bluuebunny/crossref_metadata_2025 huggingface dataset:
Chinese Cabbage BrCAP Has Potential Resistance against Plasmodiophora brassicae
Authors: Zou Jiawei , Gao Shiqi , Zhang Bo , Ge Wenjie , Zhang Jing , Ji Ruiqin
Published: April 2023

Clubroot disease caused by Plasmodiophora brassicae Woronin (P. brassicae) has seriously influenced the production of Brassica rapa crops, but the interaction mechanism between P. brassicae and Brassica rapa is not clear. In our previous study, a differentially expressed protein, G15, was found between the roots of Chinese cabbage inoculated and un-inoculated with P. brassicae through two-dimensional electrophoresis (2-DE) and mass spectrometry, and G15 was matched with Bra011464. In this study, Bra011464 was found to have a 94% percent identity with Arabidopsis thaliana CAP, named BrCAP. The expression of BrCAP was the highest in the root compared with the stems and leaves of Chinese cabbage, and its expression in the roots of Chinese cabbage inoculated with P. brassicae was significantly higher than that in the control. The results were verified by real-time quantitative polymerase chain reaction (RT-qPCR) and in situ hybridization. Subcellular localization showed that BrCAP was localized on chloroplasts of leaf epidermal cells. To verify the function of BrCAP, it was found that the Arabidopsis thaliana mutant cap was more susceptible to infection with P. brassicae than the wild type (WT), which suggested that BrCAP has a potential role in the resistance progress of Chinese cabbage to P. brassicae.
Systematic Observation of Emotional Regulation in the School Classroom: A Contribution to the Mental Health of New Generations
Authors: Alarcón-Espinoza Marina , Samper-Garcia Paula , Anguera M. Teresa
Published: April 2023

Emotional regulation is a developmental milestone, as it promotes well-being throughout life. Children between 10 and 12 years old are expected to reach capacities that allow them to regulate themselves emotionally, the school context being a natural setting for this challenge. With the objective of analyzing the forms of expression and regulation of emotions that are observed in the school classroom, this research was conducted through a mixed methods study that systematically observed nine classes during five sessions each. The design was Nomothetic, Follow-up and Multidimensional; the observations were recorded on audio and in person and were later transformed into data by coding them according to an ad hoc designed instrument. The concordance of the records was evaluated, a sequential analysis of delays (GSEQ5) was performed to detect regularities and existing sequences, and a polar coordinate analysis (HOISAN) observed the relationships between the categories. Finally, the presence of multiple cases was detected. The results detail the ways in which different actors express emotions and interact, regulating the emotions of other people. The results are discussed based on the need to foster educational intentionality and allow students’ emotional self-regulation.
Social-Emotional Skills, Career Adaptability, and Agentic School Engagement of First-Year High School Students
Authors: Oliveira Íris M. , de Castro Inês , Silva Ana Daniela , Taveira Maria do Céu
Published: April 2023

The transition to the first year of high school constitutes a critical moment because it corresponds to the implementation of a career choice, which can impact students’ satisfaction and psychosocial adjustment. The career construction model of adaptation holds potential to explain how students adapt to high school, by suggesting linkages among adaptive readiness, resources, responses, and results. However, research applying the career construction model to school transitions, combining social-emotional, career, and academic variables is still needed. This study explores the roles that social-emotional skills (an indicator of adaptive readiness) and career adaptability (an indicator of adaptability resources) play in explaining first-year high school students’ agentic school engagement (an indicator of adapting responses). Measures of social-emotional skills, career adaptability, and school engagement were completed by 136 students (63.2% girls; M age = 15.68). Results from the hierarchical linear regression analysis suggest that social-emotional skills and career adaptability explain 32% of the variance and significantly contribute to explaining agentic school engagement. These findings seem illustrative of the potential of the career construction model of adaptation to deepen knowledge and understanding about the transition to high school and the implementation of career choices. Aligned with the literature, this study supports the calls for integrative psychological practices that acknowledge social-emotional, career, and academic variables when fostering students’ psychosocial adjustment.
The Different Roles of Structural and Cognitive Social Capital on Oral Health-Related Quality of Life among Adolescents
Authors: Knorst Jessica Klöckner , Vettore Mario Vianna , Brondani Bruna , Emmanuelli Bruno , Ardenghi Thiago Machado
Published: April 2023

This study evaluated the relationship of structural and cognitive dimensions of social capital with oral health-related quality of life (OHRQoL) among adolescents. This was a cross-sectional study nested in a cohort of adolescents from southern Brazil. OHRQoL was evaluated using the short version of the Child Perceptions Questionnaire 11-14 (CPQ11-14). Structural social capital was measured by attendance of religious meetings and social networks from friends and neighbours. Cognitive social capital was evaluated through trust in friends and neighbours, perception of relationships in the neighbourhood, and social support during hard times. Multilevel Poisson regression analysis was performed to estimate the association between social capital dimensions and overall CPQ11-14 scores; higher scores corresponded to worse OHRQoL. The sample comprised 429 adolescents with a mean age of 12 years. Adolescents who attended religious meetings less than once a month or never presented higher overall CPQ11-14 scores. Adolescents who did not trust their friends and neighbours, those who believe that their neighbours did not have good relationships, and those reporting no support during hard times also presented higher overall CPQ11-14 scores. OHRQoL was poorer in individuals who presented lower structural and cognitive social capital, with the greatest impact related to the cognitive dimension.
Behavior Change in Chronic Health: Reviewing What We Know, What Is Happening, and What Is Next for Hearing Loss
Authors: Brice Sophie , Almond Helen
Published: April 2023

Untreated age-related sensorineural hearing loss is challenged by low adoption and adherence to hearing aids for treatment. Hearing care has evolved from traditional clinic-controlled treatment to online consumer-centered hearing care, supported by the increasingly person-centered design of hearing aid technology. Greater evidence and a more nuanced understanding of the personal need for adoption versus adherence to the use of consumer hearing care devices are required. Research considering consumer hearing aid acceptance behavior rests on behavior modification theories to guide clinical approaches to increasing hearing aid adoption and adherence. However, in the context of complex chronic health management, there may be a gap in how these theories effectively align with the needs of consumers. Similarly, market data indicates evolving consumer behavior patterns have implications for hearing care theory and implementation, particularly in terms of sustained behavior change. This essay proposes that evidence, including theory and application, be strengthened by revising basic theoretical premises of personal experience with complex chronic health, in addition to considering recent changes in commercial contexts.
SNP in PTPN22, PADI4, and STAT4 but Not TRAF1 and CD40 Increase the Risk of Rheumatoid Arthritis in Polish Population
Authors: Budlewski Tomasz , Sarnik Joanna , Galita Grzegorz , Dragan Grzegorz , Brzezińska Olga , Popławska Marta , Popławski Tomasz , Makowska Joanna
Published: April 2023

Single nucleotide polymorphisms in non-HLA genes are involved in the development of rheumatoid arthritis (RA). SNPS in genes: PADI4 (rs2240340), STAT4 (rs7574865), CD40 (rs4810485), PTPN22 (rs2476601), and TRAF1 (rs3761847) have been described as risk factors for the development of autoimmune diseases, including RA. This study aimed to assess the prevalence of polymorphisms of these genes in the Polish population of patients with rheumatoid arthritis as compared to healthy controls. 324 subjects were included in the study: 153 healthy subjects and 181 patients from the Department of Rheumatology, Medical University of Lodz who fulfilled the criteria of rheumatoid arthritis diagnosis. Genotypes were determined by Taqman SNP Genotyping Assay. rs2476601 (G/A, OR = 2.16, CI = 1.27–3.66; A/A, OR = 10.35, CI = 1.27–84.21), rs2240340 (C/T, OR = 4.35, CI = 2.55–7.42; T/T, OR = 2.80, CI = 1.43–4.10) and rs7574865 (G/T, OR = 1.97, CI = 1.21–3.21; T/T, OR = 3.33, CI = 1.01–11.02) were associated with RA in the Polish population. Rs4810485 was also associated with RA, however after Bonferroni’s correction was statistically insignificant. We also found an association between minor alleles of rs2476601, rs2240340, and rs7574865 and RA (OR = 2.32, CI = 1.47–3.66; OR = 2.335, CI = 1.64–3.31; OR = 1.88, CI = 1.27–2.79, respectively). Multilocus analysis revealed an association between CGGGT and rare (below 0.02 frequency) haplotypes (OR = 12.28, CI = 2.65–56.91; OR = 3.23, CI = 1.63–6.39). In the Polish population, polymorphisms of the PADI4, PTPN22, and STAT4 genes have been detected, which are also known risk factors for RA in various other populations.
Comparative Physiological and Transcriptomic Mechanisms of Defoliation in Cotton in Response to Thidiazuron versus Ethephon
Authors: Liao Baopeng , Li Fangjun , Yi Fei , Du Mingwei , Tian Xiaoli , Li Zhaohu
Published: April 2023

Thidiazuron (TDZ) is a widely used chemical defoliant in cotton and can stimulate the production of ethylene in leaves, which is believed to be the key factor in inducing leaf abscission. Ethephon (Eth) can also stimulate ethylene production in leaves, but it is less effective in promoting leaf shedding. In this study, the enzyme-linked immunosorbent assays (ELISA) and RNA-seq were used to determine specific changes at hormonal levels as well as transcriptomic mechanisms induced by TDZ compared with Eth. The TDZ significantly reduced the levels of auxin and cytokinin in cotton leaves, but no considerable changes were observed for Eth. In addition, TDZ specifically increased the levels of brassinosteroids and jasmonic acid in the leaves. A total of 13 764 differentially expressed genes that specifically responded to TDZ were identified by RNA-seq. The analysis of KEGG functional categories suggested that the synthesis, metabolism, and signal transduction of auxin, cytokinin, and brassinosteroid were all involved in the TDZ-induced abscission of cotton leaves. Eight auxin transport genes (GhPIN1-c_D, GhPIN3_D, GhPIN8_A, GhABCB19-b_A, GhABCB19-b_D, GhABCB2-b_D, GhLAX6_A, and GhLAX7_D) specifically responded to TDZ. The pro35S::GhPIN3a::YFP transgenic plants showed lower defoliation than the wild type treated with TDZ, and YFP fluorescence in leaves was almost extinguished after treatment with TDZ rather than Eth. This provides direct evidence that GhPIN3a is involved in the leaf abscission induced by TDZ. We found that 959 transcription factors (TFs) specifically responded to TDZ, and a co-expression network analysis (WGCNA) showed five hub TFs (GhNAC72, GhWRKY51, GhWRKY70, GhWRKY50, and GhHSF24) during chemical defoliation with TDZ. Our work sheds light on the molecular basis of TDZ-induced leaf abscission in cotton.
Glutamine Deficiency Promotes Immune and Endothelial Cell Dysfunction in COVID-19
Authors: Durante William
Published: April 2023

The coronavirus disease 2019 (COVID-19) pandemic has caused the death of almost 7 million people worldwide. While vaccinations and new antiviral drugs have greatly reduced the number of COVID-19 cases, there remains a need for additional therapeutic strategies to combat this deadly disease. Accumulating clinical data have discovered a deficiency of circulating glutamine in patients with COVID-19 that associates with disease severity. Glutamine is a semi-essential amino acid that is metabolized to a plethora of metabolites that serve as central modulators of immune and endothelial cell function. A majority of glutamine is metabolized to glutamate and ammonia by the mitochondrial enzyme glutaminase (GLS). Notably, GLS activity is upregulated in COVID-19, favoring the catabolism of glutamine. This disturbance in glutamine metabolism may provoke immune and endothelial cell dysfunction that contributes to the development of severe infection, inflammation, oxidative stress, vasospasm, and coagulopathy, which leads to vascular occlusion, multi-organ failure, and death. Strategies that restore the plasma concentration of glutamine, its metabolites, and/or its downstream effectors, in conjunction with antiviral drugs, represent a promising therapeutic approach that may restore immune and endothelial cell function and prevent the development of occlusive vascular disease in patients stricken with COVID-19.
Antiepileptic Properties of Scyllo-Inositol on Pentylenetetrazol-Induced Seizures
Authors: Wiśniewski Karol , Antonowski Tomasz , Juranek Judyta , Podlasz Piotr , Wojtkiewicz Joanna
Published: April 2023

Epilepsy, with about 70 million affected people worldwide, is one of the biggest challenges of medicine today. It is estimated that about one-third of epileptic patients receive inadequate treatment. Inositols have proved effective in many disorders; hence, in the current study, we tested potential antiepileptic properties of scyllo-inositol (SCI)—one of the most common commercially available inositols—in zebrafish larvae with pentylenetetrazol-induced seizures. First, we studied the general effect of SCI on zebrafish motility, and then we tested SCI antiepileptic properties over short (1 h) and long (120 h) exposure protocols. Our results demonstrated that SCI alone does not reduce zebrafish motility regardless of the dose. We also observed that short-term exposure to SCI groups reduced PTZ-treated larva motility compared to controls (p < 0.05). In contrast, prolonged exposure did not produce similar results, likely due to the insufficient concentration of SCI given. Our results highlight the potential of SCI use in epilepsy treatment and warrant further clinical studies with inositols as potential seizure-reducing drugs.
Singlet Oxygen Generation Driven by Sulfide Ligand Exchange on Porphyrin–Gold Nanoparticle Conjugates
Authors: Shinohara Akira , Shinmori Hideyuki
Published: April 2023

Here, we report a switching method of singlet oxygen (1O2) generation based on the adsorption/desorption of porphyrins to gold nanoparticles driven by sulfide (thiol or disulfide) compounds. The generation of 1O2 by photosensitization is effectively suppressed by the gold nanoparticles and can be restored by a sulfide ligand exchange reaction. The on/off ratio of 1O2 quantum yield (ΦΔ) reached 7.4. By examining various incoming sulfide compounds, it was found that the ligand exchange reaction on the gold nanoparticle surface could be thermodynamically or kinetically controlled. The remaining gold nanoparticles in the system still suppress the generation of 1O2, which can be precipitated out simultaneously with porphyrin desorption by the proper polarity choice of the incoming sulfide to restore the 1O2 generation.
Treatment of Diet-Induced Obese Rats with CB2 Agonist AM1241 or CB2 Antagonist AM630 Reduces Leptin and Alters Thermogenic mRNA in Adipose Tissue
Authors: O’Keefe Lannie , Vu Teresa , Simcocks Anna C. , Jenkin Kayte A. , Mathai Michael L. , McAinch Andrew J. , Hutchinson Dana S. , Hryciw Deanne H.
Published: April 2023

Diet-induced obesity (DIO) is a contributor to co-morbidities, resulting in alterations in hormones, lipids, and low-grade inflammation, with the cannabinoid type 2 receptor (CB2) contributing to the inflammatory response. The effects of modulating CB2 with pharmacological treatments on inflammation and adaptations to the obese state are not known. Therefore, we aimed to investigate the molecular mechanisms in adipose tissue of CB2 agonism and CB2 antagonism treatment in a DIO model. Male Sprague Dawley rats were placed on a high-fat diet (HFD) (21% fat) for 9 weeks, then received daily intraperitoneal injections with a vehicle, AM630 (0.3 mg/kg), or AM1241 (3 mg/kg), for a further 6 weeks. AM630 or AM1241 treatment in DIO rats did not alter their body weight, food intake, or liver weight, and it had no effect on their numerous circulating cytokines or peri-renal fat pad mass. AM1241 decreased heart weight and BAT weight; both treatments (AM630 or AM1241) decreased plasma leptin levels, while AM630 also decreased plasma ghrelin and GLP-1 levels. Both treatments decreased Adrb3 and TNF-α mRNA levels in eWAT and TNF-α levels in pWAT. AM630 treatment also decreased the mRNA levels of Cnr2, leptin, and Slc2a4 in eWAT. In BAT, both treatments decreased leptin, UCP1, and Slc2a4 mRNA levels, with AM1241 also decreasing Adrb3, IL1β, and PRDM16 mRNA levels, and AM630 increasing IL6 mRNA levels. In DIO, CB2 agonist and CB2 antagonist treatment reduces circulating leptin in the absence of weight loss and modulates the mRNA responsible for thermogenesis.
Overview of the Molecular Mechanism of Bacterial Environmental Adaptation by Comprehensive Analysis
Authors: Shimada Tomohiro , Yoshida Hideji
Published: April 2023

So far, the genome sequences of more than tens of thousands of organisms have been determined, and the overall picture of the genes that make up one organism has been clarified [https://www [...]
Prenatal Exposure to Metabolism-Disrupting Chemicals, Cord Blood Transcriptome Perturbations, and Birth Weight in a Belgian Birth Cohort
Authors: Cai Anran , Portengen Lützen , Ertaylan Gökhan , Legler Juliette , Vermeulen Roel , Lenters Virissa , Remy Sylvie
Published: April 2023

Prenatal exposure to metabolism-disrupting chemicals (MDCs) has been linked to birth weight, but the molecular mechanisms remain largely unknown. In this study, we investigated gene expressions and biological pathways underlying the associations between MDCs and birth weight, using microarray transcriptomics, in a Belgian birth cohort. Whole cord blood measurements of dichlorodiphenyldichloroethylene (p,p’-DDE), polychlorinated biphenyls 153 (PCB-153), perfluorooctanoic acid (PFOA), perfluorooctane sulfonic acid (PFOS), and transcriptome profiling were conducted in 192 mother–child pairs. A workflow including a transcriptome-wide association study, pathway enrichment analysis with a meet-in-the-middle approach, and mediation analysis was performed to characterize the biological pathways and intermediate gene expressions of the MDC–birth weight relationship. Among 26,170 transcriptomic features, we successfully annotated five overlapping metabolism-related gene expressions associated with both an MDC and birth weight, comprising BCAT2, IVD, SLC25a16, HAS3, and MBOAT2. We found 11 overlapping pathways, and they are mostly related to genetic information processing. We found no evidence of any significant mediating effect. In conclusion, this exploratory study provides insights into transcriptome perturbations that may be involved in MDC-induced altered birth weight.
Enzymatic Synthesis of a Novel Coumarin Aminophosphonates: Antibacterial Effects and Oxidative Stress Modulation on Selected E. coli Strains
Authors: Koszelewski Dominik , Kowalczyk Paweł , Brodzka Anna , Hrunyk Anastasiia , Kramkowski Karol , Ostaszewski Ryszard
Published: April 2023

The objective of the present study was to evaluate the synergistic effect of two important pharmacophores, coumarin and α-amino dimethyl phosphonate moieties, on antimicrobial activity toward selected LPS-varied E. coli strains. Studied antimicrobial agents were prepared via a Kabachnik–Fields reaction promoted by lipases. The products were provided with an excellent yield (up to 92%) under mild, solvent- and metal-free conditions. A preliminary exploration of coumarin α-amino dimethyl phosphonate analogs as novel antimicrobial agents was carried out to determine the basic features of the structure responsible for the observed biological activity. The structure–activity relationship revealed that an inhibitory activity of the synthesized compounds is strongly related to the type of the substituents located in the phenyl ring. The collected data demonstrated that coumarin-based α-aminophosphonates can be potential antimicrobial drug candidates, which is particularly crucial due to the constantly increasing resistance of bacteria to commonly used antibiotics.
Waiting Time Distributions in Hybrid Models of Motor–Bead Assays: A Concept and Tool for Inference
Authors: Ertel Benjamin , van der Meer Jann , Seifert Udo
Published: April 2023

In single-molecule experiments, the dynamics of molecular motors are often observed indirectly by measuring the trajectory of an attached bead in a motor–bead assay. In this work, we propose a method to extract the step size and stalling force for a molecular motor without relying on external control parameters. We discuss this method for a generic hybrid model that describes bead and motor via continuous and discrete degrees of freedom, respectively. Our deductions are solely based on the observation of waiting times and transition statistics of the observable bead trajectory. Thus, the method is non-invasive, operationally accessible in experiments and can, in principle, be applied to any model describing the dynamics of molecular motors. We briefly discuss the relation of our results to recent advances in stochastic thermodynamics on inference from observable transitions. Our results are confirmed by extensive numerical simulations for parameters values of an experimentally realized F1-ATPase assay.
Transcriptomic Analysis Reveals Key Roles of (p)ppGpp and DksA in Regulating Metabolism and Chemotaxis in Yersinia enterocolitica
Authors: Huang Can , Li Wenqian , Chen Jingyu
Published: April 2023

The stringent response is a rapid response system that is ubiquitous in bacteria, allowing them to sense changes in the external environment and undergo extensive physiological transformations. However, the regulators (p)ppGpp and DksA have extensive and complex regulatory patterns. Our previous studies demonstrated that (p)ppGpp and DksA in Yersinia enterocolitica positively co-regulated motility, antibiotic resistance, and environmental tolerance but had opposite roles in biofilm formation. To reveal the cellular functions regulated by (p)ppGpp and DksA comprehensively, the gene expression profiles of wild-type, ΔrelA, ΔrelAΔspoT, and ΔdksAΔrelAΔspoT strains were compared using RNA-Seq. Results showed that (p)ppGpp and DksA repressed the expression of ribosomal synthesis genes and enhanced the expression of genes involved in intracellular energy and material metabolism, amino acid transport and synthesis, flagella formation, and the phosphate transfer system. Additionally, (p)ppGpp and DksA inhibited amino acid utilization (such as arginine and cystine) and chemotaxis in Y. enterocolitica. Overall, the results of this study unraveled the link between (p)ppGpp and DksA in the metabolic networks, amino acid utilization, and chemotaxis in Y. enterocolitica and enhanced the understanding of stringent responses in Enterobacteriaceae.
Synthesis of Ammonium-Based ILs with Different Lengths of Aliphatic Chains and Organic Halogen-Free Anions as Corrosion Inhibitors of API X52 Steel
Authors: Cornejo Robles Emiliano , Olivares-Xometl Octavio , Likhanova Natalya V. , Arellanes-Lozada Paulina , Lijanova Irina V. , Díaz-Jiménez Víctor
Published: April 2023

In the present work, synthesis and characterization of 15 ionic liquids (ILs) derived from quaternary ammonium and carboxylates were carried out in order to proceed to their evaluation as corrosion inhibitors (CIs) of API X52 steel in 0.5 M HCl. Potentiodynamic tests confirmed the inhibition efficiency (IE) as a function of the chemical configuration of the anion and cation. It was observed that the presence of two carboxylic groups in long linear aliphatic chains reduced the IE, whereas in shorter chains it was increased. Tafel-polarization results revealed the ILs as mixed-type CIs and that the IE was directly proportional to the CI concentration. The compounds with the best IE were 2-amine-benzoate of N,N,N-trimethyl-hexadecan-1-ammonium ([THDA+][−AA]), 3-carboxybut-3-enoate of N,N,N-trimethyl-hexadecan-1-ammonium ([THDA+][−AI]), and dodecanoate of N,N,N-trimethyl-hexadecan-1-ammonium ([THDA+][−AD]) within the 56–84% interval. Furthermore, it was found that the ILs obeyed the Langmuir adsorption isotherm model and inhibited the corrosion of steel through a physicochemical process. Finally, the surface analysis by scanning electron microscopy (SEM) confirmed less steel damage in the presence of CI due to the inhibitor–metal interaction.
Influence of Simulated Microgravity on Mammary Epithelial Cells Grown as 2D and 3D Cultures
Authors: Winkelmaier Garrett , Jabbari Kosar , Chien Lung-Chang , Grabham Peter , Parvin Bahram , Pluth Janice
Published: April 2023

During space travel, astronauts will experience a unique environment that includes continuous exposure to microgravity and stressful living conditions. Physiological adaptation to this is a challenge and the effect of microgravity on organ development, architecture, and function is not well understood. How microgravity may impact the growth and development of an organ is an important issue, especially as space flight becomes more commonplace. In this work, we sought to address fundamental questions regarding microgravity using mouse mammary epithelial cells in 2D and 3D tissue cultures exposed to simulated microgravity. Mouse mammary HC11 cells contain a higher proportion of stem cells and were also used to investigate how simulated microgravity may impact mammary stem cell populations. In these studies, we exposed mouse mammary epithelial cells to simulated microgravity in 2D and then assayed for changes in cellular characteristics and damage levels. The microgravity treated cells were also cultured in 3D to form acini structures to define if simulated microgravity affects the cells’ ability to organize correctly, a quality that is of key importance for mammary organ development. These studies identify changes occurring during exposure to microgravity that impact cellular characteristics such as cell size, cell cycle profiles, and levels of DNA damage. In addition, changes in the percentage of cells revealing various stem cell profiles were observed following simulated microgravity exposure. In summary, this work suggests microgravity may cause aberrant changes in mammary epithelial cells that lead to an increase in cancer risk.
PTHrP Modulates the Proliferation and Osteogenic Differentiation of Craniofacial Fibrous Dysplasia-Derived BMSCs
Authors: Shen Lihang , He Yang , Chen Shuo , He Linhai , Zhang Yi
Published: April 2023

Fibrous dysplasia (FD) is a skeletal stem cell disease caused by mutations in the guanine nucleotide-binding protein, alpha-stimulating activity polypeptide (GNAS) gene, which results in the abnormal accumulation of cyclic adenosine monophosphate (cAMP) and hyperactivation of downstream signaling pathways. Parathyroid hormone-related protein (PTHrP) is secreted by the osteoblast lineage and is involved in various physiological and pathological activities of bone. However, the association between the abnormal expression of PTHrP and FD, as well as its underlying mechanism, remains unclear. In this study, we discovered that FD patient-derived bone marrow stromal cells (FD BMSCs) expressed significantly higher levels of PTHrP during osteogenic differentiation and exhibited greater proliferation capacity but impaired osteogenic ability compared to normal control patient-derived BMSCs (NC BMSCs). Continuous exogenous PTHrP exposure on the NC BMSCs promoted the FD phenotype in both in vitro and in vivo experiments. Through the PTHrP/cAMP/PKA axis, PTHrP could partially influence the proliferation and osteogenesis capacity of FD BMSCs via the overactivation of the Wnt/β-Catenin signaling pathway. Furthermore, PTHrP not only directly modulated cAMP/PKA/CREB transduction but was also demonstrated as a transcriptional target of CREB. This study provides novel insight into the possible pathogenesis involved in the FD phenotype and enhances the understanding of its molecular signaling pathways, offering theoretical evidence for the feasibility of potential therapeutic targets for FD.
Knockdown of BAP31 Overcomes Hepatocellular Carcinoma Doxorubicin Resistance through Downregulation of Survivin
Authors: Liu Jingjing , Zhang Qi , Wang Changli , Yang Jiaying , Yang Sheng , Wang Tianyi , Wang Bing
Published: April 2023

The expression of B-cell receptor associated protein 31 (BAP31) is increased in many tumor types, and it is reported to participate in proliferation, migration, and apoptosis. However, the relationship between BAP31 and chemoresistance is uncertain. This study investigated the role of BAP31 in regulating the doxorubicin (Dox) resistance of hepatocellular carcinoma (HCC). The expression of proteins was assessed by Western blotting. The correlation between BAP31 expression and Dox resistance was examined by MTT and colony formation assays. Apoptosis was analyzed by flow cytometry and TdT-mediated dUTP nick end labeling assays. Western blot and immunofluorescence analyses were performed in the knockdown cell lines to explore the possible mechanisms. In this study, BAP31 was strongly expressed, and knockdown of BAP31 increased Dox chemosensitivity in cancer cells. Furthermore, the expression of BAP31 was higher in the Dox-resistant HCC cells than that in their parental cells; knockdown of BAP31 reduced the half maximal inhibitory concentration value and overcame Dox resistance in Dox-resistant HCC cells. In HCC cells, knockdown of BAP31 increased Dox-induced apoptosis and enhanced Dox chemosensitivity in vitro and in vivo. The potential mechanism by which BAP31 increased Dox-induced apoptosis is that BAP31 inhibited survivin expression by promoting FoxO1 nucleus–cytoplasm translocation. Knockdown of BAP31 and survivin had a synergistic effect on Dox chemosensitivity by enhancing the apoptosis of HCC cells. These findings reveal that BAP31 knockdown enhances Dox chemosensitivity through the downregulation of survivin, suggesting that BAP31 is a potential therapeutic target for improving the treatment response of HCC with resistance to Dox.
Mutated Flt3Lg Provides Reduced Flt3 Recycling Compared to Wild-Type Flt3Lg and Retains the Specificity of Flt3Lg-Based CAR T-Cell Targeting in AML Models
Authors: Maiorova Varvara , Mollaev Murad D. , Vikhreva Polina , Chudakov Dmitriy M. , Kibardin Alexey , Maschan Michael A. , Larin Sergey
Published: April 2023

The cells of acute myeloid leukemia are defined by clonal growth and heterogenous immunophenotypes. Chimeric antigen receptors (CARs) commonly recognize molecular targets by single-chain antibody fragments (scFvs) specific to a tumor-associated antigen. However, ScFvs may form aggregates, thus stimulating tonic CAR T-cell activation and reducing CAR T-cell functioning in vivo. Harnessing natural ligands as recognition parts of CARs, specific targeting of membrane receptors can be achieved. Previously, we presented ligand-based Flt3-CAR T-cells targeting the Flt3 receptor. The extracellular part of Flt3-CAR consisted of full-size Flt3Lg. Meanwhile, upon recognition, Flt3-CAR may potentially activate Flt3, triggering proliferative signaling in blast cells. Moreover, the long-lasting presence of Flt3Lg may lead to Flt3 downregulation. In this paper, we present mutated Flt3Lg-based Flt3m-CAR (‘m’—for ‘mutant’) T-cells targeting Flt3. The extracellular part of Flt3m-CAR consists of full-length Flt3Lg-L27P. We have determined that ED50 for recombinant Flt3Lg-L27P produced in CHO cells is at least 10-fold higher than for the wild-type Flt3Lg. We show that the mutation in the recognizing domain of Flt3m-CAR did not affect the specificity of Flt3m-CAR T-cells when compared to Flt3-CAR T-cells. Flt3m-CAR T-cells combine the specificity of ligand–receptor recognition with reduced Flt3Lg-L27P bioactivity, leading to potentially safer immunotherapy.
Impact of Senolytic Treatment on Gene Expression in Aged Lung
Authors: Cho Soo Jung , Pronko Alexander , Yang Jianjun , Stout-Delgado Heather
Published: April 2023

Cellular senescence plays a key role in mediating tissue remodeling and modulation of host responses to pathogenic stimuli. Our current study was designed to gain a better understanding of the impact of short-term senolytic treatment or inflammatory stimulation on lung senescence. The results of our study demonstrate that short term treatment of aged adult mice (20 months of age) with senolytics, quercetin, and dasatinib decreases p16 and p21 expression in lung tissue. Short-term treatment with senolytics also significantly improved the expression of genes associated with genomic instability, telomere attrition, mitochondrial dysfunction, DNA binding, and the inflammatory response. In contrast, in response to low-dose LPS administration, there was increased expression of genes associated with genomic instability, mitochondrial dysfunction, and heightened inflammatory responses in young adult murine lung (3 months of age). Taken together, the results of our current study illustrate the efficacy of senolytic treatment on modulating responses in aged lung and the potential role of chronic low dose inflammation on senescence induction in the lung.
A Concerted Vision to Advance the Knowledge of Diabetes Mellitus Related to Immune Checkpoint Inhibitors
Authors: Deligiorgi Maria V. , Trafalis Dimitrios T.
Published: April 2023

The rubric of immune-related (ir) diabetes mellitus (DM) (irDM) encompasses various hyperglycemic disorders related to immune checkpoint inhibitors (ICPis). Beyond sharing similarities with conventional DM, irDM is a distinct, yet important, entity. The present narrative review provides a comprehensive overview of the literature regarding irDM published in major databases from January 2018 until January 2023. Initially considered rare, irDM is increasingly being reported. To advance the knowledge of irDM, the present review suggests a concerted vision comprising two intertwined aspects: a scientific-centered and a patient-centered view. The scientific-centered aspect addresses the pathophysiology of irDM, integrating: (i) ICPi-induced pancreatic islet autoimmunity in genetically predisposed patients; (ii) altered gut microbiome; (iii) involvement of exocrine pancreas; (iv) immune-related acquired generalized lipodystrophy. The patient-centered aspect is both nurtured by and nurturing the four pillars of the scientific-centered aspect: awareness, diagnosis, treatment, and monitoring of irDM. The path forward is a multidisciplinary initiative towards: (i) improved characterization of the epidemiological, clinical, and immunological profile of irDM; (ii) standardization of reporting, management, and surveillance protocols for irDM leveraging global registries; (iii) patient stratification according to personalized risk for irDM; (iv) new treatments for irDM; and (v) uncoupling ICPi efficacy from immunotoxicity.
Proteomic Profiling of Mouse Brain Pyruvate Kinase Binding Proteins: A Hint for Moonlighting Functions of PKM1?
Authors: Buneeva Olga , Kopylov Arthur , Gnedenko Oksana , Medvedeva Marina , Veselovsky Alexander , Ivanov Alexis , Zgoda Victor , Medvedev Alexei
Published: April 2023

Affinity-based proteomic profiling is widely used for the identification of proteins involved in the formation of various interactomes. Since protein–protein interactions (PPIs) reflect the role of particular proteins in the cell, identification of interaction partners for a protein of interest can reveal its function. The latter is especially important for the characterization of multifunctional proteins, which can play different roles in the cell. Pyruvate kinase (PK), a classical glycolytic enzyme catalyzing the last step of glycolysis, exists in four isoforms: PKM1, PKM2, PKL, and PKR. The enzyme isoform expressed in actively dividing cells, PKM2, exhibits many moonlighting (noncanonical) functions. In contrast to PKM2, PKM1, predominantly expressed in adult differentiated tissues, lacks well-documented moonlighting functions. However, certain evidence exists that it can also perform some functions unrelated to glycolysis. In order to evaluate protein partners, bound to PKM1, in this study we have combined affinity-based separation of mouse brain proteins with mass spectrometry identification. The highly purified PKM1 and a 32-mer synthetic peptide (PK peptide), sharing high sequence homology with the interface contact region of all PK isoforms, were used as the affinity ligands. This proteomic profiling resulted in the identification of specific and common proteins bound to both affinity ligands. Quantitative affinity binding to the affinity ligands of selected identified proteins was validated using a surface plasmon resonance (SPR) biosensor. Bioinformatic analysis has shown that the identified proteins, bound to both full-length PKM1 and the PK peptide, form a protein network (interactome). Some of these interactions are relevant for the moonlighting functions of PKM1. The proteomic dataset is available via ProteomeXchange with the identifier PXD041321.
An Agrobacterium-Mediated Transient Expression Method for Functional Assay of Genes Promoting Disease in Monocots
Authors: Xu Haijiao , Chang Qingle , Huang Luli , Wei Peiyao , Song Yulu , Guo Zejian , Peng You-Liang , Fan Jun
Published: April 2023

Agrobacterium-mediated transient expression (AMTE) has been widely used for high-throughput assays of gene function in diverse plant species. However, its application in monocots is still limited due to low expression efficiency. Here, by using histochemical staining and a quantitative fluorescence assay of β-glucuronidase (GUS) gene expression, we investigated factors affecting the efficiency of AMTE on intact barley plants. We found prominent variation in GUS expression levels across diverse vectors commonly used for stable transformation and that the vector pCBEP produced the highest expression. Additionally, concurrent treatments of plants with one day of high humidity and two days of darkness following agro-infiltration also significantly increased GUS expression efficiency. We thus established an optimized method for efficient AMTE on barley and further demonstrated its efficiency on wheat and rice plants. We showed that this approach could produce enough proteins suitable for split-luciferase assays of protein-protein interactions on barley leaves. Moreover, we incorporated the AMTE protocol into the functional dissection of a complex biological process such as plant disease. Based on our previous research, we used the pCBEP vector to construct a full-length cDNA library of genes upregulated during the early stage of rice blast disease. A subsequent screen of the library by AMTE identified 15 candidate genes (out of ~2000 clones) promoting blast disease on barley plants. Four identified genes encode chloroplast-related proteins: OsNYC3, OsNUDX21, OsMRS2-9, and OsAk2. These genes were induced during rice blast disease; however, constitutive overexpression of these genes conferred enhanced disease susceptibility to Colletotrichum higginsianum in Arabidopsis. These observations highlight the power of the optimized AMTE approach on monocots as an effective tool for facilitating functional assays of genes mediating complex processes such as plant-microbe interactions.
SOX2-Sensing: Insights into the Role of SOX2 in the Generation of Sensory Cell Types in Vertebrates
Authors: Mercurio Sara
Published: April 2023

The SOX2 transcription factor is a key regulator of nervous system development, and its mutation in humans leads to a rare disease characterized by severe eye defects, cognitive defects, hearing defects, abnormalities of the CNS and motor control problems. SOX2 has an essential role in neural stem cell maintenance in specific regions of the brain, and it is one of the master genes required for the generation of induced pluripotent stem cells. Sox2 is expressed in sensory organs, and this review will illustrate how it regulates the differentiation of sensory cell types required for hearing, touching, tasting and smelling in vertebrates and, in particular, in mice.
Biodegradation Studies of Polyhydroxybutyrate and Polyhydroxybutyrate-co-Polyhydroxyvalerate Films in Soil
Authors: Kim Jihyeon , Gupta Nevin S. , Bezek Lindsey B. , Linn Jacqueline , Bejagam Karteek K. , Banerjee Shounak , Dumont Joseph H. , Nam Sang Yong , Kang Hyun Woo , Park Chi Hoon , Pilania Ghanshyam , Iverson Carl N. , Marrone Babetta L. , Lee Kwan-Soo
Published: April 2023

Due to increased environmental pressures, significant research has focused on finding suitable biodegradable plastics to replace ubiquitous petrochemical-derived polymers. Polyhydroxyalkanoates (PHAs) are a class of polymers that can be synthesized by microorganisms and are biodegradable, making them suitable candidates. The present study looks at the degradation properties of two PHA polymers: polyhydroxybutyrate (PHB) and polyhydroxybutyrate-co-polyhydroxyvalerate (PHBV; 8 wt.% valerate), in two different soil conditions: soil fully saturated with water (100% relative humidity, RH) and soil with 40% RH. The degradation was evaluated by observing the changes in appearance, chemical signatures, mechanical properties, and molecular weight of samples. Both PHB and PHBV were degraded completely after two weeks in 100% RH soil conditions and showed significant reductions in mechanical properties after just three days. The samples in 40% RH soil, however, showed minimal changes in mechanical properties, melting temperatures/crystallinity, and molecular weight over six weeks. By observing the degradation behavior for different soil conditions, these results can pave the way for identifying situations where the current use of plastics can be replaced with biodegradable alternatives.
Specific Activation of T Cells by an ACE2-Based CAR-Like Receptor upon Recognition of SARS-CoV-2 Spike Protein
Authors: Gonzalez-Garcia Pablo , Muñoz-Miranda Juan P. , Fernandez-Cisnal Ricardo , Olvera Lucia , Moares Noelia , Gabucio Antonio , Fernandez-Ponce Cecilia , Garcia-Cozar Francisco
Published: April 2023

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the causative agent of the Coronavirus Disease 2019 (COVID-19) pandemic, which is still a health issue worldwide mostly due to a high rate of contagiousness conferred by the high-affinity binding between cell viral receptors, Angiotensin-Converting Enzyme 2 (ACE2) and SARS-CoV-2 Spike protein. Therapies have been developed that rely on the use of antibodies or the induction of their production (vaccination), but despite vaccination being still largely protective, the efficacy of antibody-based therapies wanes with the advent of new viral variants. Chimeric Antigen Receptor (CAR) therapy has shown promise for tumors and has also been proposed for COVID-19 treatment, but as recognition of CARs still relies on antibody-derived sequences, they will still be hampered by the high evasion capacity of the virus. In this manuscript, we show the results from CAR-like constructs with a recognition domain based on the ACE2 viral receptor, whose ability to bind the virus will not wane, as Spike/ACE2 interaction is pivotal for viral entry. Moreover, we have developed a CAR construct based on an affinity-optimized ACE2 and showed that both wild-type and affinity-optimized ACE2 CARs drive activation of a T cell line in response to SARS-CoV-2 Spike protein expressed on a pulmonary cell line. Our work sets the stage for the development of CAR-like constructs against infectious agents that would not be affected by viral escape mutations and could be developed as soon as the receptor is identified.
Interaction Network Construction and Functional Analysis of the Plasma Membrane H+-ATPase in Bangia fuscopurpurea (Rhodophyta)
Authors: Yao Haiqin , Wang Wenjun , Cao Yuan , Liang Zhourui , Zhang Pengyan
Published: April 2023

Salinity is a serious threat to most land plants. Although seaweeds adapt to salty environments, intertidal species experience wide fluctuations in external salinities, including hyper- and hypo-saline stress. Bangia fuscopurpurea is an economic intertidal seaweed with a strong tolerance to hypo-salinity. Until now, the salt stress tolerance mechanism has remained elusive. Our previous study showed that the expression of B. fuscopurpurea plasma membrane H+-ATPase (BfPMHA) genes were the most upregulated under hypo-salinity. In this study, we obtained the complete sequence of BfPMHA, traced the relative expression of this BfPMHA gene in B. fuscopurpurea under hypo-salinity, and analyzed the protein structure and properties based on the gene’s sequence. The result showed that the expression of BfPMHA in B. fuscopurpurea increased significantly with varying hypo-salinity treatments, and the higher the degree of low salinity stress, the higher the expression level. This BfPMHA had typical PMHA structures with a Cation-N domain, an E1-E2 ATPase domain, a Hydrolase domain, and seven transmembrane domains. In addition, through the membrane system yeast two-hybrid library, three candidate proteins interacting with BfPMHA during hypo-saline stress were screened, fructose–bisphosphate aldolase (BfFBA), glyceraldehyde 3-phosphate dehydrogenase (NADP+) (phosphorylating) (BfGAPDH), and manganese superoxide dismutase (BfMnSOD). The three candidates and BfPMHA genes were successfully transferred and overexpressed in a BY4741 yeast strain. All of them significantly enhanced the yeast tolerance to NaCl stress, verifying the function of BfPMHA in salt stress response. This is the first study to report the structure and topological features of PMHA in B. fuscopurpurea and its candidate interaction proteins in response to salt stress.
HD-ZIP Transcription Factors and Brassinosteroid Signaling Play a Role in Capitulum Patterning in Chrysanthemum
Authors: Castricum Annemarie , Bakker Erin H. , de Vetten Nick C. M. H. , Weemen Mieke , Angenent Gerco C. , Immink Richard G. H. , Bemer Marian
Published: April 2023

Chrysanthemum is a genus in the Asteraceae family containing numerous cut flower varieties with high ornamental value. It owes its beauty to the composite flower head, which resembles a compact inflorescence. This structure is also known as a capitulum, in which many ray and disc florets are densely packed. The ray florets are localized at the rim, are male sterile, and have large colorful petals. The centrally localized disc florets develop only a small petal tube but produce fertile stamens and a functional pistil. Nowadays, varieties with more ray florets are bred because of their high ornamental value, but, unfortunately, this is at the expense of their seed setting. In this study, we confirmed that the disc:ray floret ratio is highly correlated to seed set efficiency, and therefore, we further investigated the mechanisms that underlie the regulation of the disc:ray floret ratio. To this end, a comprehensive transcriptomics analysis was performed in two acquired mutants with a higher disc:ray floret ratio. Among the differentially regulated genes, various potential brassinosteroid (BR) signaling genes and HD-ZIP class IV homeodomain transcription factors stood out. Detailed follow-up functional studies confirmed that reduced BR levels and downregulation of HD-ZIP IV gene Chrysanthemum morifolium PROTODERMAL FACTOR 2 (CmPDF2) result in an increased disc:ray floret ratio, thereby providing ways to improve seed set in decorative chrysanthemum varieties in the future.