bluuebunny/crossref_metadata_2025 huggingface dataset:
Linear-in-dB Logarithmic Signal Strength Sensor Circuit for Wireless Power Transfer Receivers
Authors: Quadir Nasir , Alawar Fatma S. , Albasha Lutfi , Mir Hasan
Published: November 2023

Tracking systems for wireless power transfer are becoming a necessity. The received target signals are sometimes weak, which is why it is critical to have a dedicated received–signal–strength indicator (RSSI) for signal detection. It can also be used for transmitter localization and automatic gain control (AGC) to ensure continuous coverage. In this paper, a logarithmic detector coupled with a high-speed full-wave rectifier is designed for an RSSI system. The aim is to place the RSSI on the receiver side of unmanned aerial vehicles used, for example, as agricultural drones. This system is intended to operate in a scenario where multiple drones in a smart farm are charged wirelessly whilst airborne. An RSSI is placed in the receiver to detect and track wireless signals and to ensure the drone is charged while in motion. The RSSI system operates at 5.8 GHz, reported for the first time, and is capable of detecting signal strengths from −60 dBm to 0 dBm with a sensitivity level of 17 mV/dBm. A logarithmic error of 0.4 dB with a dynamic range of 34 dB was achieved. The proposed RSSI system was designed using 65 nm TSMC CMOS technology, and it exhibits high sensitivity, better efficiency, and lower power consumption than those in other reported works.
Repurposing End-of-Life Coal Mines with Business Models Based on Renewable Energy and Circular Economy Technologies
Authors: Krzemień Alicja , Frejowski Aleksander , Fidalgo Valverde Gregorio , Riesgo Fernández Pedro , Garcia-Cortes Silverio
Published: November 2023

This paper presents a methodology to select the most exciting business models based on renewable energy and circular economy technologies within end-of-life coal mines to help develop a renewable-based energy sector, promote sustainable local economic growth, and maximise the number of green and quality jobs. To achieve this goal, first, a structural analysis was developed to select the technical variables that better identify this complex system. Second, a morphological analysis allowed the construction of the scenario space. Third, a multicriteria assessment was developed to achieve this goal, based on the previously assessed relevant scenarios, considering the European Green Deal policies, technical variables that characterise end-of-life coal mine environments, technology readiness level, the European taxonomy, synergistic potentials, contributions to the circular economy, and sector coupling. Finally, result indicators were selected to analyse the alternative options derived from the justification approach, considering the targets set by the European Green Deal and related taxonomy and the regional policy indicators for the Just Transition Fund. The results show that eco-industrial parks with virtual power plants represent the most appropriate business model choice, according to the scoring given to the different aspects. They may be complemented by a hydrogen production plant, provided that specific economic subventions are obtained to achieve balanced financial results.
Series Architecture for the Reduction of the DC-DC Converter in a Hybrid Energy Storage System for Electric Vehicles
Authors: Fonseca de Freitas Caio Augusto , Bartholomeus Patrick , Margueron Xavier , Le Moigne Philippe
Published: November 2023

A Hybrid Energy Storage System (HESS) uses DC-DC converters to couple its energy sources. However, this device represents a “dead weight” in the system and must be reduced to a minimum in order to maximize the HESS’ performance. This work proposes a new coupling architecture to reduce the converter’s volume and mass. Not yet addressed in the literature, this architecture is based on a series coupling of the sources. In this case, a DC-DC converter is used to control the current difference between the two sources. If this difference is zero, so is the power processed by the converter. By reducing the power processed by the converter, its mass and volume can be reduced. Simulation and experimental tests were carried out to validate the architecture concept. For the latter, power supplies were used to emulate the batteries and the load, and a 2 kg, 3.3 kW evaluation board served as the DC-DC converter. The results show that, compared to a conventional solution that is usually adopted in the literature, with the series architecture, it is possible to reduce the converter sizing power by almost 3.7.
Correction: Siddiqui, Z.A.; Park, U. A Drone Based Transmission Line Components Inspection System with Deep Learning Technique. Energies 2020, 13, 3348
Authors: Siddiqui Zahid Ali , Park Unsang
Published: November 2023

The authors wish to make the following corrections to their paper [...]
Reference Curves for Central Appalachian Red Spruce
Authors: Gray Joseph M. , Schuler Jamie L. , Thomas-Van Gundy Melissa , Chhin Sophan
Published: November 2023

Red spruce (Picea rubens Sarg.) was a prized timber species in West Virginia during the era of resource exploitation in the late 1800s and early 1900s. As a result, central Appalachian red spruce comprise a much smaller component of high-elevation stand composition and a greatly constricted presence across the region. Widespread restoration efforts are underway to re-establish red spruce across this landscape. However, without benchmarks to gauge growth rates and stand developmental patterns, it is unclear whether these efforts are successful. Our goal was to develop reference curves predicting centile height growth for understory red spruce (≤7.6 m) across the region. We reconstructed the height growth patterns of over 250 randomly selected red spruce seedlings and saplings from 22 high-elevation stands in West Virginia. We also harvested 24 mature red spruce from the same stands to develop juvenile growth curves up to 7.6 m to compare understory growth rates of historical to contemporary rates from the reference curves. Our constructed reference curves showed height growth tended to peak between 10 and 30 years of age. Total heights ranged from 0.95 m to 6.85 m after 50 years. We identified two demographic populations in the mature red spruce trees. All the mature red spruce trees that established after 1890 exceeded the 97% growth centile by age 80. By contrast, only two trees from the pre-1890 population reached the same level by age 80. This work highlights the varied ascension pathways to the overstory for red spruce.
Association between Variation in Body Fat Mass Magnitude and Intake of Nutrients, including Carbohydrates, Fat, and B Vitamins, in a Cohort of Highly Trained Female Athletes
Authors: Baranauskas Marius , Kupčiūnaitė Ingrida , Lieponienė Jurgita , Stukas Rimantas
Published: November 2023

The most common sports nutrition strategies were constructed not only for maximizing musculoskeletal adaptations to exercise, but also to minimize health risks in athletes. Given the lack of research highlighting the potential effects of the intake of carbohydrates, fats, and B vitamins on body fat percentage in a population of female athletes, this study aimed to elucidate whether the intake of macronutrients and B vitamins could be associated with the variation in body fat percentage in a cohort of professional female athletes. This cross-sectional study was weighted to represent Lithuanian elite female athletes (n = 89). The dietary assessment of the female athletes was carried out using a 3-day dietary recall method. Their body composition was assessed using the bioelectrical impedance analysis method. For females, the reported average intakes of energy, carbohydrates, protein, and fat were 2475 kcal/day, 5.1 g/kg/day, 1.5 g/kg/day, and 36.7%, respectively. Excess B vitamin intake was revealed, ranging within plus 1–2 standard deviations (SDs) around the mean requirements. As a consequence, excessive body fat percentage was potentially factored as a negative outcome in maintaining optimal body composition in female athletes. Multivariate logistic regression analysis of a sample of female athletes revealed that, despite a slightly positive energy balance (∆ 95 kcal/day), the carbohydrate-deficient diet (adjusted odds ratio (aOR) 0.3, 95% confidence interval (CI) 0.1; 0.7), along with higher intakes of vitamin B1 (aOR 2.9, 95% CI 2.6; 7.8), vitamin B2 (aOR 6.7, 95% CI 1.1; 8.3), and vitamin B3 (aOR 1.8, 95% CI 1.4; 7.8) from food, was associated with a lower percentage of body fat. Therefore, more attention should be given to the intake of B vitamins in professional athletes with a range of body fat mass percentages for the purpose of achieving long-term goals of maintaining body composition and fitness.
Sustainable Recovery of Anthocyanins and Other Polyphenols from Red Cabbage Byproducts
Authors: Stoica Rusăndica , Ganciarov Mihaela , Constantinescu-Aruxandei Diana , Capră Luiza , Șuică-Bunghez Ioana-Raluca , Senin Raluca-Mădălina , Pricope Georgiana , Ivan Georgeta-Ramona , Călin Costin , Oancea Florin
Published: November 2023

The objective of this work was to develop a sustainable process for the extraction of anthocyanins from red cabbage byproducts using, for the first time, apple vinegar in extractant composition. Our results showed that the mixture 50% (v/v) ethanol–water, acidified with apple vinegar, used in the proportion of 25 g of red cabbage by-products per 100 mL of solvent, was the best solvent for the preparation of an anthocyanin extract with good stability for food applications. The chemical characterization of this extract was performed by FTIR, UV-VIS, HPLC-DAD, and ICP-OES. The stability was evaluated by determining the dynamics of the total polyphenol content (TPC) and the total monomeric anthocyanin pigment content (TAC) during storage. On the basis of the statistical method for analysis of variance (ANOVA), the standard deviation between subsamples and the repeatability standard deviation were determined. The detection limit of the stability test of TPC was 3.68 mg GAE/100 g DW and that of TAC was 0.79 mg Cyd-3-Glu/100 g DW. The red cabbage extract has high TPC and TAC, good stability, and significant application potential. The extracted residues, depleted of anthocyanins and polyphenols with potential allelopathic risks, fulfill the requirements for a fertilizing product and could be used for soil treatment.
Linear Polyethyleneimine-Based and Metal Organic Frameworks (DUT-67) Composite Hydrogels as Efficient Sorbents for the Removal of Methyl Orange, Copper Ions, and Penicillin V
Authors: Araque Luis M. , Fernández de Luis Roberto , Fidalgo-Marijuan Arkaitz , Infantes-Molina Antonia , Rodríguez-Castellón Enrique , Pérez Claudio J. , Copello Guillermo J. , Lázaro-Martínez Juan M.
Published: November 2023

This research explores the integration of DUT-67 metal organic frameworks into polyethyleneimine-based hydrogels to assemble a composite system with enough mechanical strength, pore structure and chemical affinity to work as a sorbent for water remediation. By varying the solvent-to-modulator ratio in a water-based synthesis path, the particle size of DUT-67 was successfully modulated from 1 μm to 200 nm. Once DUT-67 particles were integrated into the polymeric hydrogel, the composite hydrogel exhibited enhanced mechanical properties after the incorporation of the MOF filler. XPS, NMR, TGA, FTIR, and FT Raman studies confirmed the presence and interaction of the DUT-67 particles with the polymeric chains within the hydrogel network. Adsorption studies of methyl orange, copper(II) ions, and penicillin V on the composite hydrogel revealed a rapid adsorption kinetics and monolayer adsorption according to the Langmuir’s model. The composite hydrogel demonstrated higher adsorption capacities, as compared to the pristine hydrogel, showcasing a synergistic effect, with maximum adsorption capacities of 473 ± 21 mg L−1, 86 ± 6 mg L−1, and 127 ± 4 mg L−1, for methyl orange, copper(II) ions, and penicillin V, respectively. This study highlights the potential of MOF-based composite hydrogels as efficient adsorbents for environmental pollutants and pharmaceuticals.
Lithuanian Feminine Surname Debates from a Central European Perspective
Authors: Walkowiak Justyna B.
Published: November 2023

Contemporary Lithuania remains the only European country in which official feminine surnames indicate their bearers’ marital status, and this has been the object of fierce public debates over the past decade. Czechia and Slovakia grapple with surprisingly similar issues, even though Czech and Slovak feminine surnames do not reveal marital status. Similar debates in Poland took place a century earlier, a fact which may indicate the possible direction of the changes in the three countries studied. The aim of this article is to present debates concerning feminine surnames in Lithuania from a wider perspective, regarding contemporary Czechia and Slovakia, as well as Poland in the interwar period, and to show from a wider Central and Eastern European perspective that, despite the obvious differences in naming patterns, Lithuanian discussions are not exceptional, and they are part of a larger tendency towards more freedom in the choice of official surname forms for women. It is evident that, although female surnames are inexorably embedded in the language systems of the countries in which they function, their future largely depends on extralinguistic factors such as societal attitudes. While feminine surnames in European states generally seem to be on the decline, the most controversial remain those types that reveal marital status or imply male possession of women, though pragmatic factors might play some role as well, particularly in the case of minorities.
NGS Custom Panel Implementation in Patients with Non-Syndromic Autism Spectrum Disorders in the Clinical Routine of a Tertiary Hospital
Authors: Sandoval-Talamantes Ana Karen , Tenorio-Castaño Jair Antonio , Santos-Simarro Fernando , Adán Carmen , Fernández-Elvira María , García-Fernández Laura , Muñoz Yolanda , Lapunzina Pablo , Nevado Julián
Published: November 2023

Autism spectrum disorder (ASD) is a set of neurodevelopmental disorders characterized by deficiencies in communication, social interaction, and repetitive and restrictive behaviors. The discovery of genetic involvement in the etiology of ASD has made this condition a strong candidate for genome-based diagnostic tests. Next-generation sequencing (NGS) is useful for the detection of variants in the sequence of different genes in ASD patients. Herein, we present the implementation of a personalized NGS panel for autism (AutismSeq) for patients with essential ASD over a prospective period of four years in the clinical routine of a tertiary hospital. The cohort is composed of 48 individuals, older than 3 years, who met the DSM-5 (The Diagnostic and Statistical Manual of Mental Disorders) diagnostic criteria for ASD. The NGS customized panel (AutismSeq) turned out to be a tool with good diagnostic efficacy in routine clinical care, where we detected 12 “pathogenic” (including pathogenic, likely pathogenic, and VUS (variant of uncertain significance) possibly pathogenic variations) in 11 individuals, and 11 VUS in 10 individuals, which had previously been negative for chromosomal microarray analysis and other previous genetic studies, such as karyotype, fragile-X, or MLPA/FISH (Multiplex Ligation dependent Probe Amplification/Fluorescence in situ hybridization) analysis. Our results demonstrate the high genetic and clinical heterogeneity of individuals with ASD and the current difficulty of molecular diagnosis. Our study also shows that an NGS-customized panel might be useful for diagnosing patients with essential/primary autism and that it is cost-effective for most genetic laboratories.
Global Frequency Analyses of Canine Progressive Rod-Cone Degeneration–Progressive Retinal Atrophy and Collie Eye Anomaly Using Commercial Genetic Testing Data
Authors: Clark Jessica , Anderson Heidi , Donner Jonas , Pearce-Kelling Susan , Ekenstedt Kari
Published: November 2023

Hundreds of genetic variants associated with canine traits and disorders have been identified, with commercial tests offered. However, the geographic distributions and changes in allele and genotype frequencies over prolonged, continuous periods of time are lacking. This study utilized a large set of genotypes from dogs tested for the progressive rod-cone degeneration–progressive retinal atrophy (prcd-PRA) G>A missense PRCD variant (n = 86,667) and the collie eye anomaly (CEA)-associated NHEJ1 deletion (n = 33,834) provided by the commercial genetic testing company (Optigen/Wisdom Panel, Mars Petcare Science & Diagnostics). These data were analyzed using the chi-square goodness-of-fit test, time-trend graphical analysis, and regression modeling in order to evaluate how test results changed over time. The results span fifteen years, representing 82 countries and 67 breeds/breed mixes. Both diseases exhibited significant differences in genotype frequencies (p = 2.7 × 10−152 for prcd-PRA and 0.023 for CEA) with opposing graphical trends. Regression modeling showed time progression to significantly affect the odds of a dog being homozygous or heterozygous for either disease, as do variables including breed and breed popularity. This study shows that genetic testing informed breeding decisions to produce fewer affected dogs. However, the presence of dogs homozygous for the disease variant, especially for prcd-PRA, was still observed fourteen years after test availability, potentially due to crosses of unknown carriers. This suggests that genetic testing of dog populations should continue.
The Landscape of Monogenic Parkinson’s Disease in Populations of Non-European Ancestry: A Narrative Review
Authors: Koros Christos , Bougea Anastasia , Simitsi Athina Maria , Papagiannakis Nikolaos , Angelopoulou Efthalia , Pachi Ioanna , Antonelou Roubina , Bozi Maria , Stamelou Maria , Stefanis Leonidas
Published: November 2023

Introduction: There has been a bias in the existing literature on Parkinson’s disease (PD) genetics as most studies involved patients of European ancestry, mostly in Europe and North America. Our target was to review published research data on the genetic profile of PD patients of non-European or mixed ancestry. Methods: We reviewed articles published during the 2000–2023 period, focusing on the genetic status of PD patients of non-European origin (Indian, East and Central Asian, Latin American, sub-Saharan African and Pacific islands). Results: There were substantial differences regarding monogenic PD forms between patients of European and non-European ancestry. The G2019S Leucine Rich Repeat Kinase 2 (LRRK2) mutation was rather scarce in non-European populations. In contrast, East Asian patients carried different mutations like p.I2020T, which is common in Japan. Parkin (PRKN) variants had a global distribution, being common in early-onset PD in Indians, in East Asians, and in early-onset Mexicans. Furthermore, they were occasionally present in Black African PD patients. PTEN-induced kinase 1 (PINK1) and PD protein 7 (DJ-1) variants were described in Indian, East Asian and Pacific Islands populations. Glucocerebrosidase gene variants (GBA1), which represent an important predisposing factor for PD, were found in East and Southeast Asian and Indian populations. Different GBA1 variants have been reported in Black African populations and Latin Americans. Conclusions: Existing data reveal a pronounced heterogeneity in the genetic background of PD. A number of common variants in populations of European ancestry appeared to be absent or scarce in patients of diverse ethnic backgrounds. Large-scale studies that include genetic screening in African, Asian or Latin American populations are underway. The outcomes of such efforts will facilitate further clinical studies and will possibly contribute to the identification of either new pathogenic mutations in already described genes or novel PD-related genes.
Calcareous Nannofossils Biostratigraphy of Late Cretaceous–Paleocene Successions from Northern Jordan and Their Implications for Basin Analysis
Authors: Hussein Albesher , Al-Tarawneh Osama M. , Alqudah Mohammad
Published: November 2023

Local geological and tectonic processes have been pivotal in shaping the diverse sedimentation patterns observed in Jordan, forming sub-basins characterized by elevated organic matter content (TOC). This study aims to characterize the Maastrichtian basin, focusing on sedimentation rates using calcareous nannofossils and understanding paleoecological and paleo-oceanic conditions. It offers insights into the paleoenvironmental factors impacting oil shale deposition in the late Maastrichtian–Paleocene period. It employs classical biostratigraphical, semi-quantitative, and statistical methodologies to achieve its objectives of age determination and paleoecological insights. A total of 116 smear slides from two sites were obtained: the first, consisting of WA-1 (23 samples), WA-2 (18 samples), and WA-3 (11 samples), and the second, with 60 samples. Notably, the sites exhibit varying topography. WA-1 and WA-2, situated at lower elevations, have the highest Total Organic Carbon (TOC) levels, while areas with higher elevations in section four are visually identified by a light color. The study revealed varying patterns of calcareous nannofossil richness in the two investigated sites. These patterns were instrumental in defining biozones, with the utilization of marker species such as Lithraphidites quadratus, Micula murus, Micula prinsii, and Cruciplacolithus tenuis. Chronologically, these sections were classified as Maastrichtian–Paleogene, encompassing the following biozones in sequential order: UC-20a, UC-20b, UC-20c, UC-20d, and NP-2. Furthermore, the study identified two hiatus intervals, observed in sections WA-1 and KAS-1. The absence of certain biozones in the analyzed sections suggests that these sections correspond to distinct geological blocks within the basin, underscoring the role of tectonic forces during the deposition period. The sedimentation rate initially commenced at low levels but gradually increased due to topographic alterations. Notably, the biozone UC-20c demonstrated a clear trend toward warming and enhanced nutrient availability. In this context, the abundance and diversity of species were associated with increased continental influx into the sub-basin, resulting in rising nutrient levels and the number of calcareous nannofossils. This study enhances the understanding of the local and global effects such as tectonic and climates of the continuity of basins by deciphering calcareous nannofossil patterns and their correlation with sedimentation factors.
Establishment of an Efficient In Vitro Propagation Protocol for Cannabis sativa L. subsp. ruderalis Janish
Authors: Basile Giuseppe N. , Tedone Luigi , Pulvento Cataldo , De Mastro Giuseppe , Ruta Claudia
Published: November 2023

Cannabis sativa L., subsp. ruderalis Janish., ‘Finola’ is a dioecious cultivar of Finnish origin. This cultivar is very interesting because its cultivation cycle lasts less than 3 months. The aim of this study was to define an efficient micropropagation protocol to ensure in vitro multiplication and rooting and in vivo acclimatization. Two different explant sources were tested: seed-derived in vitro explants and nodal segments containing axillary buds from selected mother plants. Shoot proliferation was tested on different growth media enriched with cytokinin alone or cytokinin in combination with auxins. Among all combinations, the best results were obtained by combining the Basal Medium (BM—a Murashige and Skoog modified medium) with sucrose (20 g L−1), thidiazuron (TDZ 0.4 mg L−1), and 1-naphthalenacetic acid (NAA 0.2 mg L−1). Regarding rooting induction, the plants developed an extensive root system under red/blue lights on BM enriched with sucrose (30 g L−1) and indol-3 butyric acid (0.1 mg L−1), which allowed the survival of more than 90 percent of the plantlets once transplanted into the climate-controlled greenhouse.
Identifying Research Priorities to Promote the Well-Being of Family Caregivers of Canadians with Intellectual and/or Developmental Disabilities: A Pilot Delphi Study
Authors: Fakolade Afolasade , Stone Caitlin , Bobbette Nicole
Published: November 2023

Current programming and resources aimed at supporting the well-being of family caregivers often fail to address considerations unique to those caring for people with intellectual and/or developmental disabilities (IDDs). As a result, many caregivers of people with IDD feel isolated, stressed, and burnt out. A targeted research agenda informed by key stakeholders is needed and would allow research teams to coordinate resources, talents, and efforts to progress family caregiver well-being research in this area quickly and effectively. To address this aim, this pilot study used a Delphi design based on 2 rounds of questionnaires. In round 1, 19 stakeholders (18 females, 1 male), including 12 family caregivers, 3 rehabilitation providers, 2 researchers, and 2 organizational representatives, identified broad areas for caregiver well-being research. After collating the responses from round 1, stakeholders were asked to rank whether each area was considered a research priority in round 2. Data were analyzed using descriptive statistics and conventional content analysis. Eighteen stakeholders completed the round 2 survey (1 caregiver did not complete the round 2 survey), after which a consensus was reached. Stakeholders identified nine broad priorities, including system-level programs and services, models of care, health promotion, social inclusion, equity and diversity, capacity building, care planning along the lifespan, and balancing formal and natural community-based supports. Although preliminary in nature, the research priorities generated using an inclusive and systematic process may inform future efforts to promote the well-being of caregivers of Canadians with IDD.
An Analysis of the Impact of Religious Affiliation and Strength of Religiosity on Sexual Health Practices of Sexually Active Female College Students
Authors: Glazer Emily , Valdez Emma , DeBlauw Justin A. , Ives Stephen J.
Published: November 2023

Despite great strides in the development of contraceptive technologies, the United States has one of the highest teen pregnancy rates in the world. Religion and associated values may shape the sexual health behaviors of college students, as prior studies have aimed to determine how social factors may influence the use of contraception amongst college students. Thus, we sought to examine the differences in current contraceptive methods and the age of first contraceptive usage among sexually active female college students with different religious affiliations and strengths of religiosity. It was hypothesized that there would be no difference in current contraceptive methods among different religious affiliations and strengths of religions and that there would be a difference in the age of first contraceptive usage among different religious affiliations and strengths of religiosity. Two hundred and twenty-four college-aged females completed a 20-question survey about sexual health and religious practices. Chi-squared tests were implemented to determine the frequencies of responses across religious affiliations and strengths of religiosity. Significant differences in the frequency of responses for the age of first contraceptive usage were observed across different strengths of religiosity (p = 0.016) and for the self-perceived impact of religion on sexual health across different religious affiliations (p = 0.033) and strengths of religiosity (p = 0.005). All other differences were found not to be statistically significant. It was determined that increased strengths of religiosity resulted in delayed onset of contraceptive usage and that both different religious affiliations and greater strengths of religiosity lead to different self-perceived impacts of religion on sexual health despite low levels of current practice.
Inconspicuous Yet Indispensable: The Coronavirus Spike Transmembrane Domain
Authors: Aliper Elena T. , Efremov Roman G.
Published: November 2023

Membrane-spanning portions of proteins’ polypeptide chains are commonly known as their transmembrane domains (TMDs). The structural organisation and dynamic behaviour of TMDs from proteins of various families, be that receptors, ion channels, enzymes etc., have been under scrutiny on the part of the scientific community for the last few decades. The reason for such attention is that, apart from their obvious role as an “anchor” in ensuring the correct orientation of the protein’s extra-membrane domains (in most cases functionally important), TMDs often actively and directly contribute to the operation of “the protein machine”. They are capable of transmitting signals across the membrane, interacting with adjacent TMDs and membrane-proximal domains, as well as with various ligands, etc. Structural data on TMD arrangement are still fragmentary at best due to their complex molecular organisation as, most commonly, dynamic oligomers, as well as due to the challenges related to experimental studies thereof. Inter alia, this is especially true for viral fusion proteins, which have been the focus of numerous studies for quite some time, but have provoked unprecedented interest in view of the SARS-CoV-2 pandemic. However, despite numerous structure-centred studies of the spike (S) protein effectuating target cell entry in coronaviruses, structural data on the TMD as part of the entire spike protein are still incomplete, whereas this segment is known to be crucial to the spike’s fusogenic activity. Therefore, in attempting to bring together currently available data on the structure and dynamics of spike proteins’ TMDs, the present review aims to tackle a highly pertinent task and contribute to a better understanding of the molecular mechanisms underlying virus-mediated fusion, also offering a rationale for the design of novel efficacious methods for the treatment of infectious diseases caused by SARS-CoV-2 and related viruses.
Knockdown of Simulated-Solar-Radiation-Sensitive miR-205-5p Does Not Induce Progression of Cutaneous Squamous Cell Carcinoma In Vitro
Authors: Bender Marc , Chen I-Peng , Henning Stefan , Degenhardt Sarah , Mhamdi-Ghodbani Mouna , Starzonek Christin , Volkmer Beate , Greinert Rüdiger
Published: November 2023

Solar radiation is the main risk factor for cSCC development, yet it is unclear whether the progression of cSCC is promoted by solar radiation in the same way as initial tumorigenesis. Additionally, the role of miRNAs, which exert crucial functions in various tumors, needs to be further elucidated in the context of cSCC progression and connection to solar radiation. Thus, we chronically irradiated five cSCC cell lines (Met-1, Met-4, SCC-12, SCC-13, SCL-II) with a custom-built irradiation device mimicking the solar spectrum (UVB, UVA, visible light (VIS), and near-infrared (IRA)). Subsequently, miRNA expression of 51 cancer-associated miRNAs was scrutinized using a flow cytometric multiplex quantification assay (FirePlex®, Abcam). In total, nine miRNAs were differentially expressed in cell-type-specific as well as universal manners. miR-205-5p was the only miRNA downregulated after SSR-irradiation in agreement with previously gathered data in tissue samples. However, inhibition of miR-205-5p with an antagomir did not affect cell cycle, cell growth, apoptosis, or migration in vitro despite transient upregulation of oncogenic target genes after miR-205-5p knockdown. These results render miR-205-5p an unlikely intracellular effector in cSCC progression. Thus, effects on intercellular communication in cSCC or the simultaneous examination of complementary miRNA sets should be investigated.
Gas Chromatography–Mass Spectrometry Detection of Thymoquinone in Oil and Serum for Clinical Pharmacokinetic Studies
Authors: Tekbaş A. , Bremer-Streck S. , Wissenbach D. K. , Peters F. T. , von Lilienfeld-Toal M. , Soonawalla Z. , Rauchfuß F. , Settmacher U. , Dahmen U.
Published: November 2023

Thymoquinone (TQ) is the primary component of Nigella sativa L. (NS) oil, which is renowned for its potent hepatoprotective effects attributed to its antioxidant, anti-fibrotic, anti-inflammatory, anti-carcinogenic, and both anti- and pro-apoptotic properties. The aim of this work was to establish a method of measuring TQ in serum in order to investigate the pharmacokinetics of TQ prior to a targeted therapeutic application. In the first step, a gas chromatography–mass spectrometry method for the detection and quantification of TQ in an oily matrix was established and validated according to European Medicines Agency (EMA) criteria. For the assessment of the clinical application, TQ concentrations in 19 oil preparations were determined. Second, two serum samples were spiked with TQ to determine the TQ concentration after deproteinization using toluene. Third, one healthy volunteer ingested 1 g and another one 3 g of a highly concentrated NS oil 30 and 60 min prior to blood sampling for the determination of serum TQ level. After the successful establishment and validation of the measurement method, the highest concentration of TQ (36.56 g/L) was found for a bottled NS oil product (No. 1). Since a capsule is more suitable for oral administration, the product with the third highest TQ concentration (No. 3: 24.39 g/L) was used for all further tests. In the serum samples spiked with TQ, the TQ concentration was reliably detectable in a range between 5 and 10 µg/mL. After oral intake of NS oil (No. 3), however, TQ and/or its derivatives were not detectable in human serum. This discrepancy in detecting TQ after spiking serum or following oral ingestion may be attributed to the instability of TQ in biomatrices as well as its strong protein binding properties. A pharmacokinetics study was therefore not viable. Studies on isotopically labeled TQ in an animal model are necessary to study the pharmacokinetics of TQ using alternative modalities.
Regulatory T Cells in the Pathogenesis of Graves’ Disease
Authors: Kustrimovic Natasa , Gallo Daniela , Piantanida Eliana , Bartalena Luigi , Lai Adriana , Zerbinati Nicola , Tanda Maria Laura , Mortara Lorenzo
Published: November 2023

Maintaining a delicate balance between the prompt immune response to pathogens and tolerance towards self-antigens and commensals is crucial for health. T regulatory (Treg) cells are pivotal in preserving self-tolerance, serving as negative regulators of inflammation through the secretion of anti-inflammatory cytokines, interleukin-2 neutralization, and direct suppression of effector T cells. Graves’ disease (GD) is a thyroid-specific autoimmune disorder primarily attributed to the breakdown of tolerance to the thyroid-stimulating hormone receptor. Given the limitations of currently available GD treatments, identifying potential pathogenetic factors for pharmacological targeting is of paramount importance. Both functional impairment and frequency reduction of Tregs seem likely in GD pathogenesis. Genome-wide association studies in GD have identified polymorphisms of genes involved in Tregs’ functions, such as CD25 (interleukin 2 receptor), and Forkhead box protein P3 (FOXP3). Clinical studies have reported both functional impairment and a reduction in Treg frequency or suppressive actions in GD, although their precise involvement remains a subject of debate. This review begins with an overview of Treg phenotype and functions, subsequently delves into the pathophysiology of GD and into the existing literature concerning the role of Tregs and the balance between Tregs and T helper 17 cells in GD, and finally explores the ongoing studies on target therapies for GD.
Utilizing Extraepitopic Amino Acid Substitutions to Define Changes in the Accessibility of Conformational Epitopes of the Bacillus cereus HlyII C-Terminal Domain
Authors: Rudenko Natalia V. , Nagel Alexey S. , Melnik Bogdan S. , Karatovskaya Anna P. , Vetrova Olesya S. , Zamyatina Anna V. , Andreeva-Kovalevskaya Zhanna I. , Siunov Alexander V. , Shlyapnikov Mikhail G. , Brovko Fedor A. , Solonin Alexander S.
Published: November 2023

Hemolysin II (HlyII)—one of the pathogenic factors of Bacillus cereus, a pore-forming β-barrel toxin—possesses a C-terminal extension of 94 amino acid residues, designated as the C-terminal domain of HlyII (HlyIICTD), which plays an important role in the functioning of the toxin. Our previous work described a monoclonal antibody (HlyIIC-20), capable of strain-specific inhibition of hemolysis caused by HlyII, and demonstrated the dependence of the efficiency of hemolysis on the presence of proline at position 324 in HlyII outside the conformational antigenic determinant. In this work, we studied 16 mutant forms of HlyIICTD. Each of the mutations, obtained via multiple site-directed mutagenesis leading to the replacement of amino acid residues lying on the surface of the 3D structure of HlyIICTD, led to a decrease in the interaction of HlyIIC-20 with the mutant form of the protein. Changes in epitope structure confirm the high conformational mobility of HlyIICTD required for the functioning of HlyII. Comparison of the effect of the introduced mutations on the effectiveness of interactions between HlyIICTD and HlyIIC-20 and a control antibody recognizing a non-overlapping epitope enabled the identification of the amino acid residues N339 and K340, included in the conformational antigenic determinant recognized by HlyIIC-20.
Transcriptome and miRNAs Profiles Reveal Regulatory Network and Key Regulators of Secondary Xylem Formation in “84K” Poplar
Authors: Wang Huilin , Zhao Pan , He Yumei , Su Yuting , Zhou Xinyi , Guo Huihong
Published: November 2023

Secondary xylem produced by stem secondary growth is the main source of tree biomass and possesses great economic and ecological value in papermaking, construction, biofuels, and the global carbon cycle. The secondary xylem formation is a complex developmental process, and the underlying regulatory networks and potential mechanisms are still under exploration. In this study, using hybrid poplar (Populus alba × Populus glandulosa clone 84K) as a model system, we first ascertained three representative stages of stem secondary growth and then investigated the regulatory network of secondary xylem formation by joint analysis of transcriptome and miRNAs. Notably, 7507 differentially expressed genes (DEGs) and 55 differentially expressed miRNAs (DEMs) were identified from stage 1 without initiating secondary growth to stage 2 with just initiating secondary growth, which was much more than those identified from stage 2 to stage 3 with obvious secondary growth. DEGs encoding transcription factors and lignin biosynthetic enzymes and those associated with plant hormones were found to participate in the secondary xylem formation. MiRNA-target analysis revealed that a total of 85 DEMs were predicted to have 2948 putative targets. Among them, PagmiR396d-PagGRFs, PagmiR395c-PagGA2ox1/PagLHW/PagSULTR2/PagPolyubiquitin 1, PagmiR482d-PagLAC4, PagmiR167e-PagbHLH62, and PagmiR167f/g/h-PagbHLH110 modules were involved in the regulating cambial activity and its differentiation into secondary xylem, cell expansion, secondary cell wall deposition, and programmed cell death. Our results give new insights into the regulatory network and mechanism of secondary xylem formation.
Photoperiod Induces the Epigenetic Change of the GNAQ Gene in OVX+E2 Ewes
Authors: Wang Wei , Du Xiaolong , Chu Mingxing , He Xiaoyun
Published: November 2023

GNAQ, a member of the alpha subunit encoding the q-like G protein, is a critical gene in cell signaling, and multiple studies have shown that upregulation of GNAQ gene expression ultimately inhibits the proliferation of gonadotropin-releasing hormone (GnRH) neurons and GnRH secretion, and ultimately affects mammalian reproduction. Photoperiod is a key inducer which plays an important role in gene expression regulation by affecting epigenetic modification. However, fewer studies have confirmed how photoperiod induces epigenetic modifications of the GNAQ gene. In this study, we examined the expression and epigenetic changes of GNAQ in the hypothalamus in ovariectomized and estradiol-treated (OVX+E2) sheep under three photoperiod treatments (short photoperiod treatment for 42 days, SP42; long photoperiod treatment for 42 days, LP42; 42 days of short photoperiod followed by 42 days of long photoperiod, SP-LP42). The results showed that the expression of GNAQ was significantly higher in SP-LP42 than in SP42 and LP42 (p < 0.05). Whole genome methylation sequencing (WGBS) results showed that there are multiple differentially methylated regions (DMRs) and loci between different groups of GNAQ. Among them, the DNA methylation level of DMRs at the CpG1 locus in SP42 was significantly higher than that of SP-LP42 (p < 0.01). Subsequently, we confirmed that the core promoter region of the GNAQ gene was located with 1100 to 1500 bp upstream, and the DNA methylation level of all eight CpG sites in SP42 was significantly higher than those in LP42 (p < 0.01), and significantly higher than those in SP-LP42 (p < 0.01), except site 2 and site 4 in the first sequencing fragment (p < 0.05) in the core promoter region. The expression of acetylated GNAQ histone H3 was significantly higher than that of the control group under three different photoperiods (p < 0.01); the acetylation level of sheep hypothalamic GNAQ genomic protein H3 was significantly lower under SP42 than under SP-LP42 (p < 0.05). This suggests that acetylated histone H3 binds to the core promoter region of the GNAQ gene, implying that GNAQ is epigenetically regulated by photoperiod through histone acetylation. In summary, the results suggest that photoperiod can induce DNA methylation in the core promoter region and histone acetylation in the promoter region of the GNAQ gene, and hypothesize that the two may be key factors in regulating the differential expression of GNAQ under different photoperiods, thus regulating the hypothalamus–pituitary–gonadal axis (HPGA) through the seasonal estrus in sheep. The results of this study will provide some new information to understand the function of epigenetic modifications in reproduction in sheep.
Thalamic Neuron Resilience during Osmotic Demyelination Syndrome (ODS) Is Revealed by Primary Cilium Outgrowth and ADP-ribosylation factor-like protein 13B Labeling in Axon Initial Segment
Authors: Gilloteaux Jacques , De Swert Kathleen , Suain Valérie , Nicaise Charles
Published: November 2023

A murine osmotic demyelinating syndrome (ODS) model was developed through chronic hyponatremia, induced by desmopressin subcutaneous implants, followed by precipitous sodium restoration. The thalamic ventral posterolateral (VPL) and ventral posteromedial (VPM) relay nuclei were the most demyelinated regions where neuroglial damage could be evidenced without immune response. This report showed that following chronic hyponatremia, 12 h and 48 h time lapses after rebalancing osmolarity, amid the ODS-degraded outskirts, some resilient neuronal cell bodies built up primary cilium and axon hillock regions that extended into axon initial segments (AIS) where ADP-ribosylation factor-like protein 13B (ARL13B)-immunolabeled rod-like shape content was revealed. These AIS-labeled shaft lengths appeared proportional with the distance of neuronal cell bodies away from the ODS damaged epicenter and time lapses after correction of hyponatremia. Fine structure examination verified these neuron abundant transcriptions and translation regions marked by the ARL13B labeling associated with cell neurotubules and their complex cytoskeletal macromolecular architecture. This necessitated energetic transport to organize and restore those AIS away from the damaged ODS core demyelinated zone in the murine model. These labeled structures could substantiate how thalamic neuron resilience occurred as possible steps of a healing course out of ODS.
Identification of miRNA Associated with Trichomonas gallinae Resistance in Pigeon (Columba livia)
Authors: Li Xinyi , Ni Aixin , Zhang Ran , Li Yunlei , Yuan Jingwei , Sun Yanyan , Chen Jilan , Ma Hui
Published: November 2023

Trichomonas gallinae (T. gallinae) has a great influence on the pigeon industry. Pigeons display different resistance abilities to T. gallinae, so the study of the molecular mechanism of resistance is necessary in breeding disease resistant lines. MiRNA plays important roles in the immune response, but there are still no reports of miRNA regulating trichomonosis resistance. We used small RNA sequencing technology to characterize miRNA profiles in different groups. T. gallinae was nasally inoculated in one day old squabs, and according to the infection status, the groups were divided into control (C), susceptible (S) and tolerant (T) groups. We identified 2429 miRNAs in total, including 1162 known miRNAs and 1267 new miRNAs. In a comparison among the C, S and T groups, the target genes of differentially expressed miRNAs were analyzed via GO and KEGG annotation. The results showed that the target genes were enriched in immune-response-related pathways. This indicated that the differentially expressed miRNAs had a critical influence on T. gallinae infection. Novel_miR_741, which could inhibit the expression of PRKCQ, was down-regulated in the T group compared to the C group. It was proven that a decreased novel_miR_741 expression would increase the expression of PRKCQ and increase the immune response. This study brings new insights into understanding the mechanism of trichomonosis resistance.
Ageing-Related Alterations in Renal Epithelial Glucose Transport
Authors: Lee Chien-Te , Ng Hwee-Yeong , Zhong Hua-Rong , Wang Yi , Liu Chih-Han , Lee Yuai-Ting
Published: November 2023

The kidney plays a crucial role in glucose homeostasis by regulating glucose transport. We aimed to investigate the impact of alterations in glucose transport on glucose metabolism during ageing. Adult male Sprague Dawley rats were divided into five groups: 3-month, 6-month, and 12-month control groups, and 6- and 12-month groups receiving the hydrogen sulfide donor molecule GYY4137. The study found that, as age increased, daily urinary uric acid and protein levels increased in the 12-month group. Blood sugar level and HOMA-IR index increased in the 12-month group, and were partially improved by GYY4137. The kidney tissue showed mild glomerulosclerosis in the 12-month group, which was diminished by GYY4137. Gene expression analysis showed decreased sirtuin and increased p21 expression in the aging groups. Increased SGLT1 and SGLT2 expression was observed in the 12-month group, which was reversed by GYY4137. Both GLUT1 and GLUT2 expression was increased in the 6- and 12-month groups, and reversed by GYY4137 in the 12-month group. The study concluded that aging was associated with increased blood sugar levels and the HOMA-IR index, and the abundance of renal glucose transporters increased as aging progressed. GYY4137 effectively reversed aging-related alterations in glucose homeostasis and renal epithelial transporters.