bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of the Acoustic Environment of Urban Recreational Trails
Authors: Lin Wei , Wu Yiming
Published: June 2022

The acoustic environment state of four recreational trails in Taichung was investigated. First, the basic forms, spatial proportions, characteristics of the trail interfaces, and sound sources of the recreational trail spaces were investigated. Second, sound pressure level measurement and a questionnaire survey were conducted in relation to the four trails. The measurements demonstrated that the sound levels of the four trail spaces are lower than the permissible exposure limit 90 dB(A) recommended by the Occupational Safety and Health Administration. Meanwhile, the results of the factor analysis show that the perception of the acoustic environment of recreational trails has five dimensions: likability, intensity, communication, joyfulness, and richness. Compared with the acoustic environments of other cities, the perception of the acoustic environment of the recreational trails in this city was characterized by more dimensions and a more balanced structure.
Recent Advances and Perspectives of Nanotechnology in Anaerobic Digestion: A New Paradigm towards Sludge Biodegradability
Authors: Jeyakumar Rajesh Banu , Vincent Godvin Sharmila
Published: June 2022

Anaerobic digestion (AD) is the strategy of producing environmentally sustainable bioenergy from waste-activated sludge (WAS), but its efficiency was hindered by low biodegradability. Hence, the usage of nanomaterials was found to be essential in enhancing the degradability of sludge due to its nanostructure with specific physiochemical properties. The application of nanomaterials in sludge digestion was thoroughly reviewed. This review focused on the impact of nanomaterials such as metallic nanoparticles, metal oxide nanoparticles, carbon-based nanomaterials, and nanocomposite materials in AD enhancement, along with the pros and cons. Most of the studies detailed that the addition of an adequate dosage of nanomaterial has a good effect on microbial activity. The environmental and economic impact of the AD enhancement process is also detailed, but there are still many existing challenges when it comes to designing an efficient, cost-effective AD digester. Hence, proper investigation is highly necessary to assess the potency of utilizing the nanomaterials in enhancing AD under various conditions.
Visual Thinking Strategies—Theory and Applied Areas of Insertion
Authors: Albert Carmen Narcisa , Mihai Mihaela , Mudure-Iacob Ioana
Published: June 2022

Twenty-first century learners live in a highly visual world, being constantly surrounded by visual, technologized stimuli and the educational system, be it general or specialized—as in the case of higher education—needs to creatively meet and answer these learning requirements. Among the plethora of new or updated approaches, the concept of Visual Thinking Strategies (VTS) designed by Philip Yenawine and Abigail Housen has gained extensive popularity due to its capacity of being adapted to a wide range of specialized fields with notable improvement results. The main purpose of the present review is to synthetically and critically present relevant scientific work related to the application of the VTS procedure and to further identify possible study areas that would highly benefit from the insertion of this procedure. Thus, the theoretical perspective is tackled from a cognitive-psychology standpoint, followed by a rendering of the research variety in applied VTS contexts pertaining to different study domains, as well as online VTS web clustering. These contexts have shown a predilection for VTS usage, which improves higher-education students’ specialized vocabulary and speaking skills in the same target language, but not in the case of ESP (English for specific purposes) classes.
The Landscape and Roadmap of the Research and Innovation Infrastructures in Energy: A Review of the Case Study of the UK
Authors: Pourmirza Zoya , Hosseini Seyed Hamid Reza , Walker Sara , Giaouris Damian , Taylor Philip
Published: June 2022

Research and development are critical for driving economic growth. To realise the UK government’s Industrial Strategy, we develop an energy research and innovation infrastructure roadmap and landscape for the energy sector looking to the long term (2030). This study is based on a picture of existing UK infrastructure on energy. It shows the links between the energy sector and other sectors, the distribution of energy research and innovation infrastructures, the age of these infrastructures, where most of the energy research and innovation infrastructures are hosted, and the distribution of energy research and innovation infrastructures according to their legal structure. Next, this study identifies the roadmap of energy research and innovation infrastructures by 2030, based on a categorisation of the energy sector into seven subsectors. Challenges and future requirements are explored for each of the sub-sectors, encompassing fossil fuels and nuclear energy to renewable energy sources and hydrogen, and from pure science to applied engineering. The study discusses the potential facilities to address these challenges within each sub-sector. It explores the e-infrastructure and data needs for the energy sector and provides a discussion on other sectors of the economy that energy research and innovation infrastructures contribute to. Some of the key messages identified in this study are the need for further large-scale initiative and large demonstrators of multi-vector energy systems, the need for multi-disciplinary research and innovation, and the need for greater data sharing and cyber-physical demonstrators. Finally, this work will serve as an important study to provide guidance for future investment strategy for the energy sector.
The Impact of Carbon Emission Trading Policies on Enterprises’ Green Technology Innovation—Evidence from Listed Companies in China
Authors: Tian Hong , Lin Jiaen , Jiang Chunyuan
Published: June 2022

At present, the Chinese government has successively launched various policies to control the emission standards of greenhouse gases. As one of the most important standards, carbon emission trading policies were implemented in some provinces and regions in China in 2013, aiming to restrict the carbon emissions of enterprises. However, the government’s control of corporate carbon emissions restricts their rapid economic growth to some extent. Enterprises’ green technology innovation can be an effective means to ensure the implementation of low-carbon policies and promote sustainable economic growth simultaneously. The Porter hypothesis holds that reasonable environmental regulations can stimulate enterprises’ green technology innovation. Based on the Porter hypothesis, this paper examines the impact of China’s carbon emission trading policies on local enterprises’ green technology innovation from a micro perspective, taking China’s listed companies from 2007 to 2020 as samples and adopting the differential method. The differences in the impact of carbon emission trading policies on green technology innovation in the context of different corporate environmental strategies are also studied. Our study found that China’s carbon emissions trading policies can effectively stimulate green technology innovation, as carbon emissions trading policies under different environmental strategies have a positive influence on the technical innovation of enterprises and, compared with reactive environmental strategies, promote a greater role for enterprises’ proactive environmental strategies. The conclusions of this study not only provide relevant suggestions for the Chinese government to enact environmental regulation policies but also provide references for enterprises to choose appropriate environmental strategies and achieve sustainable development under the constraints of environmental regulation.
Predicting the Suitable Current and Future Potential Distribution of the Native Endangered Tree Tecomella undulata (Sm.) Seem. in Pakistan
Authors: Arshad Fahim , Waheed Muhammad , Fatima Kaneez , Harun Nidaa , Iqbal Muhammad , Fatima Kaniz , Umbreen Shaheena
Published: June 2022

The burgeoning human population exhibited a rapid amplification in demand for timber and fuelwood and as a result, the natural population of the native tree Tecomella undulata reduced rapidly due to its high economic and medicinal significance. The recognition of appropriate regions for threatened plants in the climate change scenario is a fundamental step for the restoration and conservation of biodiversity. The current study predicts the potentially suitable areas in Pakistan for T. undulata restoration. This research identifies the highly appropriate regions for vulnerable T. undulata through the maximum entropy model from MaxEnt software. The model’s Area Under Curve 0.968 suggested its accuracy. The mean temperature of the wettest quarter, precipitation of the warmest quarter, and mean temperature in the driest quarter significantly shaped the T. undulata distribution. Future suitable areas for T. undulata were made by using RCP (4.5 and 8.5) for the years 2050 and 2070 through 19 bioclimatic variables and 66 occurrence points. The current highly suitable area for T. undulata is approximately 135,749 km2 (15.4%) while the unsuitable area identified is approximately 404,917 km2 (45.91%). The highly suitable area for T. undulata increases by 3.6–7% under climate change regimes (RCP 4.5 and RCP 8.5). The Central Punjab (District Faisalabad, Nankana sahib, Jhang, Kasur, and Okara), Salt Range, Western Khayber Pakhtunkhwa (KPK), FATA area, Eastern Balochistan, and Thar and Tharparker in Sindh are the current appropriate habitats for T. undulata. Under all future climatic circumstances, the extremely appropriate area for T. undulata was anticipated to expand, whereas the unsuitable zones would all shrink. The research would be significant for the further development of T. undulata management and conservation techniques.
System Resilience Evaluation and Optimization Considering Epistemic Uncertainty
Authors: Dong Qiang , Li Ruiying , Kang Rui
Published: June 2022

Epistemic uncertainties, caused by data asymmetry and deficiencies, exist in resilience evaluation. Especially in the system design process, it is difficult to obtain enough data for system resilience evaluation and improvement. Mathematics methods, such as evidence theory and Bayesian theory, have been used in the resilience evaluation for systems with epistemic uncertainty. However, these methods are based on subjective information and may lead to an interval expansion problem in the calculation. Therefore, the problem of how to quantify epistemic uncertainty in the resilience evaluation is not well solved. In this paper, we propose a new resilience measure based on uncertainty theory, a new branch of mathematics that is viewed as appropriate for modeling epistemic uncertainty. In our method, resilience is defined as an uncertainty measure that is the belief degree of a system’s behavior after disruptions that can achieve the predetermined goal. Then, a resilience evaluation method is provided based on the operation law in uncertainty theory. To design a resilient system, an uncertain programming model is given, and a genetic algorithm is applied to find an optimal design to develop a resilient system with the minimal cost. Finally, road networks are used as a case study. The results show that our method can effectively reduce cost and ensure network resilience.
Trap Parameters Optimization Based on Metal Particle Dynamic Simulation Method
Authors: Ping An , Wang Jian , Xiao Ruofan , Liu Renying , Chang Yanan , Li Qingmin
Published: June 2022

Insulation failure usually occurs in AC gas-insulated transmission (AC GIL) in field operation, in which the primary cause is the charged motion of metal particles in the electric filed. At present, the particle inhibition method applied is to design particle traps on the inner wall of the GIL shell. However, due to the large randomness of the charged motion for metallic particles and the limitations of field test methods, a particle trap has not yet been designed from the perspective of particle trapping effectiveness. In this paper, firstly, referring to the size of a running 252 kV AC GIL, a 1:1 scaled 3-D similarity simulation model is established to obtain the dynamic characteristics of particles with different sizes under the operating voltage level. This model can form symmetry between the real equipment, and its simulated simulation trajectory can achieve symmetry with the actual one. Secondly, an experimental platform that can easily capture the motion of the particles is set up to experimentally verify the symmetry between the field operating equipment and the simulation model. Finally, the particle traps are set on both sides of the concave and convex surface of the basin insulator, and an optimization scheme for the design of the particle trap is proposed from three aspects: the electric field regulation of the trap, the captured probability of particles, and the trap location. The proposed research shows that, with respect to the motion characteristics of the particles, this paper selects circular hole-shaped trap and its thickness, slot spacing, and slot width are 10 mm, 6 mm, and 8 mm, respectively. When the traps are arranged, one at the bottom of the shell at 70 mm and 80 mm from each side of the concave and convex insulator, the capture probability of the traps on both sides can be as high as 78% and 70%, respectively. Therefore, the analysis and optimization method in this paper has important reference value according to similarity concepts for optimizing particle traps in AC GIL at a certain voltage level.
A Novel Approach to the Fuzzification of Fields
Authors: Zeng Mingyi , Wang Lan , Shi Fu-Gui
Published: June 2022

There are many symmetries in L-fuzzy algebras. In this paper, a novel approach to the fuzzification of a field is introduced. We define a mapping F:LX→L from the family of all the L-fuzzy sets on a field X to L such that each L-fuzzy set is an L-fuzzy subfield to some extent. Some equivalent characterizations F(μ) are given by means of cut sets. It is proved that F is L-fuzzy convex structure on X, hence (X,F) forms an L-fuzzy convexity space. A homomorphism between fields is exactly an L-fuzzy convexity preserving mapping and an L-fuzzy convex-to-convex mapping. Finally, we discuss some operations of L-fuzzy subsets.
The Metallic Ratio of Pulsating Fibonacci Sequences
Authors: Laipaporn Kittipong , Phibul Kiattiyot , Khachorncharoenkul Prathomjit
Published: June 2022

The golden ratio and the Fibonacci sequence (Fn) are well known, as is the fact that the ratio Fn+1Fn converges to the golden ratio for sufficiently large n. In this paper, we investigate the metallic ratio—a generalized version of the golden ratio—of pulsating Fibonacci sequences in three forms. Two of these forms are considered in the sense of pulsating recurrence relations, and their diagrams can be represented by symmetry, which is one of their distinguishing characteristics. The third form is the Fibonacci sequence in bipolar quantum linear algebra (BQLA), which also pulsates.
The Landweber Iterative Regularization Method for Solving the Cauchy Problem of the Modified Helmholtz Equation
Authors: Chen Yong-Gang , Yang Fan , Ding Qian
Published: June 2022

In this manuscript, the Cauchy problem of the modified Helmholtz equation is researched. This inverse problem is a serious ill-posed problem. The classical Landweber iterative regularization method is designed to find the regularized solution of this inverse problem. The error estimations between the exact solution and the regularization solution are all obtained under the a priori and the a posteriori regularization parameter selection rule. The Landweber iterative regularization method can also be applied to solve the Cauchy problem of the modified Helmholtz equation on the spherically symmetric and cylindrically symmetric regions.
Revealing the Symmetry of Materials through Neutron Diffraction
Authors: Ratcliff William
Published: June 2022

Magnetic materials are used in many devices in everyday life. To control their properties, we must first understand how they are ordered. This can be accomplished through neutron diffraction measurements. However, in many cases, there are too many parameters to determine the structure uniquely. Fortunately, symmetry can greatly constrain the number of parameters. Symmetry can also allow us to determine which physical properties are possible. In this review, I discuss the role of symmetry in magnetic structure determination using neutron diffraction. In this review, I will discuss both representational analysis as well as the magnetic superspace formalism. I will also discuss where the magnetic structure has been critical to understanding the fundamental science of the problem.
Convolutions for Bernoulli and Euler–Genocchi Polynomials of Order (r,m) and Their Probabilistic Interpretation
Authors: Frontczak Robert , Tomovski Živorad
Published: June 2022

The main purpose of this article is to derive several convolutions for generalized Bernoulli and Euler–Genocchi polynomials of order (r,m), Bn(r,m)(x) and An(r,m)(x), respectively. These polynomials have been introduced recently and contain the generalized Bernoulli, Euler and Genocchi polynomials as special members. Some of our results extend the results of M. Merca and others concerning Bernoulli numbers and polynomials. Probabilistic interpretations of the presented results are also given.
Simultaneous Removal of Mycotoxins by a New Feed Additive Containing a Tri-Octahedral Smectite Mixed with Lignocellulose
Authors: Greco Donato , D’Ascanio Vito , Abbasciano Mariagrazia , Santovito Elisa , Garbetta Antonella , Logrieco Antonio F. , Avantaggiato Giuseppina
Published: June 2022

Simultaneous removal of mycotoxins has been poorly addressed, and a limited number of studies have reported the efficacy of feed additives in sequestering a large spectrum of mycotoxins. In this study, a new mycotoxin-adsorbing agent was obtained by properly mixing a tri-octahedral smectite with a lignocellulose-based material. At a dosage of 1 mg mL−1, these materials simultaneously adsorbed frequently occurring mycotoxins and did not exert a cytotoxic effect on intestinal cells. Chyme samples obtained by a simulated GI digestion did not affect the viability of Caco-2TC7 cells as measured by the MTT test. In addition, the chyme of the lignocellulose showed a high content of polyphenols (210 mg mL−1 catechin equivalent) and good antioxidant activity. The properties of the individual constituents were maintained in the final composite, and were unaffected by their combination. When tested with a pool of seven mycotoxins at 1 µg mL−1 each and pH 5, the composite (5 mg mL−1) simultaneously sequestered AFB1 (95%), FB1 (99%), ZEA (93%), OTA (80%), T-2 (63%), and DON (22%). HT-2 adsorption did not occur. Mycotoxin adsorption increased exponentially as dosage increased, and occurred at physiological pH values. AFB1, ZEA and T-2 adsorption was not affected by pH in the range 3–9, whereas OTA and FB1 were adsorbed at pH values of 3–5. The adsorbed amount of AFB1, ZEA and T-2 was not released when pH rose from 3 to 7. FB1 and OTA desorption was less than 38%. Langmuir adsorption isotherms revealed high capacity and affinity for adsorption of the target mycotoxins. Results of this study are promising and show the potential of the new composite to remove mycotoxins in practical scenarios where several mycotoxins can co-occur.
RNA Interference-Mediated Knockdown of Bombyx mori Haemocyte-Specific Cathepsin L (Cat L)-Like Cysteine Protease Gene Increases Bacillus thuringiensis kurstaki Toxicity and Reproduction in Insect Cadavers
Authors: Yang Linlin , Sun Yanyan , Chang Meiling , Zhang Yun , Qiao Huili , Huang Siliang , Kan Yunchao , Yao Lunguang , Li Dandan , Ayra-Pardo Camilo
Published: June 2022

The silkworm’s Cat L-like gene, which encodes a lysosomal cathepsin L-like cysteine protease, is thought to be part of the insect’s innate immunity via an as-yet-undetermined mechanism. Assuming that the primary function of Cat L-like is microbial degradation in mature phagosomes, we hypothesise that the suppression of the Cat L-like gene expression would increase Bacillus thuringiensis (Bt) bacteraemia and toxicity in knockdown insects. Here, we performed a functional analysis of Cat L-like in larvae that were fed mulberry leaves contaminated with a commercial biopesticide formulation based on Bt kurstaki (Btk) (i.e., Dipel) to investigate its role in insect defence against a known entomopathogen. Exposure to sublethal doses of Dipel resulted in overexpression of the Cat L-like gene in insect haemolymph 24 and 48 h after exposure. RNA interference (RNAi)-mediated suppression of Cat L-like expression significantly increased the toxicity of Dipel to exposed larvae. Moreover, Btk replication was higher in RNAi insects, suggesting that Cat L-like cathepsin may be involved in a bacterial killing mechanism of haemocytes. Finally, our results confirm that Cat L-like protease is part of the antimicrobial defence of insects and suggest that it could be used as a target to increase the insecticidal efficacy of Bt-based biopesticides.
In Vitro Metabolism of Phenylspirodrimanes Derived from the Indoor Fungus Stachybotrys
Authors: Lindemann Viktoria , Jagels Annika , Behrens Matthias , Hübner Florian , Humpf Hans-Ulrich
Published: June 2022

Fungi belonging to the genus Stachybotrys are frequently detected in water-damaged indoor environments, and a potential correlation between emerging health problems of inhabitants of affected housing and the fungi is controversially discussed. Secondary metabolites (i.e., mycotoxins) produced by Stachybotrys, such as the highly toxic macrocyclic trichothecenes (MCTs), are of potential concern to human health. The present study, however, focused on the potential effects of the more broadly and abundantly formed group of phenylspirodrimanes (PSDs). The phase I and II metabolism of four structurally different PSDs were investigated in vitro using hepatic models in combination with high-performance liquid chromatography high-resolution mass spectrometry (HPLC-HRMS) analysis. In addition to metabolite detection by HRMS, isolation and structure elucidation by nuclear magnetic resonance spectroscopy (NMR) was part of the conducted study as well.
Reductive Amination for LC–MS Signal Enhancement and Confirmation of the Presence of Caribbean Ciguatoxin-1 in Fish
Authors: Kryuchkov Fedor , Robertson Alison , Mudge Elizabeth M. , Miles Christopher O. , Van Gothem Soetkien , Uhlig Silvio
Published: June 2022

Ciguatera poisoning is a global health concern caused by the consumption of seafood containing ciguatoxins (CTXs). Detection of CTXs poses significant analytical challenges due to their low abundance even in highly toxic fish, the diverse and in-part unclarified structures of many CTX congeners, and the lack of reference standards. Selective detection of CTXs requires methods such as liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS) or high-resolution MS (LC–HRMS). While HRMS data can provide greatly improved resolution, it is typically less sensitive than targeted LC–MS/MS and does not reliably comply with the FDA guidance level of 0.1 µg/kg CTXs in fish tissue that was established for Caribbean CTX-1 (C-CTX-1). In this study, we provide a new chemical derivatization approach employing a fast and simple one-pot derivatization with Girard’s reagent T (GRT) that tags the C-56-ketone intermediate of the two equilibrating C-56 epimers of C-CTX-1 with a quaternary ammonium moiety. This derivatization improved the LC–MS/MS and LC–HRMS responses to C-CTX-1 by approximately 40- and 17-fold on average, respectively. These improvements in sensitivity to the GRT-derivative of C-CTX-1 are attributable to: the improved ionization efficiency caused by insertion of a quaternary ammonium ion; the absence of adduct-ions and water-loss peaks for the GRT derivative in the mass spectrometer, and; the prevention of on-column epimerization (at C-56 of C-CTX-1) by GRT derivatization, leading to much better chromatographic peak shapes. This C-CTX-1–GRT derivatization strategy mitigates many of the shortcomings of current LC–MS analyses for C-CTX-1 by improving instrument sensitivity, while at the same time adding selectivity due to the reactivity of GRT with ketones and aldehydes.
Proper Selection of In Vitro Cell Model Affects the Characterization of the Neutralizing Antibody Response against SARS-CoV-2
Authors: Criscuolo Elena , Giuliani Benedetta , Ferrari Davide , Ferrarese Roberto , Diotti Roberta A. , Clementi Massimo , Mancini Nicasio , Clementi Nicola
Published: June 2022

(1) Background: Our aim is the evaluation of the neutralizing activity of BNT162b2 mRNA vaccine-induced antibodies in different in vitro cellular models, as this still represents one of the surrogates of protection against SARS-CoV-2 viral variants. (2) Methods: The entry mechanisms of SARS-CoV-2 in three cell lines (Vero E6, Vero E6/TMPRSS2 and Calu-3) were evaluated with both pseudoviruses and whole virus particles. The neutralizing capability of sera collected from vaccinated subjects was characterized through cytopathic effects and Real-Time RT PCR. (3) Results: In contrast to Vero E6 and Vero E6/TMPRSS2, Calu-3 allowed the evaluation of both viral entry mechanisms, resembling what occurs during natural infection. The choice of an appropriate cellular model can decisively influence the determination of the neutralizing activity of antibodies against SARS-CoV-2 variants. Indeed, the lack of correlation between neutralizing data in Calu-3 and Vero E6 demonstrated that testing the antibody inhibitory activity by using a single cell model possibly results in an inaccurate characterization. (4) Conclusions: Cellular systems allowing only one of the two viral entry pathways may not fully reflect the neutralizing activity of vaccine-induced antibodies moving increasingly further away from possible correlates of protection from SARS-CoV-2 infection.
Feline Panleukopenia Outbreaks and Risk Factors in Cats in Animal Shelters
Authors: Rehme Teresa , Hartmann Katrin , Truyen Uwe , Zablotski Yury , Bergmann Michèle
Published: June 2022

(1) Background: This study aimed to determine the risk factors for outbreaks of feline panleukopenia in shelters. (2) Methods: Four shelters (A−D) with 150 cats were included. Fecal samples were analyzed by parvovirus real-time polymerase chain reaction (qPCR), including culture and sequencing of qPCR-positive samples. Information on cats, husbandry, hygiene, and infection management was evaluated to determine risk factors for feline panleukopenia and parvovirus shedding by logistic regression. (3) Results: Feline panleukopenia occurred in 28.0% (42/150) of cats (0 in shelter D). Shedding was found in 48.7% (73/150) (A: 21/73; B: 29/73; C: 7/73; D: 16/73). Of 73 qPCR-positive fecal samples, 65.8% (48/73) were culture-positive; sequencing revealed feline panleukopenia virus (FPV) isolates in 34/48 samples and vaccine virus isolate in 14/48; canine parvovirus was not detected. Presence of feline panleukopenia was significantly more likely in cats from shelter A (p < 0.05), unvaccinated cats (p < 0.001), and young cats (4 weeks to 2 years; p = 0.008). Parvovirus shedding was significantly more common in young cats (p < 0.001), cats with feline panleukopenia (p = 0.033), and group-housed cats (p = 0.025). (4) Conclusions: Vaccination is the most important measure to reduce the risk of feline panleukopenia in shelters. Risk of parvovirus shedding is especially high in young, group-housed cats.
Construction of a Dengue NanoLuc Reporter Virus for In Vivo Live Imaging in Mice
Authors: Fang Enyue , Liu Xiaohui , Li Miao , Liu Jingjing , Zhang Zelun , Liu Xinyu , Li Xingxing , Li Wenjuan , Peng Qinhua , Yu Yongxin , Li Yuhua
Published: June 2022

Since the first isolation in 1943, the dengue virus (DENV) has spread throughout the world, but effective antiviral drugs or vaccines are still not available. To provide a more stable reporter DENV for vaccine development and antiviral drug screening, we constructed a reporter DENV containing the NanoLuc reporter gene, which was inserted into the 5′ untranslated region and capsid junction region, enabling rapid virus rescue by in vitro ligation. In addition, we established a live imaging mouse model and found that the reporter virus maintained the neurovirulence of prototype DENV before engineering. DENV-4 exhibited dramatically increased neurovirulence following a glycosylation site-defective mutation in the envelope protein. Significant mice mortality with neurological onset symptoms was observed after intracranial infection of wild-type (WT) mice, thus providing a visualization tool for DENV virulence assessment. Using this model, DENV was detected in the intestinal tissues of WT mice after infection, suggesting that intestinal lymphoid tissues play an essential role in DENV pathogenesis.
High Incidence of SARS-CoV-2 Variant of Concern Breakthrough Infections Despite Residual Humoral and Cellular Immunity Induced by BNT162b2 Vaccination in Healthcare Workers: A Long-Term Follow-Up Study in Belgium
Authors: Calcoen Bas , Callewaert Nico , Vandenbulcke Aline , Kerstens Winnie , Imbrechts Maya , Vercruysse Thomas , Dallmeier Kai , Van Weyenbergh Johan , Maes Piet , Bossuyt Xavier , Zapf Dorinja , Dieckmann Kersten , Callebaut Kim , Thibaut Hendrik Jan , Vanhoorelbeke Karen , De Meyer Simon F. , Maes Wim , Geukens Nick
Published: June 2022

To mitigate the massive COVID-19 burden caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), several vaccination campaigns were initiated. We performed a single-center observational trial to monitor the mid- (3 months) and long-term (10 months) adaptive immune response and to document breakthrough infections (BTI) in healthcare workers (n = 84) upon BNT162b2 vaccination in a real-world setting. Firstly, serology was determined through immunoassays. Secondly, antibody functionality was analyzed via in vitro binding inhibition and pseudovirus neutralization and circulating receptor-binding domain (RBD)-specific B cells were assessed. Moreover, the induction of SARS-CoV-2-specific T cells was investigated by an interferon-γ release assay combined with flowcytometric profiling of activated CD4+ and CD8+ T cells. Within individuals that did not experience BTI (n = 62), vaccine-induced humoral and cellular immune responses were not correlated. Interestingly, waning over time was more pronounced within humoral compared to cellular immunity. In particular, 45 of these 62 subjects no longer displayed functional neutralization against the delta variant of concern (VoC) at long-term follow-up. Noteworthily, we reported a high incidence of symptomatic BTI cases (17.11%) caused by alpha and delta VoCs, although vaccine-induced immunity was only slightly reduced compared to subjects without BTI at mid-term follow-up.
PIMGAVir and Vir-MinION: Two Viral Metagenomic Pipelines for Complete Baseline Analysis of 2nd and 3rd Generation Data
Authors: Mastriani Emilio , Bienes Kathrina Mae , Wong Gary , Berthet Nicolas
Published: June 2022

The taxonomic classification of viral sequences is frequently used for the rapid identification of pathogens, which is a key point for when a viral outbreak occurs. Both Oxford Nanopore Technologies (ONT) MinION and the Illumina (NGS) technology provide efficient methods to detect viral pathogens. Despite the availability of many strategies and software, matching them can be a very tedious and time-consuming task. As a result, we developed PIMGAVir and Vir-MinION, two metagenomics pipelines that automatically provide the user with a complete baseline analysis. The PIMGAVir and Vir-MinION pipelines work on 2nd and 3rd generation data, respectively, and provide the user with a taxonomic classification of the reads through three strategies: assembly-based, read-based, and clustering-based. The pipelines supply the scientist with comprehensive results in graphical and textual format for future analyses. Finally, the pipelines equip the user with a stand-alone platform with dedicated and various viral databases, which is a requirement for working in field conditions without internet connection.
Selection and Validation of Reference Genes for Gene Expression Studies Using Quantitative Real-Time PCR in Prunus Necrotic Ringspot Virus-Infected Cucumis sativus
Authors: Dong Zhenfei , Zhan Binhui , Li Shifang
Published: June 2022

Several members of the genus Ilarvirus infect fruit trees and are distributed worldwide. Prunus necrotic ringspot virus (PNRSV) is one of the most prevalent viruses, causing significant losses. Cucumissativus can be infected by several ilarviruses, leading to obvious symptoms, including PNRSV, which suggests that cucumbers could be good hosts for the study of the pathogenesis of ilarviruses. Real-time quantitative PCR is an optimal choice for studying gene expression because of its simplicity and its fast and high sensitivity, while its accuracy is highly dependent on the stability of the reference genes. In this study, we assessed the stability of eleven reference genes with geNorm, NormFinder, ΔCt method, BestKeeper, and the ranking software, RefFinder. The results indicated that the combined use of EF1α and F-BOX was the most accurate normalization method. In addition, the host genes AGO1, AGO4, and RDR6 were selected to test the reliability of the reference genes. This study provides useful information for gene expression analysis during PNRSV infection and will facilitate gene expression studies associated with ilarvirus infection.
Targeted Virome Sequencing Enhances Unbiased Detection and Genome Assembly of Known and Emerging Viruses—The Example of SARS-CoV-2
Authors: Pogka Vasiliki , Papadopoulou Gethsimani , Valiakou Vaia , Sgouras Dionyssios N. , Mentis Andreas F. , Karamitros Timokratis
Published: June 2022

Targeted virome enrichment and sequencing (VirCapSeq-VERT) utilizes a pool of oligos (baits) to enrich all known—up to 2015—vertebrate-infecting viruses, increasing their detection sensitivity. The hybridisation of the baits to the target sequences can be partial, thus enabling the detection and genomic reconstruction of novel pathogens with <40% genetic diversity compared to the strains used for the baits’ design. In this study, we deploy this method in multiplexed mixes of viral extracts, and we assess its performance in the unbiased detection of DNA and RNA viruses after cDNA synthesis. We further assess its efficiency in depleting various background genomic material. Finally, as a proof-of-concept, we explore the potential usage of the method for the characterization of unknown, emerging human viruses, such as SARS-CoV-2, which may not be included in the baits’ panel. We mixed positive samples of equimolar DNA/RNA viral extracts from SARS-CoV-2, coronavirus OC43, cytomegalovirus, influenza A virus H3N2, parvovirus B19, respiratory syncytial virus, adenovirus C and coxsackievirus A16. Targeted virome enrichment was performed on a dsDNA mix, followed by sequencing on the NextSeq500 (Illumina) and the portable MinION sequencer, to evaluate its usability as a point-of-care (PoC) application. Genome mapping assembly was performed using viral reference sequences. The untargeted libraries contained less than 1% of total reads mapped on most viral genomes, while RNA viruses remained undetected. In the targeted libraries, the percentage of viral-mapped reads were substantially increased, allowing full genome assembly in most cases. Targeted virome sequencing can enrich a broad range of viruses, potentially enabling the discovery of emerging viruses.
Transcriptomic Analysis of MDBK Cells Infected with Cytopathic and Non-Cytopathic Strains of Bovine Viral Diarrhea Virus (BVDV)
Authors: Mirosław Paweł , Rola-Łuszczak Marzena , Kuźmak Jacek , Polak Mirosław P.
Published: June 2022

Bovine viral diarrhea virus (BVDV) belongs to the Flaviviridae family and the Pestivirus genus. Infection with BVDV causes a disease with a wide spectrum of clinical symptoms, most often mild, although infections with this virus constitute a serious economic problem all over the world. The virus is characterized by a high genetic variability, while the accumulation of single mutations leads to the formation of its new variants. The aim of this study was to better understand the complicated pathogenesis of this disease at the molecular level via the analysis of the transcriptome of cells infected with this virus. The bovine kidney cell line (MDBK), the cytopathic (cp) reference strain, and two non-cytopathic (ncp) BVD virus field strains were used in transcriptomic studies. The cell transcriptome was tested 24 and 72 h after infection. The results of the microarray analysis revealed changes in the expression levels of numerous genes. Genes with changed expression as a result of infection with the cp strain caused changes in the expression levels of a large number of genes and enriched a number of pathways. Genes with increased expression levels were enriched among other pathways involved in the cell cycle, while genes with reduced expression levels enriched pathways mostly related to metabolism. Genes with increased expression levels as a result of infection with ncp strains enriched a much smaller number of pathways, among them, pathways related to signaling activity 24 h post-infection and serine biosynthetic pathways both 24 and 72 h post-infection. Pathways enriched by genes with reduced expression levels were related to the innate immune response (72 h post-infection) or metabolism (24 and 72 h post-infection). The results of microarray studies can help us to better understand the host’s response to BVDV infection.
Embryotoxicity of Polystyrene Microspheres of Different Sizes to the Marine Medaka Oryzias melastigma (McClelland, 1839)
Authors: Chen Jincan , Fang Chao , Zheng Ronghui , Bo Jun
Published: June 2022

Polystyrene microplastics (PS-MPs) are potentially harmful to marine organisms, especially during the early developmental stages, although the underlying mechanism remains unclear. The present study evaluated the growth and morphological characteristics of marine medaka Oryzias melastigma (McClelland, 1839) embryos exposed to PS-MP. PS-MPs of three different sizes (0.05, 0.5, and 6.0 μm with a concentration of 106 particles/L) were subjected to waterborne exposure for 19 d. The hatching time and rate of embryos exposed to 0.5 and 6.0 μm PS-MPs were significantly lower than those of the control, while no significant difference was observed in the 0.05 μm treatment. No significant differences were observed in the mortality rate of the embryos, embryo diameter, and relevant gene expression levels, including il6, il8, il-1β, jak, stat-3, nf-κb, hif-1α, epo, cyp1a1, ahr, sod, cat, and gpx, but with the exception of vtg. Fluorescent PS-MPs were found on the embryo surfaces when the embryos were exposed to 0.5 and 6.0 μm PS-MPs, but no signals were detected inside embryos using confocal microscopy. Therefore, the results indicate that PS-MPs having a diameter of 6.0 μm can only attach to the surface or villus of embryos and not enter the embryos through the membrane pores, whereas PS-MPs with diameters of 0.05 and 0.5 μm cannot enter the embryos.
The Impacts of Hydrology and Climate on Hydrological Connectivity in a Complex River–Lake Floodplain System Based on High Spatiotemporal Resolution Images
Authors: Yang Suhang , Liang Jie , Li Xiaodong , Yi Yuru , Zhu Ziqian , Li Xin , Chen Xuwu , Li Shuai , Zhai Yeqing , Pei Ziming
Published: June 2022

The drivers that determine the hydrological connectivity (HC) are complex and interrelated, and disentangling this complexity will improve the administration of the river–lake interconnection system. Dongting Lake, as a typical river–lake interconnected system, is freely connected with the Yangtze River and their HC plays a major role in keeping the system healthy. Climate, hydrology, and anthropogenic activities are associated with the HC. In this study, hydrological drivers were divided into the total flow of three inlets (T-flow) and the total flow of four tributaries (F-flow). To elucidate the HC of the Dongting Lake, HC was calculated by geostatistical methods in association with Sentinel-2 remote sensing images. Then, the structural equation model (SEM) was used to quantify the impacts of hydrology (F-flow, and T-flow) and meteorology (precipitation, evaporation, and temperature) on HC. The geostatistical analysis results demonstrated that the HC showed apparent seasonal change. For East and West Dongting Lake, the dominant element was north–south hydrological connectivity (N–S HC), and the restricted was west–east hydrological connectivity (W-E HC), but the dominant element was E–W HC and the restricted was N–S HC in South Dongting Lake. The results of SEM showed that N–S HC was mainly explained by T-flow (r = 0.49, p < 0.001) and F-flow (r = 0.28, p < 0.05). T-flow, temperature (r = 0.33, p < 0.05), and F-flow explained E–W HC. The finding of this work supports the management of both the Dongting Lake floodplain and other similar river–lake floodplain systems.
Groundwater Potential Assessment Using GIS and Remote Sensing Techniques: Case Study of West Arsi Zone, Ethiopia
Authors: Kabeto Julla , Adeba Dereje , Regasa Motuma Shiferaw , Leta Megersa Kebede
Published: June 2022

Groundwater is a crucial source of water supply due to its continuous availability, reasonable natural quality, and being easily diverted directly to the poor community more cheaply and quickly. The West Arsi Zone residents remain surface water dependent due to traditional exploration of groundwater, which is a tedious approach in terms of resources and time. This study uses remote sensing data and geographic information system techniques to evaluate the groundwater potential of the study area. This technique is a fast, accurate, and feasible technique. Groundwater potential and recharge zone influencing parameters were derived from Operational Land Imager 8, digital elevation models, soil data, lithological data, and rainfall data. Borehole data were used for results validation. With spatial analysis tools, the parameters affecting groundwater potential (LULC, soil, lithology, rainfall, drainage density, lineament density, slope, and elevation) were mapped and organized. The weight of the parameters according to percent of influence on groundwater potential and recharge was determined by Analytical Hierarchy Process according to their relative influence. For weights allocated to each parameter, the consistency ratio obtained was 0.033, which is less than 0.1, showing the weight allocated to each parameter is acceptable. In the weighted overlay analysis, from a percent influence point of view, slope, land use/cover, and lithology are equally important and account for 24% each, while the soil group has the lowest percent of influence, which accounts only 2% according to this study. The generated groundwater potential map has four ranks, 2, 3, 4, and 5, in which its classes are Low, Moderate, High, and Very High, respectively, based on its groundwater potential availability rank and class. The area coverage is 9825.84 ha (0.79%), 440,726.49 ha (35.46%), 761,438.61 ha (61.27%), and 30,748.68 ha (2.47%) of the study area, respectively. Accordingly, the western part of district is expected to have very high groundwater potential. High groundwater potential is concentrated in the central and western parts whereas moderate groundwater potential distribution is dominant in the eastern part of the area. The validation result of 87.61% confirms the very good agreement among the groundwater record data and groundwater potential classes delineated.
Effects of Light-Emitting Diode Illumination on Sediment Surface Biological Activities and Releases of Nutrients and Metals to Overlying Water in Eutrophic Lake Microcosms
Authors: Hao Aimin , Yu Hong , Kobayashi Sohei , Xia Dong , Zhao Min , Iseri Yasushi
Published: June 2022

The release of nutrients and metals from the sediment to the overlying water induced by oxygen depletion is an important issue in eutrophic aquatic systems. Effects of light-emitting diode (LED) illumination on oxygen conditions and release of nutrients and metals from the sediment were examined by comparing with those effects of aeration in microcosms using water and sediment of Lake Taihu, China. Periphyton with filamentous algae developed on the sediment surface in the LED (blue wavelength) treatment. Dissolved oxygen became rapidly saturated and gradually supersaturated in the aeration and LED treatments, respectively, but remained low in the control. A thicker oxic layer developed on the sediment for the LED than aeration but was poorly developed with a blackened surface in the control. Invertebrate burrows were distributed deeper and the bacterial community was more dominated by aerobic species in the LED, indicating deeper penetration of oxygen into the sediment. Nutrients (e.g., N and P) and some metals (e.g., Hg, As, and Mn) in water were lower for the LED and aeration than in the control; nutrients and other solutes that increased electric conductivity (e.g., Ca, Mg) were lower for the LED than aeration. These results suggest that LED can effectively oxygenate the bottom water by stimulating algal photosynthesis and benthic invertebrate activity, resulting in greater retention of nutrients and metals in/on sediment compared to aeration.
Estimating Suspended Sediment Fluxes from the Largest Glacial Lake in Svalbard to Fjord System Using Sentinel-2 Data: Trebrevatnet Case Study
Authors: Kavan Jan , Wieczorek Iwo , Tallentire Guy D. , Demidionov Mihail , Uher Jakub , Strzelecki Mateusz C.
Published: June 2022

Glacier-fed hydrological systems in high latitude regions experience high seasonal variation in meltwater runoff. The peak in runoff usually coincides with the highest air temperatures which drive meltwater production. This process is often accompanied by the release of sediments from within the glacier system that are transported and suspended in high concentrations as they reach the proglacial realm. Sediment-laden meltwater is later transported to the marine environment and is expressed on the surface of fjords and coastal waters as sediment plumes. Direct monitoring of these processes requires complex and time-intensive fieldwork, meaning studies of these processes are rare. This paper demonstrates the seasonal dynamics of the Trebrevatnet lake complex and evolution of suspended sediment in the lake and sediment plumes in the adjacent Ekmanfjorden. We use the Normalized Difference Suspended Sediment Index (NDSSI) derived from multi-temporal Sentinel-2 images for the period between 2016–2021. We propose a new SSL index combining the areal extent of the sediment plume with the NDSSI for quantification of the sediment influx to the marine environment. The largest observed sediment plume was recorded on 30 July 2018 and extended to more than 40 km2 and a SSL index of 10.4. We identified the greatest sediment concentrations in the lake in the beginning of August, whereas the highest activity of the sediment plumes is concentrated at the end of July. The temporal pattern of these processes stays relatively stable throughout all ablation seasons studied. Sediment plumes observed with the use of optical satellite remote sensing data may be used as a proxy for meltwater runoff from the glacier-fed Trebrevatnet system. We have shown that remote-sensing-derived suspended sediment indexes can (after proper in situ calibration) serve for large scale quantification of sediment flux to fjord and coastal environments.