bluuebunny/crossref_metadata_2025 huggingface dataset:
The Optimal Size of a Heterogeneous Air Taxi Fleet in Advanced Air Mobility: A Traffic Demand and Flight Scheduling Approach
Authors: Lindner Martin , Brühl Robert , Berger Marco , Fricke Hartmut
Published: February 2024

Introducing Advanced Air Mobility (AAM) as a novel transportation mode poses unique challenges due to limited practical and empirical data. One of these challenges involves accurately estimating future passenger demand and the required number of air taxis, given uncertainties in modal shift dynamics, induced traffic patterns, and long-term price elasticity. In our study, we use mobility data obtained from a Dresden traffic survey and modal shift rates to estimate the demand for AAM air taxi operations for this regional use case. We organize these operations into an air taxi rotation schedule using a Mixed Integer Linear Programming (MILP) optimization model and set a tolerance for slight deviations from the requested arrival times for higher productivity. The resulting schedule aids in determining the AAM fleet size while accounting for flight performance, energy consumption, and battery charging requirements tailored to three distinct types of air taxi fleets. According to our case study, the methodology produces feasible and high-quality air taxi flight rotations within an efficient computational time of 1.5 h. The approach provides extensive insights into air taxi utilization, charging durations at various locations, and assists in fleet planning that adapts to varying, potentially uncertain, traffic demands. Our findings reveal an average productivity of 12 trips per day per air taxi, covering distances from 13 to 99 km. These outcomes contribute to a sustainable, business-focused implementation of AAM while highlighting the interaction between operational parameters and overall system performance and contributing to vertiport capacity considerations.
The Relationship between Serum Zonulin and Innate Immunity in Patients with Inflammatory Bowel Disease
Authors: Khusainova Gusel , Genkel Vadim , Kuznetsova Alla , Nikushkina Karina , Saenko Anna , Abramovskikh Olga , Dolgushina Anastasiya
Published: February 2024

The aim of the study was to investigate the relationship between the serum zonulin and the cellular immunity in patients with ulcerative colitis (UC) and Crohn’s disease (CD). The study included 97 patients, 13 (13.4%) patients with CD and 84 (86.6%) patients with UC. The concentration of zonulin in the serum was studied using the ELISA kits. The investigation of the circulated leukocyte subpopulation was carried out through flow cytometry. The functional activity of the circulating neutrophils was studied using the absorption capacity of the monodisperse polystyrene latex particles and indicators of oxygen-dependent metabolism in the nitroblue tetrazolium test. The serum zonulin concentration in CD patients was significantly higher compared with UC patients (p = 0.003). The zonulin concentration directly correlated with the functional activity of the circulating neutrophils. Patients with a zonulin concentration > 472.4 pg/mL had a significantly higher number of band neutrophils (p = 0.0104), CD3+CD8+ cells (p = 0.0212), NK cells (p = 0.0161), and lower–CD19+ cells (p = 0.0034). Among the IBD patients, zonulin was associated with IBD severity. An increase in the serum concentration of zonulin was associated with an increase in the functional activity of circulating neutrophils and an increase in the number of CD3+CD8+ cells, NK cells, and a decrease in the number of CD19+ cells.
Performance of a 74-Microhaplotype Assay in Kinship Analyses
Authors: Tomas Carmen , Rodrigues Pedro , Jønck Carina G. , Barekzay Zohal , Simayijiang Halimureti , Pereira Vania , Børsting Claus
Published: February 2024

Microhaplotypes (MHs) consisting of multiple SNPs and indels on short stretches of DNA are new and interesting loci for forensic genetic investigations. In this study, we analysed 74 previously defined MHs in two of the populations that our laboratory provides with forensic genetic services, Danes and Greenlanders. In addition to the 229 SNPs that originally made up the 74 MHs, 66 SNPs and 3 indels were identified in the two populations, and 45 of these variants were included in new definitions of the MHs, whereas 24 SNPs were considered rare and of little value for case work. The average effective number of alleles (Ae) was 3.2, 3.0, and 2.6 in Danes, West Greenlanders, and East Greenlanders, respectively. High levels of linkage disequilibrium were observed in East Greenlanders, which reflects the characteristics of this population that has a small size, and signs of admixture and substructure. Pairwise kinship simulations of full siblings, half-siblings, first cousins, and unrelated individuals were performed using allele frequencies from MHs, STRs and SNPs from Danish and Greenlandic populations. The MH panel outperformed the currently used STR and SNP marker sets and was able to differentiate siblings from unrelated individuals with a 0% false positive rate and a 1.1% false negative rate using an LR threshold of 10,000 in the Danish population. However, the panel was not able to differentiate half-siblings or first cousins from unrelated individuals. The results generated in this study will be used to implement MHs as investigative markers for relationship testing in our laboratory.
Comparative Analyses of Complete Chloroplast Genomes of Microula sikkimensis and Related Species of Boraginaceae
Authors: Gao Yunqing , Chen Zhenjiang , Li Xiuzhang , Malik Kamran , Li Chunjie
Published: February 2024

The present study provides a detailed analysis of the chloroplast genome of Microula sikkimensis. The genome consisted of a total of 149,428 bp and four distinct regions, including a large single-copy region (81,329 bp), a small single-copy region (17,261 bp), and an inverted repeat region (25,419 bp). The genome contained 112 genes, including 78 protein-coding genes, 30 tRNA genes, and 4 rRNA genes, and some exhibited duplication in the inverted repeat region. The chloroplast genome displayed different GC content across regions, with the inverted repeat region exhibiting the highest. Codon usage analysis and the identification of simple sequence repeats (SSRs) offer valuable genetic markers. Comparative analysis with other Boraginaceae species highlighted conservation and diversity in coding and noncoding regions. Phylogenetic analysis placed M. sikkimensis within the Boraginaceae family, revealing its distinct relationship with specific species.
Frequencies of BCR::ABL1 Transcripts in Patients with Chronic Myeloid Leukemia: A Meta-Analysis
Authors: Romero-Morelos Pablo , González-Yebra Ana , Muñoz-López Daniela , Lara-Lona Elia , González-Yebra Beatriz
Published: February 2024

Chronic myeloid leukemia (CML) is associated with the Philadelphia chromosome and distinct BCR::ABL1 gene transcripts. We assessed the frequencies of these transcripts in Mexico, Latin America, and worldwide. We determined the prevalence of BCR::ABL1 transcripts in CML patients and intercontinental or regional variations using specialized databases and keywords. We analyzed 34 studies from 20 countries, encompassing 5795 patients. Keyword-based searches in specialized databases guided data collection. ANOVA was employed for transcript distribution analysis. The b3a2 transcript was most prevalent globally, followed by b2a2, with e1a2 being the least frequent. Interestingly, Mexico City exhibited a higher incidence of b2a2, while b3a2 predominated in the remaining country. Overall, no significant intercontinental or regional variations were observed. b3a2 was the most common BCR::ABL1 transcript worldwide, with b2a2 following closely; e1a2 was infrequent. Notably, this trend remained consistent in Mexico. Evaluating transcript frequencies holds clinical relevance for CML management. Understanding the frequency of transcript informs personalized CML treatments.
Serial Dual Mediating Effects of Parenting Stress on Life Satisfaction among Parents of School-Aged Children with Chronic Conditions
Authors: Han Jeong-Won , Yang Boeun , Lee Hanna
Published: February 2024

This study examines the serial dual mediating effects of marital conflict and self-esteem on the relationship between parenting stress and life satisfaction in parents of school-aged children with chronic conditions. This study aims to present foundational data for developing nursing interventions for parents caring for children with chronic illnesses. Of the 2150 parents who participated in the 13th Panel Study on Korean Children (PSKC), 271 raising a child with a chronic illness were enrolled in the study. The serial dual mediating effect was analyzed using PROCESS macro Model 6. The serial dual mediating effects of parenting stress, marital conflict, and self-esteem on parents’ life satisfaction were analyzed. Marital conflict and self-esteem had significant serial multiple mediating effects on the relationship between stress and life satisfaction in fathers (B = −0.11, bootstrap 95% CI = −0.16–−0.06) and mothers (B = −0.06, bootstrap 95% CI = −0.09–−0.03). Our results suggest that marital conflict increases with increasing parenting stress and that increased marital conflict sequentially reduces self-esteem, ultimately diminishing life satisfaction in parents raising a child with a chronic condition. Thus, relevant nursing interventions and social support are essential to boost the life satisfaction of parents raising children with chronic conditions.
Compatibility Assessment in the Replacement of Damaged Sandstone Used in the Cathedral of Huesca (Spain)
Authors: Lapuente Mercadal María Pilar , Cuchí Oterino José Antonio , Auqué Sanz Luis Francisco
Published: February 2024

In order to manage problems arising from rainwater/rock interaction in Miocene sandstones (calcareous litharenites) widely used in various monuments of the Ebro Valley (NE of Spain), a survey has been conducted with particular application to the building and architectural decorative materials of the Cathedral of Huesca. Once the current state of decay was diagnosed and the processes of alteration (enhanced by certain intrinsic factors and their particular exposure to the environmental conditions) were detected, a pre-restoration experimental assay was conducted. On the one hand, to propose the best stone replacement, this study evaluates the compatibility of the available sandstones in the local market based on their intrinsic features, especially those related to hydric behaviour. Once the most suitable sandstone was selected, pore size distributions were determined along with accelerated ageing cycles to show the importance of selecting properly the potential replacement sandstone. In a second step, to determine the effectiveness and long-term efficacy of four water-repellent products, several on-site and laboratory tests were performed. From the experimental results obtained, remedial works have been proposed which will be useful not only for the restoration programme of this monument, but also for other emblematic architectural Heritage in the Ebro Valley.
Pre-Service Teachers’ Perceptions on the Use of Heritage in Secondary Education and Their Reception of Educational Materials from the Heritage and Museum Sector: A Case Study in Flanders (Belgium)
Authors: Van Doorsselaere Joris
Published: February 2024

There has been a growing policy interest in establishing connections between heritage and education. Nevertheless, there seems to be very little evaluation or critical reflection on what actually happens in practice, and it remains unclear how heritage education is employed in countries throughout Europe. The aim of this paper is to assess the current status of heritage education in Flanders (the Dutch-speaking northern part of Belgium) via a literature review and an exploratory case study. The perceptions and opinions of pre-service teachers (n = 17) were investigated using three instruments: a questionnaire, document analysis, and the think aloud method. The results show that the pre-service teachers had a traditional interpretation of heritage, mainly relying on well-known and monumental examples and following a rather historical–artistic conception. However, it was found that their opinions were positive towards the use of heritage as an educational resource, and their evaluations of educational materials from the heritage and museum sector provided detailed information concerning teachers’ desires and needs in this regard. The implications of this study should encourage initial teacher training in Flanders to further consider the epistemological and methodological challenges in the emerging field of heritage education.
Techno-Economic Analysis of Cement Decarbonization Techniques: Oxygen Enrichment vs. Hydrogen Fuel
Authors: Domingues Bruno C. , Santos Diogo M. F. , Mateus Margarida , Cecílio Duarte
Published: February 2024

The Paris Agreement aims to limit global warming, and one of the most polluting sectors is heavy industry, where cement production is a significant contributor. This work briefly explores some alternatives, recycling, reducing clinker content, waste heat recovery, and carbon capture, discussing their advantages and drawbacks. Then, it examines the economic viability and benefits of increasing oxygen concentration in the primary burning air from 21 to 27 vol.%, which could improve clinker production by 7%, and the production of hydrogen through PEM electrolysis to make up 5% of the fuel thermal fraction, considering both in a cement plant producing 3000 tons of clinker per day. This analysis used reference values from Secil, an international company for cement and building materials, to determine the required scale of the oxygen and hydrogen production, respectively, and calculate the CAPEX of each approach. It is concluded that oxygen enrichment can provide substantial fuel savings for a relatively low cost despite a possible significant increase in NOx emissions. However, hydrogen production at this scale is not currently economically viable.
Perspectives and Attitudes of Newer New Jersey High School Teachers towards Cleaning, Sanitizing, and Disinfecting Consumer Products Used in School Classrooms
Authors: Aggarwal Juhi , Campbell Maryanne L. , Rehman Midhat , Nguyen Kimberly T. , Shendell Derek G.
Published: February 2024

During the COVID-19 pandemic, there was an increased reported use of chemical cleaning, sanitizing, and disinfecting products (CSDPs), which created public concerns about negative health consequences for both children and adults in public schools. A subset of newer teachers shared experiences regarding safety and health (S&H) while working in school-based settings through a series of online surveys. Surveys were provided to teachers who completed work-based learning supervisory trainings provided by the New Jersey Safe Schools Program between October 2021 and June 2023. The participants answered questions focusing on CSDPs purchased for school use, their attitudes towards CSDPs, their use of personal protective equipment, and symptoms employees may have had due to CSDPs. A total of 205 teacher participants successfully completed the surveys. Over 25% of the teachers did not know where their CSDPs originated from, as they were provided by the school. Most participants “sometimes”, “not often”, or “never” read labels for CSDP ingredients or looked them up on healthy product apps. The participants (60%) tended to wear gloves while cleaning/disinfecting but did not wear masks. A third of the participants experienced respiratory health problems after working at school. Overall, the data suggest that more education on S&H regarding CSDPs needs to be provided to New Jersey teachers.
SARS-CoV-2 and Adolescent Psychiatric Emergencies at the Tübingen University Hospital: Analyzing Trends, Diagnoses, and Contributing Factors
Authors: Schneider Priska S. , Pantis Michelle , Preiser Christine , Hagmann Daniela , Barth Gottfried M. , Renner Tobias J. , Allgaier Katharina
Published: February 2024

Psychiatric emergencies have increased in recent decades, particularly with the onset of the SARS-CoV-2 pandemic, and far too little is known about the backgrounds of these emergencies. In this study, we investigated the extent to which the number of psychiatric emergencies changed during and in the aftermath of the SARS-CoV-2 pandemic at the Child and Adolescent Psychiatry (CAP) Tübingen. We considered age and psychiatric diagnoses. Additionally, we evaluated the backgrounds of the emergencies. We applied a mixed- (quantitative and qualitative) methods approach to data on emergency presentations at the CAP Tübingen from the pre-SARS-CoV-2 pandemic period (October 2019–January 2020) to Restriction Phase 1 (October 2020–January 2021), Restriction Phase 2 (October 2021–January 2022), and endemic phase (October 2022–January 2023). The total number of emergencies and those with eating disorders and affective disorders increased, while obsessive–compulsive disorders, expansive disorders and anxiety disorders decreased. The patients presenting in the pre-SARS-CoV-2 pandemic period were younger than those in the subsequent periods. We content-coded the reasons behind the emergency presentations. We also identified four areas of stressors and personality characteristics associated with the emergency presentations. In light of the increasing number of psychiatric emergencies, the long-term aim should be to meet the growing demands and create options for prevention.
Historical Heritage Maintenance via Scan-to-BIM Approaches: A Case Study of the Lisbon Agricultural Exhibition Pavilion
Authors: Rocha Gustavo , Mateus Luís , Ferreira Victor
Published: February 2024

Building Information Modeling (BIM) has emerged as a revolutionary tool in the domain of architectural conservation and documentation. When combined with terrestrial 3D laser scanning, it presents a powerful method to capture and represent the intricate details and nuances of historic structures. Such buildings, with their unique architectural lineage, often exude a geometric complexity unparalleled by standard designs. Consequently, the transition from scan data to a BIM framework, or the scan-to-BIM process, becomes intricate and time-intensive. Beyond the challenge of digital translation, the true essence of these historic buildings lies not only in their geometric form but also in understanding and preserving their design logic, formal composition rules, and primitive geometry. It then becomes imperative that the resulting model maintains fidelity in terms of proportion, shape, symmetry, and spatial rationale. Considering these challenges and potentials, this article delves into the process of digitalizing and BIM modeling of the Lisbon Agricultural Exhibition Pavilion located in Portugal. Our study proceeds in a tripartite structure: initiating with an in-depth terrestrial 3D laser scanning of the pavilion, followed by a comprehensive registration, processing, and alignment of the acquired scans, and culminating in a detailed BIM model using the industry-standard Revit 2020 software.
Evaluation of the Accessibility of Children’s Spaces at the Community Scale: The Case Study of Hangzhou
Authors: Cui Yuanzheng , Wang Qiuting , Zha Guixiang , Dang Yunxiao , Duan Xuejun , Wang Lei , Luo Ming
Published: February 2024

The safety, inclusivity, accessibility, and green communities emphasized in the United Nations’ Sustainable Development Goals (SDGs) play a vital role in the establishment of child-friendly cities. The governments are actively promoting the development of sustainable, child-friendly cities that prioritize people’s needs and aim to enhance the well-being of residents, from children to families. However, there is limited research utilizing GIS analysis techniques and internet big data to analyze spatial equity in children’s spatial accessibility. Therefore, this study introduces an innovative approach focusing on the community level. Drawing on data from the popular social networking platform mobile application “Xiaohongshu” and employing network analysis methods based on walking and driving modes, this study analyzed and investigated the accessibility of children’s spaces in the city of Hangzhou, China. Regarding spatial characteristics, the distribution of children’s space resources in the main urban area of Hangzhou exhibited a “peripheral low and central high” trend, which was closely linked to the distribution of population space. This pattern indicates potential significant disparities in the allocation of children’s space resources. Notably, the core area of Hangzhou demonstrated the highest level of accessibility to children’s spaces, with Gongshu District exhibiting the best accessibility. Conversely, non-core urban areas generally had relatively poor accessibility. Furthermore, different types of children’s spaces, such as indoor cultural spaces, indoor entertainment spaces, outdoor parks, and outdoor nature areas, all exhibited the highest accessibility in the city center, which gradually decreased towards the periphery. Additionally, this study evaluated the convenience of children’s spaces in various communities by combining population size and accessibility levels. The findings revealed that communities in the core area had higher accessibility levels in the northwest–southeast direction, while accessibility decreased towards the northeast–southwest direction. Consequently, the relative convenience of these communities tended to be lower. By examining spatial equity, this study provides valuable insights into the promotion of sustainable, child-friendly cities that prioritize people’s needs and contribute to the well-being of residents, from children to families.
The Role of Mechanotransduction in Contact Inhibition of Locomotion and Proliferation
Authors: Nakamura Fumihiko
Published: February 2024

Contact inhibition (CI) represents a crucial tumor-suppressive mechanism responsible for controlling the unbridled growth of cells, thus preventing the formation of cancerous tissues. CI can be further categorized into two distinct yet interrelated components: CI of locomotion (CIL) and CI of proliferation (CIP). These two components of CI have historically been viewed as separate processes, but emerging research suggests that they may be regulated by both distinct and shared pathways. Specifically, recent studies have indicated that both CIP and CIL utilize mechanotransduction pathways, a process that involves cells sensing and responding to mechanical forces. This review article describes the role of mechanotransduction in CI, shedding light on how mechanical forces regulate CIL and CIP. Emphasis is placed on filamin A (FLNA)-mediated mechanotransduction, elucidating how FLNA senses mechanical forces and translates them into crucial biochemical signals that regulate cell locomotion and proliferation. In addition to FLNA, trans-acting factors (TAFs), which are proteins or regulatory RNAs capable of directly or indirectly binding to specific DNA sequences in distant genes to regulate gene expression, emerge as sensitive players in both the mechanotransduction and signaling pathways of CI. This article presents methods for identifying these TAF proteins and profiling the associated changes in chromatin structure, offering valuable insights into CI and other biological functions mediated by mechanotransduction. Finally, it addresses unanswered research questions in these fields and delineates their possible future directions.
Transcriptomic Analyses Reveal Insights into the Shared Regulatory Network of Phenolic Compounds and Steviol Glycosides in Stevia rebaudiana
Authors: Simoni Samuel , Vangelisti Alberto , Clemente Clarissa , Usai Gabriele , Santin Marco , Ventimiglia Maria , Mascagni Flavia , Natali Lucia , Angelini Luciana G. , Cavallini Andrea , Tavarini Silvia , Giordani Tommaso
Published: February 2024

Stevia rebaudiana (Bertoni) is a highly valuable crop for the steviol glycoside content in its leaves, which are no-calorie sweeteners hundreds of times more potent than sucrose. The presence of health-promoting phenolic compounds, particularly flavonoids, in the leaf of S. rebaudiana adds further nutritional value to this crop. Although all these secondary metabolites are highly desirable in S. rebaudiana leaves, the genes regulating the biosynthesis of phenolic compounds and the shared gene network between the regulation of biosynthesis of steviol glycosides and phenolic compounds still need to be investigated in this species. To identify putative candidate genes involved in the synergistic regulation of steviol glycosides and phenolic compounds, four genotypes with different contents of these compounds were selected for a pairwise comparison RNA-seq analysis, yielding 1136 differentially expressed genes. Genes that highly correlate with both steviol glycosides and phenolic compound accumulation in the four genotypes of S. rebaudiana were identified using the weighted gene co-expression network analysis. The presence of UDP-glycosyltransferases 76G1, 76H1, 85C1, and 91A1, and several genes associated with the phenylpropanoid pathway, including peroxidase, caffeoyl-CoA O-methyltransferase, and malonyl-coenzyme A:anthocyanin 3-O-glucoside-6″-O-malonyltransferase, along with 21 transcription factors like SCL3, WRK11, and MYB111, implied an extensive and synergistic regulatory network involved in enhancing the production of such compounds in S. rebaudiana leaves. In conclusion, this work identified a variety of putative candidate genes involved in the biosynthesis and regulation of particular steviol glycosides and phenolic compounds that will be useful in gene editing strategies for increasing and steering the production of such compounds in S. rebaudiana as well as in other species.
The Effects of Aging on Sarcoplasmic Reticulum-Related Factors in the Skeletal Muscle of Mice
Authors: Kanazawa Yuji , Takahashi Tatsuo , Nagano Mamoru , Koinuma Satoshi , Shigeyoshi Yasufumi
Published: February 2024

The pathogenesis of sarcopenia includes the dysfunction of calcium homeostasis associated with the sarcoplasmic reticulum; however, the localization in sarcoplasmic reticulum-related factors and differences by myofiber type remain unclear. Here, we investigated the effects of aging on sarcoplasmic reticulum-related factors in the soleus (slow-twitch) and gastrocnemius (fast-twitch) muscles of 3- and 24-month-old male C57BL/6J mice. There were no notable differences in the skeletal muscle weight of these 3- and 24-month-old mice. The expression of Atp2a1, Atp2a2, Sln, and Pln increased with age in the gastrocnemius muscles, but not in the soleus muscles. Subsequently, immunohistochemical analysis revealed ectopic sarcoplasmic reticulum calcium ion ATPase (SERCA) 1 and SERCA2a immunoreactivity only in the gastrocnemius muscles of old mice. Histochemical and transmission electron microscope analysis identified tubular aggregate (TA), an aggregation of the sarcoplasmic reticulum, in the gastrocnemius muscles of old mice. Dihydropyridine receptor α1, ryanodine receptor 1, junctophilin (JPH) 1, and JPH2, which contribute to sarcoplasmic reticulum function, were also localized in or around the TA. Furthermore, JPH1 and JPH2 co-localized with matrix metalloproteinase (MMP) 2 around the TA. These results suggest that sarcoplasmic reticulum-related factors are localized in or around TAs that occur in fast-twitch muscle with aging, but some of them might be degraded by MMP2.
Roles of Interferon Regulatory Factor 1 in Tumor Progression and Regression: Two Sides of a Coin
Authors: Perevalova Alina M. , Gulyaeva Lyudmila F. , Pustylnyak Vladimir O.
Published: February 2024

IRF1 is a transcription factor well known for its role in IFN signaling. Although IRF1 was initially identified for its involvement in inflammatory processes, there is now evidence that it provides a function in carcinogenesis as well. IRF1 has been shown to affect several important antitumor mechanisms, such as induction of apoptosis, cell cycle arrest, remodeling of tumor immune microenvironment, suppression of telomerase activity, suppression of angiogenesis and others. Nevertheless, the opposite effects of IRF1 on tumor growth have also been demonstrated. In particular, the “immune checkpoint” molecule PD-L1, which is responsible for tumor immune evasion, has IRF1 as a major transcriptional regulator. These and several other properties of IRF1, including its proposed association with response and resistance to immunotherapy and several chemotherapeutic drugs, make it a promising object for further research. Numerous mechanisms of IRF1 regulation in cancer have been identified, including genetic, epigenetic, transcriptional, post-transcriptional, and post-translational mechanisms, although their significance for tumor progression remains to be explored. This review will focus on the established tumor-suppressive and tumor-promoting functions of IRF1, as well as the molecular mechanisms of IRF1 regulation identified in various cancers.
A Dinuclear Copper(II) Complex Electrochemically Obtained via the Endogenous Hydroxylation of a Carbamate Schiff Base Ligand: Synthesis, Structure and Catalase Activity
Authors: Fernández-Fariña Sandra , Velo-Heleno Isabel , Rodríguez-Silva Laura , Maneiro Marcelino , González-Noya Ana M. , Pedrido Rosa
Published: February 2024

In the present work, we report a neutral dinuclear copper(II) complex, [Cu2(L1)(OH)], derived from a new [N,O] donor Schiff base ligand L1 that was formed after the endogenous hydroxylation of an initial carbamate Schiff base H2L coordinated with copper ions in an electrochemical cell. The copper(II) complex has been fully characterized using different techniques, including X-ray diffraction. Direct current (DC) magnetic susceptibility measurements were also performed at variable temperatures, showing evidence of antiferromagnetic behavior. Its catalase-like activity was also tested, demonstrating that this activity is affected by temperature.
Interferon-γ as a Potential Inhibitor of SARS-CoV-2 ORF6 Accessory Protein
Authors: Krachmarova Elena , Petkov Peicho , Lilkova Elena , Stoynova Dayana , Malinova Kristina , Hristova Rossitsa , Gospodinov Anastas , Ilieva Nevena , Nacheva Genoveva , Litov Leandar
Published: February 2024

The ORF6 protein of the SARS-CoV-2 virus plays a crucial role in blocking the innate immune response of the infected cells by inhibiting interferon pathways. Additionally, it binds to and immobilises the RAE1 protein on the cytoplasmic membranes, thereby blocking mRNA transport from the nucleus to the cytoplasm. In all these cases, the host cell proteins are tethered by the flexible C-terminus of ORF6. A possible strategy to inhibit the biological activity of ORF6 is to bind its C-terminus with suitable ligands. Our in silico experiments suggest that hIFNγ binds the ORF6 protein with high affinity, thus impairing its interactions with RAE1 and, consequently, its activity in viral invasion. The in vitro studies reported here reveal a shift of the localisation of RAE1 in ORF6 overexpressing cells upon treatment with hIFNγ from predominantly cytoplasmic to mainly nuclear, resulting in the restoration of the export of mRNA from the nucleus. We also explored the expression of GFP in transfected-with-ORF6 cells by means of fluorescence microscopy and qRT-PCR, finding that treatment with hIFNγ unblocks the mRNA trafficking and reinstates the GFP expression level. The ability of the cytokine to block ORF6 is also reflected in minimising its negative effects on DNA replication by reducing accumulated RNA-DNA hybrids. Our results, therefore, suggest hIFNγ as a promising inhibitor of the most toxic SARS-CoV-2 protein.
MicroRNAs in Testicular Germ Cell Tumors: The Teratoma Challenge
Authors: Yodkhunnatham Nuphat , Pandit Kshitij , Puri Dhruv , Yuen Kit L. , Bagrodia Aditya
Published: February 2024

Testicular germ cell tumors (TGCTs) are relatively common in young men, making accurate diagnosis and prognosis assessment essential. MicroRNAs (miRNAs), including microRNA-371a-3p (miR-371a-3p), have shown promise as biomarkers for TGCTs. This review discusses the recent advancements in the use of miRNA biomarkers in TGCTs, with a focus on the challenges surrounding the noninvasive detection of teratomas. Circulating miR-371a-3p, which is expressed in undifferentiated TGCTs but not in teratomas, is a promising biomarker for TGCTs. Its detection in serum, plasma, and, potentially, cystic fluid could be useful for TGCT diagnosis, surveillance, and monitoring of therapeutic response. Other miRNAs, such as miR-375-3p and miR-375-5p, have been investigated to differentiate between TGCT subtypes (teratoma, necrosis/fibrosis, and viable tumors), which can aid in treatment decisions. However, a reliable marker for teratoma has yet to be identified. The clinical applications of miRNA biomarkers could spare patients from unnecessary surgeries and allow for more personalized therapeutic approaches. Particularly in patients with residual masses larger than 1 cm following chemotherapy, it is critical to differentiate between viable tumors, teratomas, and necrosis/fibrosis. Teratomas, which mimic somatic tissues, present a challenge in differentiation and require a comprehensive diagnostic approach. The combination of miR-371 and miR-375 shows potential in enhancing diagnostic precision, aiding in distinguishing between teratomas, viable tumors, and necrosis. The implementation of miRNA biomarkers in TGCT care could improve patient outcomes, reduce overtreatment, and facilitate personalized therapeutic strategies. However, a reliable marker for teratoma is still lacking. Future research should focus on the clinical validation and standardization of these biomarkers to fully realize their potential.
Effect of Temperature on the Development of Stages of Spermatogenesis and the Functionality of Sertoli Cells In Vitro
Authors: Jorban Areej , Lunenfeld Eitan , Huleihel Mahmoud
Published: February 2024

Spermatogenesis is the process of proliferation and differentiation of spermatogonial cells to meiotic and post-meiotic stages and sperm generation. Normal spermatogenesis occurs in vivo at 34 °C to 35 °C, and high temperatures are known to cause male infertility. The aim of the present study was to examine the effect of temperature (35 °C compared to 37 °C) on the viability/apoptosis of developed cells, on the development of different stages of spermatogenesis in 3D in vitro culture conditions, and the functionality of Sertoli cells under these conditions. We used isolated cells from seminiferous tubules of sexually immature mice. The cells were cultured in methylcellulose (as a three-dimensional (3D) in vitro culture system) and incubated in a CO2 incubator at 35 °C or 37 °C. After two to six weeks, the developed cells and organoids were collected and examined for cell viability and apoptosis markers. The development of different stages of spermatogenesis was evaluated by immunofluorescence staining or qPCR analysis using specific antibodies or primers, respectively, for cells at each stage. Factors that indicate the functionality of Sertoli cells were assessed by qPCR analysis. The developed organoids were examined by a confocal microscope. Our results show that the percentages and/or the expression levels of the developed pre-meiotic, meiotic, and post-meiotic cells were significantly higher at 35 °C compared to those at 37 °C, including the expression levels of the androgen receptor, the FSH receptor, transferrin, the androgen-binding protein (ABP), and the glial-derived nerve growth factor (GDNF) which were similarly significantly higher at 35 °C than at 37 °C. The percentages of apoptotic cells (according to acridine orange staining) and the expression levels of BAX, FAS, and CASPAS 3 were significantly higher in cultures incubated at 37 °C compared to those incubated at 35 °C. These findings support the in vivo results regarding the negative effect of high temperatures on the process of spermatogenesis and suggest a possible effect of high temperatures on the viability/apoptosis of spermatogenic cells. In addition, increasing the temperature in vitro also impaired the functionality of Sertoli cells. These findings may deepen our understanding of the mechanisms behind optimal conditions for normal spermatogenesis in vivo and in vitro.
Protective Effect of Betulin on Streptozotocin–Nicotinamide-Induced Diabetes in Female Rats
Authors: Adepoju Feyisayo O. , Sokolova Ksenia V. , Gette Irina F. , Danilova Irina G. , Tsurkan Mikhail V. , Mondragon Alicia C. , Kovaleva Elena G. , Miranda Jose Manuel
Published: February 2024

Type 2 diabetes is characterized by hyperglycemia and a relative loss of β–cell function. Our research investigated the antidiabetic potential of betulin, a pentacyclic triterpenoid found primarily in birch bark and, intriguingly, in a few marine organisms. Betulin has been shown to possess diverse biological activities, including antioxidant and antidiabetic activities; however, no studies have fully explored the effects of betulin on the pancreas and pancreatic islets. In this study, we investigated the effect of betulin on streptozotocin–nicotinamide (STZ)-induced diabetes in female Wistar rats. Betulin was prepared as an emulsion, and intragastric treatments were administered at doses of 20 and 50 mg/kg for 28 days. The effect of treatment was assessed by analyzing glucose parameters such as fasting blood glucose, hemoglobin A1C, and glucose tolerance; hepatic and renal biomarkers; lipid peroxidation; antioxidant enzymes; immunohistochemical analysis; and hematological indices. Administration of betulin improved the glycemic response and decreased α–amylase activity in diabetic rats, although insulin levels and homeostatic model assessment for insulin resistance (HOMA–IR) scores remained unchanged. Furthermore, betulin lowered the levels of hepatic biomarkers (aspartate aminotransferase, alanine aminotransferase, and alpha-amylase activities) and renal biomarkers (urea and creatine), in addition to improving glutathione levels and preventing the elevation of lipid peroxidation in diabetic animals. We also found that betulin promoted the regeneration of β–cells in a dose-dependent manner but did not have toxic effects on the pancreas. In conclusion, betulin at a dose of 50 mg/kg exerts a pronounced protective effect against cytolysis, diabetic nephropathy, and damage to the acinar pancreas and may be a potential treatment option for diabetes.
The Role of the Vascular System in Degenerative Diseases: Mechanisms and Implications
Authors: Sheikh Abdullah Md. , Yano Shozo , Tabassum Shatera , Nagai Atsushi
Published: February 2024

Degenerative diseases, encompassing a wide range of conditions affecting various organ systems, pose significant challenges to global healthcare systems. This comprehensive review explores the intricate interplay between the vascular system and degenerative diseases, shedding light on the underlying mechanisms and profound implications for disease progression and management. The pivotal role of the vascular system in maintaining tissue homeostasis is highlighted, as it serves as the conduit for oxygen, nutrients, and immune cells to vital organs and tissues. Due to the vital role of the vascular system in maintaining homeostasis, its dysfunction, characterized by impaired blood flow, endothelial dysfunction, and vascular inflammation, emerges as a common denominator of degenerative diseases across multiple systems. In the nervous system, we explored the influence of vascular factors on neurodegenerative diseases such as Alzheimer’s and Parkinson’s, emphasizing the critical role of cerebral blood flow regulation and the blood–brain barrier. Within the kidney system, the intricate relationship between vascular health and chronic kidney disease is scrutinized, unraveling the mechanisms by which hypertension and other vascular factors contribute to renal dysfunction. Throughout this review, we emphasize the clinical significance of understanding vascular involvement in degenerative diseases and potential therapeutic interventions targeting vascular health, highlighting emerging treatments and prevention strategies. In conclusion, a profound appreciation of the role of the vascular system in degenerative diseases is essential for advancing our understanding of degenerative disease pathogenesis and developing innovative approaches for prevention and treatment. This review provides a comprehensive foundation for researchers, clinicians, and policymakers seeking to address the intricate relationship between vascular health and degenerative diseases in pursuit of improved patient outcomes and enhanced public health.
The Human Mutation K237_V238del in a Putative Lipid Binding Motif within the V-ATPase a2 Isoform Suggests a Molecular Mechanism Underlying Cutis Laxa
Authors: Chu Anh , Yao Yeqi , Glibowicka Miroslawa , Deber Charles M. , Manolson Morris F.
Published: February 2024

Vacuolar ATPases (V-ATPases), proton pumps composed of 16 subunits, are necessary for a variety of cellular functions. Subunit “a” has four isoforms, a1–a4, each with a distinct cellular location. We identified a phosphoinositide (PIP) interaction motif, KXnK(R)IK(R), conserved in all four isoforms, and hypothesize that a/PIP interactions regulate V-ATPase recruitment/retention to different organelles. Among the four isoforms, a2 is enriched on Golgi with a2 mutations in the PIP motif resulting in cutis laxa. We hypothesize that the hydrophilic N-terminal (NT) domain of a2 contains a lipid-binding domain, and mutations in this domain prevent interaction with Golgi-enriched PIPs, resulting in cutis laxa. We recreated the cutis laxa-causing mutation K237_V238del, and a double mutation in the PIP-binding motif, K237A/V238A. Circular dichroism confirmed that there were no protein structure alterations. Pull-down assays with PIP-enriched liposomes revealed that wildtype a2NT preferentially binds phosphatidylinositol 4-phosphate (PI(4)P), while mutants decreased binding to PI(4)P. In HEK293 cells, wildtype a2NT was localized to Golgi and co-purified with microsomal membranes. Mutants reduced Golgi localization and membrane association. Rapamycin depletion of PI(4)P diminished a2NT-Golgi localization. We conclude that a2NT is sufficient for Golgi retention, suggesting the lipid-binding motif is involved in V-ATPase targeting and/or retention. Mutational analyses suggest a molecular mechanism underlying how a2 mutations result in cutis laxa.
Identification and Regulation of Hypoxia-Tolerant and Germination-Related Genes in Rice
Authors: Yuan Hongyan , Zheng Zhenzhen , Bao Yaling , Zhao Xueyu , Lv Jiaqi , Tang Chenghang , Wang Nansheng , Liang Zhaojie , Li Hua , Xiang Jun , Qian Yingzhi , Shi Yingyao
Published: February 2024

In direct seeding, hypoxia is a major stress faced by rice plants. Therefore, dissecting the response mechanism of rice to hypoxia stress and the molecular regulatory network is critical to the development of hypoxia-tolerant rice varieties and direct seeding of rice. This review summarizes the morphological, physiological, and ecological changes in rice under hypoxia stress, the discovery of hypoxia-tolerant and germination-related genes/QTLs, and the latest research on candidate genes, and explores the linkage of hypoxia tolerance genes and their distribution in indica and japonica rice through population variance analysis and haplotype network analysis. Among the candidate genes, OsMAP1 is a typical gene located on the MAPK cascade reaction for indica–japonica divergence; MHZ6 is involved in both the MAPK signaling and phytohormone transduction pathway. MHZ6 has three major haplotypes and one rare haplotype, with Hap3 being dominated by indica rice varieties, and promotes internode elongation in deep-water rice by activating the SD1 gene. OsAmy3D and Adh1 have similar indica–japonica varietal differentiation, and are mainly present in indica varieties. There are three high-frequency haplotypes of OsTPP7, namely Hap1 (n = 1109), Hap2 (n = 1349), and Hap3 (n = 217); Hap2 is more frequent in japonica, and the genetic background of OsTPP7 was derived from the japonica rice subpopulation. Further artificial selection, natural domestication, and other means to identify more resistance mechanisms of this gene may facilitate future research to breed superior rice cultivars. Finally, this study discusses the application of rice hypoxia-tolerant germplasm in future breeding research.
Theoretical Analysis and Expression Profiling of 17β-Hydroxysteroid Dehydrogenase Genes in Gonadal Development and Steroidogenesis of Leopard Coral Grouper (Plectropomus leopardus)
Authors: Liu Mingjian , Ding Hui , Jin Chaofan , Wang Mingyi , Li Peiyu , Bao Zhenmin , Wang Bo , Hu Jingjie
Published: February 2024

The differentiation and developmental trajectory of fish gonads, significantly important for fish breeding, culture, and production, has long been a focal point in the fields of fish genetics and developmental biology. However, the mechanism of gonadal differentiation in leopard coral grouper (Plectropomus leopardus) remains unclear. This study investigates the 17β-Hydroxysteroid Dehydrogenase (Hsd17b) gene family in P. leopardus, with a focus on gene characterization, expression profiling, and functional analysis. The results reveal that the P. leopardus’s Hsd17b gene family comprises 11 members, all belonging to the SDR superfamily. The amino acid similarity is only 12.96%, but conserved motifs, such as TGxxxGxG and S-Y-K, are present in these genes. Hsd17b12a and Hsd17b12b are unique homologs in fish, and chromosomal localization has confirmed that they are not derived from different transcripts of the same gene, but rather are two independent genes. The Hsd17b family genes, predominantly expressed in the liver, heart, gills, kidneys, and gonads, are involved in synthesizing or metabolizing sex steroid hormones and neurotransmitters, with their expression patterns during gonadal development categorized into three distinct categories. Notably, Hsd17b4 and Hsd17b12a were highly expressed in the testis and ovary, respectively, suggesting their involvement in the development of reproductive cells in these organs. Fluorescence in situ hybridization (FISH) further indicated specific expression sites for these genes, with Hsd17b4 primarily expressed in germ stem cells and Hsd17b12a in oocytes. This comprehensive study provides foundational insights into the role of the Hsd17b gene family in gonadal development and steroidogenesis in P. leopardus, contributing to the broader understanding of fish reproductive biology and aquaculture breeding.