XPR1: a regulator of cellular phosphate homeostasis rather than a Pi exporter

\ua9 The Author(s) 2024. Phosphate (Pi) is an essential nutrient, and its plasma levels are under tight hormonal control. Uphill transport of Pi into cells is mediated by the two Na-dependent Pi transporter families SLC34 and SLC20. The molecular identity of a potential Pi export pathway is controversial, though XPR1 has recently been suggested by Giovannini and coworkers to mediate Pi export. We expressed XPR1 in Xenopus oocytes to determine its functional characteristics. Xenopus isoforms of proteins were used to avoid species incompatibility. Protein tagging confirmed the localization of XPR1 at the plasma membrane. Efflux experiments, however, failed to detect translocation of Pi attributable to XPR1. We tested various counter ions and export medium compositions (pH, plasma) as well as potential protein co-factors that could stimulate the activity of XPR1, though without success. Expression of truncated XPR1 constructs and individual domains of XPR1 (SPX, transmembrane core, C-terminus) demonstrated downregulation of the uptake of Pi mediated by the C-terminal domain of XPR1. Tethering the C-terminus to the transmembrane core changed the kinetics of the inhibition and the presence of the SPX domain blunted the inhibitory effect. Our observations suggest a regulatory role of XPR1 in cellular Pi handling rather than a function as Pi exporter. Accordingly, XPR1 senses intracellular Pi levels via its SPX domain and downregulates cellular Pi uptake via the C-terminal domain. The molecular identity of a potential Pi export protein remains therefore elusive

PD-1 Inhibitory Receptor Downregulates Asparaginyl Endopeptidase and Maintains Foxp3 Transcription Factor Stability in Induced Regulatory T Cells

CD4+ T cell differentiation into multiple T helper (Th) cell lineages is critical for optimal adaptive immune responses. This report identifies an intrinsic mechanism by which programmed death-1 receptor (PD-1) signaling imparted regulatory phenotype to Foxp3+ Th1 cells (denoted as Tbet+iTregPDL1 cells) and inducible regulatory T (iTreg) cells. Tbet+iTregPDL1 cells prevented inflammation in murine models of experimental colitis and experimental graft versus host disease (GvHD). Programmed death ligand-1 (PDL-1) binding to PD-1 imparted regulatory function to Tbet+iTregPDL1 cells and iTreg cells by specifically downregulating endo-lysosomal protease asparaginyl endopeptidase (AEP). AEP regulated Foxp3 stability and blocking AEP imparted regulatory function in Tbet+iTreg cells. Also, Aep−/− iTreg cells significantly inhibited GvHD and maintained Foxp3 expression. PD-1-mediated Foxp3 maintenance in Tbet+ Th1 cells occurred both in tumor infiltrating lymphocytes (TILs) and during chronic viral infection. Collectively, this report has identified an intrinsic function for PD-1 in maintaining Foxp3 through proteolytic pathway.Bio-organic Synthesi

Terapia cognitiva : aplicações de uma técnica para qualidade de vida e saúde

Esta pesquisa teve por objetivo geral aplicar e avaliar uma técnica específica de terapia cognitiva - organizada em 12 sessões grupais e denominada Tomada de Decisão e Qualidade de Vida -, destinada a promover saúde e incrementar qualidade de vida. No total, participaram 18 servidores de uma instituição pública de ensino superior. Nas etapas de admissão e de encerramento, aplicaram-se : Questionário de Qualidade de Vida, Inventário Beck de Ansiedade e Inventário Beck de Depressão. Foram identificadas melhoras significativas nos domínios físico, psicológico, meio ambiente, geral e saúde, relacionados à qualidade de vida. Não se verificaram alterações significantes nos escores de ansiedade (p=0,26). Em contrapartida, os escores de depressão indicaram melhora (p=0,02). Os resultados sugerem que a técnica pode ser empregada para promover saúde e qualidade de vida.In this study we implemented and assessed a specific cognitive therapy technique - Decision Making and Quality of Life, which is used to promote health and improve quality of life. Eighteen employees from a higher education institution participated in the study, which was organized into 12 group sessions. At the admission and concluding phases, we asked participants to complete the World Health Organization Quality of Life - Bref Questionnaire, the Beck Anxiety Inventory and the Beck Depression Inventory. Results showed significant improvement in five of the domains that measure quality of life: physical, psychological, environmental, general, and health. There were no significant changes (p=0.26) in anxiety scores. In contrast, the depression scores got significantly better (p=0.02). The results suggest that the proposed technique is conducive to health promotion and quality of life

Impacto de un programa intervención em alunos del segundo ciclo

O objetivo do presente estudo consistiuem avaliar um programa de intervenção junto a alunos do 2º ciclo de escolaridade nas seguintes dimensões: tomada de decisão, conhecimentos sobre sexualidade, competências sociais, assertividade e autoconceito. Metodologia: Participaram 145 alunos, distribuídos pelos grupos controle e experimental. Os instrumentos utilizados foram: TCU Decision-Making; Questionário de Conhecimentos sobre Sexualidade; Assertion Self-Statement Test- Revised; Questionário de Competências Sociais; Piers-Harris Children’s Self-Concept Scale. Os resultados revelaram diferenças no pós-teste entre os grupos ao nível da sexualidade. Verificaram-se diferenças do pré-teste para o pós-teste no grupo experimental nos níveis da sexualidade, da assertividade e das competências sociais. No grupo experimental encontraram-se associações positivas entre tomada de decisão, competências sociais e assertividade, bem como entre sexualidade, competências sociais e autoconceito, no pós-teste. Os preditores da assertividade no pós-teste foram tomada de decisão, sexualidade e competências sociais. Como conclusão, os resultados enfatizam a importância de intervenção junto a adolescentes, particularmente na tomada de decisão, na sexualidade e nas competências sociais. Palavras-chave: Habilidades sociais, sexualidade, autoconceito.In this study we evaluate an intervention program in the following dimensions: Decision Making, Knowledge on Sexuality, Social Skills, Assertiveness and Self-Concept with students in 5th and 6th grade. Methodology: 145 students participated in the study divided by control and experimental group. The instruments used were: Decision-Making TCU, Knowledge on Sexuality Questionnaire; Assertion Self-Statement Test-Revised;Social Skills Questionnaire, and Piers-Harris Children's Self- Concept Scale. The results indicate differences at post-test between the groups on knowledge regarding sexuality. There were also differences from pre-test to post-test in the experimental group on knowledge on sexuality, assertiveness and social skills. Positive associations among decision making, social skills and assertiveness were found as well as among knowledge on sexuality, social skills and self-concept, in the experimental group, in the pos-test. Finally, the predictors of assertiveness regarding health behaviors, in the pos-test were: decision making, knowledge regarding sexuality and social skills. The results emphasize the importance of intervention for adolescents in terms of health promotion particularly in decision making, sexuality and social skills.El objetivo del presente estudio fue evaluar un programa de intervención con alumnos del 2º ciclo de escolaridad en las siguientes dimensiones: Toma de Decisión, Conocimientos sobre Sexualidad, Habilidades Sociales, Asertividad y Autoconcepto. Metodología: Participaron 145 alumnos, distribuidos en grupo control y experimental. Los instrumentos utilizados fueron: TCU Decision-Making; Cuestionario de Conocimientos sobre Sexualidad; AssertionSelf-Statement Test-Revised; Cuestionario de Habilidades Sociais; Piers-Harris Children'sSelf-Concept Scale. Los resultados mostraron diferencias en el post-test entre los grupos en cuanto a la sexualidad. Se verificaron diferencias del pre-test para el post-teste en el grupo experimental, cuanto a sexualidad, asertividad y habilidades sociales. Se encontraron asociaciones positivas entre toma de decisión, habilidades sociales y asertividad, así como entre sexualidad,habilidades sociales yautoconcepto, en el post-test en el grupo experimental. Los predictores de la asertividad en el post-test fueron toma de decisión, sexualidad y habilidades sociales. Los resultados destacan la importancia de la intervención con adolescentes particularmente en la toma de decisiones, sexualidad y habilidades sociales.(undefined

Establishment of Cancer Stem Cell Cultures from Human Conventional Osteosarcoma

The current improvements in therapy against osteosarcoma (OS) have prolonged the lives of cancer patients, but the survival rate of five years remains poor when metastasis has occurred. The Cancer Stem Cell (CSC) theory holds that there is a subset of tumor cells within the tumor that have stem-like characteristics, including the capacity to maintain the tumor and to resist multidrug chemotherapy. Therefore, a better understanding of OS biology and pathogenesis is needed in order to advance the development of targeted therapies to eradicate this particular subset and to reduce morbidity and mortality among patients. Isolating CSCs, establishing cell cultures of CSCs, and studying their biology are important steps to improving our understanding of OS biology and pathogenesis. The establishment of human-derived OS-CSCs from biopsies of OS has been made possible using several methods, including the capacity to create 3-dimensional stem cell cultures under nonadherent conditions. Under these conditions, CSCs are able to create spherical floating colonies formed by daughter stem cells; these colonies are termed "cellular spheres". Here, we describe a method to establish CSC cultures from primary cell cultures of conventional OS obtained from OS biopsies. We clearly describe the several passages required to isolate and characterize CSCs

High-resolution simulations of chromatin folding at genomic rearrangements in malignant B cells provide mechanistic insights into proto-oncogene deregulation

Genomic rearrangements are known to result in proto-oncogene deregulation in many cancers, but the link to 3D genome structure remains poorly understood. Here, we used the highly predictive heteromorphic polymer (HiP-HoP) model to predict chromatin conformations at the proto-oncogene CCND1 in healthy and malignant B cells. After confirming that the model gives good predictions of Hi-C data for the nonmalignant human B cell–derived cell line GM12878, we generated predictions for two cancer cell lines, U266 and Z-138. These possess genome rearrangements involving CCND1 and the immunoglobulin heavy locus (IGH), which we mapped using targeted genome sequencing. Our simulations showed that a rearrangement in U266 cells where a single IGH super-enhancer is inserted next to CCND1 leaves the local topologically associated domain (TAD) structure intact. We also observed extensive changes in enhancer-promoter interactions within the TAD, suggesting that it is the downstream chromatin remodeling which gives rise to the oncogene activation, rather than the presence of the inserted super-enhancer DNA sequence per se. Simulations of the IGH-CCND1 reciprocal translocation in Z-138 cells revealed that an oncogenic fusion TAD is created, encompassing CCND1 and the IGH super-enhancers. We predicted how the structure and expression of CCND1 changes in these different cell lines, validating this using qPCR and fluorescence in situ hybridization microscopy. Our work demonstrates the power of polymer simulations to predict differences in chromatin interactions and gene expression for different translocation breakpoints

Functional and Molecular Analysis of Human Osteoarthritic Chondrocytes Treated with Bone Marrow-Derived MSC-EVs

\ua9 2024 by the authors.Osteoarthritis (OA) is a degenerative joint disease, causing impaired mobility. There are currently no effective therapies other than palliative treatment. Mesenchymal stromal cells (MSCs) and their secreted extracellular vesicles (MSC-EVs) have shown promise in attenuating OA progression, promoting chondral regeneration, and modulating joint inflammation. However, the precise molecular mechanism of action driving their beneficial effects has not been fully elucidated. In this study, we analyzed MSC-EV-treated human OA chondrocytes (OACs) to assess viability, proliferation, migration, cytokine and catabolic protein expression, and microRNA and mRNA profiles. We observed that MSC-EV-treated OACs displayed increased metabolic activity, proliferation, and migration compared to the controls. They produced decreased proinflammatory (Il-8 and IFN-γ) and increased anti-inflammatory (IL-13) cytokines, and lower levels of MMP13 protein coupled with reduced expression of MMP13 mRNA, as well as negative microRNA regulators of chondrogenesis (miR-145-5p and miR-21-5p). In 3D models, MSC-EV-treated OACs exhibited enhanced chondrogenesis-promoting features (elevated sGAG, ACAN, and aggrecan). MSC-EV treatment also reversed the pathological impact of IL-1β on chondrogenic gene expression and extracellular matrix component (ECM) production. Finally, MSC-EV-treated OACs demonstrated the enhanced expression of genes associated with cartilage function, collagen biosynthesis, and ECM organization and exhibited a signature of 24 differentially expressed microRNAs, associated with chondrogenesis-associated pathways and ECM interactions. In conclusion, our data provide new insights on the potential mechanism of action of MSC-EVs as a treatment option for early-stage OA, including transcriptomic analysis of MSC-EV-treated OA, which may pave the way for more targeted novel therapeutics

Optogenetic Multiphysical Fields Coupling Model for Implantable Neuroprosthetic Probes

AuthorsOptogenetic-based neuroprosthetic therapies are increasingly being considered for human trials. However, the optoelectronic design of clinical-grade optogenetic-based neuroprosthetic probes still requires some thought. Design constraints include light penetration into the brain, stimulation efficacy, and probe/tissue heating. Optimisation can be achieved through experimental iteration. However, this is costly, time-consuming and ethically problematic. Hence it is highly desirable to have an alternative to excessive animal trials. Thus, a simulation tool for optimising probe design can be an important benefit for the community. The challenge is to understand the interplay between the optical, neural and thermal aspects in the interaction of probe and living neural tissue. In this work, we propose a model which combines these aspects to allow clinically orientated neuroprosthetic teams to design neuroprosthetic probes for optogenetic therapies. Our model provides analyses for optical, thermal and optogenetic electrophysiological processes based on the energy equivalence and exchange among different physical fields. To validate and calibrate the model, optogenetic implantable neuroprosthetic arrayed probes based on miniature LEDs were developed. Then, optical, thermal measurement and neural photocurrent recording experiments were implemented on the probes. We can then provide analysis on exemplar arrayed neural probes

Analysis of a preliminary microRNA expression signature in a human telangiectatic osteogenic sarcoma cancer cell line

Telangiectatic osteosarcoma (TOS) is an aggressive variant of osteosarcoma (OS) with distinctive radiographic, gross, microscopic features, and prognostic implications. Despite several studies on OS, we are still far from understanding the molecular mechanisms of TOS. In recent years, many studies have demonstrated not only that microRNAs (miRNAs) are involved in OS tumorigenesis, development, and metastasis, but also that the presence in high-grade types of OS of cancer stem cells (CSCs) plays an important role in tumor progression. Despite these findings, nothing has been described previously about the expression of miRNAs and the presence of CSCs in human TOS. Therefore, we have isolated/characterized a putative CSC cell line from human TOS (TOS-CSCs) and evaluated the expression levels of several miRNAs in TOS-CSCs using real-time quantitative assays. We show, for the first time, the existence of CSCs in human TOS, highlighting the in vitro establishment of this unique stabilized cell line and an identification of a preliminary expression of the miRNA profile, characteristic of TOS-CSCs. These findings represent an important step in the study of the biology of one of the most aggressive variants of OS and the role of miRNAs in TOS-CSC behavior