Background: Copper (Cu) binds to amyloid beta (Aβ), generating reactive oxygen species (ROS) and altering cholesterol metabolism, leading to accumulation that worsens Alzheimer’s disease (AD). Cu near amyloid plaques can mediate inflammation via the NOD-like receptor protein 3 (NLRP3) inflammasome, which activates caspase-1 and interleukin-1β (IL-1β). Whether Cu exacerbates Aβ-induced inflammasome activation and microglial dysfunction remains unclear. Methods: Mouse microglia (SIM-A9) were treated with 1 µM Aβ and 100 µM Cu for 24 hours. ROS production was measured using dihydroethidium (DHE). Cholesterol levels were assessed via confocal microscopy and the Amplex Red kit. Inflammasome and phagocytosis proteins were analyzed by Western blot and PCR. Phagocytosis was measured using fluorescent microbeads, while inflammasome assembly was visualized with ASC-GFP speckle formation. Results: Cu increased IL-1β and NLRP3 expression, with a synergistic effect observed during Cu+Aβ treatment. Cu and Cu+Aβ elevated total and mitochondrial cholesterol levels while reducing glutathione (GSH). ROS production increased but was mitigated by glutathione ester (GSHee) and mitochondrial antioxidants (MitoTEMPO and MitoQ). Microglial phagocytosis, impaired by Cu and Cu+Aβ, was partially restored by GSHee. Cu also reduced phagocytosis proteins (ABCA7 and CD36) and caused their cytosolic redistribution, leading to decreased phagocytic activity. Conclusions: Elevated Cu levels exacerbate Aβ-induced microglial dysfunction by increasing mitochondrial oxidative stress, cholesterol accumulation—both at the cellular and mitochondrial level—and inflammasome activation. These effects impair phagocytosis, contributing to Aβ plaque accumulation and worsening AD pathology.

This research was funded by MCIN/AEI/10.13039/501100011033 and co-funded by NextGenera-tionEU/PRTR and “ERDF A way of making Europe”; grants PID2022-143279OB-100 and RED2022-134786-T, AGAUR, and grant 2021-SGR00490, respectively; and by the CIBERNED (Convocatoria 2022 Proyectos Colaborativos 2020/21).

Introduction: Modified citrus pectin (MCP) exhibits diverse biological activities, including anti-proliferative, anti-metastatic, and immunomodulatory effects. Additionally, MCP acts as an antagonist of galectin-3 (Gal-3), a key regulator of inflammation. This study aimed to investigate MCP's therapeutic potential in a murine colitis model induced using dextran sulfate sodium (DSS), focusing on its impact on inflammation and disease severity. Methods: Male C57BL/6 mice were divided into four groups: a control; DSS (colitis induced through the oral administration of 1.25% DSS in water for 7 days ad libitum); MCP (administered via orogastric gavage for 7 days at 100 mg/kg/day); and MCP+DSS. C57BL/6 Gal-3^-/- mice were divided into two groups: control and DSS groups. All of the procedures were approved by the Ethics Committee in Animal Experimentation of UNIFESP (CEUA nº4973090123).

Results: The DSS-treated mice presented significant weight loss on the eighth day and elevated disease activity index scores compared to their respective controls (p<0.0001). The DSS and MCP+DSS animals showed significant shortening of the intestines compared to the controls (p<0.0001), with increased production of IL-17 and TNF-α. A lack of Gal-3 abolished these effects, showing no changes in gut length or these cytokines between the control and DSS groups. Systemically, the spleens of the Gal-3^-/- DSS and MCP+DSS animals were longer than those of the DSS group (p<0.01). However, the plasma KC levels increased significantly in the DSS and MCP+DSS groups; again, the lack of Gal-3 abolished these differences between the DSS and control groups.

Conclusion: MCP therapy as a preventive treatment in the development of DSS-induced colitis was not effective. On the other hand, the lack of endogenous Gal-3 mitigates the deleterious effects of SSD-induced colitis, possibly by preventing an increase in IL-17 and TNF-α. Funding: CAPES, CNPq, and FAPESP.

Regular exercise reduces the risk of cardiovascular diseases, stroke, hypertension, and diabetes and enhances bone, muscle, and mental health. Recent evidence strongly suggests that aerobic exercise promotes adult neurogenesis in the hippocampus, improving synaptic plasticity. This study aimed to identify target miRNA candidates regulated by aerobic exercise that may contribute to enhanced hippocampal neurogenesis.

Male and female C57BL/6 mice (13 weeks old) were divided into two groups: a control group housed in locked-wheel cages and an exercise group housed in wheel-equipped cages for two weeks. The mice in the exercise group had free access to the running wheel in each cage, while the control mice were unable to use the wheel, as it was secured with a cable tie. Each group consisted of four mice. Total RNA was extracted from the dissected mouse hippocampi and used for RNA sequencing. Differentially expressed miRNAs were then identified based on gender and/or the exercise conditions.

A total of 698 miRNAs were identified as differentially expressed across the groups. A two-way ANOVA revealed that 67 miRNAs were differentially expressed by sex and 87 miRNAs by exercise (p < 0.05). Additionally, 55 miRNAs were influenced by both factors. The expression levels of miR-7b-3p, miR-212-3p, and miR-132-3p were significantly increased by exercise in both the male and female groups, while miR-12200-5p expression was reduced by exercise (p = 0.0135) but was lower in the male group overall.

This study shows that aerobic exercise alters the expression of specific miRNAs in the hippocampus, with some differences between males and females. miR-7b-3p, miR-212-3p, and miR-132-3p were significantly increased by exercise in both sexes, while miR-122000-5p was reduced. These miRNAs may play important roles in exercise-induced neurogenesis and brain health. Future research will focus on examining the effects of these target miRNAs on hippocampal neurogenesis.

Background

Telomere biology, even during fetal development, is influenced by parental manifestations that can imprint parental epigenetic marks onto the newborn. The interplay between genes, immune biology, and oxidative stress contributes to age-related non-communicable diseases (NCDs), and the telomere length (TL) dynamics in utero are not fully known. This study investigated the impact of parental biomedical factors, including NCDs, on newborn TL, as well as telomerase genes and immune biology.

Method
Blood samples (n=612) were collected from 204 parent–newborn pairs. Their demographics, socioeconomic status (SES), education, and NCD exposure were assessed. TL was quantified using the T/S ratio via qPCR; telomerase gene (TERC, TERT) polymorphisms were identified through Sanger sequencing; and immune senescence was analyzed using flow cytometry. Multivariate regression was used to analyze the paternal–newborn LTL associations, with p<0.05 as the significance threshold.

Results
The mean ages of the mothers and fathers were 27±5.12 and 34±6.36 years, respectively. The newborns of parents aged >30 years old had longer TLs (2.31±1.45; p=0.034). A low SES and blue-collar occupations were correlated with a shorter TL in the parent–newborn pairs (1.5±1.14; p<0.05), with higher frequencies of the telomerase CC genotype (TERC: 28%, 41%; TERT: 22%, 37%) compared to those in high-SES and white-collar groups. NCDs and viral pathologies significantly influenced telomere biology (p<0.05). The analysis of immune senescence markers showed decreased CD57+ KLRG1+ expression in the newborn T-cells (2.45±4.34; p<0.05) compared to those of their parents (3.5±5.49; p=0.04).

Conclusion
Parental biomedical factors and diseases influence newborn telomere biology and immune development in utero, highlighting the importance of mitigating prenatal risk factors.

Background: Hypoxia, common in high-grade serous ovarian cancer (HGSOC) microenvironments, contributes to resistance to platinum-based treatments and PARP inhibitors by activating AP-1 transcription factors. AP-1 promotes proliferation, invasion, metastasis, and angiogenesis, but its role in DNA damage signaling, repair, and resistance mechanisms remains unclear.

Methods: TF activity was assessed in PARP-sensitive and -resistant HGSOC cells (PEO1, PEO1R, OVCAR4) using a luciferase reporter assay. AP-1 subunit (c-JUN, JUND, JUNB, cFOS, FOSL2) expression was analyzed under normoxic and hypoxic (1% O₂) conditions. Immunohistochemistry for JUNB, FOSL2, MRE11, CA-9, and CD-31 was conducted on PEO1 and PEO1R tumor xenografts. CRISPR knockouts of JUNB and FOSL2 were studied for effects on DNA repair gene expression, proliferation, invasion, and cisplatin sensitivity using DNA repair profiling, RNA sequencing, and functional assays. Protein stability and co-immunoprecipitation were analyzed. The clinical significance of FOSL2, JUNB, and MRE11 expression was evaluated in a cohort of 331 ovarian cancer patients.

Results: In platinum/PARP-resistant PEO1R cells, AP-1 transcription activity was elevated compared to PEO1 cells, with JUNB and FOSL2 overexpressed under normoxic and hypoxic (1% O₂) conditions. Tumor xenografts showed high JUNB and FOSL2 in hypoxic regions. Knockout (KO) of JUNB and FOSL2 reduced proliferation and increased cisplatin and olaparib sensitivity, linked to higher double-strand breaks (DSBs), G2/M arrest, and apoptosis. KO cells showed a downregulation of DNA repair genes, including MRE11, which JUNB and FOSL2 interact with to enhance stability. RNA sequencing revealed enriched pathways in platinum response, oxidative phosphorylation, and translation. Clinically, higher FOSL2, JUNB, and MRE11 expression correlated with shorter progression-free survival (PFS) and poorer overall survival (OS).

Conclusion: JUNB and FOSL2 are key players at the intersection of hypoxia and DNA damage response (DDR) in high-grade serous ovarian cancer (HGSOC). They have predictive and prognostic value and may serve as therapeutic targets in platinum/PARP-resistant HGSOC.

Introduction. Pancreatic β-cells, by releasing insulin, play a critical role in the control of glucose homeostasis. In vivo, they reside in the islet niche, which provides a myriad of stimuli derived from the extracellular matrix (ECM) and the neighbouring cells. This multifaceted environment plays a pivotal role in the regulation of pancreas development and in the control of β-cell function. Even though the contribution of chemical stimuli has been widely investigated, the mechanical signals are poorly known. Therefore, the aim of the proposed research was to explore the role of nanotopography in the regulation of β-cell functionality and to investigate the underlying molecular mechanisms.

Methods. Human islets of Langerhans were grown on cluster-assembled zirconia substrates with a tailored roughness mimicking the ECM nanotopography, and flat zirconia substrates were used as controls. The β-cell functionality was evaluated by means of super-resolution fluorescence microscopy, Western blot, and ELISAs and confirmed by shot-gun proteomics.

Results. Quantitative immunofluorescence revealed that β-cells are mechanosensitive and respond to nanotopography through mechanotransduction, which impacts on focal adhesions and cytoskeletal and nuclear organization. These modifications are paralleled by a profound gene reprogramming which promotes the expression of pro-survival and pro-differentiation factors and proteins involved in the regulation of granule trafficking in the islets grown on the nanostructure. In line with these observations, we found that the nanotopography preserves β-cell differentiation and function in long-term cultured islets, as suggested by increased β-cell number, reduced β-cell death, and potentiated glucose-stimulated insulin secretion.

Conclusions. This study provides a better understanding of how mechanical forces contribute to β-cell fate, offering the possibility to harness these mechanisms for promoting β-cell function in physiological and pathological conditions.

Introduction: Dystonia is a movement disorder characterized by involuntary twisting and sustained postures. Impaired torsinA function causes the neurodevelopmental disorder of early-onset dystonia (DYT-TOR1A). TorsinA is highly expressed during development, and its loss of function during neural maturation impacts circuit development, leading to abnormal movements. Motor symptoms can be improved by lesioning or stimulating the globus pallidus interna (GPi) or administering antimuscarinic medications, implicating basal ganglia and cholinergic neuron involvement.

Methods: We conditionally deleted Tor1a from forebrain GABAergic and cholinergic neurons (using Dlx5/6-Cre) to generate overtly symptomatic “Dlx-CKO” mice. Like the neurodevelopmental onset of human DYT-TOR1A, motor abnormalities in Dlx-CKO mice emerge during juvenile development. Electrophysiological alterations found in developing Dlx-CKO striata and cholinergic interneurons (ChIs) motivate our anatomical studies. Using immunohistochemistry, confocal imaging, and neuroanatomical analyses, we characterized striatal ChI morphology and connectivity during development and maturation, with or without cholinergic torsinA re-expression using a novel ChAT-IRES-Tor1a construct.

Results: We find that maturing ChIs in Dlx-CKO striata exhibit transient defects in neurite outgrowth and persistent alterations in afferent connectivity that coincide with the onset of abnormal motor behavior. TorsinA re-expression in developing ChIs supports neuronal survival and morphology and promotes proper physiological striatal connectivity.

Conclusions: TorsinA loss-of-function causes morphological and synaptic alterations to maturing ChIs, consistent with a contribution to abnormal motor behavior.

Mitochondrial dysfunction is one contributor to aging, and it is implicated in many neurodegenerative diseases, including Alzheimer’s Disease (AD). Additionally, age-related vascular and neuronal pathologies may precede AD development. Studies showed that aging correlated with alterations in mitochondrial function and structure. However, how mitochondrial dysfunction affects cellular decline in the brain is still unclear. The mitochondria are regulated by the cristae organizing system (MICOS) complex and mitochondria–ER contact sites (MERCSs) in various organs. In the hippocampus of an AD mice model, researchers have also observed decreases in a MICOS protein, CHCHD6. Therefore, in other vulnerable regions of AD, such as the hypothalamus, dysregulation of the MICOS and MERCSs may lead to mitochondrial dysfunction in the aging process and eventually accelerate AD. Using quantitative polymerase chain reaction, serial block-face scanning electron microscopy, light microscopy, and 3D reconstruction, we found that the gene expression of the MICOS and MERCSs decreased in the aged amygdala and hypothalamus. The mitochondria also had significant morphological changes in the aged hypothalamus. Understanding the importance of the MICOS complex, we investigated mitochondrial homeostasis and observed that the inactivation of MICOS genes disrupted the synaptic transmission in crucial hypothalamic circuits. These results suggested that aging abates mitochondrial dynamics and physiology in the amygdala and the hypothalamus, and the MICOS is necessary for mitochondrial calcium regulation. This study provides targetable therapeutic strategies for aging in the brain, preventing the progression of AD at the early stages.

Autophagy is a crucial cellular process that maintains homeostasis by degrading and recycling intracellular components. A network of autophagy-related proteins orchestrates this process, many of which exhibit intrinsic disorder. Intrinsically disordered proteins (IDPs), lacking stable three-dimensional structures, possess structural flexibility, which enables dynamic interactions and diverse biological functions.

In this study, we analyzed 95 autophagy-related proteins from the Human Autophagy Database (HADb) and UniProt using sets of bioinformatics tools like ESpritz and leveraging datasets trained on X-ray, NMR, and DisProt. Our findings revealed that these proteins are significantly enriched with intrinsically disordered protein regions (IDPRs), particularly in key functional roles such as cargo recognition (e.g., SQSTM1, NBR1), autophagosome formation (e.g., ATG8, ATG12, WIPI1), and lysosomal degradation. Remarkably, proteins such as Beclin-1, LC3, and ATG9 exhibited high levels of intrinsic disorder, underscoring their critical regulatory roles.

The statistical analysis demonstrated that 80.21% of autophagy-related proteins contain at least one disordered region longer than 30 amino acids, and 65.97% have regions exceeding 50 amino acids. A total of 159 long disordered regions (greater than 30 amino acids) and 100 very long disordered regions (greater than 50 amino acids) were identified, emphasizing their functional relevance. Proteins like SPO95817 and SPP35638 showed extreme levels of disorder, with a mean percentage disorder above 90%, while others, such as SPP51809, exhibited minimal disorder, highlighting the variability within the autophagy-related proteome.

These results reinforce the critical role of IDPs in mediating transient and dynamic interactions that are essential for autophagy. This study advances our understanding of the molecular dynamics of autophagy-related proteins and provides a foundation for developing disorder-targeted therapeutic strategies. Such strategies hold potential for addressing neurodegenerative diseases, cancers, and other disorders linked to autophagic dysregulation.

The emergence of SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) variants continues to challenge global healthcare systems, necessitating the exploration of innovative therapeutic modalities. This study proposes a novel nasal treatment approach leveraging the antiviral properties of Beta-escin (B-escin), a natural compound, in conjunction with gold and silver nanoparticles. B-escin, extracted from horse chestnut seeds, exhibits notable efficacy against coronaviruses, while nanoparticles offer advantageous features for targeted delivery and stability enhancement. Through integrated computational analyses encompassing molecular docking, molecular dynamics simulations, and quantum mechanical calculations, we assess the potential of this combined strategy against prevalent COVID-19 variants, including Alpha, Beta, Gamma, Delta, and Omicron. Specifically tailored for nasal administration, our investigation emphasizes the synergistic interactions between B-escin and nanoparticles, elucidating their collective impact on variant-specific viral targets within the nasal mucosa. The computational results indicate augmented antiviral activity and target specificity when combining B-escin with gold (Au) and silver (Ag) nanoparticles, surpassing individual treatment efficacy. Moreover, we investigate the influence of nanoparticle characteristics, such as size, morphology, and surface functionalization, on the observed synergistic effects. These findings underscore the promise of developing a nasal treatment utilizing natural compounds and nanotechnology, offering a potential frontline defense against the diverse spectrum of SARS-CoV-2 variants.