bluuebunny/crossref_metadata_2025 huggingface dataset:
The Phylogenetic Characterization of Balantioides coli Isolated in the Pavlova Culture Medium Supplemented with Coconut Water and Animal Serum
Authors: Class Camila Souza Carvalho , Corrêa Laís Lisboa , Knackfuss Fabiana Batalha , Amendoeira Maria Regina Reis , Gordo Francisco Ponce , Barbosa Alynne da Silva
Published: June 2024

Balantioides coli is a ciliated protist that can cause dysentery in humans, pigs and nonhuman primates and may have the potential for zoonotic transmission. Its diagnosis is routinely performed through conventional parasitological techniques, and few studies have used culturing techniques to isolate it, applying molecular tools for the characterization of this protozoan. Thus, the objective of this study was to confirm B. coli diagnosis using molecular tools and to characterize the genetic variants of this parasite isolated from pigs kept on family farms in Brazil using three different culture media that differed in the serum added. Fecal samples from pigs were inoculated in Pavlova medium plus coconut water (PC), fetal bovine serum (PB) and horse serum (PH). Of the 127 samples positive for forms compatible with the phylum Ciliophora, 31 were selected for isolation. The most successful medium for isolation was PB 19/31 (61.3%), followed by PH 18/31 (58.1%) and PC 11/31 (35.5%). Of the nucleotide sequences generated, 20 were classified as genetic variant type B0, two as A1 and 15 as A0. The results indicated that PC, despite having allowed the isolation of B. coli for a short period, was not an adequate medium for the maintenance of this parasite in vitro, therefore requiring improvement.
Therapeutic Applications of Nanoformulated Resveratrol and Quercetin Phytochemicals in Colorectal Cancer—An Updated Review
Authors: Unnikrishnan Meenakshi Dhanalekshmi , Narde Gurpreet Kaur , Ahuja Alka , Al Balushi Khalid , Francis Arul Prakash , Khan Shah Alam
Published: June 2024

Natural compounds such as polyphenols play several positive roles in maintaining the oxidative and inflammatory capacity of cells, which leads to their potential use as anticancer therapeutics. There is promising evidence for the in vitro and in vivo anticancer activity of many polyphenols, including resveratrol and quercetin, specifically in the treatment of colorectal cancer (CRC). There is a clear association between resveratrol and quercetin in interfering with the mechanistic pathways involved in CRC, such as Wnt, P13K/AKT, caspase-3, MAPK, NF-κB, etc. These molecular pathways establish the role of resveratrol and quercetin in controlling cancer cell growth, inducing apoptosis, and inhibiting metastasis. The major bottleneck in the progression of the use of resveratrol and quercetin as anticancer therapeutics is their reduced bioavailability in vivo because of their rapid metabolism in humans. Recent advancements in various nanotechnological formulations are promising for overcoming these bioavailability issues. Various nanoformulations of resveratrol and quercetin have shown an optimistic impact on reducing the solubility and improving the stability of resveratrol and quercetin in vivo. A combinatorial approach using nanoformulations of resveratrol with quercetin could potentially increase the impact of resveratrol in controlling CRC cell proliferation. This review discusses the mechanism of resveratrol and quercetin, the two bioactive polyphenolics, in colon cancer, with an emphasis on various types of nanoformulations of the two molecules targeting colon cancer. It also explores the synergistic effect of combining resveratrol and quercetin in various nanoformulations, targeting colon cancer. This research delves into the enhanced pharmacokinetics and potential chemotherapeutic benefits of these bioactive polyphenolics when used together in innovative ways.
Solitary and Synergistic Effects of Different Hydrophilic and Hydrophobic Phospholipid Moieties on Rat Behaviors
Authors: Kikuchi Shuhei , Iwasaki Yugo , Yoshioka Mina , Hino Kodai , Morita Shin-ya , Tada Ryu , Uchimura Yasuhiro , Kubo Yoshinori , Kobayashi Tomoya , Kinoshita Yusuke , Hayashi Masahiro , Furusho Yoshio , Tamiaki Hitoshi , Ishiyama Hiroaki , Kuroda Minoru , Udagawa Jun
Published: June 2024

Glycerophospholipids have hydrophobic and hydrophilic moieties. Previous studies suggest that phospholipids with different moieties have different effects on rodent behavior; however, the relationship between chemical structures and behavioral effects remains unclear. To clarify the functions of phospholipid moieties, we injected male rats with phospholipids with different moieties and conducted behavioral tests. Exploratory activity was reduced by phosphatidylethanolamine (PE)(18:0/22:6) but not PE(18:0/18:0) or PE(18:0/20:4). Conversely, exploratory activity was increased by plasmanyl PE(16:0/22:6), which harbors an alkyl–ether linkage, but not by phosphatidylcholine (PC)(16:0/22:6) or plasmanyl PC(16:0/22:6). Docosahexaenoic acid (DHA)(22:6) and an alkyl–ether linkage in PE were thus postulated to be involved in exploratory activity. Anxiety-like behavior was reduced by plasmenyl PC(18:0/20:4), which harbors a vinyl–ether linkage, but not by PC(18:0/20:4) or plasmanyl PC(18:0/20:4), suggesting the anxiolytic effects of vinyl–ether linkage. The activation of social interaction was suppressed by PE(18:0/18:0), PE(18:0/22:6), PC(16:0/22:6), plasmanyl PE(16:0/22:6), and plasmanyl PC(16:0/22:6) but not by PE(18:0/20:4), plasmenyl PE(18:0/20:4), or plasmanyl PC(18:0/22:6). DHA may suppress social interaction, whereas arachidonic acid(20:4) or a combination of alkyl–ether linkage and stearic acid(18:0) may restore social deficits. Our findings indicate the characteristic effects of different phospholipid moieties on rat behavior, and may help to elucidate patterns between chemical structures and their effects.
Effects of Jasmonic Acid on Stress Response and Quality Formation in Vegetable Crops and Their Underlying Molecular Mechanisms
Authors: Wu Jiaqi , Chen Yangyang , Xu Yujie , An Yahong , Hu Zhenzhu , Xiong Aisheng , Wang Guanglong
Published: June 2024

The plant hormone jasmonic acid plays an important role in plant growth and development, participating in many physiological processes, such as plant disease resistance, stress resistance, organ development, root growth, and flowering. With the improvement in living standards, people have higher requirements regarding the quality of vegetables. However, during the growth process of vegetables, they are often attacked by pests and diseases and undergo abiotic stresses, resulting in their growth restriction and decreases in their yield and quality. Therefore, people have found many ways to regulate the growth and quality of vegetable crops. In recent years, in addition to the role that JA plays in stress response and resistance, it has been found to have a regulatory effect on crop quality. Therefore, this study aims to review the jasmonic acid accumulation patterns during various physiological processes and its potential role in vegetable development and quality formation, as well as the underlying molecular mechanisms. The information provided in this manuscript sheds new light on the improvements in vegetable yield and quality.
Improving Pore Characteristics, Mechanical Properties and Thermal Performances of Near-Net Shape Manufacturing Phenolic Resin Aerogels
Authors: Sha Ruyi , Dai Jixiang , Wang Bingzhu , Sha Jianjun
Published: June 2024

Thermally stable high-performance phenolic resin aerogels (PRAs) are of great interest for thermal insulation because of their light weight, fire retardancy and low thermal conductivity. However, the drawbacks of PRA synthesis, such as long processing time, inherent brittleness and significant shrinkage during drying, greatly restrict their wide applications. In this work, PRAs were synthesized at ambient pressure through a near-net shape manufacturing technique, where boron-containing thermosetting phenolic resin (BPR) was introduced into the conventional linear phenolic resin (LPR) to improve the pore characteristics, mechanical properties and thermal performances. Compared with the traditional LPR-synthesized aerogel, the processing time and the linear shrinkage rate during the drying of the PRAs could be significantly reduced, which was attributed to the enhanced rigidity and the unique bimodal pore size distribution. Furthermore, no catastrophic failure and almost no mechanical degradation were observed on the PRAs, even with a compressive strain of up to 60% at temperatures ranging from 25 to 200 °C, indicating low brittleness and excellent thermo-mechanical stability. The PRAs also showed outstanding fire retardancy. On the other hand, the PRAs with a density of 0.194 g/cm3 possessed a high Young’s modulus of 12.85 MPa and a low thermal conductivity of 0.038 W/(m·K).
‘The Last Bastion of Evangelicalism in Europe?’ Evangelicalism and Religiosity in Northern Ireland
Authors: Ganiel Gladys , Soye Emma
Published: June 2024

This article explores whether or to what extent Northen Ireland—long-noted for its unusually high levels of religiosity—remains, as the late preacher-politician Rev Ian Paisley (d. 2014) described it: the last bastion of evangelicalism in Europe. It presents the results of two major polls conducted in 2023, which together provide the most comprehensive picture of religion in Northern Ireland in two decades. The polls were a representative survey of Northern Ireland, carried out by a professional research company, and a self-selecting online questionnaire distributed by the Evangelical Alliance in Northern Ireland. The data confirm continued high levels of religiosity, with 50 percent of the general population reporting that they are practising Christians. Surprisingly, 38 percent of practising Catholics self-identify as evangelical—up from six percent in a 2004 survey. Men are more likely to identify as evangelical than women, and young practising Christians (18–34) are more likely to identify as evangelical than other age groups. As expected, evangelicals hold more morally/socially conservative views on a range of issues. We also develop a new four-fold typology to describe evangelicals in Northern Ireland: broad-church evangelicals, classic evangelicals, Catholic evangelicals, and ex-vangelicals (those who were once evangelical but no longer identify as such).
Islamophobia beyond Explicit Hate Speech: Analyzing the Coverage of Muslims in Slovenia’s Public Broadcasting
Authors: Jurekovič Igor
Published: June 2024

Religion in Europe has been undergoing two fundamental changes in the past four decades. As a side effect of secularization, religious fields have been pluralizing. On the other hand, religions themselves have taken a qualitative shift towards lived, material characteristics. Focusing exclusively on the diversification of European religious fields, we are interested in the concept of religious literacy as a tool for competent engagement in contemporary religious plural societies. To better understand the role of public media in fostering religious literacy, we offer an analysis of the public broadcaster’s coverage of smaller religious communities in Slovenia. Focusing particularly on Muslims as the largest religious minority in Slovenia, we provide an analysis of 245 episodes, consisting of 540 items, in the 2015–2020 period. We show that the coverage given to smaller religious communities is unevenly spread amongst the communities, with disproportional airtime given to Christian churches and communities. Furthermore, we pinpoint the key qualitative difference in portrayals of Slovenian Muslims and non-Catholic Christians, explaining how the process of racialized Islamophobia may continue beyond explicit hate speech. In conclusion we set out limitations of our study and provide guidelines for future research.
Observations, Remote Sensing, and Model Simulation to Analyze Southern Brazil Antarctic Ozone Hole Influence
Authors: Peres Lucas Vaz , Pinheiro Damaris Kirsh , Bencherif Hassan , Begue Nelson , Bageston José Valentin , Bittencourt Gabriela Dorneles , Portafaix Thierry , Schuch Andre Passaglia , Anabor Vagner , da Silva Rodrigo , Neves Theomar Trindade de Araujo Tiburtino , Silva Raphael Pablo Tapajós , dos Reis Gabriela Cacilda Godinho , dos Reis Marco Antônio Godinho , Martins Maria Paulete Pereira , Toihir Mohamed Abdoulwahab , Mbatha Nkanyiso , Steffenel Luiz Angelo , Mendes David
Published: June 2024

This paper presents the observational, remote sensing, and model simulation used to analyze southern Brazil Antarctic ozone hole influence (SBAOHI) events that occurred between 2005 and 2014. To analyze it, we use total ozone column (TOC) data provided by a Brewer spectrophotometer (BS) and the OMI (Ozone Monitoring Instrument). In addition to the AURA/MLS (Microwave Limb Sounder) instrument, satellite ozone profiles were utilized with DYBAL (Dynamical Barrier Localization) code in the MIMOSA (Modélisation Isentrope du Transport Mésoéchelle de l’Ozone Stratosphérique par Advection) model Potential Vorticity (PV) fields. TOC has 7.0 ± 2.9 DU reductions average in 62 events. October has more events (30.7%). Polar tongue events are 19.3% in total, being more frequently observed in October (50% of cases), with medium intensity (58.2%), and in the stratosphere medium levels (55.0%). Already, polar filament events (80.7%) are more frequent in September (32.0%), with medium intensity (42.0%), and stratosphere medium levels (40.7%).
Revealing the Eco-Environmental Quality of the Yellow River Basin: Trends and Drivers
Authors: Zhou Meiling , Li Zhenhong , Gao Meiling , Zhu Wu , Zhang Shuangcheng , Ma Jingjing , Ta Liangyu , Yang Guijun
Published: June 2024

The Yellow River Basin (YB) acts as a key barrier to ecological security and is an important experimental region for high-quality development in China. There is a growing demand to assess the ecological status in order to promote the sustainable development of the YB. The eco-environmental quality (EEQ) of the YB was assessed at both the regional and provincial scales utilizing the remote sensing-based ecological index (RSEI) with Landsat images from 2000 to 2020. Then, the Theil–Sen (T-S) estimator and Mann–Kendall (M-K) test were utilized to evaluate its variation trend. Next, the optimal parameter-based geodetector (OPGD) model was used to examine the drivers influencing the EEQ in the YB. Finally, the geographically weighted regression (GWR) model was utilized to further explore the responses of the drivers to RSEI changes. The results suggest that (1) a lower RSEI value was found in the north, while a higher RSEI value was found in the south of the YB. Sichuan (SC) and Inner Mongolia (IM) had the highest and the lowest EEQ, respectively, among the YB provinces. (2) Throughout the research period, the EEQ of the YB improved, whereas it deteriorated in both Henan (HA) and Shandong (SD) provinces. (3) The soil-available water content (AWC), annual precipitation (PRE), and distance from impervious surfaces (IMD) were the main factors affecting the spatial differentiation of RSEI in the YB. (4) The influence of meteorological factors (PRE and TMP) on RSEI changes was greater than that of IMD, and the influence of IMD on RSEI changes showed a significant increasing trend. The research results provide valuable information for application in local ecological construction and regional development planning.
A GPU-Based Integration Method from Raster Data to a Hexagonal Discrete Global Grid
Authors: Zheng Senyuan , Zhou Liangchen , Lu Chengshuai , Lv Guonian
Published: June 2024

This paper proposes an algorithm for the conversion of raster data to hexagonal DGGSs in the GPU by redevising the encoding and decoding mechanisms. The researchers first designed a data structure based on rhombic tiles to convert the hexagonal DGGS to a texture format acceptable for GPUs, thus avoiding the irregularity of the hexagonal DGGS. Then, the encoding and decoding methods of the tile data based on space-filling curves were designed, respectively, so as to reduce the amount of data transmission from the CPU to the GPU. Finally, the researchers improved the algorithmic efficiency through thread design. To validate the above design, raster integration experiments were conducted based on the global Aster 30 m digital elevation dataDEM, and the experimental results showed that the raster integration accuracy of this algorithms was around 1 m, while its efficiency could be improved to more than 600 times that of the algorithm for integrating the raster data to the hexagonal DGGS data, executed in the CPU. Therefore, the researchers believe that this study will provide a feasible method for the efficient and stable integration of massive raster data based on a hexagonal grid, which may well support the organization of massive raster data in the field of GIS.
Sensors Layout Optimization Design of Rocket Sled Test System
Authors: Qian Hongjun , Wang Wenjie , Zhao Xu , Jiang Yi
Published: June 2024

The rocket sled, as a ground dynamic test system, combines the characteristics of the wind tunnel test and the flight test. However, some practical factors, such as shock wave interference, ground effect, and high-intensity aerodynamic noise will cause serious interference and even failure of the uniformly distributed sensors during horizontal sliding in a wide speed range. The AGARD HB-2 standard model is employed as the payload to simulate the aerodynamic and aeroacoustic characteristics during the variable acceleration period, aiming to optimize the test sensors layout. It is observed that in the high Mach number flow fields, strong coupling behaviors among complex waves will occur. The peak of wake vortex strength will appear at 1.5 s and gradually diminish over time. In addition, when the vortex between the load and the booster is monitored, its position shifts forward in the subsonic stage, then gradually moves backward and expands in the supersonic stage. Acoustic directivity is pronounced at subsonic and transonic speeds, pointing towards 75° and 135° relative to the sliding speed, respectively. These results can provide technical support for sensor layout and high-precision testing in rocket sled tests.
Potential of Modified Reduced Tillage with Cover/Green Manure Crop for Climate Change Mitigation in a Smallholder Rainfed Farming System
Authors: Javed Nabeeha , Ijaz Shahzada Sohail , Hussain Qaiser , Ansar Muhammad , Alrefaei Abdulwahed Fahad , Almutairi Bader O. , Zaman Wajid , Yousra Munazza
Published: June 2024

Soil can function as a reservoir and a source of greenhouse gases (GHGs), contingent on its management. This study assesses the potential of a modified reduced tillage (MRT) approach involving the use of cover or green manure crops as a substitute for crop residues to mitigate GHG emissions from soil within smallholder rainfed farming systems. A two-year field experiment with different tillage techniques (moldboard plow, tine cultivator, and modified reduced tillage) and crop rotations (summer fallow–wheat and cover/green manure–wheat) was conducted at Rawalpindi, Pakistan. The results showed that MRT reduced carbon dioxide (CO2) and nitrous oxide (N2O) emissions by 8% and 15.3%, respectively, from soil while maintaining consistently higher soil moisture than conventional tillage techniques. The modified reduced tillage reduced the global warming potential (GWP) and greenhouse gas intensity (GHGI) by 15.8% and 20.7%, respectively. The net ecosystem exchange (NEE) was unaffected by the tillage systems. Therefore, adopting the MRT technique and incorporating green manure is a viable strategy for curtailing GHG emissions from soil, particularly in the context of smallholder rainfed farming systems. Extended, multi-year studies under various climate scenarios and agronomic practices are needed to understand the long-term impacts of MRT and crop rotations on GHG emissions.
An Emissions Offset Strategy to Accomplish 2 °C Long-Term Mitigation Goals in the European Union
Authors: Perissi Ilaria , Jones Aled
Published: June 2024

Regional carbon budgeting in policymaking is underutilized despite its importance for achieving global climate goals, particularly the Paris Agreement’s target of limiting global warming to 2 °C by 2050. In this work, we present the model PLEDGES, a novel system dynamic-based simulation tool that focuses on the European Union region to equitably distribute carbon budgets among the Member States and activate emissions offset strategies to manage unexpected deviations from the EU27 carbon budget. The emissions trading dynamic is based on the “Gains from Trade” approach. The tool also calculates the cost of the offset strategies based on the use of the abatement cost curves for the Member States. Using a case study of the recent increase in carbon emissions in Germany in response to reduced Russian gas supplies, different emissions scenarios for Germany’s quota redistribution among the Member States are explored. The study reveals varied cost implications of between 30–60 Eur/ton CO2eq to offset the emissions increase across other Member States. Final recommendations include promoting cross-border collaboration at the EU27 level.
An Innovative, Lightweight, and Sustainable Solution for the Integrated Seismic Energy Retrofit of Existing Masonry Structures
Authors: Longobardi Giovanna , Moşoarca Marius , Gruin Aurelian , Ion Alexandru , Formisano Antonio
Published: June 2024

A large percentage of existing building stock in Italy and throughout Europe is ageing and no longer complies with current regulations, particularly in terms of sustainability. For these reasons, an urgent consolidation plan is needed to ensure an increase in both seismic response and energy response. Indeed, these constructions were built before the actual technical codes, and currently, they are not able to withstand seismic actions. Meanwhile, they are subject to thermal dispersions that could be due to the use of materials with poor properties or construction errors. Among the numerous consolidation techniques, an innovative solution consisting of a coating system has appeared on the construction market in recent decades. It is an integrated solution that simultaneously improves the seismic and energy behaviour of the building. The paper proposes the evaluation of this lightweight and sustainable solution through some experimental tests which were performed at the National Institute for Research and Development in Constructions, located in the city of Timişoara (Romania). The tests were aimed to investigate the out-of-plane behaviour of a masonry wall (1.20 m × 2.40 m × 0.60 m) obtained by combining two smaller panels with mortar and subjecting them to constant vertical force and pushing by an increasing horizontal one. Its response was assessed before and after the application of extruded aluminium alloy base profiles belonging to the system under study.
Biodiversity Characteristics and Carbon Sequestration Potential of Successional Woody Plants versus Tree Plantation under Different Reclamation Treatments on Hard-Coal Mine Heaps––A Case Study from Upper Silesia
Authors: Woś Bartłomiej , Misebo Amisalu Milkias , Ochał Wojciech , Klamerus-Iwan Anna , Pająk Marek , Sierka Edyta , Kompała-Bąba Agnieszka , Bujok Michał , Bierza Wojciech , Józefowska Agnieszka , Bujak Julita , Pietrzykowski Marcin
Published: June 2024

In the discussion about sustainable forestry, a key role is played by the development of ecosystem services, including ecological, social, and economic ones, in which biodiversity and carbon (C) sequestration are among the most important. Afforestation of disturbed and post-mining sites is one of the ways to minimize the negative impact of civilization on the environment. Optimizing C sequestration strategies at post-mining sites plays a crucial role in promoting ecosystem recovery, supporting climate change mitigation, and enabling C offsetting. In this study, we compared the C storage in the soil and plant biomass of forest ecosystems developed on coal-mine heaps for different scenarios of reclamation and succession. We tested combinations of sites (i.e., non-reclaimed sites on bare carboniferous rock [BR] and sites reclaimed by applying topsoil [TS]) and successional woodland and tree plantation. The estimated potential for total C storage (in the soil + biomass) for TS sites ranged from 68.13 to 121.08 Mg ha−1, of which 52.20–102.89 Mg ha−1 was stored in the soil and 12.09–20.15 Mg ha−1 in the biomass. In the non-reclaimed sites on BR, the total C storage was much higher, amounting to 523.14 Mg ha−1 (507.66 Mg ha−1 being in the soil), which was due to the geogenic coal content in the BR. However, the C storage in the biomass (15.48 Mg ha−1) and litter (5.91 Mg ha−1) was similar to the amounts obtained from the reclaimed sites. The number of species did not differ statistically significantly between the analyzed variants. On average, 14 species were recorded in the plots. The average Shannon–Wiener index (H’) value was higher for sites with BR (1.99) than TS variants on reclaimed plots (1.71). The lowest H’ value was for those plots with Robinia pseudacacia in the stand. One of the main implications of the obtained results for sustainable forestry is the perspective of using succession in the recovery of a disturbed ecosystem. We noted that woodlands from succession on BR are highly biodiverse, have high C sequestration potential, and do not require time-consuming reclamation treatments.
A Novel Constraint-Aware Flexible Model with Ant Colony Optimization for Symmetrical Travel Recommendation
Authors: Alatiyyah Mohammed
Published: June 2024

This paper proposes a flexible travel recommender model (FTRM) that emphasizes the symmetry between user preferences and travel constraints, addressing key challenges in the field such as the integration of diverse constraint types and the customization of travel itineraries. The key contribution of the proposed model lies in its integration with the item constraints data model (ICDM), which effectively manages a plethora of constraint types. Additionally, this study develops a novel algorithm inspired by ant colony optimization (ACO) principles, demonstrating performance metrics that are comparable to state-of-the-art algorithms in this field. A comprehensive set of systematic experimental analyses is conducted, employing various models across diverse situational contexts, with the primary goal of illustrating the capabilities of the proposed symmetrical FTRM using real-world data from the Durham dataset. The obtained results highlight the model’s ability to accommodate diverse constraint types, facilitating the customization of travel itineraries to suit individual user preferences and achieve a balanced and symmetrical travel experience. Specifically, our model outperforms existing models in terms of flexibility and customization, showing significant improvements in user satisfaction and itinerary efficiency.
Chronic Lead Exposure in Adult Mice: Associations with miR-671/CDR1as Regulation, NF-κB Signaling, and Alzheimer’s Disease-like Pathology
Authors: Qiao Mengyun , Yang Haitao , Liu Li , Yu Tao , Wang Haihua , Chen Xiao , Zhang Yi , Duan Airu , Lyu Shujun , Wu Siyu , Xiao Jingwei , Li Bin
Published: June 2024

Long-term exposure to lead (Pb) can result in chronic damage to the body through accumulation in the central nervous system (CNS) leading to neurodegenerative diseases, such as Alzheimer’s disease (AD). This study delves into the intricate role of miR-671/CDR1as regulation in the etiology of AD-like lesions triggered by chronic Pb exposure in adult mice. To emulate the chronic effects of Pb, we established a rodent model spanning 10 months of controlled Pb administration, dividing 52 C57BL/6J mice into groups receiving varying concentrations of Pb (1, 2, or 4 g/L) alongside an unexposed control. Blood Pb levels were monitored using serum samples to ensure accurate dosing and to correlate with observed toxicological outcomes. Utilizing the Morris water maze, a robust behavioral assay for assessing cognitive functions, we documented a dose-dependent decline in learning and memory capabilities among the Pb-exposed mice. Histopathological examination of the hippocampal tissue revealed tell-tale signs of AD-like neurodegeneration, characterized by the accumulation of amyloid plaques and neurofibrillary tangles. At the molecular level, a significant upregulation of AD-associated genes, namely amyloid precursor protein (APP), β-secretase 1 (BACE1), and tau, was observed in the hippocampal tissue of Pb-exposed mice. This was accompanied by a corresponding surge in the protein levels of APP, BACE1, amyloid-β (Aβ), and phosphorylated tau (p-tau), further implicating Pb in the dysregulation of these key AD markers. The expression of CDR1as, a long non-coding RNA implicated in AD pathogenesis, was found to be suppressed in Pb-exposed mice. This observation suggests a potential mechanistic link between Pb-induced neurotoxicity and the dysregulation of the CDR1as/miR-671 axis, which warrants further investigation. Moreover, our study identified a dose-dependent alteration in the intracellular and extracellular levels of the transcription factor nuclear factor-kappa B (NF-κB). This finding implicates Pb in the modulation of NF-κB signaling, a pathway that plays a pivotal role in neuroinflammation and neurodegeneration. In conclusion, our findings underscored the deleterious effects of Pb exposure on the CNS, leading to the development of AD-like pathology. The observed modulation of NF-κB signaling and miR-671/CDR1as regulation provides a plausible mechanistic framework for understanding the neurotoxic effects of Pb and its potential contribution to AD pathogenesis.
Tetrodotoxin Derivatization with a Newly Designed Boron Reagent Leads to Conventional Reversed-Phase Liquid Chromatography
Authors: Kawasue Shimba , Kuniyoshi Kyoko , Uema Masashi , Oshiro Naomasa
Published: June 2024

Tetrodotoxin (TTX) is a representative natural toxin causing pufferfish food poisoning, which is especially prominent in East and Southeast Asia, including Japan. TTX has been analyzed through post-column derivatization high-performance liquid chromatography (HPLC), ion-pair LC-MS(/MS), and hydrophilic interaction liquid chromatography (HILIC)-MS(/MS) as alternatives to the mouse bioassay method. However, post-column derivatization requires a system for online derivatization reactions, and with the ion-pair LC-MS approach, it is difficult to remove residual ion-pair reagents remaining in the equipment. Moreover, HILIC-MS provides poor separation compared to reversed-phase (RP) HPLC and requires a long time to reach equilibration. Therefore, we decided to develop a TTX analytical method using pre-column derivatization and RP HPLC for the rapid assessment of outbreak samples, including food remnants. In this study, we focused on the vic-diol moiety of TTX and designed a new derivatization reagent coded as NBD-H-DAB. This NBD-H-DAB was synthesized from 4-hydrazino-7-nitro-2,1,3-benzoxadiazole (NBD-H) and 3-fluoro-2-formylphenylboronic acid (FFPBA) with a simple reaction system and rapidly converted to its boronate form, coded NBD-H-PBA, in an aqueous reaction solution. The NBD-H-PBA demonstrated appropriate hydrophobicity to be retained on the RP analytical column and successfully detected with a UV spectrometer. It was easily reacted with the vic-diol moiety of TTX (C6 and C11) to synthesized a boronic ester. The derivatized TTX could be detected using the RP HPLC-UV, and the limit of detection in the fish flesh samples was 0.06 mg/kg. This novel pre-column derivatization of TTX with NBD-H-PBA proves capable for the analysis of TTX.
Embracing the Versatility of Botulinum Neurotoxins in Conventional and New Therapeutic Applications
Authors: Rasetti-Escargueil Christine , Palea Stefano
Published: June 2024

Botulinum neurotoxins (BoNTs) have been used for almost half a century in the treatment of excessive muscle contractility. BoNTs are routinely used to treat movement disorders such as cervical dystonia, spastic conditions, blepharospasm, and hyperhidrosis, as well as for cosmetic purposes. In addition to the conventional indications, the use of BoNTs to reduce pain has gained increased recognition, giving rise to an increasing number of indications in disorders associated with chronic pain. Furthermore, BoNT-derived formulations are benefiting a much wider range of patients suffering from overactive bladder, erectile dysfunction, arthropathy, neuropathic pain, and cancer. BoNTs are categorised into seven toxinotypes, two of which are in clinical use, and each toxinotype is divided into multiple subtypes. With the development of bioinformatic tools, new BoNT-like toxins have been identified in non-Clostridial organisms. In addition to the expanding indications of existing formulations, the rich variety of toxinotypes or subtypes in the wild-type BoNTs associated with new BoNT-like toxins expand the BoNT superfamily, forming the basis on which to develop new BoNT-based therapeutics as well as research tools. An overview of the diversity of the BoNT family along with their conventional therapeutic uses is presented in this review followed by the engineering and formulation opportunities opening avenues in therapy.
Specific Amino Acid Residues in the Three Loops of Snake Cytotoxins Determine Their Membrane Activity and Provide a Rationale for a New Classification of These Toxins
Authors: Dubovskii Peter V. , Utkin Yuri N.
Published: June 2024

Cytotoxins (CTs) are three-finger membrane-active toxins present mainly in cobra venom. Our analysis of the available CT amino acid sequences, literature data on their membrane activity, and conformational equilibria in aqueous solution and detergent micelles allowed us to identify specific amino acid residues which interfere with CT incorporation into membranes. They include Pro9, Ser28, and Asn/Asp45 within the N-terminal, central, and C-terminal loops, respectively. There is a hierarchy in the effect of these residues on membrane activity: Pro9 > Ser28 > Asn/Asp45. Taking into account all the possible combinations of special residues, we propose to divide CTs into eight groups. Group 1 includes toxins containing all of the above residues. Their representatives demonstrated the lowest membrane activity. Group 8 combines CTs that lack these residues. For the toxins from this group, the greatest membrane activity was observed. We predict that when solely membrane activity determines the cytotoxic effects, the activity of CTs from a group with a higher number should exceed that of CTs from a group with a lower number. This classification is supported by the available data on the cytotoxicity and membranotropic properties of CTs. We hypothesize that the special amino acid residues within the loops of the CT molecule may indicate their involvement in the interaction with non-lipid targets.
Mutations in the Receptor Binding Domain of Severe Acute Respiratory Coronavirus-2 Omicron Variant Spike Protein Significantly Stabilizes Its Conformation
Authors: Peters Michael H.
Published: June 2024

The Omicron variant and its sub-lineages are the only current circulating SARS-CoV-2 viruses worldwide. In this study, the conformational stability of the isolated Receptor Binding Domain (RBD) of Omicron’s spike protein is examined in detail. The parent Omicron lineage has over ten mutations in the ACE2 binding region of the RBD that are specifically associated with its β hairpin loop domain. It is demonstrated through biophysical molecular computations that the mutations in the β hairpin loop domain significantly increase the intra-protein interaction energies of intra-loop and loop–RBD interactions. The interaction energy increases include the formation of new hydrogen bonds in the β hairpin loop domain that help stabilize this critical ACE2 binding region. Our results also agree with recent experiments on the stability of Omicron’s core β barrel domain, outside of its loop domain, and help demonstrate the overall conformational stability of the Omicron RBD. It is further shown here through dynamic simulations that the unbound state of the Omicron RBD remains closely aligned with the bound state configuration, which was not observed for the wild-type RBD. Overall, these studies demonstrate the significantly increased conformational stability of Omicron over its wild-type configuration and raise a number of questions on whether conformational stability could be a positive selection feature of SARS-CoV-2 viral mutational changes.
ISKNV Triggers AMPK/mTOR-Mediated Autophagy Signaling through Oxidative Stress, Inducing Antioxidant Enzyme Expression and Enhancing Viral Replication in GF-1 Cells
Authors: Hsueh Tsai-Ching , Chen Pin-Han , Hong Jiann-Ruey
Published: June 2024

Infectious spleen and kidney necrosis virus (ISKNV) infections can induce the process of host cellular autophagy but have rarely been identified within the molecular autophagy signaling pathway. In the present study, we demonstrated that ISKNV induces ROS-mediated oxidative stress signals for the induction of 5′AMP-activated protein kinase/mechanistic target of rapamycin kinase (AMPK/mTOR)-mediated autophagy and upregulation of host antioxidant enzymes in fish GF-1 cells. We also examined ISKNV-induced oxidative stress, finding that reactive oxidative species (ROS) increased by 1.5-fold and 2.5-fold from day 2 to day 3, respectively, as assessed by the H2DCFDA assay for tracing hydrogen peroxide (H2O2), which was blocked by NAC treatment in fish GF-1 cells. Furthermore, ISKNV infection was shown to trigger oxidative stress/Nrf2 signaling from day 1 to day 3; this event was then correlated with the upregulation of antioxidant enzymes such as Cu/ZnSOD and MnSOD and was blocked by the antioxidant NAC. Using an MDC assay, TEM analysis and autophagy marker LC3-II/I ratio, we found that ROS stress can regulate autophagosome formation within the induction of autophagy, which was inhibited by NAC treatment in GF-1 cells. Through signal analysis, we found that AMPK/mTOR flux was modulated through inhibition of mTOR and activation of AMPK, indicating phosphorylation levels of mTOR Ser 2448 and AMPK Thr 172 from day 1 to day 3; however, this process was reversed by NAC treatment, which also caused a reduction in virus titer (TCID50%) of up to 1000 times by day 3 in GF-1 cells. Thus, ISKNV-induced oxidative stress signaling is blocked by antioxidant NAC, which can also either suppress mTOR/AMPK autophagic signals or reduce viral replication. These findings may provide the basis for the creation of DNA control and treatment strategies.
Reference Material Production and Milk Protein Concentration as Elements to Improve Bluetongue Serological Diagnosis in Bulk Tank Milk
Authors: Romero-Trancón David , Valero-Lorenzo Marta , Agüero Montserrat , Villalba Rubén
Published: June 2024

The serological surveillance of bluetongue in bulk tank milk is an efficient and cost-effective method for the early detection of bluetongue virus incursions in unvaccinated free areas of the disease. In addition, the availability of standardized and reliable reagents and refined diagnostic procedures with high sensitivity and specificity are essential for surveillance purposes. However, no available reference materials for bluetongue virus serological surveillance in bulk tank milk exist. This study shows the production and characterization of reference material for the implementation of a commercially available bluetongue milk ELISA test in official laboratories, as well as the evaluation of a procedure to increase the sensitivity in samples with low levels of antibodies. This procedure, based on milk protein concentration, allowed us to notably increase the ELISA test’s analytical sensitivity, which is useful for milk samples from farms with low within-herd prevalence or pools of bulk tank milk samples. The standardized milk reference material produced here, together with the evaluated procedure to improve analytical sensitivity, could be applied as tools to ensure an accurate diagnosis by official laboratories in bluetongue unvaccinated free areas.
Recombinant Viruses from the Picornaviridae Family Occurring in Racing Pigeons
Authors: Łukaszuk Ewa , Dziewulska Daria , Stenzel Tomasz
Published: June 2024

Viruses from Picornaviridae family are known pathogens of poultry, although the information on their occurrence and pathogenicity in pigeons is scarce. In this research, efforts are made to broaden the knowledge on Megrivirus B and Pigeon picornavirus B prevalence, phylogenetic relationship with other avian picornaviruses and their possible connection with enteric disease in racing pigeons. As a result of Oxford Nanopore Sequencing, five Megrivirus and two pigeon picornavirus B-like genome sequences were recovered, among which three recombinant strains were detected. The recombinant fragments represented an average of 10.9% and 25.5% of the genome length of the Pigeon picornavirus B and Megrivirus B reference strains, respectively. The phylogenetic analysis revealed that pigeons are carriers of species-specific picornaviruses. TaqMan qPCR assays revealed 7.8% and 19.0% prevalence of Megrivirus B and 32.2% and 39.7% prevalence of Pigeon picornavirus B in the group of pigeons exhibiting signs of enteropathy and in the group of asymptomatic pigeons, respectively. In turn, digital droplet PCR showed a considerably higher number of genome copies of both viruses in sick than in asymptomatic pigeons. The results of quantitative analysis leave the role of picornaviruses in enteropathies of pigeons unclear.
Zero-Dose Childhood Vaccination Status in Rural Democratic Republic of Congo: Quantifying the Relative Impact of Geographic Accessibility and Attitudes toward Vaccination
Authors: Mbunga Branly Kilola , Liu Patrick Y. , Bangelesa Freddy , Mafuta Eric , Dalau Nkamba Mukadi , Egbende Landry , Hoff Nicole A. , Kasonga Jean Bosco , Lulebo Aimée , Manirakiza Deogratias , Mudipanu Adèle , Mvuama Nono , Ouma Paul , Wong Kerry , Lusamba Paul , Burstein Roy
Published: June 2024

Despite efforts to increase childhood vaccination coverage in the Democratic Republic of the Congo (DRC), approximately 20% of infants have not started their routine immunization schedule (zero-dose). The present study aims to evaluate the relative influence of geospatial access to health facilities and caregiver perceptions of vaccines on the vaccination status of children in rural DRC. Pooled data from two consecutive nationwide immunization surveys conducted in 2022 and 2023 were used. Geographic accessibility was assessed based on travel time from households to their nearest health facility using the AccessMod 5 model. Caregiver attitudes to vaccination were assessed using the survey question “How good do you think vaccines are for your child?” We used logistic regression to assess the relationship between geographic accessibility, caregiver attitudes toward vaccination, and their child’s vaccination status. Geographic accessibility to health facilities was high in rural DRC, with 88% of the population living within an hour’s walk to a health facility. Responding that vaccines are “Bad, Very Bad, or Don’t Know” relative to “Very Good” for children was associated with a many-fold increased odds of a zero-dose status (ORs 69.3 [95%CI: 63.4–75.8]) compared to the odds for those living 60+ min from a health facility, relative to <5 min (1.3 [95%CI: 1.1–1.4]). Similar proportions of the population fell into these two at-risk categories. We did not find evidence of an interaction between caregiver attitude toward vaccination and travel time to care. While geographic access to health facilities is crucial, caregiver demand appears to be a more important driver in improving vaccination rates in rural DRC.
Therapeutic Effects of Propionibacterium acnes and Lipopolysaccharide from Escherichia coli in Cats with Feline Panleukopenia
Authors: Chanachaivirada Rattanakhon , Chuammitri Phongsakorn , Lampa Kannika Na , Prachasilchai Worapat , Sodarat Chollada
Published: June 2024

The objective of this study was to investigate the therapeutic effects of inactivated Propionibacterium acnes and lipopolysaccharide derived from Escherichia coli cells in cats affected by feline panleukopenia virus (FPV). A retrospective study of 80 FPV-positive cats was divided into two groups: a treatment group receiving inactivated Propionibacterium acnes and lipopolysaccharide derived from Escherichia coli cells along with supportive treatment and a no-treatment group receiving only supportive treatment. There was no significant difference in the total white blood cell counts between the two groups. However, the total white blood cell counts of both groups were low on day 0 and increased significantly on days 3 and 6 of treatment. Additionally, the white blood cell counts in the treatment group significantly increased during days 3 to 6 compared with those of the no-treatment group (p < 0.01). The mortality rate was not significantly different between the two groups. In a prospective study, the serum and fecal immunoglobulin A (IgA) levels were measured in both groups. There were no significant differences in IgA levels between the two groups in either the serum or feces.
Canine Leishmaniosis Associated with Acute Pleural Effusion and Sudden Death in a Dog
Authors: Brito Maria Caroline Pereira , Sousa Maria de Fátima , Sampaio Rubia Avlade Guedes , Linhares Markyson Tavares , Riquelme Lourdes Fernandez , Lacerda Wellida Karinne , Lucena Ricardo Barbosa
Published: June 2024

A two-year-old female crossbreed dog, previously a stray with no known owner, was adopted and subsequently spayed. The dog exhibited weight loss over a period of two months and died suddenly during a leashed walk. Upon necropsy, enlargement of the submandibular, prescapular, and popliteal lymph nodes was noted. The intrathoracic cavity contained a substantial volume of yellowish-white fluid. Lymph nodes in the mediastinal and ventral thoracic centers were also enlarged, hemorrhagic, and friable. Microscopic examination revealed significant architectural changes in the lymph nodes, characterized by a pronounced cellular infiltrate consisting of lymphocytes and histiocytes, along with macrophages containing intracytoplasmic Leishmania amastigotes. Immunohistochemical analysis of the lymph nodes confirmed positive staining for Leishmania amastigotes. This case represents the first report of canine leishmaniasis associated with acute pleural effusion and sudden death.
Subjective Affective Responses to Natural Scenes Require Understanding, Not Spatial Frequency Bands
Authors: Mastria Serena , Codispoti Maurizio , Tronelli Virginia , De Cesarei Andrea
Published: June 2024

It is debated whether emotional processing and response depend on semantic identification or are preferentially tied to specific information in natural scenes, such as global features or local details. The present study aimed to further examine the relationship between scene understanding and affective response while manipulating visual content. To this end, we presented affective and neutral natural scenes which were progressively band-filtered to contain global features (low spatial frequencies) or local details (high spatial frequencies) and assessed both affective response and scene understanding. We observed that, if scene content was correctly reported, subjective ratings of arousal and valence were modulated by the affective content of the scene, and this modulation was similar across spatial frequency bands. On the other hand, no affective modulation of subjective ratings was observed if picture content was not correctly reported. The present results indicate that subjective affective response requires content understanding, and it is not tied to a specific spatial frequency range.