bluuebunny/crossref_metadata_2025 huggingface dataset:
Long-Term Forecasting Framework for Renewable Energy Technologies’ Installed Capacity and Costs for 2050
Authors: Rozon Francois , McGregor Craig , Owen Michael
Published: September 2023

Published forecasts underestimate renewable energy capacity growth and potential cost reductions, creating uncertainty around investment decisions and slowing progress. Scenario-based projections diverge widely, driven by variations in modelling techniques and underlying assumptions, with policy-based models typically being overly conservative. With historical generation capacity and cost data readily available, this research demonstrates that data-driven approaches can be leveraged to improve long-term capacity and cost forecasts of solar, wind, and battery storage technologies. Unlike exponential growth models prevailing over shorter time scales, logistic curves requiring asymptotic limits, or machine learning algorithms dependent on extensive datasets, this analysis demonstrates that temporal quadratic regressions are a better starting point to represent capacity growth trends over two to three decades. When coupled with published learning rates, trend-based capacity forecasts provided tighter and lower capital and levelized cost of energy outlooks than most reviewed scenarios, with photovoltaics global average levelized cost of energy reducing from 0.057/kWh to below USD 0.03/kWh by 2030 and below USD 0.02/kWh by 2040. Greater transparency on manufacturing ecosystems is proposed so that more advanced analytical techniques can be utilized. This analysis indicates that without direct interventions to accelerate the growth in wind power generation, global renewable energy technology deployment will fall short of the generation capacities required to meet climate change objectives.
A Comprehensive Review in Microwave Pyrolysis of Biomass, Syngas Production and Utilisation
Authors: Al-Qahtani Ali Mubarak
Published: September 2023

Lignocellulosic and waste materials, such as sewage sludge, can be broken down into its useful constituents and converted into fuel for engines. This paper investigates microwave pyrolysis to decompose biomass into H2 and CO (syngas), which may be catalysed in the Fischer–Tropsch (F-T) process to liquid biofuels. Using microwave radiation as the heat source for pyrolysis proves to yield large quantities of gas with higher concentrations of H2 and CO compared to conventional heating methods. This is largely due to the energy transfer mechanism of microwaves. Pyrolysis parameters such as temperature (which increases with input power), feedstock type, microwave absorber, and biomass moisture content influence syngas yield. Several papers reviewed for this study showed differing optimal conditions for microwave pyrolysis, all being heavily dependent on the biomass used and its composition. However, all researchers agreed on the thermal efficiency of microwave heating and how its material-selective nature can increase syngas yield. Compared to diesel fuels (while processing a similar efficiency and a higher cetane number), FT fuels and specifically pyrolysis may yield the benefit of reduced nitric oxides (NOx), particulate matter (PM), unburnt hydrocarbons (HC) and carbon monoxide (CO) emissions.
Model-Based Optimization of Spiral Coils for Improving Wireless Power Transfer
Authors: Ben Fadhel Yosra , Bouattour Ghada , Bouchaala Dhouha , Derbel Nabil , Kanoun Olfa
Published: September 2023

Inductive wireless power transfer is a promising technology for powering smart wearable devices. The spiral coil shape is widely used in wireless power transfer applications. Nevertheless, during the coil design process, there are many challenges to overcome considering all the design constraints. The most important is to determine the optimal coil parameters (internal radius, external radius, spacing, wire width, and conductive wire) with the aim of obtaining the highest coil quality factor. Coil modeling is very important for the wireless power transfer system’s efficiency. Indeed, it is challenging because it requires a high computational effort and has convergence problems. In this paper, we propose a new approach for the approximation of spiral coils through concentric circular turns to reduce the computational effort. The mathematical model determines the optimal coil parameters to obtain the highest coil quality factor. We have chosen the smart textile as an application. The system operates at a frequency of 100 Khz considering the Qi guidelines. To validate this approach, we compared the approximated circular coil model with the spiral coil model through a finite element method simulation using the COMSOL software. The obtained results show that the proposed approximation reduces the complexity of the coil design process and performs well compared to the model corresponding to the spiral shape, without significantly modifying the coil inductance. For a wire width smaller than 1 mm, the total deviation is around 4% in terms of the coil quality factor in a predetermined domain of its parameters.
A Review on Testing of Electrochemical Cells for Aging Models in BESS
Authors: Pakjoo Mehrshad , Piegari Luigi , Rancilio Giuliano , Colnago Silvia , Mengou Joseph Epoupa , Bresciani Federico , Gorni Giacomo , Mandelli Stefano , Merlo Marco
Published: September 2023

The use of electrochemical cells is becoming more widespread, especially in the energy industry and battery energy storage systems (BESSs). As we continue to deploy BESSs, it becomes increasingly important for us to understand how these systems age and accurately predict their performance over time. This knowledge is essential for ensuring that the systems operate optimally and can be properly maintained. Since the structure of a BESS is different from a single electrochemical cell, the existing models at the cell level cannot predict and estimate the life of the BESS with suitable accuracy. Furthermore, the test protocols available at the cell level mostly cannot be executed at the BESS level for many reasons. Therefore, in this paper, a review of test protocols for building aging models for BESSs has been performed. After reviewing the protocols for a single electrochemical cell and addressing the differences between BESSs and cells, a review of the works performed on a larger scale has been carried out, and the possible ways for testing the BESS for aging models were investigated.
Wood Modification Using Imidazole and Succinimide: Effects on Dimensional Stability and Bending Properties
Authors: Scharf Alexander , Dernegård Henric , Oja Johan , Sandberg Dick , Jones Dennis
Published: September 2023

The modification of Scots pine sapwood (Pinus sylvestris L.) with the heterocyclic compounds imidazole and succinimide was investigated. Pressure-impregnation with aqueous solutions containing imidazole, imidazole + citric acid, succinimide, succinimide + citric acid, and citric acid + sorbitol (CIOL®) with solid contents of 5%, 10%, and 15% was followed by oven-curing at 220 °C for 1 h. During the treatment steps, the changes in mass, bending properties, and anti-swelling efficiency (ASE) were examined. The results indicate that solid concentrations within the range of 5% to 10% were optimal. The results seem to show that there are two differing mechanisms in the modification of imidazole and succinimide, respectively. Mass loss due to heat treatment was highest in the imidazole-treated specimens, whereas it remained low and concentration-independent in the succinimide-treated specimens. After three cycles, the ASE reached 31% for the imidazole-treated specimens and improved to 38% with the addition of citric acid. For succinimide, the ASE increased from 17% to 41%. The bending properties generally showed improvement, except for succinimide + citric acid and CIOL®, which displayed a reduced modulus of rupture. Chemical analyses are warranted to fully understand the reaction mechanisms of these treatments. The positive effects of imidazole treatment are suggested to stem from a thermal reaction between the chemical and the wood, indicated by substantial mass loss during leaching and specimen darkening. Succinimide and citric acid might exhibit polymerization with each other and with wood components, which is akin to the CIOL® process. Further research should delve into the reaction mechanisms and the impact of imidazole and succinimide on biological durability.
Mitigation of Climate Change Impact on Bioclimatic Conditions Using Different Green Space Scenarios: The Case of a Hospital in Gorgan Subtropical Climates
Authors: Borna Reza , Roshan Gholamreza , Moghbel Masoumeh , Szabó György , Ata Behnam , Attia Shady
Published: September 2023

Urban development and its climatic consequences have caused urban decision-makers to establish strategies to mitigate climate change. The implementation of different green spaces is one of the main strategies to reduce the environmental and climatic consequences of urbanization. Therefore, the main objective of this research is to reveal the effect of different green space scenarios on micro-bioclimatic conditions of a hospital located in Gorgan city, Golestan province. Therefore, in order to determine the position of the hospital building relative to Gorgan’s urban heat island (UHI), the location and changes in UHI intensity of Gorgan were determined as evidence of urban expansion. Since 27 July was determined as the hottest day in Gorgan city based on historical data analysis, the climatic conditions during 27 July 2021 were measured using an AR847 data logger installed in the hospital environment. Additionally, four different conditions, including actual environmental conditions of the hospital (actual conditions), along with the application of cypress trees (scenario A), plane trees (scenario B), and Buxus shrubs (scenario C), have been used to analyze the impact of different vegetation species on the bioclimatic conditions of 5 Azar Hospital during two time intervals, including observational periods (1970–2020) and the decade of the 2040s. Finally, spatiotemporal patterns of the predicted mean vote (PMV) thermal index were calculated for the observational period and during the 2040s using the ENVI-met micro-scale model. Results showed that the study site is in the UHI, which can affect the micro-bioclimatic conditions and the patient’s thermal perception. For all designed scenarios, results indicate that the average PMV index will increase by the 2040s. However, implementing different green space scenarios showed that the minimum and maximum values of PMV were found in scenario B, of 2.7. The actual PMV conditions of the studied site increased by 3.5. The scenario introduction of green spaces during the 2040s indicates that the average PMV at the hospital site will be decreased by 0.9 compared to the actual conditions. The study proves that appropriate green space strategies can reduce thermal loads occurring due to global climate change and improve the thermal conditions in the study area.
Dynamic Risk Assessment in Cybersecurity: A Systematic Literature Review
Authors: Cheimonidis Pavlos , Rantos Konstantinos
Published: September 2023

Traditional information security risk assessment (RA) methodologies and standards, adopted by information security management systems and frameworks as a foundation stone towards robust environments, face many difficulties in modern environments where the threat landscape changes rapidly and new vulnerabilities are being discovered. In order to overcome this problem, dynamic risk assessment (DRA) models have been proposed to continuously and dynamically assess risks to organisational operations in (near) real time. The aim of this work is to analyse the current state of DRA models that have been proposed for cybersecurity, through a systematic literature review. The screening process led us to study 50 DRA models, categorised based on the respective primary analysis methods they used. The study provides insights into the key characteristics of these models, including the maturity level of the examined models, the domain or application area in which these models flourish, and the information they utilise in order to produce results. The aim of this work is to answer critical research questions regarding the development of dynamic risk assessment methodologies and provide insights on the already developed methods as well as future research directions.
Investigating IPTV Malware in the Wild
Authors: Lockett Adam , Chalkias Ioannis , Yucel Cagatay , Henriksen-Bulmer Jane , Katos Vasilis
Published: September 2023

Technologies providing copyright-infringing IPTV content are commonly used as an illegal alternative to legal IPTV subscriptions and services, as they usually have lower monetary costs and can be more convenient for users who follow content from different sources. These infringing IPTV technologies may include websites, software, software add-ons, and physical set-top boxes. Due to the free or low cost of illegal IPTV technologies, illicit IPTV content providers will often resort to intrusive advertising, scams, and the distribution of malware to increase their revenue. We developed an automated solution for collecting and analysing malware from illegal IPTV technologies and used it to analyse a sample of illicit IPTV websites, application (app) stores, and software. Our results show that our IPTV Technologies Malware Analysis Framework (IITMAF) classified 32 of the 60 sample URLs tested as malicious compared to running the same test using publicly available online antivirus solutions, which only detected 23 of the 60 sample URLs as malicious. Moreover, the IITMAF also detected malicious URLs and files from 31 of the sample’s websites, one of which had reported ransomware behaviour.
Anti-Obesity Effects of the Larval Powder of Steamed and Lyophilized Mature Silkworms in a Newly Designed Adult Mouse Model
Authors: Kim Min Woo , Ham Yu-Jin , Kim Hyun-Bok , Lee Ji young , Lim Jung-Dae , Lee Hyun-Tai
Published: September 2023

Recently, “mature” silkworms (MS) of Bombix mori have been considered a potential nutraceutical, with a number of health benefits reported for steamed and lyophilized MS powder (SMSP). However, no obesity-related effects have been reported for SMSP. In the present study, anti-obesity effects of SMSP were investigated in adult mice in vivo, aged 12 weeks at the onset of SMSP treatment, fed a normal diet (ND) and a high-fat diet (HFD), respectively, for 12 weeks. SMSP significantly suppressed body weight gain, intra-abdominal adipose tissue, and food efficiency in both ND-fed and HFD-fed adult mice. In addition, SMSP significantly decreased food intake and liver weight in HFD-fed mice, indicating that SMSP suppressed appetite and simultaneously reduced the conversion of feed into body weight in HFD-fed mice. SMSP also significantly lowered the serum levels of glucose, triglyceride, total cholesterol, low-density lipoprotein cholesterol, asparagine transaminase, alanine transaminase, and alkaline phosphatase. However, SMSP had no significant effect on the weights of the kidney, spleen, or thymus or the serum levels of blood urea nitrogen and creatinine. Taken together, the above results suggest that SMSP has potent anti-obesity effects and is safe for long-term use as a potential therapeutic and/or nutraceutical in both obese patients and non-obese individuals.
Combining Vis-NIR and NIR Spectral Imaging Techniques with Data Fusion for Rapid and Nondestructive Multi-Quality Detection of Cherry Tomatoes
Authors: Tan Fei , Mo Xiaoming , Ruan Shiwei , Yan Tianying , Xing Peng , Gao Pan , Xu Wei , Ye Weixin , Li Yongquan , Gao Xiuwen , Liu Tianxiang
Published: September 2023

Firmness, soluble solid content (SSC) and titratable acidity (TA) are characteristic substances for evaluating the quality of cherry tomatoes. In this paper, a hyper spectral imaging (HSI) system using visible/near-infrared (Vis-NIR) and near-infrared (NIR) was proposed to detect the key qualities of cherry tomatoes. The effects of individual spectral information and fused spectral information in the detection of different qualities were compared for firmness, SSC and TA of cherry tomatoes. Data layer fusion combined with multiple machine learning methods including principal component regression (PCR), partial least squares regression (PLSR), support vector regression (SVR) and back propagation neural network (BP) is used for model training. The results show that for firmness, SSC and TA, the determination coefficient R2 of the multi-quality prediction model established by Vis-NIR spectra is higher than that of NIR spectra. The R2 of the best model obtained by SSC and TA fusion band is greater than 0.9, and that of the best model obtained by the firmness fusion band is greater than 0.85. It is better to use the spectral bands after information fusion for nondestructive quality detection of cherry tomatoes. This study shows that hyperspectral imaging technology can be used for the nondestructive detection of multiple qualities of cherry tomatoes, and the method based on the fusion of two spectra has a better prediction effect for the rapid detection of multiple qualities of cherry tomatoes compared with a single spectrum. This study can provide certain technical support for the rapid nondestructive detection of multiple qualities in other melons and fruits.
Inhibition Mechanism of Lactiplantibacillus plantarum on the Growth and Biogenic Amine Production in Morganella morganii
Authors: Sun Zhenxiao , Zhang Yi , Lin Xinping , Zhang Sufang , Chen Yingxi , Ji Chaofan
Published: September 2023

Morganella morganii, a spoilage bacterium in fermented foods, produces harmful biogenic amines (BAs). Although Lactiplantibacillus plantarum is widely used to inhibit spoilage bacteria, the inhibition pattern and inhibition mechanism of M. morganii by Lpb. plantarum are not well studied. In this study, we analysed the effects of the addition of Lpb. plantarum cell-free supernatant (CFS) on the growth and BA accumulation of M. morganii and revealed the mechanisms of changes in different BAs by using RNA sequencing transcriptome analysis. The results showed that Lpb. plantarum CFS could significantly inhibit M. morganii BAs in a weak acid environment (pH 6), and the main changes were related to metabolism. Carbohydrate and energy metabolism were significantly down-regulated, indicating that Lpb. plantarum CFS inhibited the growth activity and decreased the BA content of M. morganii. In addition, the change in histamine content is also related to the metabolism of its precursor amino acids, the change in putrescine content may also be related to the decrease in precursor amino acid synthesis and amino acid transporter, and the decrease in cadaverine content may also be related to the decrease in the cadaverine transporter. The results of this study help to inhibit the accumulation of harmful metabolites in fermented foods.
Design of a Modified MIMO Antenna Based on Tweaked Spherical Fractal Geometry for 5G New Radio (NR) Band N258 (24.25–27.25 GHz) Applications
Authors: Bisht Nitasha , Malik Praveen Kumar , Das Sudipta , Islam Tanvir , Asha Sivaji , Alathbah Moath
Published: September 2023

This article describes a fractal-based MIMO antenna for 5G mm-wave mobile applications with micro-strip feeding. The proposed structure is a fractal-based spherical configuration that incorporates spherical slots of different iterations on the patch, as well as rectangular slots on the ground plane. These additions are meant to reduce patch isolation. The two-element MIMO antenna has closely spaced antenna elements that resonate at multiple frequencies, 9.5 GHz, 11.1 GHz, 13.4 GHz, 15.8 GHz, 21.1 GHz, and 26.6 GHz, in the frequency range of 8 to 28 GHz. The antenna’s broadest operational frequency range spans from 17.7 GHz to 28 GHz, encompassing a bandwidth of 10,300 MHz. Consequently, it is well-suited for utilization within the millimeter wave (mm wave) application, specifically for the 5G new radio frequency band n258, and partially covers some other bands X (8.9–9.9 GHz, 10.4–11.4 GHz), and Ku (13.1–13.7 GHz, 15.4–16.2 GHz). All the resonating bands have isolation levels below the acceptable range of (|S12| > −16 dB). The proposed antenna utilizes a FR4 material with dimension of 28.22 mm × 44 mm. An investigation is conducted to analyze the effectiveness of parameters of the antenna, including radiation pattern, surface current distributions and S parameters. Furthermore, an examination and assessment are conducted on the efficacy of the diversity system inside the multiple input multiple output (MIMO) framework. This evaluation encompasses the analysis of key performance metrics such as the envelope correlation coefficient (ECC), diversity gain (DG), and mean effective gain (MEG). All antenna characteristics are determined to be within a suitable range for this suggested MIMO arrangement. The antenna design underwent experimental validation and the simulated outcomes were subsequently verified.
Comparative Evaluation of Empirical Approaches and Artificial Intelligence Techniques for Predicting Uniaxial Compressive Strength of Rock
Authors: Li Chuanqi , Zhou Jian , Dias Daniel , Du Kun , Khandelwal Manoj
Published: September 2023

The uniaxial compressive strength (UCS) of rocks is one of the key parameters for evaluating the safety and stability of civil and mining structures. In this study, 386 rock samples containing four properties named the load strength (PLS), the porosity (Pn), the P-wave velocity (Vp), and the Schmidt hardness rebound number (SHR) are utilized to predict the UCS using several typical empirical equations (EA) and artificial intelligence (AI) methods, i.e., 16 single regression (SR) equations, 2 multiple regression (MR) equations, and the random forest (RF) models optimized by grey wolf optimization (GWO), moth flame optimization (MFO), lion swarm optimization (LSO), and sparrow search algorithm (SSA). The root mean square error (RMSE), determination coefficient (R2), Willmott’s index (WI), and variance accounted for (VAF) are used to evaluate the predictive performance of all developed models. The evaluation results show that the overall performance of AI models is superior to empirical approaches, especially the LSO-RF model. In addition, the most important input variable is the Pn for predicting the UCS. Therefore, AI techniques are considered as more efficient and accurate approaches to replace the empirical equations for predicting the UCS of these collected rock samples, which provides a reliable and effective idea to predict the rock UCS in the filed site.
Caring for People with Rare Diseases: A Systematic Review of the Challenges of, and Strategies for Dealing with, COVID-19
Authors: Faccio Elena , Bottecchia Matteo , Rocelli Michele
Published: September 2023

The COVID-19 pandemic took a toll on everyone’s lives, and patients with rare diseases (RDs) had to pay an even higher price. In this systematic review, we explored the impact of the COVID-19 pandemic on individuals with RDs from a psychological perspective. Using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, we retrieved articles from the Google Scholar, Scopus, and PubMed databases focusing on ‘COVID-19,’ ‘psychology,’ and ‘rare diseases.’ Seventeen primary articles were identified (mainly from continental Europe). The results revealed the psychological effects of the pandemic on rare disease patients, including increased anxiety, stress, and depressive moods. This review also highlighted the increased vulnerability and reduced quality of life of rare disease patients during the pandemic, as well as the importance of telecare and psychological support as critical interventions for improving their well-being. There is an urgent need for multidisciplinary research and stronger healthcare systems to meet the unique challenges of rare disease patients, who represent 3.5–5.9% of the global population.
MucR from Sinorhizobium meliloti: New Insights into Its DNA Targets and Its Ability to Oligomerize
Authors: Slapakova Martina , Sgambati Domenico , Pirone Luciano , Russo Veronica , D’Abrosca Gianluca , Valletta Mariangela , Russo Rosita , Chambery Angela , Malgieri Gaetano , Pedone Emilia Maria , Dame Remus Thei , Pedone Paolo Vincenzo , Baglivo Ilaria
Published: September 2023

Proteins of the MucR/Ros family play a crucial role in bacterial infection or symbiosis with eukaryotic hosts. MucR from Sinorhizobium meliloti plays a regulatory role in establishing symbiosis with the host plant, both dependent and independent of Quorum Sensing. Here, we report the first characterization of MucR isolated from Sinorhizobium meliloti by mass spectrometry and demonstrate that this protein forms higher-order oligomers in its native condition of expression by SEC-MALS. We show that MucR purified from Sinorhizobium meliloti can bind DNA and recognize the region upstream of the ndvA gene in EMSA, revealing that this gene is a direct target of MucR. Although MucR DNA binding activity was already described, a detailed characterization of Sinorhizobium meliloti DNA targets has never been reported. We, thus, analyze sequences recognized by MucR in the rem gene promoter, showing that this protein recognizes AT-rich sequences and does not require a consensus sequence to bind DNA. Furthermore, we investigate the dependence of MucR DNA binding on the length of DNA targets. Taken together, our studies establish MucR from Sinorhizobium meliloti as a member of a new family of Histone-like Nucleoid Structuring (H-NS) proteins, thus explaining the multifaceted role of this protein in many species of alpha-proteobacteria.
Meeting at the Crossroad between Obesity and Hepatic Carcinogenesis: Unique Pathophysiological Pathways Raise Expectations for Innovative Therapeutic Approaches
Authors: Arvanitakis Konstantinos , Papadakos Stavros P. , Lekakis Vasileios , Koufakis Theocharis , Lempesis Ioannis G. , Papantoniou Eleni , Kalopitas Georgios , Georgakopoulou Vasiliki E. , Stergiou Ioanna E. , Theocharis Stamatios , Germanidis Georgios
Published: September 2023

The escalating global prevalence of obesity and its intricate association with the development of hepatocellular carcinoma (HCC) pose a substantial challenge to public health. Obesity, acknowledged as a pervasive epidemic, is linked to an array of chronic diseases, including HCC, catalyzing the need for a comprehensive understanding of its molecular underpinnings. Notably, HCC has emerged as a leading malignancy with rising incidence and mortality. The transition from viral etiologies to the prominence of metabolic dysfunction-associated fatty liver disease (MAFLD)-related HCC underscores the urgent need to explore the intricate molecular pathways linking obesity and hepatic carcinogenesis. This review delves into the interwoven landscape of molecular carcinogenesis in the context of obesity-driven HCC while also navigating using the current therapeutic strategies and future prospects for combating obesity-related HCC. We underscore the pivotal role of obesity as a risk factor and propose an integrated approach encompassing lifestyle interventions, pharmacotherapy, and the exploration of emerging targeted therapies. As the obesity-HCC nexus continues to challenge healthcare systems globally, a comprehensive understanding of the intricate molecular mechanisms and innovative therapeutic strategies is imperative to alleviate the rising burden of this dual menace.
Electrophoretic Determination of L-Carnosine in Health Supplements Using an Integrated Lab-on-a-Chip Platform with Contactless Conductivity Detection
Authors: Pukleš Iva , Páger Csilla , Sakač Nikola , Šarkanj Bojan , Matasović Brunislav , Samardžić Mirela , Budetić Mateja , Marković Dean , Jozanović Marija
Published: September 2023

The health supplement industry is one of the fastest growing industries in the world, but there is a lack of suitable analytical methods for the determination of active compounds in health supplements such as peptides. The present work describes an implementation of contactless conductivity detection on microchip technology as a new strategy for the electrophoretic determination of L-carnosine in complex health supplement formulations without pre-concentration and derivatization steps. The best results were obtained in the case of +1.00 kV applied for 20 s for injection and +2.75 kV applied for 260 s for the separation step. Under the selected conditions, a linear detector response of 5 × 10−6 to 5 × 10−5 M was achieved. L-carnosine retention time was 61 s. The excellent reproducibility of both migration time and detector response confirmed the high precision of the method. The applicability of the method was demonstrated by the determination of L-carnosine in three different samples of health supplements. The recoveries ranged from 91 to 105%. Subsequent analysis of the samples by CE-UV-VIS and HPLC-DAD confirmed the accuracy of the obtained results.
Standardized Computer-Assisted Analysis of 5-hmC Immunoreactivity in Dysplastic Nevi and Superficial Spreading Melanomas
Authors: Koch Elias A. T. , Berking Carola , Erber Ramona , Erdmann Michael , Kiesewetter Franklin , Schliep Stefan , Heppt Markus V.
Published: September 2023

5-Hydroxymethylcytosine (5-hmC) is an important intermediate of DNA demethylation. Hypomethylation of DNA is frequent in cancer, resulting in deregulation of 5-hmC levels in melanoma. However, the interpretation of the intensity and distribution of 5-hmC immunoreactivity is not very standardized, which makes its interpretation difficult. In this study, 5-hmC-stained histological slides of superficial spreading melanomas (SSM) and dysplastic compound nevi (DN) were digitized and analyzed using the digital pathology and image platform QuPath. Receiver operating characteristic/area under the curve (ROCAUC) and t-tests were performed. A p-value of <0.05 was used for statistical significance, and a ROCAUC score of >0.8 was considered a “good” result. In total, 92 5-hmC-stained specimens were analyzed, including 42 SSM (45.7%) and 50 DN (54.3%). The mean of 5-hmC-positive cells/mm2 for the epidermis and dermo-epidermal junction and the entire lesion differed significantly between DN and SSM (p = 0.002 and p = 0.006, respectively) and showed a trend towards higher immunoreactivity in the dermal component (p = 0.069). The ROCAUC of 5-hmC-positive cells of the epidermis and dermo-epidermal junction was 0.79, for the dermis 0.74, and for the entire lesion 0.76. These results show that the assessment of the epidermal with junctional expression of 5-hmC is slightly superior to dermal immunoreactivity in distinguishing between DN and SSM.
Comparison of Two Human Skin Cell Isolation Protocols and Their Influence on Keratinocyte and Fibroblast Culture
Authors: Sierra-Sánchez Álvaro , Barbier Martin A. , Magne Brice , Larouche Danielle , Arias-Santiago Salvador , Germain Lucie
Published: September 2023

For the development of advanced therapies, the use of primary cells instead of cell lines is preferred. The manufacture of human tissue-engineered skin substitutes requires efficient isolation and culture protocols allowing a massive expansion of the cells in culture from an initial specimen of a minimal size. This study compared two skin cell isolation protocols, routinely applied in two clinical laboratories. Epithelial (keratinocytes) and dermal (fibroblasts) cells were isolated and cultured from three human skin biopsies (N = 3). The two-step digestion protocol (LOEX-Protocol) firstly used thermolysin to enzymatically disrupt the dermal–epidermal junction while, for the one-step digestion protocol (UPCIT-Protocol), mechanical detachment with scissors was applied. Then, the epidermal and dermal layers were digested, respectively, to achieve cell isolation. The cell size, viability, yield and growth were analyzed over five passages (P). The colony-forming efficiency (CFE) and Keratin 19 (K19) expression of epithelial cells were also assessed after P0 and P1. Regarding the dermal cells, no significant differences were observed in the tested parameters of isolation and culture. However, for the epithelial cells, viability was higher (93% vs. 85%) and the number of cells extracted per cm2 of skin was 3.4 times higher using the LOEX-Protocol compared to the UPCIT-Protocol. No significant difference was observed for any parameter once the keratinocytes were cultured from P1 to P4. The CFE and K19 expression decreased from P0 to P1 in both protocols, probably due to the culture process. This study shows that both protocols enable the efficient isolation of skin dermal and epithelial cells and subsequent culture to produce grafts destined for the treatment of patients.
Two-Dimensional Polyacrylamide Gel Electrophoresis Coupled with Nanoliquid Chromatography–Tandem Mass Spectrometry-Based Identification of Differentially Expressed Proteins and Tumorigenic Pathways in the MCF7 Breast Cancer Cell Line Transfected for Jumping Translocation Breakpoint Protein Overexpression
Authors: Jayathirtha Madhuri , Jayaweera Taniya , Whitham Danielle , Petre Brîndușa Alina , Neagu Anca-Narcisa , Darie Costel C.
Published: September 2023

The identification of new genes/proteins involved in breast cancer (BC) occurrence is widely used to discover novel biomarkers and understand the molecular mechanisms of BC initiation and progression. The jumping translocation breakpoint (JTB) gene may act both as a tumor suppressor or oncogene in various types of tumors, including BC. Thus, the JTB protein could have the potential to be used as a biomarker in BC, but its neoplastic mechanisms still remain unknown or controversial. We previously analyzed the interacting partners of JTBhigh protein extracted from transfected MCF7 BC cell line using SDS-PAGE complemented with in-solution digestion, respectively. The previous results suggested the JTB contributed to the development of a more aggressive phenotype and behavior for the MCF7 BC cell line through synergistic upregulation of epithelial–mesenchymal transition (EMT), mitotic spindle, and fatty acid metabolism-related pathways. In this work, we aim to complement the previously reported JTB proteomics-based experiments by investigating differentially expressed proteins (DEPs) and tumorigenic pathways associated with JTB overexpression using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Statistically different gel spots were picked for protein digestion, followed by nanoliquid chromatography–tandem mass spectrometry (nLC-MS/MS) analysis. We identified six DEPs related to the JTBhigh condition vs. control that emphasize a pro-tumorigenic (PT) role. Twenty-one proteins, which are known to be usually overexpressed in cancer cells, emphasize an anti-tumorigenic (AT) role when low expression occurs. According to our previous results, proteins that have a PT role are mainly involved in the activation of the EMT process. Interestingly, JTB overexpression has been correlated here with a plethora of significant upregulated and downregulated proteins that sustain JTB tumor suppressive functions. Our present and previous results sustain the necessity of the complementary use of different proteomics-based methods (SDS-PAGE, 2D-PAGE, and in-solution digestion) followed by tandem mass spectrometry to avoid their limitations, with each method leading to the delineation of specific clusters of DEPs that may be merged for a better understanding of molecular pathways and neoplastic mechanisms related to the JTB’s role in BC initiation and progression.
Exome Sequencing and Optical Genome Mapping in Molecularly Unsolved Cases of Duchenne Muscular Dystrophy: Identification of a Causative X-Chromosomal Inversion Disrupting the DMD Gene
Authors: Erbe Leoni S. , Hoffjan Sabine , Janßen Sören , Kneifel Moritz , Krause Karsten , Gerding Wanda M. , Döring Kristina , Güttsches Anne-Katrin , Roos Andreas , Buena Atienza Elena , Gross Caspar , Lücke Thomas , Nguyen Hoa Huu Phuc , Vorgerd Matthias , Köhler Cornelia
Published: September 2023

Duchenne muscular dystrophy (DMD) is a severe progressive muscle disease that mainly affects boys due to X-linked recessive inheritance. In most affected individuals, MLPA or sequencing-based techniques detect deletions, duplications, or point mutations in the dystrophin-encoding DMD gene. However, in a small subset of patients clinically diagnosed with DMD, the molecular cause is not identified with these routine methods. Evaluation of the 60 DMD patients in our center revealed three cases without a known genetic cause. DNA samples of these patients were analyzed using whole-exome sequencing (WES) and, if unconclusive, optical genome mapping (OGM). WES led to a diagnosis in two cases: one patient was found to carry a splice mutation in the DMD gene that had not been identified during previous Sanger sequencing. In the second patient, we detected two variants in the fukutin gene (FKTN) that were presumed to be disease-causing. In the third patient, WES was unremarkable, but OGM identified an inversion disrupting the DMD gene (~1.28 Mb) that was subsequently confirmed with long-read sequencing. These results highlight the importance of reanalyzing unsolved cases using WES and demonstrate that OGM is a useful method for identifying large structural variants in cases with unremarkable exome sequencing.
Biodegradable Water-Soluble Matrix for Immobilization of Biocidal 4-Hexylresorcinol
Authors: Novoskoltseva Olga A. , Litmanovich Ekaterina A. , Loiko Nataliya G. , Nikolaev Yury A. , Yaroslavov Alexander A.
Published: September 2023

Biocidal coatings have been used in biomedicine, cosmetology and the food industry. In this article, the coatings are described as being composed of non-stoichiometric polycomplexes, products of electrostatic coupling of two commercial biodegradable ionic polymers, anionic sodium alginate and cationic quaternized hydroxyethyl cellulose ethoxylate. Non-stoichiometric polycomplexes with a 5-fold excess of the cationic polymer were used for immobilizing hydrophobic biocidal 4-hexylresorcinol (HR). Being dispersed in water, the polycomplex particles were capable of absorbing a tenfold excess of HR in relation to the polycation. After deposition onto the plastic surface and drying, the aqueous polycomplex–HR composite formulation forms a transparent homogeneous coating, which swells slightly in water. The interpolyelectrolyte complex (IPEC) is substantially non-toxic. The incorporation of HR in the IPEC imparts antimicrobial activity to the resulting composite, in both aqueous solutions and coatings, against Gram-negative and Gram-positive bacteria and yeast. The polysaccharide-based polycomplexes with embedded HR are promising for the fabrication of biocidal films and coatings.
Contribution of ADD3 and the HLA Genes to Biliary Atresia Risk in Chinese
Authors: Cui Meng-Meng , Gong Yi-Ming , Pan Wei-Hua , Pei Hao-Yue , Bai Mei-Rong , Song Huan-Lei , Han Xin-Ru , Wu Wen-Jie , Yu Wen-Wen , Gu Bei-Lin , Cai Wei , Zhou Ying , Chu Xun
Published: September 2023

Nonsyndromic biliary atresia (BA) is a rare polygenic disease, with autoimmunity, virus infection and inflammation thought to play roles in its pathogenesis. We conducted a genome-wide association study in 336 nonsyndromic BA infants and 8900 controls. Our results validated the association of rs17095355 in ADD3 with BA risk (odds ratio (OR) = 1.70, 95% confidence interval (95% CI) = 1.49–1.99; p = 4.07 × 10−11). An eQTL analysis revealed that the risk allele of rs17095355 was associated with increased expression of ADD3. Single-cell RNA-sequencing data and immunofluorescence analysis revealed that ADD3 was moderately expressed in cholangiocytes and weakly expressed in hepatocytes. Immuno-fluorescent staining showed abnormal deposition of ADD3 in the cytoplasm of BA hepatocytes. No ADD3 auto-antibody was observed in the plasma of BA infants. In the HLA gene region, no variants achieved genome-wide significance. HLA-DQB1 residue Ala57 is the most significant residue in the MHC region (OR = 1.44, 95% CI = 1.20–1.74; p = 1.23 × 10−4), and HLA-DQB1 was aberrantly expressed in the bile duct cells. GWAS stratified by cytomegalovirus (CMV) IgM status in 87 CMV IgM (+) BA cases versus 141 CMV IgM (−) BA cases did not yield genome-wide significant associations. These findings support the notion that common variants of ADD3 account for BA risk. The HLA genes might have a minimal role in the genetic predisposition of BA due to the weak association signal. CMV IgM (+) BA patients might not have different genetic risk factor profiles compared to CMV IgM (−) subtype.
Exploring the Role of Different Cell-Death-Related Genes in Sepsis Diagnosis Using a Machine Learning Algorithm
Authors: Wang Xuesong , Wang Ziyi , Guo Zhe , Wang Ziwen , Chen Feng , Wang Zhong
Published: September 2023

Sepsis, a disease caused by severe infection, has a high mortality rate. At present, there is a lack of reliable algorithmic models for biomarker mining and diagnostic model construction for sepsis. Programmed cell death (PCD) has been shown to play a vital role in disease occurrence and progression, and different PCD-related genes have the potential to be targeted for the treatment of sepsis. In this paper, we analyzed PCD-related genes in sepsis. Implicated PCD processes include apoptosis, necroptosis, ferroptosis, pyroptosis, netotic cell death, entotic cell death, lysosome-dependent cell death, parthanatos, autophagy-dependent cell death, oxeiptosis, and alkaliptosis. We screened for diagnostic-related genes and constructed models for diagnosing sepsis using multiple machine-learning models. In addition, the immune landscape of sepsis was analyzed based on the diagnosis-related genes that were obtained. In this paper, 10 diagnosis-related genes were screened for using machine learning algorithms, and diagnostic models were constructed. The diagnostic model was validated in the internal and external test sets, and the Area Under Curve (AUC) reached 0.7951 in the internal test set and 0.9627 in the external test set. Furthermore, we verified the diagnostic gene via a qPCR experiment. The diagnostic-related genes and diagnostic genes obtained in this paper can be utilized as a reference for clinical sepsis diagnosis. The results of this study can act as a reference for the clinical diagnosis of sepsis and for target discovery for potential therapeutic drugs.
Derivation of Arbas Cashmere Goat Induced Pluripotent Stem Cells in LCDM with Trophectoderm Lineage Differentiation and Interspecies Chimeric Abilities
Authors: Liu Fang , Wang Jing , Yue Yongli , Li Chen , Zhang Xuemin , Xiang Jinzhu , Wang Hanning , Li Xueling
Published: September 2023

The Arbas cashmere goat is a unique biological resource that plays a vital role in livestock husbandry in China. LCDM is a medium with special small molecules (consisting of human LIF, CHIR99021, (S)-(+)-dimethindene maleate, and minocycline hydrochloride) for generation pluripotent stem cells (PSCs) with bidirectional developmental potential in mice, humans, pigs, and bovines. However, there is no report on whether LCDM can support for generation of PSCs with the same ability in Arbas cashmere goats. In this study, we applied LCDM to generate goat induced PSCs (giPSCs) from goat fetal fibroblasts (GFFs) by reprogramming. The derived giPSCs exhibited stem cell morphology, expressing pluripotent markers, and could differentiate into three germ layers. Moreover, the giPSCs differentiated into the trophectoderm lineage by spontaneous and directed differentiation in vitro. The giPSCs contributed to embryonic and extraembryonic tissue in preimplantation blastocysts and postimplantation chimeric embryos. RNA-sequencing analysis showed that the giPSCs were very close to goat embryos at the blastocyst stage and giPSCs have similar properties to typical extended PSCs (EPSCs). The establishment of giPSCs with LCDM provides a new way to generate PSCs from domestic animals and lays the foundation for basic and applied research in biology and agriculture.
The Genome-Wide Identification, Characterization, and Expression Analysis of the Strictosidine Synthase-like Family in Maize (Zea mays L.)
Authors: Gu Lei , Cao Yongyan , Chen Xuanxuan , Wang Hongcheng , Zhu Bin , Du Xuye , Sun Yiyue
Published: September 2023

Maize is often subjected to various environmental stresses. The strictosidine synthase-like (SSL) family is thought to catalyze the key step in the monoterpene alkaloids synthesis pathway in response to environmental stresses. However, the role of ZmSSL genes in maize growth and development and its response to stresses is unknown. Herein, we undertook the systematic identification and analysis of maize SSL genes. Twenty SSL genes were identified in the maize genome. Except for chromosomes 3, 5, 6, and 10, they were unevenly distributed on the remaining 6 chromosomes. A total of 105 SSL genes from maize, sorghum, rice, Aegilops tauschii, and Arabidopsis were divided into five evolutionary groups, and ZmSSL gene structures and conserved protein motifs in the same group were similar. A collinearity analysis showed that tandem duplication plays an important role in the evolution of the SSL family in maize, and ZmSSL genes share more collinear genes in crops (maize, sorghum, rice, and Ae. tauschii) than in Arabidopsis. Cis-element analysis in the ZmSSL gene promoter region revealed that most genes contained many development and stress response elements. We evaluated the expression levels of ZmSSL genes under normal conditions and stress treatments. ZmSSL4–9 were widely expressed in different tissues and were positively or negatively regulated by heat, cold, and infection stress from Colletotrichum graminicola and Cercospora zeina. Moreover, ZmSSL4 and ZmSSL5 were localized in the chloroplast. Taken together, we provide insight into the evolutionary relationships of the ZmSSL genes, which would be useful to further identify the potential functions of ZmSSLs in maize.
How Does cGAS Avoid Sensing Self-DNA under Normal Physiological Conditions?
Authors: Zheng Wangli , Chen Nanhua , Meurens François , Zheng Wanglong , Zhu Jianzhong
Published: September 2023

cGAS is a cytosolic DNA sensor that activates innate immune responses by producing the second messenger 2′3′-cGAMP, which activates the adaptor STING. cGAS senses dsDNA in a length-dependent but sequence-independent manner, meaning it cannot discriminate self-DNA from foreign DNA. In normal physiological conditions, cellular DNA is sequestered in the nucleus by a nuclear envelope and in mitochondria by a mitochondrial membrane. When self-DNA leaks into the cytosol during cellular stress or mitosis, the cGAS can be exposed to self-DNA and activated. Recently, many studies have investigated how cGAS keeps inactive and avoids being aberrantly activated by self-DNA. Thus, this narrative review aims to summarize the mechanisms by which cGAS avoids sensing self-DNA under normal physiological conditions.
New Insights into Acylhydrazones E/Z Isomerization: An Experimental and Theoretical Approach
Authors: Fernández-Palacios Sara , Matamoros Esther , Morato Rojas Isabel , López Navarrete Juan T. , Ruiz Delgado M. Carmen , Vida Yolanda , Perez-Inestrosa Ezequiel
Published: September 2023

A family of acylhydrazones have been prepared and characterized with the aim of investigating their potential as information storage systems. Their well-established synthetic methodologies allowed for the preparation of seven chemically stable acylhydrazones in excellent yields that have been photophysically and photochemically characterized. In addition, DFT and TD-DFT calculations have been performed to gain more insights into the structural, energetic and photophysical properties of the E/Z isomers. Our results reveal that E/Z configurational isomerization upon irradiation is highly dependent on the stabilization of the E or Z isomers due to the formation of intramolecular H bonds and the electronic/steric effects intrinsically related to their structures. In addition, Raman spectroscopy is also used to confirm the molecular structural changes after the formation of hydrogen bonds in the isomers.