Repetitive transcranial magnetic stimulation (rTMS) has been widely used in both clinical and research settings to modulate brain activity and assess brain plasticity. The effects of rTMS are generally believed to involve modulation of cortical excitability; protocols such as 10 Hz and iTBS increase cortical excitability, while 1 Hz and cTBS are considered inhibitory. However, recent meta-analyses and studies have shown a wide array of interindividual responses to each protocol and poor reliability of the results. Here, we applied two sessions of the four most commonly used rTMS protocols (iTBS, cTBS, 10 Hz, 1 Hz) and sham to the primary motor cortex and assessed their neurophysiological and behavioral effects. We hypothesized that relative to sham, 10 Hz and iTBS would increase motor excitability and modulate motor response times, while 1 Hz and cTBS would have opposite effects. We recorded single-pulse TMS-EEG-EMG pre-rTMS and 5 and 25 minutes post-rTMS. Participants performed a sequential finger-tapping task (SFTT) 20 minutes pre-rTMS and 15 minutes post-rTMS. Our main outcome measures were P30 amplitude, early LMFP (15-75 ms) area under-the-curve, MEP, and SFTT completion time. We calculated the Intra-Class Correlation between each session for each measure. For each protocol, we found no consistent protocol-dependent group effect on MEPs, TEPs, or behavior, with high intra-individual variability across sessions. An increase in SFTT completion time occurred for all active protocols but also sham. The test-retest reliability of rTMS effects was low (ICC < 0.5) for almost all protocols and metrics. This study is the first comprehensive assessment of the effects and reliability of the most commonly used rTMS protocols on cortical excitability and behavior. Our results suggest that rTMS effects are not reliable, at least at the single-session level, and that the inclusion of sham rTMS is essential for identifying protocol-specific effects. These results also suggest that a fundamental and unbiased re-evaluation of potential rTMS mechanisms is necessary.

While the field of imaging transcriptomics is evolving rapidly, several methodological challenges persist in functional enrichment analysis. Here, we introduce BrainEnrich, an R package that integrates whole-brain gene expression profiles from the Allen Human Brain Atlas (AHBA) with in vivo imaging-derived phenotypes (IDPs). By offering a suite of flexible association methods, aggregation strategies, comprehensive lists of predefined gene sets, and both competitive and self-contained null models, the package enables researchers to examine the spatial coupling between molecular profiles and IDPs at both group and individual levels. A novel feature of BrainEnrich is its individual-level enrichment analysis, which mapped individual IDPs onto a molecular coordinate framework, capturing the molecular signature of individual IDPs and enabling a deeper exploration of inter-individual variability. Its statistical power was examined through extensive simulation studies based on linear regression with different combinations of test statistics and null models. The results suggest that with appropriate null models, this approach effectively controlled Type 1 error while retaining sensitivity to detect associations between molecular profiles and phenotypic data. Two case studies were performed to demonstrate the utility of the package. In the group-level enrichment analysis, the effect size map of case-control comparison in cortical thickness of major depressive disorder patients was associated with molecular pathways such as synaptic signaling, lipid regulation, and steroid hormone balance, providing candidate molecular annotations for group-level IDPs. A separate case study found that synaptic gene set scores showed nominal associations with multiple cognitive measures, demonstrating its utility for individual-level molecular annotations to explore associations with phenotypic variables. Collectively, BrainEnrich provides a flexible framework for integrating macro-level IDPs with micro-level transcriptomic profiles for molecular contextualization of IDPs.

Trimethyl chitosan (TMC)-coated alginate/dextran sulfate (ADS) nanoparticles were developed as mucoadhesive nanocarriers for oral insulin delivery using a Quality-by-Design strategy. In a first screening step, a two-level factorial design was applied to evaluate the influence of ADS concentration, TMC concentration, insulin concentration, and poloxamer® concentration on particle size and encapsulation efficiency. The screening design identified the ADS-TMC pair as the main formulation parameter for particle size, while TMC and poloxamer® were the most influential factors for encapsulation efficiency. In a second step, formulation optimization was performed using a three-factor, three-level Box-Behnken design in which ADS concentration, TMC concentration, and the degree of quaternization (DQ) of TMC were investigated as critical material attributes. Particle size, zeta potential, and in vitro mucoadhesion were selected as critical quality attributes. Across the Box-Behnken design, the experimental formulations showed particle sizes ranging from 316 to 1340 nm, zeta potentials between +17 and +39 mV, and mucin-binding values from 7 to 87%. Numerical optimization by Design-Expert® desirability analysis identified an optimal formulation composed of 0.096% (w/v) ADS and 0.700% (w/v) TMC with 60% DQ. The model predicted a particle size of 316.24 nm, a zeta potential of +38.43 mV, and an in vitro mucoadhesion of 87.14%. Experimental confirmation yielded values of 330.79 nm, +37.09 mV, and 84.61%, respectively, with prediction errors below 5% for all responses. In simulated gastric medium, partial insulin leakage was observed during the first 120 min, whereas cumulative insulin release reached 54% after 5 h in simulated intestinal medium. These results demonstrate the usefulness of a QbD framework combined with desirability-based optimization for defining robust formulation conditions for mucoadhesive TMC-coated ADS nanoparticles intended for oral insulin delivery.

Shift work sleep disorder (SWSD) is prevalent among female nurses and is associated with significant health morbidities. While inflammation is implicated in SWSD, how it relates to brain network alterations and clinical symptoms remains underexplored. This study aimed to investigate the associations among peripheral inflammation, brain functional network topological disruptions, clinical symptoms, and transcriptomic signatures in female nurses with SWSD.

Anhedonia is a core feature of major depressive disorder (MDD), yet links between peripheral molecular signatures and cortical network architecture remain poorly defined. We enrolled 210 participants, including 56 unmedicated MDD patients with high-anhedonia (HA), 61 with low-anhedonia (LA), and 93 healthy controls (HC). Morphometric similarity networks (MSNs) from structural MRI were compared between HA and LA. MSNs index individual-level network organization by quantifying inter-regional morphometric similarity. Regional MSN patterns were linked to Allen Human Brain Atlas using Spearman correlations with spin tests and a multi-K stability screen. Whole-blood RNA-seq (n = 199) was integrated with MSN features via sparse partial least squares-canonical correlation (sPLS-C), with key blood analyses repeated after leukocyte-composition adjustment. Gene Ontology over-representation and MAGMA gene-level analyses provided pathway context. HA showed greater MSN integration than LA, particularly within default-mode and somatomotor networks. MSN maps were negatively correlated with dopamine-transporter and kappa-opioid-receptor densities. Imaging-derived gene associations were enriched for regulation of Toll-like-receptor-3 signaling. In blood, sPLS-C revealed coupling between MSN features and a transcriptomic signature enriched for T-cell activation/differentiation and lymphocyte-apoptosis pathways. After composition adjustment, the pre-specified blood signature did not differ between HA and LA, indicating that between-group differences were largely composition-driven. As supportive genetic context, over-representation on MAGMA FDR-significant genes suggested protocadherin-mediated homophilic adhesion. Peripheral immune-redox pathway enrichment aligns with anhedonia-related cortical network alterations, whereas between-group blood differences are chiefly composition-driven. Adjusting for blood-cell composition is essential, this multimodal framework nominates immune-modulatory/redox targets and synaptic-adhesion biology for precision stratification and intervention.

BACKGROUND: White matter hyperintensities (WMHs) are a hallmark imaging sign of cerebral small vessel disease (CSVD) and are closely linked to motor and cognitive deficits. Using voxel‑based morphometry (VBM) together with spatial transcriptomics, this study explored the altered molecular pathways underlying the reduction in gray matter volume (GMV) associated with WMHs. METHODS: We included 25 patients with WMHs and 26 healthy controls. The VBM analysis was used to identify brain regions with reduced GMV. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were then performed using spatial transcriptomic data from the Allen Human Brain Atlas (AHBA) to explore the underlying molecular pathways. RESULTS: Compared to healthy controls, patients with WMHs exhibited significantly reduced GMV in several key brain regions (p < 0.01, cluster-level corrected). These GMV reductions were widely distributed across the frontal, parietal, temporal, and occipital lobes, as well as the cingulate cortex and cerebellum. A total of 597 genes significantly associated with GMV loss were enriched in core neurobiological pathways, including protein synthesis, RNA metabolism, intracellular protein transport, and protein homeostasis. GO analysis revealed ribosome- and translation-related terms as the most significantly enriched across all three domains—biological process (BP), molecular function (MF), and cellular component (CC)—with structural constituent of ribosome and cytosolic ribosome being the most prominent in the MF and CC categories, respectively. Consistently, KEGG pathway analysis highlighted significant enrichment of both the ribosome and spliceosome pathways, further supporting the central role of cellular stress responses, DNA repair, and lysosomal function in the observed GMV-associated genes. CONCLUSIONS: WMHs-related gray matter atrophy may be spatially associated with coordinated perturbations in RNA metabolism and ribosome-mediated translation, together with dysregulated DNA metabolic processes and altered cellular stress responses.

Development of gray and white matter tissue microstructure is critical for the emergence of sensory and cognitive functions. However, it is unknown how microstructural tissue properties of the human visual system develop in the first year of human life. Across 85 infant sessions, we used tissue relaxation rate (R1) obtained using quantitative MRI to measure the longitudinal development of gray and white matter in brain areas spanning three visual processing streams: dorsal, lateral, and ventral, during the first year of life. R1 in gray and white matter of all visual regions in the three processing streams increases postnatally, indicating microstructural tissue growth. R1 increases faster between 0-6 months than 6-12 months, and faster in white matter than gray matter, with white matter R1 surpassing that of gray matter after two months of age. Notably, this microstructural growth is hierarchical: across all streams, early visual areas are more mature at birth than higher-level areas but develop more slowly postnatally than higher-level areas. The exception is TO1 (MT), which is similar to V1: it is microstructurally more mature at birth and develops more slowly than neighboring areas. Overall, our findings provide the first comprehensive measurement of microstructural tissue growth in infancy across three visual processing streams and propose a new hypothesis that functional development of the visual cortex may follow the hierarchical pattern of microstructural development.