Voltage‐gated calcium channels (VGCC) are abundant in the central nervous system and serve a broad spectrum of functions, either directly in cellular excitability or indirectly to regulate Ca²⁺ homeostasis. Ca²⁺ ions act as one of the main connections in excitation–transcription coupling, muscle contraction and excitation–exocytosis coupling, including synaptic transmission. In recent years, many genes encoding VGCCs main α or additional auxiliary subunits have been associated with epilepsy. This review sums up the current state of knowledge on disease mechanisms and provides guidance on disease‐specific therapies where applicable. image

Scedosporium (S.) apiospermum is a typical mold causing cerebral abscesses, often after near-drowning. Infections are associated with high morbidity and mortality due to diagnostic challenges including the need for prolonged incubation of cultures. In addition, histopathological differentiation from other filamentous fungi, including Aspergillus fumigatus, may not be possible, excluding early specific diagnosis and targeted therapy. Polymerase chain reaction (PCR) on tissue samples can rapidly identify fungi, leading to an earlier adequate treatment. Due to an extensive spectrum of causative fungi, broad-range PCRs with amplicon sequencing have been endorsed as the best DNA amplification strategy. We herein describe a case with brain abscesses due to S. apiospermum in a 66-year-old immunocompromised female patient. While broad-range PCR failed to identify a fungal pathogen from a cerebral biopsy demonstrating hyaline mold hyphae, specific quantitative PCR (qPCR) identified Scedosporium and ruled out Aspergillus, the most prevalent agent of central nervous system mold infection. A panel of specific qPCR assays, guided by the morphology of fungal elements in tissue or as a multiplex assay, may be a successful molecular approach to identify fungal agents of brain abscesses. This also applies in the presence of negative broad-range fungal PCR, therefore providing diagnostic and therapeutic potential for early specific management and improvement of patient clinical outcome.

Background and Objectives: Developmental and Epileptic Encephalopathies (DEEs) encompass a clinically and molecularly heterogeneous group of mainly genetic disorders. Despite standard genetic testing, approximately two-thirds of patients remain undiagnosed. Pathogenic noncoding and structural variants (SVs), particularly complex rearrangements that are difficult to detect or resolve using short-read sequencing, may contribute to this diagnostic gap. To evaluate the diagnostic contribution of long-read genome sequencing (lrGS), we recruited a cohort of 124 families with sporadic DEE who remained unsolved by prior ES. Methods: Trio lrGS (Oxford Nanopore or PacBio HiFi) was performed, complemented zby short-read genome sequencing (srGS) and mRNA sequencing (blood or fibroblast) of index cases, where available. Comprehensive variant analysis included small nucleotide variants (SNVs), indels, SVs, and short tandem repeats. To identify the diagnostic yield of each method, we compared lrGS, srGS and ES re-analysis, and evaluated the additional value of RNA sequencing. Results: We established a diagnosis in 21 of 124 families (16.9%). Fourteen variants were retrospectively detectable in prior ES., Two cases with non-coding variants in SCN1A and RNU2-2 were detectable by both srGS and lrGS. Five SVs also required genome sequencing (5.6% of the total cohort), three of which could be detected, but not confidently resolved by srGS due to breakpoint complexity, repeat-mediated rearrangements, and large copy-neutral events: LrGS enabled accurate characterization of Alu-mediated CDKL5 and STXBP1 deletions and a MEF2C deletion with large inversion. A copy-neutral SCN1A inversion was well characterizable by both srGS and lrGS. One balanced translocation t(X;17) disrupting CDKL5 was resolved by lrGS (srGS unavailable). RNA sequencing confirmed pathogenicity of two SVs including t(X;17), the MEF2C deletion with inversion, and two CNVs. In addition, ZNF569 and ZNF570 emerged as candidate epilepsy genes from our cohort. Discussion: In ES-negative DEE, the incremental diagnostic yield of genome sequencing was modest, while lrGS provided particular value for the structural resolution of complex rearrangements, especially those involving repetitive sequences. The finding that two-thirds of diagnoses were retrospectively detectable in prior ES highlights the substantial value of systematic reanalysis. Further gains from genome sequencing may increasingly depend on improved interpretation of non-coding variation and integration of long-read-specific information. Our findings indicate that complex SVs represent the class for which lrGS currently yields the most distinctive diagnostic information.