bluuebunny/crossref_metadata_2025 huggingface dataset:
Alginate Gel Encapsulated with Enzybiotics Cocktail Is Effective against Multispecies Biofilms
Authors: Vasina Daria V. , Antonova Nataliia P. , Shidlovskaya Elena V. , Kuznetsova Nadezhda A. , Grishin Alexander V. , Akoulina Elizaveta A. , Trusova Ekaterina A. , Lendel Anastasiya M. , Mazunina Elena P. , Kozlova Sofia R. , Dudun Andrei A. , Bonartsev Anton P. , Lunin Vladimir G. , Gushchin Vladimir A.
Published: January 2024

The development of new and effective antibacterials for pharmaceutical or cosmetic skin care that have a low potential for the emergence and expansion of bacterial resistance is of high demand in scientific and applied research. Great hopes are placed on alternative agents such as bactericidal peptidoglycan hydrolases, depolymerases, etc. Enzybiotic-based preparations are being studied for the treatment of various infections and, among others, can be used as topical formulations and dressings with protein-polysaccharide complexes. Here, we investigate the antibiofilm properties of a novel enzybiotic cocktail of phage endolysin LysSi3 and bacteriocin lysostaphin, formulated in the alginate gel matrix and its ability to control the opportunistic skin-colonizing bacteria Staphylococcus aureus, Pseudomonas aeruginosa, and Klebsiella pneumoniae, as well as mixed-species biofilms. Our results propose that the application of SiL-gel affects different components of biofilm extracellular polymeric substances, disrupts the matrix, and eliminates the bacteria embedded in it. This composition is highly effective against biofilms composed of Gram-negative and Gram-positive species and does not possess significant cytotoxic effects. Our data form the basis for the development of antibacterial skin care products with a gentle but effective mode of action.
Correction: Nordin et al. The State of the Art of Natural Polymer Functionalized Fe3O4 Magnetic Nanoparticle Composites for Drug Delivery Applications: A Review. Gels 2023, 9, 121
Authors: Nordin Abu Hassan , Ahmad Zuliahani , Husna Siti Muhamad Nur , Ilyas Rushdan Ahmad , Azemi Ahmad Khusairi , Ismail Noraznawati , Nordin Muhammad Luqman , Ngadi Norzita , Siti Nordin Hawa , Nabgan Walid , Norfarhana Abd Samad , Azami Mohammad Saifulddin Mohd
Published: January 2024

In the original publication [...]
A New Method for the Production of High-Concentration Collagen Bioinks with Semiautonomic Preparation
Authors: Matejkova Jana , Kanokova Denisa , Supova Monika , Matejka Roman
Published: January 2024

It is believed that 3D bioprinting will greatly help the field of tissue engineering and regenerative medicine, as live patient cells are incorporated into the material, which directly creates a 3D structure. Thus, this method has potential in many types of human body tissues. Collagen provides an advantage, as it is the most common extracellular matrix present in all kinds of tissues and is, therefore, very natural for cells and the organism. Hydrogels with highly concentrated collagen make it possible to create 3D structures without additional additives to crosslink the polymer, which could negatively affect cell proliferation and viability. This study established a new method for preparing highly concentrated collagen bioinks, which does not negatively affect cell proliferation and viability. The method is based on two successive neutralizations of the prepared hydrogel using the bicarbonate buffering mechanisms of the 2× enhanced culture medium and pH adjustment by adding NaOH. Collagen hydrogel was used in concentrations of 20 and 30 mg/mL dissolved in acetic acid with a concentration of 0.05 and 0.1 wt.%. The bioink preparation process is automated, including colorimetric pH detection and adjustment. The new method was validated using bioprinting and subsequent cultivation of collagen hydrogels with incorporated stromal cells. After 96 h of cultivation, cell proliferation and viability were not statistically significantly reduced.
Can First Parents Speak? A Spivakean Reading of First Parents’ Agency and Resistance in Transnational Adoption
Authors: Cawayu Atamhi , Sacré Hari Prasad
Published: January 2024

This article analyses the search strategies of first families in Bolivia contesting the separation of their children through transnational adoption. These first parents’ claims to visibility and acknowledgement have remained largely ignored by adoption policy and scholarship, historically privileging the perspectives of actors in adoptive countries, such as adoptive parents and adoption professionals. Filling in this gap, we discuss the search strategies employed by first families in Bolivia who desire a reunion with their child. Drawing on Gayatri Chakravorty Spivak’s feminist postcolonial theory, we analyse ethnographic fieldwork with fourteen first families in Bolivia. We read how the agency of first parents, severely limited by the loss of legal rights through the adoption system, is caught in a double bind of dependency and possibility. While hegemonic adoption discourse portrays first parents as passive and consenting to the adoption system, the results of our study complicate this picture. Moreover, we argue that the search activity of the first parents can be read as a claim and request to revise and negotiate their consent to transnational adoption. Ultimately, we read first parents’ search efforts as resistance to the closed nature of the adoption system, which restricts them in their search for their children.
AcMYB10 Involved in Anthocyanin Regulation of ‘Hongyang’ Kiwifruit Induced via Fruit Bagging and High-Postharvest-Temperature Treatments
Authors: Yu Min , Xiong Jinyu , Dong Kun , Quan Xin , Guo Hao , Huo Junwei , Qin Dong , Wang Yanchang , Lu Xuemei , Zhu Chenqiao
Published: January 2024

Light and temperature are key factors influencing the accumulation of anthocyanin in fruit crops. To assess the effects of fruit bagging during development and high post-ripening temperature on ‘Hongyang’ kiwifruit, we compared the pigmentation phenotypes and expression levels of anthocyanin-related genes between bagged and unbagged treatments, and between 25 °C and 37 °C postharvest storage temperatures. Both the bagging and 25 °C treatments showed better pigmentation phenotypes with higher anthocyanin concentrations. The results of the qRT-PCR analysis revealed that the gene expression levels of LDOX (leucoanthocyanidin dioxygenase), F3GT (UDP-flavonoid 3-O-glycosyltransferase ), AcMYB10, and AcbHLH42 were strongly correlated and upregulated by both the bagging treatment and 25 °C storage. The results of bimolecular fluorescence complementation and luciferase complementation imaging assays indicated an interaction between AcMYB10 and AcbHLH42 in plant cells, whereas the results of a yeast one-hybrid assay further demonstrated that AcMYB10 activated the promoters of AcLODX and AcF3GT. These results strongly suggest that enhanced anthocyanin synthesis is caused by the promoted expression of AcLODX and AcF3GT, regulated by the complex formed by AcMYB10–AcbHLH42.
Deciphering the Immune Microenvironment at the Forefront of Tumor Aggressiveness by Constructing a Regulatory Network with Single-Cell and Spatial Transcriptomic Data
Authors: Xu Kun , Yu Dongshuo , Zhang Siwen , Chen Lanming , Liu Zhenhao , Xie Lu
Published: January 2024

The heterogeneity and intricate cellular architecture of complex cellular ecosystems play a crucial role in the progression and therapeutic response of cancer. Understanding the regulatory relationships of malignant cells at the invasive front of the tumor microenvironment (TME) is important to explore the heterogeneity of the TME and its role in disease progression. In this study, we inferred malignant cells at the invasion front by analyzing single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics (ST) data of ER-positive (ER+) breast cancer patients. In addition, we developed a software pipeline for constructing intercellular gene regulatory networks (IGRNs), which help to reduce errors generated by single-cell communication analysis and increase the confidence of selected cell communication signals. Based on the constructed IGRN between malignant cells at the invasive front of the TME and the immune cells of ER+ breast cancer patients, we found that a high expression of the transcription factors FOXA1 and EZH2 played a key role in driving tumor progression. Meanwhile, elevated levels of their downstream target genes (ESR1 and CDKN1A) were associated with poor prognosis of breast cancer patients. This study demonstrates a bioinformatics workflow of combining scRNA-seq and ST data; in addition, the study provides the software pipelines for constructing IGRNs automatically (cIGRN). This strategy will help decipher cancer progression by revealing bidirectional signaling between invasive frontline malignant tumor cells and immune cells, and the selected signaling molecules in the regulatory network may serve as biomarkers for mechanism studies or therapeutic targets.
Differential Interferon Signaling Regulation and Oxidative Stress Responses in the Cerebral Cortex and Cerebellum Could Account for the Spatiotemporal Pattern of Neurodegeneration in Niemann–Pick Disease Type C
Authors: Tolan Andrew J. , Sanchez Kayla L. , Shin Samuel D. , White Jacob B. , Currais Antonio , Soriano-Castell David , Wilson Christopher G. , Maher Pamela , Soriano Salvador
Published: January 2024

Niemann–Pick disease type C (NPC) is a fatal neurodegenerative condition caused by genetic mutations of the NPC1 or NPC2 genes that encode the NPC1 and NPC2 proteins, respectively, which are believed to be responsible for cholesterol efflux from late-endosomes/lysosomes. The pathogenic mechanisms that lead to neurodegeneration in NPC are not well understood. There are, however, well-defined spatiotemporal patterns of neurodegeneration that may provide insight into the pathogenic process. For example, the cerebellum is severely affected from early disease stages, compared with cerebral regions, which remain relatively spared until later stages. Using a genome-wide transcriptome analysis, we have recently identified an aberrant pattern of interferon activation in the cerebella of pre-symptomatic Npc1−/− mice. Here, we carried out a comparative transcriptomic analysis of cerebral cortices and cerebella of pre-symptomatic Npc1−/− mice and age-matched controls to identify differences that may help explain the pathological progression within the NPC brain. We report lower cerebral expression of genes within interferon signaling pathways, and significant differences in the regulation of oxidative stress, compared with the cerebellum. Our findings suggest that a delayed onset of interferon signaling, possibly linked to lower oxidative stress, may account for the slower onset of cerebral cortical pathology in the disease.
Predictive Model of the Relationship between Positive and Negative Affect, Self-Consciousness of Appearance, and Positive Body Image in Physical Exercise Practice
Authors: Mendes José , Alexandre-Sousa Pedro , Tavares Márcio
Published: January 2024

The biopsychosocial development of individuals is influenced by body image and physical exercise. This study aimed to evaluate seven hypotheses regarding the impact of positive affect, negative affect, self-consciousness of appearance, and positive body image on the practice of physical exercise. The data were processed using the statistical package SmartPLS 4.0.9.5. Using an exploratory methodology based on structural equation modeling (SEM) of structural equations applied to small sample sizes, a tentative model has been generated to define the drivers of physical activity. The survey used the Body Mass Index, Body Appreciation Scale—Revised, and Derriford Appearance Scale—14 items. The study involved 129 men and 279 women (N = 408). The model demonstrated a distinct internal consistency in that, out of the seven hypotheses analyzed, only one was rejected. The findings of the multigroup analysis indicate that there are no statistically significant discrepancies between the constructed measures and the practice of physical activity. The conclusions of this study showed that, although positive and negative affect, and self-consciousness influence positive body image, there were no statistically significant differences between those who exercise and those who do not exercise. Various factors influence biopsychosocial development. Future studies should explore the influence of psychological and social variables on understanding body image and physical exercise.
Physical Leisure Activities in Adolescents with Disabilities: Data from National Survey of Disabled Persons
Authors: Ryu Mikyung , Seo Kawoun , Song Youngshin
Published: January 2024

The impact of activities of daily living (ADLs) on the life satisfaction of adolescents with disabilities and the potential role of physical leisure activities as a moderator in this context have received attention. However, little is known about the influence of physical leisure activities on the relationship between ADLs and life satisfaction in adolescents with disabilities. This cross-sectional study aimed to investigate the moderating effect of physical leisure activities on the relationship between ADLs and life satisfaction among adolescents with disabilities. Secondary data analysis was conducted by extracting data from 5364 adolescents aged 12–18 years with disabilities from the 2020 National Survey of Disabled Persons of Korea. The potential moderating effect of physical leisure activities on the relationship between ADLs and life satisfaction was investigated using Pearson’s correlation analysis and hierarchical multiple regression. Significant positive correlations were found for living, life satisfaction, and physical leisure activities. Physical leisure activities were found to play a moderating role in the relationship between ADLs and life satisfaction. This study concluded that increasing physical leisure activities, even with dependent ADLs, promotes life satisfaction, especially if ADLs are low, highlighting the necessity of increasing leisure activities in adolescents with disabilities.
Patterns of Blood Transfusion in Sickle Cell Disease Hospitalizations
Authors: Sharma Aditi , Dahiya Amit , Alavi Asif , Woldie Indryas , Sharma Aditya , Karson Jeffrey , Singh Vijendra
Published: January 2024

Background: Transfusional iron overload causes significant morbidity and mortality in sickle cell disease (SCD). Nevertheless, red blood cell transfusions continue to be essential in its management. This study describes the transfusion patterns among SCD hospitalizations. Methods: Hospitalizations for SCD in the 2017–2018 Nationwide Readmissions Database were divided into two groups based on whether they received transfusions. Descriptive analysis was performed to compare their demographics and complications. Multivariable logistic regression was performed to determine the factors associated with transfusions. Results: Out of 109,783 hospitalizations, 28,300 were transfused, and 81,483 were not transfused. Females and older individuals were higher in the transfused category than the non-transfused category (59.49% vs. 53.52% and 28.86% vs. 21.27%, respectively; p < 0.001 for both). The wealthiest population was more likely to be in the transfused category (11.27% vs. 8.34%; p < 0.001). Admissions to teaching hospitals, large metropolitan hospitals, and highest-volume hospitals were higher in the non-transfused category vs. transfused category (79.89% vs. 72.17%; p < 0.001, 69.26% vs. 65.35%; p 0.003 and 74.71% vs. 63.51%; p < 0.001, respectively). Most admissions were transfused once, with three or more transfusions being given more in the non-teaching hospitals than the teaching hospitals (1.27% vs. 0.41%; p 0.01). Furthermore, a higher proportion of early transfusions occurred in the non-teaching hospitals (65.6% vs. 57.82% for admission days 1 and 2; p < 0.001). Admission to a teaching hospital was associated with lower blood transfusion odds than a non-teaching hospital. Conclusion: A quarter of admissions for SCD receive a blood transfusion. In addition to performing more frequent and early transfusions, the odds of being transfused are higher in non-teaching hospitals.
Diversity of Crithmum maritimum L. from Salento Coastal Area: A Suitable Species for Domestication
Authors: Accogli Rita , Nutricati Eliana , De Bellis Luigi , Renna Massimiliano , Luvisi Andrea , Negro Carmine
Published: January 2024

Crithmum maritimum L., known as sea fennel, is an aromatic halophyte typical of the cliffs and coastal areas of the Mediterranean Sea and Atlantic Ocean. Their phytochemicals have been of great interest in the food and pharmaceutical industry. In this work, we analyzed, by SPME/gas chromatography coupled with mass spectrometry, the chemical variability of C. maritimum accessions in terms of volatile organic compounds. C. maritimum seeds were collected from different coastal sites in Salento, Southern Apulia, Italy, and subsequently cultivated ex situ. Several volatile compounds produced by C. maritimum leaves were detected, and, among them, D-limonene was found to be emitted at high levels by plants of all accessions representing the main compound, while other monoterpenes were produced at low levels. Moreover, the phenylpropene volatiles dillapiol and apiol (designated together as (dill)apiol) were emitted at variable amounts with different accessions. The correlation among groups based on volatile compounds has been analyzed using hierarchical cluster analysis, which has revealed three main groups based on (dill)apiol presence and its total amount in the cultivated plants of different geographic origins, confirming intraspecies biodiversity. Moreover, we have evaluated the seed germination and seedling development of C. maritimum in controlled conditions. We found no dormancy and a high germination rate for all samples analyzed. The chemo-diversity evidenced in cultivated plants obtained from seeds collected at different locations on the Salento peninsula is probably related to variations in climate resulting from different exposures along the coast. These findings highlighted the importance of C.maritimum as a suitable candidate for cultivation because it can tolerate harsh conditions/stresses and also has a possible use besides food and pharmaceuticals and for the restoration of coastal environments.
Recovery Barrier Characterizations by Hospitalized Patients with Substance Use Disorders: Results from a Randomized Clinical Study on Inpatient Peer Recovery Coaching
Authors: Byrne Kaileigh A. , Roth Prerana J. , Cumby Sam , Goodwin Eli , Herbert Kristin , Schmidt William Michael , Worth Samantha , Connolly Kyleigh , Uzor Onye , Eiff Brandi , Black Dominique
Published: January 2024

Patients hospitalized with medical complications from substance use disorder (SUD) encounter unique health problems that may complicate their recovery. Recovery barriers are not well understood in this population. The study objective is to characterize recovery barriers in this patient population. Participants (n = 96) in this six-month longitudinal study were randomized to a peer recovery coaching intervention or standard of care. The primary outcome measures were qualitative, open-ended questions addressing factors interfering with participants’ recovery. Data were analyzed using content analysis. Themes were identified a priori using past research on recovery capital domains; these seven barriers were (1) psychological health difficulties, (2) physical health challenges, (3) lack of social support, (4) insufficient treatment or recovery support to maintain sobriety, (5) environmental and housing concerns, (6) deficits in coping skills, and (7) lack of meaningful activities. At baseline, the most common recovery barriers were in the environment and housing (28.1%), psychological health (27.1%), and social support (22.9%) domains. At six-month follow-up, participants were asked to describe barriers they felt they had made improvement in over the last six months. The primary themes that participants reported improvements in were treatment and recovery support to maintain sobriety (52.1%), coping skills (35.4%), and social support (27.1%). Hospitalization and participation in a randomized controlled trial may be a turning point in which to address recovery barriers for patients hospitalized with complications from SUD.
Measuring Quality of Life: Incorporating Objectively Measurable Parameters within the Cross-Sectional Bern Cohort Study 2014 (BeCS-14)
Authors: Theis Susanne , Bitterlich Norman , von Wolff Michael , Stute Petra
Published: January 2024

Up until now, the measurement of Quality of Life (QoL) was based on validated subjective rating tools rather than objective measurement. To become more independent of the self-assessment of probands, a way to objectively measure QoL should be found. A monocenter, cross-sectional, observational, non-interventional trial was performed from 2012 to 2014 at Inselspital Bern to evaluate the bio-functional status (BFS), a complex, generic, non-invasive, sex- and age-validated assessment tool, in a wide range of areas. A standardized battery of assessments was performed on 464 females and 166 males, ages 18 to 65 (n = 630). In addition to the survey of the BFS, participants replied—among others—to the validated questionnaire SF-36 for health-related QoL (n = 447, subgroup 1). Since the accepted cut-off value for BFA calculation is age ≥ 35 years, subgroup 2 included 227 subjects (all participants aged ≥ 35 years out of subgroup 1). In order to be able to compare the eight SF-36 subscales to BFS parameters, a comparable score set of single BFS items had to be constructed. Subsequently, we aimed to statistically identify BFS item combinations that best represented each SF-36 subscale. All eight SF-36 subscales were significantly represented by various different combinations of BFS items. A total of 24 single BFS items significantly correlated with SF-36 subscales, of which 15 were objective and nine were subjective. All eight SF-36 subscales were significantly represented by various different combinations of BFS items leading to stronger correlations (range five to nine BFS items), and overall, sex and age did not affect these associations, but in the SF-36 subscales ‘bodily pain’ (sex) and ‘role limitations due to physical health problems’ (age in men). To our knowledge, we are the first to correlate a validated set of 34 objective and 9 subjective parameters with subjectively evaluated SF-36 subscales. This first study on the objectifiability of the SF-36 questionnaire demonstrated that questions on quality of life can be answered independently of a subjective assessment by subjects in future scientific studies.
A Containerized Service-Based Integration Framework for Heterogeneous-Geospatial-Analysis Models
Authors: Zhu Lilu , Wang Yang , Kong Yunbo , Hu Yanfeng , Huang Kai
Published: January 2024

The integration of geospatial-analysis models is crucial for simulating complex geographic processes and phenomena. However, compared to non-geospatial models and traditional geospatial models, geospatial-analysis models face more challenges owing to extensive geographic data processing and complex computations involved. One core issue is how to eliminate model heterogeneity to facilitate model combination and capability integration. In this study, we propose a containerized service-based integration framework named GeoCSIF, specifically designed for heterogeneous-geospatial-analysis models. Firstly, by designing the model-servicized structure, we shield the heterogeneity of model structures so that different types of geospatial-analysis models can be effectively described and integrated based on standardized constraints. Then, to tackle the heterogeneity in model dependencies, we devise a prioritization-based orchestration method, facilitating optimized combinations of large-scale geospatial-analysis models. Lastly, considering the heterogeneity in execution modes, we design a heuristic scheduling method that establishes optimal mappings between models and underlying computational resources, enhancing both model stability and service performance. To validate the effectiveness and progressiveness of GeoCSIF, a prototype system was developed, and its integration process for flood disaster models was compared with mainstream methods. Experimental results indicate that GeoCSIF possesses superior performance in model management and service efficiency.
Exploring Embryo-Ototoxic Effects: Insights into Deodorant-Induced Hair Cell Damage in Zebrafish
Authors: Yoon Hee Soo , Hyun Kyung Tae , Hong Sumin , Park Saemi , Han Eunjung , Baek Hyun woo , Lee Yun Kyoung , Lim Kang Hyeon , Rah Yoon Chan , Choi June
Published: January 2024

Our study investigated the embryo-ototoxic effects of deodorant2 (DA2) on zebrafish embryos, which serve as valuable model organisms due to genetic and physiological similarities to humans. We focused on understanding DA2’s impact on zebrafish hair cells, which are vital for sensory perception and balance regulation. DA2, provided by the Ministry of Environment, Republic of Korea, was used at 460 μg/mL in dimethyl sulfoxide (DMSO), with a 0.43% DMSO solvent control group. Three experiments, each using 10 zebrafish specimens from each group, showed an initial 13% hair cell count reduction in the DA2-exposed group. Subsequent experiments demonstrated reductions of 37% and 22%, each with one mortality case. Statistical analysis revealed a significant 24% hair cell count reduction in the DA2-exposed group. We also assessed DA2’s impact on zebrafish behavior. Although not statistically significant, differences in distances traveled (0.33–0.39, 95% confidence interval: −0.46–1.1, p = 0.2033) and latencies (−0.016–0.018, 95% confidence interval: −0.052–0.021, p = 0.1917) hinted at negative effects. These results highlight DA2’s ototoxic properties affecting zebrafish auditory systems and behavior. Further investigation into DA2’s effects on aquatic organisms and potential mitigation strategies are essential. These findings contribute to understanding DA2’s safety profile, benefiting aquatic ecosystems and human health assessments.
SMC5/6 Promotes Replication Fork Stability via Negative Regulation of the COP9 Signalosome
Authors: Xu Michelle J. , Jordan Philip W.
Published: January 2024

It is widely accepted that DNA replication fork stalling is a common occurrence during cell proliferation, but there are robust mechanisms to alleviate this and ensure DNA replication is completed prior to chromosome segregation. The SMC5/6 complex has consistently been implicated in the maintenance of replication fork integrity. However, the essential role of the SMC5/6 complex during DNA replication in mammalian cells has not been elucidated. In this study, we investigate the molecular consequences of SMC5/6 loss at the replication fork in mouse embryonic stem cells (mESCs), employing the auxin-inducible degron (AID) system to deplete SMC5 acutely and reversibly in the defined cellular contexts of replication fork stall and restart. In SMC5-depleted cells, we identify a defect in the restart of stalled replication forks, underpinned by excess MRE11-mediated fork resection and a perturbed localization of fork protection factors to the stalled fork. Previously, we demonstrated a physical and functional interaction of SMC5/6 with the COP9 signalosome (CSN), a cullin deneddylase that enzymatically regulates cullin ring ligase (CRL) activity. Employing a combination of DNA fiber techniques, the AID system, small-molecule inhibition assays, and immunofluorescence microscopy analyses, we show that SMC5/6 promotes the localization of fork protection factors to stalled replication forks by negatively modulating the COP9 signalosome (CSN). We propose that the SMC5/6-mediated modulation of the CSN ensures that CRL activity and their roles in DNA replication fork stabilization are maintained to allow for efficient replication fork restart when a replication fork stall is alleviated.
Pro-Inflammatory Cytokines Enhanced In Vitro Cytotoxic Activity of Clostridioides difficile Toxin B in Enteric Glial Cells: The Achilles Heel of Clostridioides difficile Infection?
Authors: Fettucciari Katia , Spaterna Andrea , Marconi Pierfrancesco , Bassotti Gabrio
Published: January 2024

Bacterial infections are characterized by an inflammatory response, which is essential for infection containment but is also responsible for negative effects on the host. The pathogen itself may have evolved molecular mechanisms to antagonize the antimicrobial effects of an inflammatory response and to enhance its pathogenicity using inflammatory response mediators, such as cytokines. Clostridioides difficile (C. difficile) infection (CDI) causes gastrointestinal diseases with markedly increasing global incidence and mortality rates. The main C. difficile virulence factors, toxin A and B (TcdA/TcdB), cause cytopathic/cytotoxic effects and inflammation. We previously demonstrated that TcdB induces enteric glial cell (EGC) apoptosis, which is enhanced by the pro-inflammatory cytokine tumor necrosis factor alpha plus interferon gamma (CKs). However, it is unknown whether CKs-enhanced TcdB cytotoxicity (apoptosis/necrosis) is affected by the timing of the appearance of the CKs. Thus, we simulated in vitro, in our experimental model with TcdB and EGCs, three main situations of possible interactions between TcdB and the timing of CK stimulation: before TcdB infection, concomitantly with infection, or at different times after infection and persisting over time. In these experimental conditions, which all represent situations of possible interactions between C. difficile and the timing of CK stimulation, we evaluated apoptosis, necrosis, and cell cycle phases. The CKs, in all of these conditions, enhanced TcdB cytotoxicity, which from apoptosis became necrosis when CK stimulation persisted over time, and was most relevant after 48 h of TcdB:EGCs interaction. Particularly, the enhancement of apoptosis by CKs was dependent on the TcdB dose and in a less relevant manner on the CK stimulation time, while the enhancement of necrosis occurred always independently of the TcdB dose and CK stimulation time. However, since in all conditions stimulation with CKs strongly enhanced the TcdB cytotoxicity, it always had a negative impact on C. difficile pathogenicity. This study might have important implications for the treatment of CDI.
Correlation between Inflammasomes and Microbiota in Peri-Implantitis
Authors: Padial-Molina Miguel , Montalvo-Acosta Saray , Martín-Morales Natividad , Pérez-Carrasco Virginia , Magan-Fernandez Antonio , Mesa Francisco , O’Valle Francisco , Garcia-Salcedo Jose Antonio , Galindo-Moreno Pablo
Published: January 2024

The activation of inflammasomes is thought to induce the inflammatory process around dental implants. No information is available on the correlation between microbiota and inflammasomes in clinical samples from patients suffering peri-implantitis. For this cross-sectional study, 30 biofilm samples were obtained from 19 patients undergoing surgical treatment for peri-implantitis because of the presence of bleeding on probing, probing depth higher than 6 mm, and radiographic bone loss higher than 3 mm. Then, soft tissue samples from around the implant were also collected. The relative abundance of bacteria and alpha-diversity indexes were calculated after analyzing the 16S rRNA gene using next-generation sequencing. The soft-tissue samples were processed for evaluation of the inflammasomes NLRP3 and AIM2 as well as caspase-1 and IL-1β. The relative abundance (mean (SD)) of specific species indicated that the most abundant species were Porphyromonas gingivalis (10.95 (14.17)%), Fusobacterium vincentii (10.93 (13.18)%), Porphyromonas endodontalis (5.89 (7.23)%), Prevotella oris (3.88 (4.94)%), Treponema denticola (2.91 (3.19)%), and Tannerella forsythia (2.84 (4.15)%). Several correlations were found between the species and the immunohistochemical detection of the inflammasomes NLRP3 and AIM2 as well as caspase-1 and IL-1β, both in the epithelium and the lamina propria. A network analysis found an important cluster of variables formed by NLRP3 in the lamina propria and AIM2, caspase-1, and IL-1β in the lamina propria and the epithelium with Prevotella dentalis, Prevotella tannerae, Tannerella forsythia, or Selenomonas timonae. Thus, it could be concluded that inflammasomes NLRP3 and AIM2 and their downstream effectors caspase-1 and interleukin-1β can be significantly associated with specific bacteria.
Proteomic Profiling Identifies Candidate Diagnostic Biomarkers of Hydrosalpinx in Endometrial Fluid: A Pilot Study
Authors: Gonzalez-Martin Roberto , de Castro Pedro , Fernandez Carmen , Quintana Fernando , Quiñonero Alicia , Ferrando Marcos , Dominguez Francisco
Published: January 2024

Hydrosalpinx is a fluid occlusion and distension of the fallopian tubes, often resulting from pelvic inflammatory disease, which reduces the success of artificial reproductive technologies (ARTs) by 50%. Tubal factors account for approximately 25% of infertility cases, but their underlying molecular mechanisms and functional impact on other reproductive tissues remain poorly understood. This proteomic profiling study applied sequential window acquisition of all theoretical fragment ion spectra mass spectrometry (SWATH-MS) to study hydrosalpinx cyst fluid and pre- and post-salpingectomy endometrial fluid. Among the 967 proteins identified, we found 19 and 17 candidate biomarkers for hydrosalpinx in pre- and post-salpingectomy endometrial fluid, respectively. Salpingectomy significantly affected 76 endometrial proteins, providing insights into the enhanced immune response and inflammation present prior to intervention, and enhanced coagulation cascades and wound healing processes occurring one month after intervention. These findings confirmed that salpingectomy reverses the hydrosalpinx-related functional impairments in the endometrium and set a foundation for further biomarker validation and the development of less-invasive diagnostic strategies for hydrosalpinx.
Role of Cannabinoids in Oral Cancer
Authors: Cretu Brigitte , Zamfir Alexandra , Bucurica Sandica , Scheau Andreea Elena , Savulescu Fiedler Ilinca , Caruntu Constantin , Caruntu Ana , Scheau Cristian
Published: January 2024

Cannabinoids have incited scientific interest in different conditions, including malignancy, due to increased exposure to cannabis. Furthermore, cannabinoids are increasingly used to alleviate cancer-related symptoms. This review paper aims to clarify the recent findings on the relationship between cannabinoids and oral cancer, focusing on the molecular mechanisms that could link cannabinoids with oral cancer pathogenesis. In addition, we provide an overview of the current and future perspectives on the management of oral cancer patients using cannabinoid compounds. Epidemiological data on cannabis use and oral cancer development are conflicting. However, in vitro studies assessing the effects of cannabinoids on oral cancer cells have unveiled promising anti-cancer features, including apoptosis and inhibition of cell proliferation. Downregulation of various signaling pathways with anti-cancer effects has been identified in experimental models of oral cancer cells exposed to cannabinoids. Furthermore, in some countries, several synthetic or phytocannabinoids have been approved as medical adjuvants for the management of cancer patients undergoing chemoradiotherapy. Cannabinoids may improve overall well-being by relieving anxiety, depression, pain, and nausea. In conclusion, the link between cannabinoid compounds and oral cancer is complex, and further research is necessary to elucidate the potential risks or their protective impact on oral cancer.
Radiation Hormesis in Barley Manifests as Changes in Growth Dynamics Coordinated with the Expression of PM19L-like, CML31-like, and AOS2-like
Authors: Kazakova Elizaveta , Gorbatova Irina , Khanova Anastasia , Shesterikova Ekaterina , Pishenin Ivan , Prazyan Alexandr , Podlutskii Mikhail , Blinova Yana , Bitarishvili Sofia , Bondarenko Ekaterina , Smirnova Alena , Lychenkova Maria , Bondarenko Vladimir , Korol Marina , Babina Daria , Makarenko Ekaterina , Volkova Polina
Published: January 2024

The stimulation of growth and development of crops using ionising radiation (radiation hormesis) has been reported by many research groups. However, specific genes contributing to the radiation stimulation of plant growth are largely unknown. In this work, we studied the impact of the low-dose γ-irradiation of barley seeds on the growth dynamics and gene expression of eight barley cultivars in a greenhouse experiment. Our findings confirmed that candidate genes of the radiation growth stimulation, previously established in barley seedlings (PM19L-like, CML31-like, and AOS2-like), are significant in radiation hormesis throughout ontogeny. In γ-stimulated cultivars, the expression of these genes was aligned with the growth dynamics, yield parameters, and physiological conditions of plants. We identified contrasting cultivars for future gene editing and found that the γ-stimulated cultivar possessed some specific abiotic stress-responsive elements in the promotors of candidate genes, possibly revealing a new level of radiation hormesis effect execution. These results can be used in creating new productive barley cultivars, ecological toxicology of radionuclides, and eustress biology studies.
CRISPR-Cas12a for Highly Efficient and Marker-Free Targeted Integration in Human Pluripotent Stem Cells
Authors: Hammad Ruba , Alzubi Jamal , Rhiel Manuel , Chmielewski Kay O. , Mosti Laura , Rositzka Julia , Heugel Marcel , Lawrenz Jan , Pennucci Valentina , Gläser Birgitta , Fischer Judith , Schambach Axel , Moritz Thomas , Lachmann Nico , Cornu Tatjana I. , Mussolino Claudio , Schäfer Richard , Cathomen Toni
Published: January 2024

The CRISPR-Cas12a platform has attracted interest in the genome editing community because the prototypical Acidaminococcus Cas12a generates a staggered DNA double-strand break upon binding to an AT-rich protospacer-adjacent motif (PAM, 5′-TTTV). The broad application of the platform in primary human cells was enabled by the development of an engineered version of the natural Cas12a protein, called Cas12a Ultra. In this study, we confirmed that CRISPR-Cas12a Ultra ribonucleoprotein complexes enabled allelic gene disruption frequencies of over 90% at multiple target sites in human T cells, hematopoietic stem and progenitor cells (HSPCs), and induced pluripotent stem cells (iPSCs). In addition, we demonstrated, for the first time, the efficient knock-in potential of the platform in human iPSCs and achieved targeted integration of a GFP marker gene into the AAVS1 safe harbor site and a CSF2RA super-exon into CSF2RA in up to 90% of alleles without selection. Clonal analysis revealed bi-allelic integration in >50% of the screened iPSC clones without compromising their pluripotency and genomic integrity. Thus, in combination with the adeno-associated virus vector system, CRISPR-Cas12a Ultra provides a highly efficient genome editing platform for performing targeted knock-ins in human iPSCs.
Rats Selected for Different Nervous Excitability: Long-Term Emotional–Painful Stress Affects the Dynamics of DNA Damage in Cells of Several Brain Areas
Authors: Shcherbinina Veronika , Pavlova Marina , Daev Eugene , Dyuzhikova Natalia
Published: January 2024

The maintenance of genome stability is critical for health, but during individual ontogenesis, different stressors affect DNA integrity, which can lead to functional and/or structural changes in the cells of target organs. In the nervous system, cell genome destabilization is associated with different neurological and psychiatric diseases, but experiments in vivo, where a link between stress and DNA instability has been demonstrated, are relatively rare. Here, we use rat strains selected for the contrast excitability of the tibialis nerve (n. tibialis) and nonselected Wistar rats to investigate the reasons for individual differences in developing post-stress pathologies. Previous research on the behavioral response of these strains to prolonged emotional–painful stress (PEPS) allows us to consider one strain as a model of post-traumatic stress disorder (PTSD) and another strain as a model of compulsive disorder (CD). We study DNA damage in the cells of the prefrontal cortex (PFC), hippocampus, and amygdala, regions involved in stress responses and the formation of post-stress dysfunctions. The evaluation of cell genome integrity via the comet assay shows different responses to PEPS in each brain area analyzed and for all strains used. This could help us to understand the reasons for individual differences in the consequences of stress and the pathophysiology of post-stress disease formation.
Anti-Obesity Effects of GABA in C57BL/6J Mice with High-Fat Diet-Induced Obesity and 3T3-L1 Adipocytes
Authors: Jin Heegu , Han Hyein , Song Gunju , Oh Hyun-Ji , Lee Boo-Yong
Published: January 2024

Obesity is the excessive accumulation of body fat resulting from impairment in energy balance mechanisms. In this study, we aimed to investigate the mechanism whereby GABA (γ-aminobutyric acid) prevents high-fat diet-induced obesity, and whether it induces lipolysis and browning in white adipose tissue (WAT), using high-fat diet (HFD)-fed obese mice and 3T3-L1 adipocytes. We demonstrated that GABA substantially inhibits the body mass gain of mice by suppressing adipogenesis and lipogenesis. Consistent with this result, histological analysis of WAT demonstrated that GABA decreases adipocyte size. Moreover, we show that GABA administration decreases fasting blood glucose and improves serum lipid profiles and hepatic lipogenesis in HFD-fed obese mice. Furthermore, Western blot and immunofluorescence analyses showed that GABA activates protein kinase A (PKA) signaling pathways that increase lipolysis and promote uncoupling protein 1 (UCP1)-mediated WAT browning. Overall, these results suggest that GABA exerts an anti-obesity effect via the regulation of lipid metabolism.
Polyamine Signal through HCC Microenvironment: A Key Regulator of Mitochondrial Preservation and Turnover in TAMs
Authors: Liu Qingqing , Yan Xiaoyu , Li Runyuan , Yuan Yuan , Wang Jian , Zhao Yuanxin , Fu Jiaying , Su Jing
Published: January 2024

Hepatocellular carcinoma (HCC) is the most common primary liver cancer, and, with increasing research on the tumor immune microenvironment (TIME), the immunosuppressive micro-environment of HCC hampers further application of immunotherapy, even though immunotherapy can provide survival benefits to patients with advanced liver cancer. Current studies suggest that polyamine metabolism is not only a key metabolic pathway for the formation of immunosuppressive phenotypes in tumor-associated macrophages (TAMs), but it is also profoundly involved in mitochondrial quality control signaling and the energy metabolism regulation process, so it is particularly important to further investigate the role of polyamine metabolism in the tumor microenvironment (TME). In this review, by summarizing the current research progress of key enzymes and substrates of the polyamine metabolic pathway in regulating TAMs and T cells, we propose that polyamine biosynthesis can intervene in the process of mitochondrial energy metabolism by affecting mitochondrial autophagy, which, in turn, regulates macrophage polarization and T cell differentiation. Polyamine metabolism may be a key target for the interactive dialog between HCC cells and immune cells such as TAMs, so interfering with polyamine metabolism may become an important entry point to break intercellular communication, providing new research space for developing polyamine metabolism-based therapy for HCC.
The Abundant Distribution and Duplication of SARS-CoV-2 in the Cerebrum and Lungs Promote a High Mortality Rate in Transgenic hACE2-C57 Mice
Authors: Li Heng , Zhao Xin , Peng Shasha , Li Yingyan , Li Jing , Zheng Huiwen , Zhang Yifan , Zhao Yurong , Tian Yuan , Yang Jinling , Wang Yibin , Zhang Xinglong , Liu Longding
Published: January 2024

Patients with COVID-19 have been reported to experience neurological complications, although the main cause of death in these patients was determined to be lung damage. Notably, SARS-CoV-2-induced pathological injuries in brains with a viral presence were also found in all fatal animal cases. Thus, an appropriate animal model that mimics severe infections in the lungs and brain needs to be developed. In this paper, we compared SARS-CoV-2 infection dynamics and pathological injuries between C57BL/6Smoc-Ace2em3(hACE2-flag-Wpre-pA)Smoc transgenic hACE2-C57 mice and Syrian hamsters. Importantly, the greatest viral distribution in mice occurred in the cerebral cortex neuron area, where pathological injuries and cell death were observed. In contrast, in hamsters, viral replication and distribution occurred mainly in the lungs but not in the cerebrum, although obvious ACE2 expression was validated in the cerebrum. Consistent with the spread of the virus, significant increases in IL-1β and IFN-γ were observed in the lungs of both animals. However, in hACE2-C57 mice, the cerebrum showed noticeable increases in IL-1β but only mild increases in IFN-γ. Notably, our findings revealed that both the cerebrum and the lungs were prominent infection sites in hACE2 mice infected with SARS-CoV-2 with obvious pathological damage. Furthermore, hamsters exhibited severe interstitial pneumonia from 3 dpi to 5 dpi, followed by gradual recovery. Conversely, all the hACE2-C57 mice experienced severe pathological injuries in the cerebrum and lungs, leading to mortality before 5 dpi. According to these results, transgenic hACE2-C57 mice may be valuable for studying SARS-CoV-2 pathogenesis and clearance in the cerebrum. Additionally, a hamster model could serve as a crucial resource for exploring the mechanisms of recovery from infection at different dosage levels.
Association of the rs1126616 and rs9138 Variants in the SPP1 Gene among Mexican Patients with Systemic Lupus Erythematosus and Lupus Nephritis
Authors: Rivera-Cameras Alicia , Gallegos-Arreola Martha Patricia , Morán-Moguel María Cristina , Salazar-Páramo Mario , Alcaraz-López Miriam Fabiola , Echeverría-González Gustavo , Topete-Reyes Jorge Fernando , Franco-Chávez Sergio Adalberto , Dávalos-Rodríguez Ingrid Patricia
Published: January 2024

Systemic lupus erythematosus (SLE) is a multisystem disease considered a prototype of the main autoimmune disease and presents serious complications, such as lupus nephritis (LN), which generates a significant impact on morbidity and mortality. The SPP1 gene encodes the osteopontin (OPN) protein, which plays a crucial role in the regulation of inflammation and immunity. The variants rs1126616 and rs9138 of this gene have been associated with the inflammatory response. The study aims to analyze the association of the rs1126616 and rs9138 variants of the SPP1 gene in SLE Mexican-Mestizo patients without LN (SLE-LN). In this cross-sectional study, a total of 171 genomic DNA samples from SLE patients were clinically confirmed, of which 111 were SLE without LN, 60 were SLE with LN, and 100 healthy individuals were included as reference group. The rs1126616 variant was genotyped using PCR-RFLPs, and the rs9138 variant was genotyped using qPCR TaqMan. The TT genotype, the recessive model [OR 2.76 (95% CI 1.31–5.82), p = 0.011], and the T allele [OR 2.0 (95% CI 1.26–3.16), p = 0.003] of the rs1126616 variant are risk factors for SLE with LN. By contrast, the rs9138 variant did not show statistically significant differences among SLE patients stratified by LN. In our study of SLE Mexican-Mestizo patients with and without NL, demographic and clinical characteristics do not differ from other SLE populations, and the TT genotype of the rs1126616 variant of the SPP1 gene confers a risk factor for the presentation of LN. Otherwise, the rs9138 variant did not show association with NL.