong , Kim Dong-Hwi , Kim Da-Yoon , Kim Jae-Hyeong , Lim Kyu-Beom , Lee Joong-Bok , Park Seung-Yong , Song Chang-Seon , Lee Sang-Won , Choi Yang-Kyu , Choi In-Soo
Published: July 2022

We investigated the cross-species transmission of rabbit hepatitis E virus (rb HEV) to pigs and evaluated the cross-protection of a swine (sw) HEV-3 virus-like particle (VLP) vaccine against rb HEV infection in pigs. Twelve 4-week-old conventional pigs were divided into negative control (n = 3), positive control (rb HEV-infected, n = 4), and vaccinated (vaccinated and rb HEV-challenged, n = 5) groups. The vaccine was administered at weeks 0 and 2, and viral challenge was conducted at week 4. Serum HEV RNA, anti-HEV antibody, cytokine, and liver enzyme levels were determined. Histopathological lesions were examined in abdominal organs. Viral RNA was detected and increased anti-HEV antibody and alanine aminotransferase (ALT) levels were observed in positive control pigs; liver fibrosis, inflammatory cell infiltration in the lamina propria of the small intestine and shortened small intestine villi were also observed. In vaccinated pigs, anti-HEV antibody and Th1 cytokine level elevations were observed after the second vaccination; viral RNA was not detected, and ALT level elevations were not observed. The results verified the cross-species transmission of rb HEV to pigs and cross-protection of the sw HEV-3 VLP vaccine against rb HEV infection in pigs. This vaccine may be used for cross-protection against HEV infection in other species.
Activation of Anti-SARS-CoV-2 Human CTLs by Extracellular Vesicles Engineered with the N Viral Protein
Authors: Manfredi Francesco , Chiozzini Chiara , Ferrantelli Flavia , Leone Patrizia , Giovannelli Andrea , Sanchez Massimo , Federico Maurizio
Published: July 2022

We propose an innovative anti-SARS-CoV-2 immune strategy based on extracellular vesicles (EVs) inducing an anti-SARS-CoV-2 N CD8+ T cytotoxic lymphocyte (CTL) immune response. We previously reported that the SARS-CoV-2 N protein can be uploaded at high levels in EVs upon fusion with Nefmut, i.e., a biologically inactive HIV-1 Nef mutant incorporating into EVs at quite high levels. Here, we analyze the immunogenic properties in human cells of EVs engineered with SARS-CoV-2 N fused at the C-terminus of either Nefmut or a deletion mutant of Nefmut referred to as NefmutPL. The analysis of in vitro-produced EVs has supported the uploading of N protein when fused with truncated Nefmut. Mice injected with DNA vectors expressed each fusion protein developed robust SARS-CoV-2 N-specific CD8+ T cell immune responses. When ex vivo human dendritic cells were challenged with EVs engineered with either fusion products, the induction of a robust N-specific CTL activity, as evaluated by both CD107a and trogocytosis assays, was observed. Through these data we achieved the proof-of-principle that engineered EVs can be instrumental to elicit anti-SARS-CoV-2 CTL immune response in human cells. This achievement represents a mandatory step towards the upcoming experimentations in pre-clinical models focused on intranasal administration of N-engineered EVs.
Predictors of Willingness of the General Public to Receive a Second COVID-19 Booster Dose or a New COVID-19 Vaccine: A Cross-Sectional Study in Greece
Authors: Galanis Petros , Vraka Irene , Katsiroumpa Aglaia , Siskou Olga , Konstantakopoulou Olympia , Katsoulas Theodoros , Mariolis-Sapsakos Theodoros , Kaitelidou Daphne
Published: July 2022

Given the concerns of waning immunity from the primary COVID-19 vaccines and the first booster dose, we conducted an online cross-sectional study in May 2022 to investigate willingness to receive a second COVID-19 booster dose or a new COVID-19 vaccine and its associated factors. Overall, 62% of the participants were willing to be vaccinated, 25.8% were unsure, and 12.3% were unwilling to be vaccinated. The main reasons against accepting a second COVID-19 booster dose/new COVID-19 vaccine were concerns about the side effects and the effectiveness and the opinion that further vaccination is unnecessary. Males, younger individuals, participants without a previous COVID-19 diagnosis, and those with good/very good self-perceived physical health were significantly more frequently willing to receive a second COVID-19 booster dose or a new COVID-19 vaccine. Additionally, increased fear of the COVID-19, increased trust in COVID-19 vaccinations, and decreased fear of a second booster dose or a new COVID-19 vaccine was associated with increased willingness. Our results show some hesitancy and unwillingness toward further COVID-19 vaccination and indicate that the fear of COVID-19 and trust in COVID-19 vaccination affects public opinion.
Differences in Behavior between Normal and Atopic Keratinocytes in Culture: Pilot Studies
Authors: Marsella Rosanna , Ahrens Kim , Wilkes Rachel
Published: July 2022

Skin barrier dysfunction is important in atopic dermatitis and can be secondary to inflammation. Observation of keratinocytes in culture may show intrinsic differences. TransEpithelial Electrical Resistance (TEER) measures epithelial permeability. We cultured normal and atopic keratinocytes and found that TEER of atopic keratinocytes was significantly lower (p < 0.0001) than that of normals. Atopic keratinocytes grew upwards, first creating isolated dome-like structures and later horizontally into a monolayer. At time of confluence (D0), atopic keratinocytes were more differentiated, with higher filaggrin gene expression than normals. No differences existed between groups for TJ proteins (claudin, occludin, and Zonula Occludens-1) on D0 and D6. On D6, claudin and occludin were higher than D0, in normal (p = 0.0296 and p = 0.0011) and atopic keratinocytes (p = 0.0348 and 0.0491). Immunofluorescent staining showed nuclear location of filaggrin on D0 and cytoplasmic on D6. ANOVA showed increased cell size from D0 to D6 in both groups (effect of time, p = 0.0076) but no differences between groups. Significant subject effect (p = 0.0022) was found, indicating that cell size was subject-dependent but not disease-dependent. No difference for continuity for TJ protein existed between groups. These observations suggest that decreased TEER in atopics is not linked to TJ differences but is possibly linked to different growth behavior.