bluuebunny/crossref_metadata_2025 huggingface dataset:
Hydrogen Hydrate Promoters for Gas Storage—A Review
Authors: Saikia Tinku , Patil Shirish , Sultan Abdullah
Published: March 2023

Clathrate and semi-clathrate hydrates have recently been gaining major interest as hydrogen storage material. The benefits of hydrates, such as reversible formation and dissociation, their environmentally friendly nature, economical costs, and lower fire risk, make them one of the most promising hydrogen storage materials. One of the major challenges when storing hydrogen in hydrate crystals is the extreme pressure and temperature conditions required for the formation of hydrogen hydrates. Solving the problems of extreme pressure and temperature through the use of promoter molecules would make these materials a promising storage medium with high potential. Through the use of efficient, economical, and green promoter molecules, hydrogen hydrate can be used to store large amounts of hydrogen economically and safely. This review aims to present a comprehensive summary of the different hydrate promoters that have been tested specifically in terms of hydrogen storage. The hydrate promoters are classed according to the structure of the hydrate crystals they form, i.e., sI, sII, sH, and semi-clathrate hydrate. This review article provides summarized information for readers about the different promoters tested and their benefits and shortcomings.
Image-Based Sexual Abuse: Online Gender-Sexual Violations
Authors: Hall Matthew , Hearn Jeff , Lewis Ruth
Published: March 2023

Image-based sexual abuse describes the offline or online non-consensual sharing of real or fake images or videos with (un)known others of a person that are either sexually explicit or sexually suggestive. New information and communication technologies (ICTs) provide many open-ended and undefined possibilities for image-based sexual abuse (IBSA), such as ‘revenge pornography’, ‘upskirting’, deepfake pornography, sexual spycamming, and cyberflashing, to name just a few. These forms of abuse refer to the online, and also at times offline, non-consensual distribution or sharing of explicit images or videos of someone else by ex-partners, partners, others, or hackers seeking revenge, entertainment, or peer group status. The vast majority of these are committed by men against women. Given the many adverse impacts on physical and psychological health and well-being it has on its victim-survivors, exploring this form of online gender-sexual abuse and violation becomes an important endeavor. Situating the discussion within debates on gender and sexuality, the entry discusses the increasing use of new technologies for online gender-sexual abuse and violation, highlighting the motivations of those perpetrating IBSA, the negative physical and psychological impacts of IBSA on victim-survivors, and what has been, and could be, done to combat image-based sexual abuses and other misuses of new technologies, notably through legal, policy, and practice interventions within and between nations.
A Novel MPPT Technique Based on Combination between the Incremental Conductance and Hysteresis Control Applied in a Standalone PV System
Authors: El Ouardi Hind , El Gadari Ayoub , Mokhlis Mohcine , Ounejjar Youssef , Bejjit Lahcen , Al-Haddad Kamal
Published: March 2023

A new Maximum Power Point Tracking (MPPT) method, consisting in combining the Incremental Conductance (INC) algorithm with the Hysteresis control, was developed and applied to a standalone photovoltaic (PV) system to generate the maximum power of the PV array. The INC allows one to search for the Maximum Power Point (MPP). The hysteresis improves the accuracy of tracking the MPP very fast even after severe changes in weather conditions and has no oscillations around the MPP. The five-level S-Packed U Cells (SPUC5) inverter is used to transform the produced DC voltage to AC voltage; it generates five-level output voltage with a small number of switches and only DC source voltage. The capacitors of the SPUC5 are controlled by the Pulse Width Modulation (PWM) in order to balance their voltages. The proposed PV system was established and trained in the MATLAB/Simulink environment under various irradiation conditions. A comparison between different MPPT methods, INC-PWM and INC-PI, was investigated in order to examine the effectiveness of the developed MPPT technique in particular, and of all the PV system components. The results of the simulation validate the effectiveness of the suggested MPPT algorithm as well as the used SPUC5 inverter.
Fucoxanthin as a Biofunctional Compound in Goat Milk Yogurt: Stability and Physicochemical Effects
Authors: Nuñez de González Maryuri T. , Attaie Rahmat , Woldesenbet Selamawit , Mora-Gutierrez Adela , Jung Yoonsung
Published: March 2023

The food industry has increasingly added nutrients and other ingredients to products to enhance their health benefits. Fucoxanthin is recognized for its benefits in mitigating obesity, diabetes, hypertension, and inflammation. Therefore, addition of fucoxanthin into goat milk yogurt, its stability, and the physicochemical properties of yogurt during processing and storage was investigated. Yogurts with and without fucoxanthin were manufactured by mixing goat whole milk (82.85%, w/w), powdered goat milk (10.68%, w/w), and sugar (6.47%, w/w). Fucoxanthin (0.052 mg/g of yogurt mix) was added to the treatment. The mix was heated at 80 °C for 30 min, cooled, inoculated with a culture, and incubated at 43 °C for 5 h. Fucoxanthin in the yogurt mix and yogurt was quantified by an HPLC method. The recoveries of fucoxanthin from the mix before and after heating were 98.25% and 98.83%, respectively. However, less fucoxanthin (90.13%) was recovered from the freshly prepared yogurt than from the mix. Heating the yogurt mix did not affect the concentration of fucoxanthin but adding the inoculum to the mix reduced its concentration during fermentation. During the storage period, the concentration of fucoxanthin in yogurt remained the same. Fucoxanthin did not adversely affect the chemical composition and physicochemical properties of yogurt, but it influenced the color, decreasing lightness (81.47 ± 0.09), and increasing redness (7.67 ± 0.09) and yellowness (38.24 ± 0.09). Thus, goat milk yogurt can be an effective food matrix to deliver fucoxanthin to human diet.
Potential and Restrictions of Food-Waste Valorization through Fermentation Processes
Authors: Ortiz-Sanchez Mariana , Inocencio-García Pablo-José , Alzate-Ramírez Andrés Felipe , Alzate Carlos Ariel Cardona
Published: March 2023

Food losses (FL) and waste (FW) occur throughout the food supply chain. These residues are disposed of on landfills producing environmental issues due to pollutants released into the air, water, and soil. Several research efforts have focused on upgrading FL and FW in a portfolio of added-value products and energy vectors. Among the most relevant research advances, biotechnological upgrading of these residues via fermentation has been demonstrated to be a potential valorization alternative. Despite the multiple investigations performed on the conversion of FL and FW, a lack of comprehensive and systematic literature reviews evaluating the potential of fermentative processes to upgrade different food residues has been identified. Therefore, this article reviews the use of FL and FW in fermentative processes considering the composition, operating conditions, platforms, fermentation product application, and restrictions. This review provides the framework of food residue fermentation based on reported applications, experimental, and theoretical data. Moreover, this review provides future research ideas based on the analyzed information. Thus, potential applications and restrictions of the FL and FW used for fermentative processes are highlighted. In the end, food residues fermentation must be considered a mandatory step toward waste minimization, a circular economy, and the development of more sustainable production and consumption patterns.
Intrinsic Smoke Properties and Prediction of Smoke Production in National Bureau of Standards (NBS) Smoke Chamber
Authors: Sonnier Rodolphe , Dumazert Loïc , Vangrevelynghe Mathieu , Brendlé Clément , Ferry Laurent
Published: March 2023

Smoke production in a smoke chamber is characterized by the accumulation of smoke and the continuous consumption of oxygen leading to a vitiated atmosphere. However, a method is proposed to predict the smoke evolution in a smoke chamber at 25 kW/m2 by using material properties calculated from a cone calorimeter, as already shown in a previous article. These properties represent the ability of a material to produce smoke at a specific mass loss rate. The influence of a flame retardant on these properties can be used as a quantitative measurement of its action on smoke production. These properties can be calculated at another heat flux than 25 kW/m2. The knowledge of the curve “mass loss rate = f(time)” in a smoke chamber is still required, but this curve is close to that measured in a cone calorimeter at the same heat flux. The results prove that the smoke production in a smoke chamber and cone calorimeter is qualitatively similar, i.e., the decrease of oxygen content in a smoke chamber has no influence on smoke (at least as long as optical density does not exceed 800).
Species-Specific Responses of Medium and Large Mammals to Fire Regime Attributes in a Fire-Prone Neotropical Savanna
Authors: Souza Clarice Vieira , Lourenço Águeda , Vieira Emerson Monteiro
Published: March 2023

Fire occurrence affects the distribution of key resources for fauna in natural ecosystems worldwide. For fire management strategies adequate for biodiversity conservation, the understanding of how species respond to fire-induced changes is essential. In this study, we investigated the role of fire regimes on spaces used by medium and large mammals at multiple spatial scales (0.8 ha to 78.5 ha) in a fire-prone savanna ecosystem (Brazilian Cerrado). We sampled mammals using 60 camera traps distributed in 30 sampling units located in grassland and typical savanna formations. We applied single-species occupancy models and AIC-based model selection to assess how mammals use the space in response to pyrodiversity (both diversity of fire frequencies and diversity of fire ages), the proportion of recently burned area, and the proportion of long-unburned area while accounting for detectability. Our results showed that fire regime variables affected the study species differently. Deer species used the space regardless of mosaic pyrodiversity and the proportion of specific fire ages. Fire-related variables, however, affected space use by tapirs and maned wolves. Tapirs preferred to use fire mosaics with lower diversity of fire frequencies, whereas maned wolves more intensively used mosaics with high fire age diversity and a high proportion of recently burned areas. Based on our findings, we recommend that fire management targeting specific mammal species should not necessarily focus on maximum pyrodiversity. Instead, we suggest a management strategy combining “patch mosaic burning” with the maintenance of specific fire-age patches suitable for different species’ requirements.
Differentiating Fire Regimes and Their Biophysical Drivers in Central Portugal
Authors: Bergonse Rafaello , Oliveira Sandra , Zêzere José Luís , Moreira Francisco , Ribeiro Paulo Flores , Leal Miguel , Santos José Manuel Lima
Published: March 2023

We characterize fire regimes in central Portugal and investigate the degree to which the differences between regimes are influenced by a set of biophysical drivers. Using civil parishes as units of analysis, we employ three complementary parameters to describe the fire regime over a reference period of 44 years (1975–2018), namely cumulative percentage of parish area burned, Gini concentration index of burned area over time, and area-weighted total number of wildfires. Cluster analysis is used to aggregate parishes into groups with similar fire regimes based on these parameters. A classification tree model is then used to assess the capacity of a set of potential biophysical drivers to discriminate between the different parish groups. The results allowed us to distinguish four types of fire regime and show that these can be significantly differentiated using the biophysical drivers, of which land use/land cover (LULC), slope, and spring rainfall are the most important. Among LULC classes, shrubland and herbaceous vegetation play the foremost role, followed by agriculture. Our results highlight the importance of vegetation type, availability, and rate of regeneration, as well as that of topography, in influencing fire regimes in the study area, while suggesting that these regimes should be subject to specific wildfire prevention and mitigation policies.
Volatile Oil in Pinus yunnanensis Potentially Contributes to Extreme Fire Behavior
Authors: Chen Feng , Si Liqing , Zhao Fengjun , Wang Mingyu
Published: March 2023

Volatile oils in forest fuel can significantly affect forest fire behavior, especially extreme fire behavior, e.g., deflagration, fire storms, blowups, eruptive fires and crown fires. However, how these oils influence fire behavior remains unclear, as few qualitative studies have been performed globally. In the present study, we compared the volatile oil contents and components in live branches and surface dead fuel of Pinus yunnanensis Franch, which is widely distributed in Southwest China, to explore their potential effects on extreme fire behavior. Fifteen samples of live branches and fifteen samples of surface dead fuel were collected. Volatile oils were extracted from the samples using steam distillation, and their components were identified and analyzed using gas chromatography–mass spectrometry (GC-MS). The results show that the volatile oil content in live branches was as high as 8.28 mL·kg−1 (dry weight) and was significantly higher than that in surface dead fuel (3.55 mL·kg−1). The volatile oil content in the P. yunnanensis forest was 126.12 kg per hectare. The main volatile oil components were terpenoids, of which monoterpenes accounted for the highest proportion based on their content (62.63%), followed by sesquiterpenes (22.44%). The terpenoid compounds in live branches were more abundant than those in surface dead fuel. Monoterpenes and sesquiterpenes in volatile oils in forest fuel have low boiling points, high calorific values and a lower explosion limit (LEL; 38.4 g·m−3), which are important characteristics in the manifestation of extreme fire behavior such as deflagration. The analysis results indicate that when heated, the oily gases from P. yunnanensis forest could fill 3284.26 m3 per hectare, with a gas concentration reaching the LEL. We conclude that volatile oil in P. yunnanensis has an important influence on the manifestation of extreme fire behavior, and live branches have a greater effect than surface dead fuel.
Simulation Test on Cooling and Fire Suppression with Liquid Nitrogen in Computer Room of Data Center
Authors: Meng Jianbing , Wang Tingrong , Li Guanghua , Kang Jianhong
Published: March 2023

With the rapid development of worldwide computer data center construction, the reliability requirements of the fire-fighting system for data center rooms are also increasing. By using the self-designed simulation platform of liquid nitrogen spray, this paper studies the liquid nitrogen cooling process in the initial heating stage of a computer data center room fire and the liquid nitrogen extinguishing effects for common combustible materials, revealing the feasibility of applying liquid nitrogen to the fire extinguishing system for data center room. The results show that the cooling and inerting effects with quarter sector fan-shaped 6520 spray nozzle are the best among seven types of spray nozzles, the relative temperature changes by more than 50% within 5 min, and the oxygen concentration in the test space drops below 10%. Compared with optical fiber, the ignition range of uninterruptible power supply com-bination during combustion is relatively small. Liquid nitrogen has a significant fire-extinguishing effect on two combustible materials, which can successfully extinguish optical fiber and UPS within 3 min and 2 min, respectively.
Application of a Lipopolysaccharide (LPS)-Stimulated Mitogenesis Assay in Smallmouth Bass (Micropterus dolomieu) to Augment Wild Fish Health Studies
Authors: Smith Cheyenne R. , Ottinger Christopher A. , Walsh Heather L. , Mazik Patricia M. , Blazer Vicki S.
Published: March 2023

The utility of a functional immune assay for smallmouth bass (Micropterus dolomieu) lymphocyte mitogenesis was evaluated. Wild populations in the Potomac River have faced disease and mortality with immunosuppression from exposure to chemical contaminants a suspected component. However, a validated set of immune parameters to screen for immunosuppression in wild fish populations is not available. Prior to use in ecotoxicology studies, ancillary factors influencing the mitogenic response need to be understood. The assay was field-tested with fish collected from three sites in West Virginia as part of health assessments occurring in spring (pre-spawn; April–May) and fall (recrudescence; October–November). Anterior kidney leukocytes were exposed to lipopolysaccharide (LPS) from E.coli O111:B4 or mitogen-free media and proliferation was measured using imaging flow cytometry with advanced machine learning to distinguish lymphocytes. An anti-smallmouth bass IgM monoclonal antibody was used to identify IgM+ lymphocytes. Lymphocyte mitogenesis, or proliferative responses, varied by site and season and positively and negatively correlated with factors such as sex, age, tissue parasites, and macrophage aggregates. Background proliferation of IgM− lymphocytes was negatively correlated to LPS-induced proliferation in both seasons at all sites, but only in spring for IgM+ lymphocytes. The results demonstrate that many factors, in addition to chemical contaminants, may influence lymphocyte proliferation.
Simulation of Particulate Matter Structure Detachment from Surfaces of Wall-Flow Filters for Elevated Velocities Applying Lattice Boltzmann Methods
Authors: Hafen Nicolas , Marquardt Jan E. , Dittler Achim , Krause Mathias J.
Published: March 2023

Rearrangement events in wall-flow filters lead to the formation of specific deposition patterns, which affect a filter’s pressure drop, its loading capacity and the separation efficiency. A universal and consistent formulation of probable causes and influence factors does not exist and appropriate calculation models that enable a quantification of respective influence factors are missing. In this work, a previously developed lattice Boltzmann method, which has been used with inflow velocities of up to 2 m s−1, is applied to elevated velocities of up to 60 m s−1. The particle-free flow, a single layer fragment and a deposition layer during break-up are investigated as three different scenarios. One goal of this work is a comprehensive quantification of the stability and accuracy of both particle-free and particle-including flows, considering static, impermeable deposition-layer fragments. A second goal is the determination of the hydrodynamic surface forces and the deduction of the local detachment likelihood of individual layer fragments. Satisfactory stability and accuracy can be shown for fluid velocity, fluid pressure and the hydrodynamic forces. When considering layer fragments, the parameter domain turns out to be limited to inflow velocities of 28 m s−1. It is shown that fragment detachment rather occurs consecutively and regions of no possible detachment are identified. The work contributes to an understanding of rearrangement events and respective deposition pattern predictions and enables potential optimizations in engine performance, fuel consumption and the service life of wall-flow filters.
GIS Models for Sustainable Urban Mobility Planning: Current Use, Future Needs and Potentials
Authors: Liu Xu , Payakkamas Peerawat , Dijk Marc , de Kraker Joop
Published: March 2023

GIS models are currently available for a broad range of applications in mobility planning. However, it is not known how widespread the current use of GIS models is among European urban mobility planners, nor what their user experiences and needs are. There is therefore a risk that the development of GIS models for urban mobility planning will be mainly driven by technical possibilities and data availability rather than by the needs of the prospective users. To inform model developers and ensure a good match between model options and user needs, we conducted a survey investigating the current application of GIS models in urban mobility planning practice in Europe as well as model data availability and the needs and priorities of European mobility planners regarding GIS models. We received 51 valid responses from the transport departments of 42 cities from 21 European countries. For developers of GIS-based traffic models, the findings indicate that in Europe there is scope for wider adoption and further improvement. The models currently used are considered useful to support urban mobility planning, but more than 60% of the surveyed cities do not yet use them. Increased user-friendliness, in particular for non-experts, appears important to promote wider adoption. Availability of non-traditional types of data, such as real-time data or data at neighborhood level, is still limited in most cities, but this may rapidly change. Finally, there is also considerable interest in traffic models that integrate social and environmental aspects.
Indoor Signs Detection for Visually Impaired People: Navigation Assistance Based on a Lightweight Anchor-Free Object Detector
Authors: Said Yahia , Atri Mohamed , Albahar Marwan Ali , Ben Atitallah Ahmed , Alsariera Yazan Ahmad
Published: March 2023

Facilitating the navigation of visually impaired people in indoor environments requires detecting indicating signs and informing them. In this paper, we proposed an indoor sign detection based on a lightweight anchor-free object detection model called FAM-centerNet. The baseline model of this work is the centerNet, which is an anchor-free object detection model with high performance and low computation complexity. A Foreground Attention Module (FAM) was introduced to extract target objects in real scenes with complex backgrounds. This module segments the foreground to extract relevant features of the target object using midground proposal and boxes-induced segmentation. In addition, the foreground module provides scale information to improve the regression performance. Extensive experiments on two datasets prove the efficiency of the proposed model for detecting general objects and custom indoor signs. The Pascal VOC dataset was used to test the performance of the proposed model for detecting general objects, and a custom dataset was used for evaluating the performance in detecting indoor signs. The reported results have proved the efficiency of the proposed FAM in enhancing the performance of the baseline model.
Effects of 3D Postural Correction and Abdominal Muscle Contraction on the Symmetry of the Transverse Abdominis and Spinal Alignment in Patients with Idiopathic Scoliosis
Authors: Yoon Sung-Young , Lee Sang-Yeol
Published: March 2023

This study aimed to investigate the effectiveness of 3D postural correction (3DPC) using corrective cushions (CCs) and abdominal muscle contraction (AMC) on the thickness symmetry of the transversus abdominis (TrA) and spinal alignment in patients with idiopathic scoliosis (IS). In the first experiment, ultrasound measurements were taken of the TrA thickness on both the convex and concave sides of the lumbar curve in the supine position during AMC and non-AMC without 3DPC, and during AMC and non-AMC with 3DPC using CCs, in 11 IS patients. In the second experiment, 37 IS patients participated in a four-week 3DPC exercise program that aimed to maintain TrA thickness symmetry based on the results of the first experiment. The study found that TrA thickness symmetry significantly increased after 3DPC using CCs and combined with AMC (p < 0.05). Additionally, the Cobb angles and trunk rotation angles showed significant decreases, and trunk expansion showed a significant increase (p < 0.05). These results indicate that the simultaneous application of 3DPC and AMC is the most effective way to achieve TrA thickness symmetry in IS patients. Therefore, 3DPC and AMC should be considered as crucial elements in exercise interventions for IS patients.
The Devastating Health Consequences of the Ohio Derailment: A Closer Look at the Effects of Vinyl Chloride Spill
Authors: Sun Wenjie
Published: March 2023

On 3 February 2023, a train derailed in the village of East Palestine, Ohio, home to approximately 4700 residents [...]
The Effect and Cost-Effectiveness of Offering a Combined Lifestyle Intervention for the Prevention of Cardiovascular Disease in Primary Care: Results of the Healthy Heart Stepped-Wedge Trial
Authors: Nieuwenhuijse Emma A. , Vos Rimke C. , van den Hout Wilbert B. , Struijs Jeroen N. , Verkleij Sanne M. , Busch Karin , Numans Mattijs E. , Bonten Tobias N.
Published: March 2023

Objective: To evaluate the effectiveness and cost-effectiveness of offering the combined lifestyle programme “Healthy Heart”, addressing overweight, diet, physical activity, smoking and alcohol, to improve lifestyle behaviour and reduce cardiovascular risk. Design: A practice-based non-randomised stepped-wedge cluster trial with two-year follow-up. Outcomes were obtained via questionnaires and routine care data. A cost–utility analysis was performed. During the intervention period, “Healthy Heart” was offered during regular cardiovascular risk management consultations in primary care in The Hague, The Netherlands. The period prior to the intervention period served as the control period. Results: In total, 511 participants (control) and 276 (intervention) with a high cardiovascular risk were included (overall mean ± SD age 65.0 ± 9.6; women: 56%). During the intervention period, 40 persons (15%) participated in the Healthy Heart programme. Adjusted outcomes did not differ between the control and intervention period after 3–6 months and 12–24 months. Intervention versus control (95% CI) 3–6 months: weight: β −0.5 (−1.08–0.05); SBP β 0.15 (−2.70–2.99); LDL-cholesterol β 0.07 (−0.22–0.35); HDL-cholesterol β −0.03 (−0.10–0.05); physical activity β 38 (−97–171); diet β 0.95 (−0.93–2.83); alcohol OR 0.81 (0.44–1.49); quit smoking OR 2.54 (0.45–14.24). Results were similar for 12–24 months. Mean QALYs and mean costs of cardiovascular care were comparable over the full study period (mean difference (95% CI) QALYs: −0.10 (−0.20; 0.002); costs: EUR 106 (−80; 293)). Conclusions: For both the shorter (3–6 months) and longer term (12–24 months), offering the Healthy Heart programme to high-cardiovascular-risk patients did not improve their lifestyle behaviour nor cardiovascular risk and was not cost-effective on a population level.
Implantable Cardioverter-Defibrillators in Poland Compared with other European Countries from the Patient’s Perspective: Insights from the EHRA Patient Survey
Authors: Januszkiewicz Łukasz , Grabowski Marcin , Farkowski Michał Mirosław , Życiński Paweł , Jędrychowski Tomasz , Pytkowski Mariusz , Chun Julian K. R. , Guerra Jose M. , Conte Giulio , Barra Sérgio , Boveda Serge , Kempa Maciej
Published: March 2023

Background: The aim of this study was to compare differences between Polish ICD recipients and ICD recipients from other European countries in terms of quality of life, information provision before ICD implantation, and end-of-life issues. Methods: This is a sub-analysis of the “Living with an ICD” patient survey (25-item questionnaire) organized by the European Heart Rhythm Association between 12 April 2021 and 5 July 2021 in ten European countries. Results: There were 410 (22.7%) patients from Poland and 1399 (77.3%) from other European countries. A total of 51.0% of Polish patients reported improvement in their quality of life compared with 44.3% in other countries (p = 0.041). Remote monitoring was three times more often utilized in other countries than in Poland (66.8% vs. 21.0%, p < 0.001). While 78.1% of Poles felt well informed before ICD implantation compared with 69.6% of subjects from other countries (p = 0.001), they were less familiar with the ICD deactivation process than others (38.9% vs. 52.5%, p < 0.001). Conclusions: Despite the less frequent use of remote monitoring and gaps in end-of-life issues, Polish ICD recipients reported more favorable quality of life and a higher level of information received before device placement than patients in other European countries.
A Study on the Interactions of Proteinase K with Myricetin and Myricitrin by Multi-Spectroscopy and Molecular Modeling
Authors: Liu Kefan , Zhang Yubo , Zhang Wei , Liu Liyan , Yu Zhan
Published: March 2023

Myricetin (MYR) and myricitrin (MYT) are well recognized for their nutraceutical value, such as antioxidant, hypoglycemic, and hypotensive effects. In this work, fluorescence spectroscopy and molecular modeling were adopted to investigate the conformational and stability changes of proteinase K (PK) in the presence of MYR and MYT. The experimental results showed that both MYR and MYT could quench fluorescence emission via a static quenching mechanism. Further investigation demonstrated that both hydrogen bonding and van der Waals forces play significant roles in the binding of complexes, which is consistent with the conclusions of molecular modeling. Synchronous fluorescence spectroscopy, Förster resonance energy transfer, and site-tagged competition experiments were performed to prove that the binding of MYR or MYT to PK could alter its micro-environment and conformation. Molecular docking results revealed that either MYR or MYT spontaneously interacted with PK at a single binding site via hydrogen bonding and hydrophobic interactions, which is consistent with the results of spectroscopic measurements. A 30 ns molecular dynamics simulation was conducted for both PK-MYR and PK-MYT complexes. The calculation results showed that no large structural distortions or interaction changes occurred during the entire simulation time span. The average RMSD changes of PK in PK-MYR and PK-MYT were 2.06 and 2.15 Å, respectively, indicating excellent stability of both complexes. The molecular simulation results suggested that both MYR and MYT could interact with PK spontaneously, which is in agreement with spectroscopic results. This agreement between experimental and theoretical results indicates that the method herein could be feasible and worthwhile for protein–ligand complex studies.
Deep Learning of Phase-Contrast Images of Cancer Stem Cells Using a Selected Dataset of High Accuracy Value Using Conditional Generative Adversarial Networks
Authors: Zhang Zaijun , Ishihata Hiroaki , Maruyama Ryuto , Kasai Tomonari , Kameda Hiroyuki , Sugiyama Tomoyasu
Published: March 2023

Artificial intelligence (AI) technology for image recognition has the potential to identify cancer stem cells (CSCs) in cultures and tissues. CSCs play an important role in the development and relapse of tumors. Although the characteristics of CSCs have been extensively studied, their morphological features remain elusive. The attempt to obtain an AI model identifying CSCs in culture showed the importance of images from spatially and temporally grown cultures of CSCs for deep learning to improve accuracy, but was insufficient. This study aimed to identify a process that is significantly efficient in increasing the accuracy values of the AI model output for predicting CSCs from phase-contrast images. An AI model of conditional generative adversarial network (CGAN) image translation for CSC identification predicted CSCs with various accuracy levels, and convolutional neural network classification of CSC phase-contrast images showed variation in the images. The accuracy of the AI model of CGAN image translation was increased by the AI model built by deep learning of selected CSC images with high accuracy previously calculated by another AI model. The workflow of building an AI model based on CGAN image translation could be useful for the AI prediction of CSCs.
Identification of CD114 Membrane Receptors as a Molecular Target in Medulloblastomas
Authors: Monteiro Jander Moreira , Reis Ramos Jaqueline Isadora , Teixeira e Sousa Ian , Bighetti-Trevisan Rayana Longo , Ribas Filho Jurandir Marcondes , Isolan Gustavo Rassier
Published: March 2023

Medulloblastomas are the most common solid tumors in children, accounting for 8–30% of pediatric brain cancers. It is a high-grade tumor with aggressive behavior and a typically b poor prognosis. Its treatment includes surgery, chemotherapy, and radiotherapy, and presents high morbidity. Significant clinical, genetic, and prognostic differences exist between its four molecular subgroups: WNT, SHH, Group 3, and Group 4. Many studies seek to develop new chemotherapeutic agents for medulloblastomas through the identification of genes whose expressions are new molecular targets for drugs, such as membrane receptors associated with cell replication. This study aimed to assess the association of CD114 expression with mortality in patients with medulloblastoma. Databases from the Medulloblastoma Advanced Genomics International Consortium (MAGIC) were analyzed, focusing on the expression of the CD114 membrane receptor in different molecular types and its possible association with mortality. Our findings showed different CD114 expressions between Group 3 and other molecular groups, as well as between the molecular subtypes SHH γ and Group 3 α and Group 3 β. There was no statistically significant difference between the other groups and subtypes. Regarding mortality, this study did not find statistical significance in the association between low and high CD114 expressions and mortality. Medulloblastoma is a heterogeneous disease with many subtype variations of its genetic and intracellular signaling pathways. Similarly to this study, which could not demonstrate different CD114 membrane receptor expression patterns between groups, others who sought to associate CD114 expression with mortality in other types of cancer failed to establish a direct association. Since many indications point to the relation of this gene with cancer stem cells (CSCs), it may be part of a more extensive cellular signaling pathway with an eventual association with tumor recurrence. This study found no direct relationship between CD114 expression and mortality in patients with medulloblastoma. Further studies are needed on the intracellular signaling pathways associated with this receptor and its gene (the CSF3R).
Knockout Mutants of OsPUB7 Generated Using CRISPR/Cas9 Revealed Abiotic Stress Tolerance in Rice
Authors: Kim Me-Sun , Ko Seo-Rin , Jung Yu Jin , Kang Kwon-Kyoo , Lee Yung-Jo , Cho Yong-Gu
Published: March 2023

Plants produce and accumulate stress-resistant substances when exposed to abiotic stress, which involves a protein conversion mechanism that breaks down stress-damaged proteins and supplies usable amino acids. Eukaryotic protein turnover is mostly driven by the ubiquitination pathway. Among the three enzymes required for protein degradation, E3 ubiquitin ligase plays a pivotal role in most cells, as it determines the specificity of ubiquitination and selects target proteins for degradation. In this study, to investigate the function of OsPUB7 (Plant U-box gene in Oryza sativa), we constructed a CRISPR/Cas9 vector, generated OsPUB7 gene-edited individuals, and evaluated resistance to abiotic stress using gene-edited lines. A stress-tolerant phenotype was observed as a result of drought and salinity stress treatment in the T2 OsPUB7 gene-edited null lines (PUB7-GE) lacking the T-DNA. In addition, although PUB7-GE did not show any significant change in mRNA expression analysis, it showed lower ion leakage and higher proline content than the wild type (WT). Protein–protein interaction analysis revealed that the expression of the genes (OsPUB23, OsPUB24, OsPUB66, and OsPUB67) known to be involved in stress increased in PUB7-GE and this, by forming a 1-node network with OsPUB66 and OsPUB7, acted as a negative regulator of drought and salinity stress. This result provides evidence that OsPUB7 will be a useful target for both breeding and future research on drought tolerance/abiotic stress in rice.
The Role of Toll-like Receptor 2 (TLR2) in the Development and Progression of Hashimoto’s Disease (HD): A Case Study on Female Patients in Poland
Authors: Klatka Maria , Polak Agnieszka , Mertowska Paulina , Mertowski Sebastian , Łyczba Jakub , Hymos Anna , Korona-Głowniak Izabela , Grywalska Ewelina
Published: March 2023

Chronic lymphocytic thyroiditis, commonly known as HD, is one of the most common thyroid disorders. Due to the diverse factors affecting the etiopathogenesis of this disease (hormonal disorders and genetic and environmental factors), as well as the direct involvement of the immune system, scientists are increasingly willing to undertake research aimed at explaining the impact of the loss of immune tolerance and reactivity of autoantigens on the development of the disease. One of the directions of research in recent years is the role of the innate immune response, particularly Toll-like receptors (TLRs), in the pathogenesis of HD. The purpose of this study was to determine the importance of Toll-like receptor 2 (TLR2) expression on selected populations of immune cells, namely, monocytes (MONs) and dendritic cells (DCs), in the course of HD. Particular attention was paid to the analysis of TLR2’s correlation with clinical parameters and the possibility its use as a potential biomarker molecule in the diagnostic process. Based on the obtained results, we found a statistically significant increase in the percentage of all analyzed populations of immune cells, i.e., mDC BDCA-1+CD19−, pDC BDCA-1+CD123, classical MONs CD14+CD16−, and non-classical MONs CD14+CD16+ showing on their surface TLR2 expression in patients diagnosed with HD compared to the healthy volunteers. Moreover, in the study group, we noted a more than 6-fold increase in the concentration of the soluble form of TLR2 in plasma compared to healthy patients. In addition, the correlation analysis showed significant positive correlations between the level of TLR2 expression on selected subpopulations of immune cells and biochemical indicators of thyroid function. Based on the obtained results, we can assume that TLR2 may be involved in the immunopathogenesis of HD.
Hepatocyte-Specific Triggering of Hepatic Stellate Cell Profibrotic Activation by Apoptotic Bodies: The Role of Hepatoma-Derived Growth Factor, HIV, and Ethanol
Authors: New-Aaron Moses , Koganti Siva Sankar , Ganesan Murali , Kanika Sharma , Kumar Vikas , Wang Weimin , Makarov Edward , Kharbanda Kusum K. , Poluektova Larisa Y. , Osna Natalia A.
Published: March 2023

Liver disease is one of the leading comorbidities in HIV infection. The risk of liver fibrosis development is potentiated by alcohol abuse. In our previous studies, we reported that hepatocytes exposed to HIV and acetaldehyde undergo significant apoptosis, and the engulfment of apoptotic bodies (ABs) by hepatic stellate cells (HSC) potentiates their pro-fibrotic activation. However, in addition to hepatocytes, under the same conditions, ABs can be generated from liver-infiltrating immune cells. The goal of this study is to explore whether lymphocyte-derived ABs trigger HSC profibrotic activation as strongly as hepatocyte-derived ABs. ABs were generated from Huh7.5-CYP2E1 (RLW) cells and Jurkat cells treated with HIV+acetaldehyde and co-culture with HSC to induce their pro-fibrotic activation. ABs cargo was analyzed by proteomics. ABs generated from RLW, but not from Jurkat cells activated fibrogenic genes in HSC. This was driven by the expression of hepatocyte-specific proteins in ABs cargo. One of these proteins is Hepatocyte-Derived Growth Factor, for which suppression attenuates pro-fibrotic activation of HSC. In mice humanized with only immune cells but not human hepatocytes, infected with HIV and fed ethanol, liver fibrosis was not observed. We conclude that HIV+ABs of hepatocyte origin promote HSC activation, which potentially may lead to liver fibrosis progression.
Induction of Antimicrobial Protein S100A15 Expression by Oral Microbial Pathogens Is Toll-like Receptors-Dependent Activation of c-Jun-N-Terminal Kinase (JNK), p38, and NF-κB Pathways
Authors: Selimovic Denis , Kharouf Naji , Carrouel Florence , Hassan Sofie-Yasmin , Flanagan Thomas W. , Hassan Sarah-Lilly , Megahed Mosaad , Haikel Youssef , Santourlidis Simeon , Hassan Mohamed
Published: March 2023

The antimicrobial protein S100A15 belongs to the S100 family, which is differentially expressed in a variety of normal and pathological tissues. Although the function of S100A15 protein has been discussed in several studies, its induction and regulation in oral mucosa, so far, are largely unknown. In this study, we demonstrate that S100A15 is induced by the stimulation of oral mucosa with gram− or gram+ bacterial pathogens, as well as with the purified membrane components, namely lipopolysaccharides (LPS) and lipoteichoic acid (LTA). The stimulation of the human gingival fibroblast (GF) and the human mouth epidermal carcinoma (KB) cell lines with either gram− or gram+ bacterial pathogens or their purified membrane components (LPS and LTA) results in the activation of NF-κB, apoptosis-regulating kinase1 (ASK1), and MAP kinase signaling pathways including, c-Jun N-terminal kinase (JNK) and p38 together with their physiological substrates AP-1 and ATF-2, respectively. Inhibition of S100A15 by antibodies-mediated Toll-like receptor 4 (TLR4) or Toll-like receptor 2 (TLR2) neutralization reveals the induction of S100A15 protein by LPS/gram− bacterial pathogens to be TLR4- dependent mechanism, whereas induction by LTA/gram+ bacterial pathogens to be TLR2- dependent mechanism. Pre-treatment of GF and KB cells with JNK (SP600125), p38 (SB-203580), or NF-κB (Bay11-7082) specific inhibitors further demonstrates the importance of JNK, p38 and NF-κB pathways in the regulation of gram−/gram+ bacterial pathogen-induced S100A15 expression. Our data provide evidence that S100A15 is induced in cancer and non-cancer oral mucosa-derived cell lines by gram−/gram+ bacterial pathogens and provide insight into the molecular mechanisms by which gram− and gram+ bacterial pathogens induce S100A15 expression in the oral mucosa.
Importance, Applications and Features of Assays Measuring SARS-CoV-2 Neutralizing Antibodies
Authors: Gattinger Pia , Ohradanova-Repic Anna , Valenta Rudolf
Published: March 2023

More than three years ago, the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) caused the unforeseen COVID-19 pandemic with millions of deaths. In the meantime, SARS-CoV-2 has become endemic and is now part of the repertoire of viruses causing seasonal severe respiratory infections. Due to several factors, among them the development of SARS-CoV-2 immunity through natural infection, vaccination and the current dominance of seemingly less pathogenic strains belonging to the omicron lineage, the COVID-19 situation has stabilized. However, several challenges remain and the possible new occurrence of highly pathogenic variants remains a threat. Here we review the development, features and importance of assays measuring SARS-CoV-2 neutralizing antibodies (NAbs). In particular we focus on in vitro infection assays and molecular interaction assays studying the binding of the receptor binding domain (RBD) with its cognate cellular receptor ACE2. These assays, but not the measurement of SARS-CoV-2-specific antibodies per se, can inform us of whether antibodies produced by convalescent or vaccinated subjects may protect against the infection and thus have the potential to predict the risk of becoming newly infected. This information is extremely important given the fact that a considerable number of subjects, in particular vulnerable persons, respond poorly to the vaccination with the production of neutralizing antibodies. Furthermore, these assays allow to determine and evaluate the virus-neutralizing capacity of antibodies induced by vaccines and administration of plasma-, immunoglobulin preparations, monoclonal antibodies, ACE2 variants or synthetic compounds to be used for therapy of COVID-19 and assist in the preclinical evaluation of vaccines. Both types of assays can be relatively quickly adapted to newly emerging virus variants to inform us about the magnitude of cross-neutralization, which may even allow us to estimate the risk of becoming infected by newly appearing virus variants. Given the paramount importance of the infection and interaction assays we discuss their specific features, possible advantages and disadvantages, technical aspects and not yet fully resolved issues, such as cut-off levels predicting the degree of in vivo protection.
Combinatorial Network of Transcriptional and miRNA Regulation in Colorectal Cancer
Authors: Kumar Rupesh , Mahmoud Maged Mostafa , Tashkandi Hanaa M. , Haque Shafiul , Harakeh Steve , Ponnusamy Kalaiarasan , Haider Shazia
Published: March 2023

Colorectal cancer is one of the leading causes of cancer-associated mortality across the worldwide. One of the major challenges in colorectal cancer is the understanding of the regulatory mechanisms of biological molecules. In this study, we aimed to identify novel key molecules in colorectal cancer by using a computational systems biology approach. We constructed the colorectal protein–protein interaction network which followed hierarchical scale-free nature. We identified TP53, CTNBB1, AKT1, EGFR, HRAS, JUN, RHOA, and EGF as bottleneck-hubs. The HRAS showed the largest interacting strength with functional subnetworks, having strong correlation with protein phosphorylation, kinase activity, signal transduction, and apoptotic processes. Furthermore, we constructed the bottleneck-hubs’ regulatory networks with their transcriptional (transcription factor) and post-transcriptional (miRNAs) regulators, which exhibited the important key regulators. We observed miR-429, miR-622, and miR-133b and transcription factors (EZH2, HDAC1, HDAC4, AR, NFKB1, and KLF4) regulates four bottleneck-hubs (TP53, JUN, AKT1 and EGFR) at the motif level. In future, biochemical investigation of the observed key regulators could provide further understanding about their role in the pathophysiology of colorectal cancer.