Monoclonal antibodies targeting the calcitonin gene-related peptide (CGRP) pathway have been tested in cluster headache with mixed results, and four systematic reviews and meta-analyses reached different conclusions, leaving clinicians without a clear answer on whether, and in whom, to use these drugs.

Sleep disturbances are frequently observed in patients with neurodegenerative diseases, but in vivo evidence linking subcortical sleep-wake structures to sleep phenotypes is limited. We aim to examine associations between MRI-derived measures of subcortical sleep-wake regulating regions and polysomnographic sleep parameters in individuals with Alzheimer's disease and cognitively healthy controls. Fifty-seven adults (45 with biomarker-confirmed Alzheimer's disease and 12 cognitively healthy controls with normal pTau217 plasma levels) were recruited at the Hospital Clínic de Barcelona. All participants completed overnight polysomnography and 3T brain MRI, including a neuromelanin-sensitive sequence. MRI-derived measures of wake-promoting structures [locus coeruleus (LC), posterior hypothalamus and basal forebrain] and sleep-promoting regions (anterior hypothalamus) were obtained. We estimated the association between nocturnal polysomnographic variables [including total sleep time, sleep efficiency, time in rapid eye movement (REM) and non-REM sleep stages, REM sleep latency, wake after sleep onset and arousals] and MRI-derived measures. Then, a principal component analysis (PCA) was used to obtain derived patterns of sleep-wake regulation. Among sleep-promoting regions, higher anterior hypothalamic volumes correlated with greater time in bed (ρ = 0.19, P < 0.05), increased sleep period time (SPT) (ρ = 0.25, P < 0.01), and decreased REM over SPT (ρ = -0.22, P < 0.05). Within wake-promoting structures, higher posterior hypothalamic volume and LC integrity were associated with higher arousals (ρ = 0.32, P < 0.05 and ρ = 0.18, P < 0.05, respectively), and higher basal forebrain volume was associated with less REM over SPT (ρ = -0.25, P < 0.05). Control anatomic regions showed no associations with sleep parameters. The first two principal components of the PCA explain 47.7% of the variance. Anterior hypothalamic volumes clustered with longer, more efficient sleep, whereas LC integrity aligned with increased wakefulness and REM sleep latency. Alzheimer's disease participants showed a shift towards the latter pattern, indicating disrupted sleep-wake regulation. MRI-derived measures of subcortical sleep-wake regions were associated with clinical sleep phenotypes in Alzheimer's disease and ageing. These in vivo findings align with prior post-mortem research, reinforcing the role of subcortical structures in sleep disturbances and highlighting these circuits as potential targets to alleviate sleep symptoms and modify disease progression.

A significant challenge in the study of neuroimmune responses is the identification and separation of resident microglia from infiltrating bone marrow-derived macrophages. Therefore, we investigated whether dual labelling of Cx3cr1 and Ccr2 is sufficient to discriminate retinal microglia from infiltrating macrophages in mice that develop spontaneous autoimmune uveoretinitis-a condition dependent on infiltration of lymphocytes and antigen presenting cells from the circulation into the retina. Mice expressing red fluorescent protein (RFP) under control of the Ccr2 promoter and green fluorescent protein (GFP) under control of the Cx3cr1 promoter (Ccr2[RFP]Cx3cr1[GFP] mice) were crossed with the uveitogenic R161H mouse line. Retinas of R161H[neg] x Ccr2[RFP+/-]Cx3cr1[GFP+/-] F1 mice, including those given an optic nerve crush injury to stimulate retinal MG expansion, and R161H[+/-] x Ccr2[RFP+/-]Cx3cr1[GFP+/-] F1 mice were analyzed for RFP and GFP expressing cells by fundoscopy, flow cytometry, fluorescence microscopy, and for Ccr2 expression by single cell RNA sequencing of retinal CD45[+] cells. Retinas from control and optic nerve crushed R161H[neg] x Ccr2[RFP+/-]Cx3cr1[GFP+/-] F1 mice contained a single population of CD45[+] cells expressing a uniquely high level of GFP but were devoid of RFP[+] cells, while retinas from R161H[+/-] x Ccr2[RFP+/-]Cx3cr1[GFP+/-] F1 mice had GFP[hi] cells and increasing amounts of GFP[neg]RFP[+] and GFP[lo-med]RFP[+] cells as uveoretinitis progressed. Thus, dual labelling of Ccr2 and Cx3cr1 can distinguish retinal microglia from infiltrating immune cells, and as GFP[+], RFP[+], and GFP[+]RFP[+] cells can be easily separated by flow cytometry, our system provides a method for their separation for downstream cellular analysis.

Paclitaxel (PTX) resistance in triple-negative breast cancer (TNBC) is closely associated with ATP-binding cassette (ABC) transporters-medicated drug efflux and hypoxia-driven upregulation of hypoxia-inducible factor-1α (HIF-1α), which together promote tumor progression and metastasis. Although blood transfusion or hyperbaric oxygen therapy can partially alleviate systemic hypoxia, neither approach effectively relieves chemoresistance driven by intratumoral hypoxia. To address this challenge, we have developed a biomimetic hybrid delivery system (red blood cell hitchhiking-liposomes encapsulate hemoglobin and paclitaxel, RBC-LEHP) that utilizes red blood cell-hitchhiking technology along with pH-responsive liposomes, to co-deliver hemoglobin (Hb) and PTX to overcome hypoxia-associated chemoresistance. This system achieves dual-stage targeting: passive sequestration in pulmonary metastatic lesions through erythrocyte hitchhiking and active tumor internalization mediated by pH-sensitive DOPE phospholipids. Our study revealed that RBC-LEHP effectively alleviated the hypoxia microenvironment through Hb-mediated oxygenation upon tumor cellular internalization and downregulated HIF-1α and glutathione peroxidase 4 (GPX4), while simultaneously enhancing reactive oxygen species (ROS)-mediated lipid peroxidation to induce ferroptosis. This amplified oxidative stress further triggered mitochondrial dysfunction, suppressed P-glycoprotein (P-gp)-mediated PTX efflux, reversed PTX resistance, and promoted apoptotic cell death. In vivo results confirmed that RBC-LEHP-mediated therapeutic cascade of "lung targeted delivery → hypoxia alleviation → ferroptosis/chemotherapy → tumor suppression" significantly enhanced antitumor efficacy against TNBC. This biomimetic platform provides a promising strategy for overcoming physiological barriers and chemoresistance in metastatic TNBC therapy.

Background: Post-traumatic stress disorder (PTSD) is prevalent among children and adolescents, with cognitive behavioral therapy (CBT) and eye movement desensitization and reprocessing (EMDR) widely used treatment options. This study compared the effects of CBT and EMDR in children and adolescents using a meta-analysis approach.Methods: PubMed, EMBASE, CENTRAL, CINAHL, ProQuest Dissertations & Theses, and PsycInfo were searched in March 2025. Randomized controlled trials that compared the effectiveness of CBT and EMDR on PTSD symptoms in children and adolescents aged 6-18 years who had been exposed to traumatic events were included. The Cochrane Risk of Bias version 2 was used to assess the methodological quality of the included studies. A random-effects model was applied to calculate the standardized mean difference (SMD) along with 95% confidence interval (CI), and the I[2] statistic was used to quantify statistical heterogeneity.Results: Five trials (248 participants) were included. There was no statistically significant difference in PTSD symptoms between the CBT and EMDR groups, whether reported by children (SMD: 0.18; 95% CI: -0.07-0.43; I[2 ]= 0%) or by parents (SMD: 0.23; 95% CI: -0.02-0.48; I[2 ]= 0%). For anxiety, the pooled estimate did not reach statistical significance (SMD: 0.44; 95% CI: -0.19-1.07; I[2 ]= 69%). Depression symptoms were higher in the CBT group than in the EMDR group (SMD: 0.44; 95% CI: 0.13-0.76; I[2 ]= 0%).Conclusion: The comparative effects of CBT and EMDR in reducing PTSD and anxiety symptoms remain inconclusive. Although CBT was associated with higher depression symptoms compared with EMDR, the current evidence is based on a small number of trials, and the pooled estimates should be interpreted with caution.

Nervous necrosis virus (NNV) is a neurotropic pathogen that causes serious losses in aquaculture worldwide. B1 and B2 proteins, nonstructural proteins of NNV, play vital roles in viral function. In this study, a plasmid-based reverse genetics system was established for red-spotted grouper nervous necrosis virus (RGNNV), and codon mutations on B1 and B2 proteins were introduced. One recombinant virus (recomb) and five mutant viruses (B1QM, B2QM, BoQM, B1JM, and B2JM) were successfully rescued. Mature virions of approximately 30 nm in diameter within intracellular vacuoles were observed under transmission electron microscopy. The rescue results were further confirmed by PCR and western blotting. Compared with wild-type virus, all recombinant viruses displayed similar infectivity in vitro and in vivo. B2QM induced enhanced vacuolation-associated cytopathic effects at 5 days post-infection (dpi). Viral genome copy number peaked at 5 dpi in B1QM, whereas other recombinant viruses peaked at 3 dpi. RNA-dependent RNA polymerase expression showed significant differences only at 5 dpi, with 0.29-fold change (recomb) and 0.19-fold change (BoQM). Capsid protein expression level decreased in the B1QM group by 0.35-fold and increased by 2.29-fold in the B2QM group at 2 dpi. In vivo challenge showed that all groups except B2QM group reached 100% mortality within 30-h post infection (hpi), with peak mortality at 24 hpi. In the B2QM group, viral genome copy number remained relatively stable in brain and heart tissues but declined over time in eye tissue. Sequencing analyzes and immunofluorescence assay revealed that all mutated codons had been reverted to the original sequence, and B1 and B2 protein expression occurred in all mutant strains. These may suggest that the comparable infectivity between mutant and wild-type viruses is related to the reversion of codon mutations in nonstructural proteins, and revertant sequences that emerged during passaging probably account for this similarity. These findings provide new insights into NNV pathogenesis, but whether these mutations remain stable during passaging requires further investigation.