Background Metachromatic leukodystrophy (MLD) is a rapidly progressive leukodystrophy that leads to severe disability and early death if untreated. Autologous hematopoietic stem and progenitor cell gene therapy (HSPC-GT, atidarsagene autotemcel, arsa-cel) for early onset subtypes and allogenic hematopoietic stem cell transplantation (HSCT) for late onset disease substantially alters disease progression for early onset disease when administered before symptom onset, creating a strong rationale for newborn screening (NBS). At the same time, NBS technique for MLD in dried blood spots has recently been demonstrated to be robust and highly accurate. The aim was to give real-world results from the world’s first NBS pilots for clinical management and treatment of identified children. Methods Between September 2021 and July 2025, 359,282 newborns underwent NBS for MLD in two different laboratories in Germany and Austria using a three-tier algorithm integrating sulfatide quantification, arylsulfatase A (ARSA) activity measurement, and ARSA sequencing. Screen-positive infants underwent a predefined care pathway including standardized confirmatory diagnostics, genotype-based and biochemical prediction of disease onset, clinical assessment and management guiding early treatment and surveillance at the qualified treatment center (QTC) in Tübingen. Results Nine newborns screened positive and all were confirmed to have MLD (detection rate approximately 1 per 40,000). Based on genotype and leukocyte ARSA enzyme activity, disease onset prediction was possible in all of them. Seven infants were classified as having pre-symptomatic early-onset MLD and were referred for HSPC-GT. All treated infants showed preserved neurological function at follow-up 30 months after treatment. Two infants predicted to develop late-onset MLD entered structured surveillance for treatment with HSCT and have remained clinically stable. No false-positive or known false-negative results were observed. Conclusion These results from our pilot programs demonstrate that NBS enables reliable early identification of MLD and support streamlined care pathways leading to timely intervention. Importantly, this study provides real-world evidence illustrating that NBS for MLD can enable timely, pre-symptomatic treatment and structured surveillance within standard national healthcare systems. These findings further substantiate the value of NBS for MLD at a critical moment as several countries consider national implementation of MLD screening.

BACKGROUND AND PURPOSE:Clinical outcomes after hematopoietic stem cell transplantation (HSCT) in juvenile metachromatic leukodystrophy (MLD) are variable. This study evaluated whether multiparametric MRI parameters at the time of HSCT can predict long-term clinical outcomes and assessed their longitudinal evolution over an extended follow-up. MATERIALS AND METHODS:In this retrospective, registry-based cohort study, 15 children with juvenile MLD underwent a comprehensive MRI protocol at HSCT including quantitative demyelination load, DWI, MRS, and volumetric analysis. Patients were followed clinically for a median of 5 years and retrospectively stratified into stabilized, moderately progressive, or severely progressive trajectories based on Gross Motor Function Classification-metachromatic leukodystrophy (GMFC-MLD) and cognitive performance. Baseline imaging metrics and their changes over time were compared across outcome groups. RESULTS:Higher baseline demyelination load, increased ADC in the posterior limb of the internal capsule, and lower MRS metabolite ratios (NAA/Cr and NAA/Cho) were statistically significantly associated with a severely progressive clinical course. In contrast, baseline MRI severity score and brain volumes at HSCT were not predictive of outcome. Longitudinal analysis demonstrated consistent group differences in visually-assessable MRI parameters, including lesion burden, gray matter volume, total brain volume, and MRI severity score, which were most pronounced in patients with unfavorable clinical trajectories. Diffusion and spectroscopy measures showed region- and parameter-specific longitudinal effects but weaker and less consistent group separation compared with volumetric and lesion-based measures. Notably, baseline MRS metrics allowed retrospective stratification of patients with mild motor impairment (GMFC-MLD = 1) into distinct outcome groups, supporting early prognostic value. CONCLUSIONS:Multiparametric MRI at the time of HSCT provides complementary prognostic and longitudinal information in juvenile MLD. Baseline microstructural and metabolic abnormalities are associated with long-term clinical trajectories, whereas longitudinal disease evolution after HSCT is more consistently captured by volumetric and lesion-based MRI markers. These findings support the use of advanced MRI biomarkers for early risk stratification and subsequent monitoring and warrant further evaluation, including in gene therapy-treated cohorts.

OBJECTIVE:The detection of subtle epileptogenic lesions such as focal cortical dysplasias (FCDs) is a clinical challenge in the management of drug-resistant focal epilepsy (DRFE). Ultra-high-field (UHF) magnetic resonance imaging (MRI) offers increased signal-to-noise ratios and spatial resolution compared to 3-T MRI and may improve diagnostic yield. METHODS:We recruited n = 21 DRFE patients (with 3-T MRI findings: two positive, three equivocal, 16 negative) undergoing presurgical workup and n = 20 healthy controls for 9.4-T MRI (.8 mm isotropic magnetization-prepared 2 rapid acquisition gradient echo [MP2RAGE], slabs of .375 × .375 × .8 mm T2*-weighted gradient echo) and 3-T MRI (magnetization prepared rapid acquisition gradient echo [MPRAGE], magnetization-prepared 2 rapid acquisition gradient echo [MP2RAGE], fluid-attenuated inversion recovery [FLAIR]) acquisitions. Visual review for possible epileptogenic lesions was performed by clinical experts. For histopathologically confirmed FCDs, we extracted surface-based quantitative features (cortical thickness, quantitative T1, FLAIR, T2*, and quantitative susceptibility mapping values) across cortical depths and distances from the lesion center and performed high-resolution cortical profiling of 9.4-T T2* values. RESULTS:In two patients with histopathologically confirmed FCD IIb, lesions were visible with distinct qualitative and quantitative features at both field strengths. One of these type IIb FCDs showed a focal cortical T2* reduction at 9.4 T that could be quantified via automated cortical profiling, consistent with the previously described "black line sign." No new epileptogenic lesions were identified at 9.4 T in 3-T MRI-negative patients, who also had no histological evidence of such lesions. SIGNIFICANCE:9.4-Tesla MRI findings in epileptogenic lesions underlying DRFE are consistent with those on 3-T MRI. UHF T2*-weighted sequences may be useful to detect the black line sign and thereby refine surgical or ablation targeting for some FCDs. Assessment of the diagnostic yield of 9.4-T MRI was limited by the lack of 3-T MRI-negative but histopathologically confirmed cases and by the unavailability of parallel transmit and FLAIR at 9.4 T. Further optimization of UHF protocols and analysis methods on larger cohorts may enhance clinically applicable diagnostic benefits.

Metachromatic leukodystrophy (MLD) is a rare lysosomal storage disorder characterized by progressive white matter demyelination. Quantification of demyelinated white matter on MRI—typically expressed as the demyelination load—serves as a key imaging biomarker of disease burden, enabling objective monitoring beyond visual rating scales. However, current semi-automated pipelines are limited by manual interaction, pediatric brain variability, and differences in MRI acquisition. This study aimed to develop and validate a self-configuring convolutional neural network (CNN) for automated segmentation of demyelinated white matter in MLD and to compare its performance with a conventional semi-automated method across heterogeneous MRI datasets. An nnU-Net was trained on 189 3D T1- and axial T2-weighted scans from 35 MLD patients using visually controlled conventional masks as ground truth. Independent testing was performed on 130 scans (73 high-resolution 3D, 57 lower-resolution 2D T1-weighted) from 49 patients. Performance was assessed by Dice coefficient, Bland-Altman bias, correlation with Gross Motor Function Classification (GMFC-MLD), MLD MRI severity score, longitudinal consistency, and qualitative review of outliers. CNN-based segmentation showed strong spatial agreement with the reference method, with a median Dice coefficient of 0.82 for 3D T1-weighted scans and 0.75 for 2D scans. Volumetric bias was minimal on Bland-Altman analysis. CNN-derived demyelination load correlated significantly with motor impairment (rS = 0.38 for 3D and r = 0.56 for 2D; both p < 0.001) and showed a stronger association with the MLD MRI severity score than conventional segmentation (3D: rS = 0.48 vs. 0.28; 2D: rS = 0.83 vs. 0.29). Correlations with clinical status were slightly lower (CNN: rS = 0.38, p < 0.001; conventional: (rS = 0.26, p < 0.025)) Longitudinal analyses demonstrated stable, monotonic changes over time, and qualitative review revealed fewer boundary misclassifications. The nnU-Net enables fast, reproducible, and clinically meaningful segmentation of demyelinated white matter in MLD. It generalizes across MRI protocols, correlates with motor function, and offers a scalable tool for standardized biomarker extraction in clinical trials and other leukodystrophies.

1. Abstract Background The detection of subtle epileptogenic lesions such as focal cortical dysplasias (FCDs) is a clinical challenge in the management of drug-resistant focal epilepsy (DRFE). Ultra-high field (UHF) MRI offers increased signal-to-noise ratios and spatial resolution compared to 3 Tesla (T) MRI and may improve diagnostic yield. Here, we present a 9.4T MRI cohort study of patients with DRFE. Methods We recruited n=21 DRFE patients (with 3T-MRI findings: 2 positive, 3 equivocal, 16 negative) undergoing presurgical workup, and n=20 healthy controls for 9.4T MRI (0.8 mm isotropic MP2RAGE, slabs of 0.375 × 0.375 × 0.8 mm T2*-weighted GRE) and 3T MRI (MP2RAGE, FLAIR) acquisitions. Visual review for possible epileptogenic lesions was performed by clinical experts. For histopathologically confirmed FCD lesions, we extracted surface-based quantitative features (cortical thickness, qT1, FLAIR, T2*, and QSM values) across cortical depths and distances from the lesion centre and performed high-resolution cortical profiling of 9.4T T2* values. Results No new epileptogenic lesions were visually identified at 9.4T in 3T MRI negative patients. In the two patients with histopathologically confirmed lesions, the FCD IIb lesions were visible with distinct qualitative and quantitative features at both field strengths. One of these FCD IIb showed a focal cortical T2* reduction at 9.4T that could here be quantified via automated cortical profiling, consistent with the previously described “black line sign”. Conclusion 9.4T MRI findings in epileptogenic lesions underlying DRFE are consistent with those on 3T MRI. While additional lesions were not identified in patients with negative 3T MRI, higher resolution T2*-weighted sequences can reveal a feature not seen at 3T: Cortical profiling of FCDs highlights the black line sign and can possibly help refine surgical or ablation targeting for some FCDs. Further optimization of UHF protocols and analysis methods on larger cohorts may reveal clinically applicable diagnostic benefits. Key Points - 9.4T MRI shows focal cortical dysplasia (FCD) IIb with distinct qualitative and quantitative features that are consistent with 3T MRI. - High-resolution quantitative T2* maps at 9.4T may provide additional information for defining resection or ablation targets in FCDs. - Without using parallel transmit technology, artefacts in the temporal lobes pose a limitation of 9.4T MRI in presurgical epilepsy workup.

Background Metachromatic leukodystrophy (MLD) is a rare, progressive neurodegenerative disorder caused by arylsulfatase A deficiency, leading to accumulation of sulfatides and widespread demyelination. Autologous haematopoietic stem and progenitor cell gene therapy (HSPC-GT; atidarsagene autotemcel, arsa-cel) has emerged as an effective treatment for early-onset MLD when administered before or at very early stages of neurological involvement. However, standardized recommendations for pre-treatment evaluation, treatment phase care, and long-term follow-up are lacking. Methods Under the auspices of the INTEGRATE-ATMP project, a European multidisciplinary expert panel conducted a multi-round consensus process. Following a literature review, key clinical questions were addressed during five structured virtual and hybrid meetings. Recommendations were developed for pre-treatment assessment, treatment-phase management, and post-treatment follow-up. Investigations were classified as “mandatory” or “optional” based on expert agreement. Results The panel defined comprehensive, phase-specific recommendations for children undergoing HSPC-GT for MLD. Pre-treatment guidance emphasizes rapid diagnostic confirmation, standardized neurological and developmental assessments, and multidisciplinary eligibility evaluation. Treatment phase recommendations address stem cell collection, busulfan conditioning, supportive care, and monitoring for acute complications. Post-treatment guidance outlines a structured long-term follow-up programme, including neurological, developmental, imaging, and laboratory surveillance. Long-term data collection and systematic biobanking are strongly encouraged for a minimum of 15 years to support safety monitoring and outcome evaluation. Conclusion These European expert recommendations provide a practical framework for standardized care of children treated with HSPC-GT for MLD. Implementation across qualified treatment centres may improve clinical consistency, facilitate real-world data collection, and support sustainable delivery of gene therapy programmes. By detailing the comprehensive monitoring and follow-up required, including assessments beyond current standards of care, this work highlights the resource-intensive nature of gene therapy. It also provides a framework for the multidisciplinary care efforts needed to support planning and appropriate reimbursement by health authorities.

The UltraCortex repository houses magnetic resonance imaging data of the human brain obtained at an ultra-high field strength of 9.4 T. It contains 86 structural MR images with spatial resolutions ranging from 0.6 to 0.8 mm. Additionally, the repository includes segmentations of 12 brains into gray and white matter compartments. These segmentations have been independently validated by two expert neuroradiologists, thus establishing them as a reliable gold standard. This resource provides researchers with access to high-quality brain imaging data and validated segmentations, facilitating neuroimaging studies and advancing our understanding of brain structure and function. Existing repositories do not accommodate field strengths beyond 7 T, nor do they offer validated segmentations, underscoring the significance of this new resource.

Background Metachromatic leukodystrophy (MLD) is a life-limiting neurodegenerative disease due to pathogenic variants in the ARSA gene. Patients experience severe neurological symptoms, developmental regression, and early death. Aim of the study was to analyze disease burden and healthcare utilization in different stages of the disease in children with late infantile and juvenile MLD in Germany. Methods Out of a nationwide total cohort study (TC) (n = 83), we undertook telephone interviews in a representative follow-up cohort (FC) defined by advanced disease stages (n = 19). The FC allowed detailed long-term data of the disease in addition to cross-sectional data of the TC. Results Nearly all patients showed spasticity, truncal hypotonia and dysphagia, and about half of the patients developed epilepsy. Most children required special education; none finished regular school. Analysis of the FC showed that neuronal intestinal burden was extensive, including obstipation (57%), micturition problems (47%), and tube feeding (63%). Gallbladder polyposis was seen in 52%. General well-being did not strongly correlate with motor function, whereas pain was associated with reduced well-being. Baclofen, Omeprazole, Vigabatrin and Polyethyleneglycol were the most frequently used drugs. Patients took up to 15 different drugs daily. Altogether, 127 hospitalisations (485 treatment days) were registered in the FC (median age 9 years, median one inpatient stay per patient per year). Diagnostic procedures were main reasons for hospitalization (29 hospitalizations, 128 treatment days), and accounted for the main burden for families (68%). The median use of 15 different devices (maximum 29) throughout life illustrated a high burden of the disease. During disease course, there was a change from more “active” devices (e.g., walker) to more “passive” devices (e.g., form seat). Physical therapy was the most relevant therapy in advanced disease stages (100%), while occupational therapy or speech therapy primarily were used in early disease stages. State welfare benefits, home- and palliative care were used broadly. Conclusion Diagnostic and treatment routine pathways and sociomedical support in MLD require extensive resources. We provide detailed cross-sectional and long-term data of MLD-associated disease burden in different stages of disease. This data may serve as a reference when analyzing disease- and healthcare burden also after gene-/stem cell-therapy.

Myasthenia gravis is a neuromuscular disease typically associated with acetylcholine receptor (AChR) autoantibodies. However, in AChR-negative cases, autoantibodies against muscle-specific receptor tyrosine kinase (MuSK) may be present. Given the rarity of MuSK autoantibodies, there is limited information on their development and significance as biomarkers of disease activity. In a retrospective 10-year study, we tested 749 serum samples from 641 patients for MuSK autoantibodies. For MuSK-positive Myasthenia gravis cases, we extended the observation period to almost 17 years and analyzed the correlation between MuSK titers and clinical severity (Besinger score). We also examined the association between elevated but formally negative MuSK titers and other autoimmune diseases. Of the 749 samples, 21 (3 %, n = 7 patients) were MuSK-positive. In these patients, MuSK titers did not correlate with disease severity or treatment changes. 78 samples (10 %, n = 73 patients) showed elevated but formally negative MuSK titers, with no link to other autoimmune diseases. Our results contradict previous studies suggesting that MuSK autoantibodies could serve as biomarkers of Myasthenia gravis severity, and they shed light on the clinical relevance of elevated but formally negative MuSK titers.

BACKGROUND AND OBJECTIVES:Temporal lobe epilepsy (TLE) is commonly associated with mesiotemporal pathology and widespread alterations of gray and white matter structures. Evidence supports a progressive condition, although the temporal evolution of TLE is poorly defined. In this ENIGMA-Epilepsy study, we aim to investigate structural alterations in gray and white matter across the adult lifespan in patients with TLE by charting both gray and white matter changes and explore the covariance of age-related alterations in both compartments. METHODS:Mega-analysis of parcellated T1-weighted and diffusion MRI data across 18 international sites for patients with TLE was compared against healthy controls. We combined median-age split groupwise comparisons with cross-sectional sliding age-window analyses to explore gray (cortical thickness, subcortical volume) and white matter microstructure (fractional anisotropy, mean diffusivity) age-related changes. Five-year range age windows were constructed from mean z scores of all patients. Covariance analyses examined the coupled correlations of gray and white matter lifespan curves for each region. RESULTS:We studied 769 patients with TLE and 885 healthy controls across an age range of 17-73 years. Robust (pFDR < 0.05) gray matter thickness/volume decline (d < -0.20) was seen across a broad cortico-subcortical territory, extending beyond the mesiotemporal lobe throughout the adult lifespan in patients with TLE. White matter changes were also widespread across multiple fiber tracts with peak effects in temporolimbic fibers in fractional anisotropy (d < -0.3, pFDR < 0.05) and mean diffusivity measures (d > 0.3, pFDR < 0.05). Changes spanned the adult time window and effects exceeded typical aging-related processes in patients at the level of cortical thickness, subcortical volume, and diffusion measures, particularly in patients older than 55 years. Covariance analyses revealed strong associations across multiple white matter tracts, subcortical structures, and cortical regions within and beyond the temporolimbic system. DISCUSSION:This study highlights that patients with TLE exhibit more pronounced and widespread gray and white matter atrophy across the lifespan. The cross-sectional nature of our study limits definitive conclusions on whether the atrophy shown is progressive but emphasizes the importance of prompt diagnosis and intervention in patients. Collectively, our results motivate future longitudinal studies to clarify consequences of drug-resistant epilepsy.

Background:Evidence for the efficacy of nusinersen in adults with 5q-associated spinal muscular atrophy (SMA) has been demonstrated up to a period of 16 months in relatively large cohorts but whereas patients reach a plateau over time is still to be demonstrated. We investigated the efficacy and safety of nusinersen in adults with SMA over 38 months, the longest time period to date in a large cohort of patients from multiple clinical sites. Methods:Our prospective, observational study included adult patients with SMA from Germany, Switzerland, and Austria (July 2017 to May 2022). All participants had genetically-confirmed, 5q-associated SMA and were treated with nusinersen according to the label. The total Hammersmith Functional Motor Scale Expanded (HFMSE) and Revised Upper Limb Module (RULM) scores, and 6-min walk test (6 MWT; metres), were recorded at baseline and 14, 26, and 38 months after treatment initiation, and pre and post values were compared. Adverse events were also recorded. Findings:Overall, 389 patients were screened for eligibility and 237 were included. There were significant increases in all outcome measures compared with baseline, including mean HFMSE scores at 14 months (mean difference 1.72 [95% CI 1.19-2.25]), 26 months (1.20 [95% CI 0.48-1.91]), and 38 months (1.52 [95% CI 0.74-2.30]); mean RULM scores at 14 months (mean difference 0.75 [95% CI 0.43-1.07]), 26 months (mean difference 0.65 [95% CI 0.27-1.03]), and 38 months (mean difference 0.72 [95% CI 0.25-1.18]), and 6 MWT at 14 months (mean difference 30.86 m [95% CI 18.34-43.38]), 26 months (mean difference 29.26 m [95% CI 14.87-43.65]), and 38 months (mean difference 32.20 m [95% CI 10.32-54.09]). No new safety signals were identified. Interpretation:Our prospective, observational, long-term (38 months) data provides further real-world evidence for the continuous efficacy and safety of nusinersen in a large proportion of adult patients with SMA. Funding:Financial support for the registry from Biogen, Novartis and Roche.

In a pilot program in which 109,259 newborns were screened for metachromatic leukodystrophy, blood-spot samples were obtained for analysis and three newborns with MLD were identified, which made presymptomatic treatment possible.

ABSTRACTObjectivesTemporal lobe epilepsy (TLE) is commonly associated with mesiotemporal pathology and widespread alterations of grey and white matter structures. Evidence supports a progressive condition although the temporal evolution of TLE is poorly defined. This ENIGMA-Epilepsy study utilized multimodal magnetic resonance imaging (MRI) data to investigate structural alterations in TLE patients across the adult lifespan. We charted both grey and white matter changes and explored the covariance of age-related alterations in both compartments.MethodsWe studied 769 TLE patients and 885 healthy controls across an age range of 17-73 years, from multiple international sites. To assess potentially non-linear lifespan changes in TLE, we harmonized data and combined median split assessments with cross-sectional sliding window analyses of grey and white matter age-related changes. Covariance analyses examined the coupling of grey and white matter lifespan curves.ResultsIn TLE, age was associated with a robust grey matter thickness/volume decline across a broad cortico-subcortical territory, extending beyond the mesiotemporal disease epicentre. White matter changes were also widespread across multiple tracts with peak effects in temporo-limbic fibers. While changes spanned the adult time window, changes accelerated in cortical thickness, subcortical volume, and fractional anisotropy (all decreased), and mean diffusivity (increased) after age 55 years. Covariance analyses revealed strong limbic associations between white matter tracts and subcortical structures with cortical regions.ConclusionsThis study highlights the profound impact of TLE on lifespan changes in grey and white matter structures, with an acceleration of aging-related processes in later decades of life. Our findings motivate future longitudinal studies across the lifespan and emphasize the importance of prompt diagnosis as well as intervention in patients.

>Recently Zhang et al.(2024) published their study entitled “Lentivirus-modified hematopoietic stem cell gene therapy for advanced symptomatic juvenile metachromatic leukodystrophy: A long-term follow-up pilot study.” The authors present three metachromatic leukodystrophy(MLD) patients treated with gene therapy and claim stabilization or even improvement, despite advanced symptomatic disease stage. The metachromatic leukodystrophy initiative(MLDi)(Schoenmakers et al., 2022), an international collaborative network and registry for MLD, urges caution in interpreting these results, as the evidence raises several critical concerns. These claims risk fostering false hope among MLD patients and their families, particularly given the significant gaps in the data provided(Fig. 1).

In drug-resistant focal epilepsy, detecting epileptogenic lesions using MRI poses a critical diagnostic challenge. Here, we assessed the utility of MP2RAGE-a T1-weighted sequence with self-bias correcting properties commonly utilized in ultra-high field MRI-for the detection of epileptogenic lesions using a surface-based morphometry pipeline based on FreeSurfer, and compared it to the common approach using T1w MPRAGE, both at 3T. We included data from 32 patients with focal epilepsy (5 MRI-positive, 27 MRI-negative with lobar seizure onset hypotheses) and 94 healthy controls from two epilepsy centres. Surface-based morphological measures and intensities were extracted and evaluated in univariate GLM analyses as well as multivariate unsupervised 'novelty detection' machine learning procedures. The resulting prediction maps were analyzed over a range of possible thresholds using alternative free-response receiver operating characteristic (AFROC) methodology with respect to the concordance with predefined lesion labels or hypotheses on epileptogenic zone location. We found that MP2RAGE performs at least comparable to MPRAGE and that especially analysis of MP2RAGE image intensities may provide additional diagnostic information. Secondly, we demonstrate that unsupervised novelty-detection machine learning approaches may be useful for the detection of epileptogenic lesions (maximum AFROC AUC 0.58) when there is only a limited lesional training set available. Third, we propose a statistical method of assessing lesion localization performance in MRI-negative patients with lobar hypotheses of the epileptogenic zone based on simulation of a random guessing process as null hypothesis. Based on our findings, it appears worthwhile to study similar surface-based morphometry approaches in ultra-high field MRI (≥ 7 T).

OBJECTIVE:The intricate neuroanatomical structure of the cerebellum is of longstanding interest in epilepsy, but has been poorly characterized within the current corticocentric models of this disease. We quantified cross-sectional regional cerebellar lobule volumes using structural magnetic resonance imaging in 1602 adults with epilepsy and 1022 healthy controls across 22 sites from the global ENIGMA-Epilepsy working group. METHODS:A state-of-the-art deep learning-based approach was employed that parcellates the cerebellum into 28 neuroanatomical subregions. Linear mixed models compared total and regional cerebellar volume in (1) all epilepsies, (2) temporal lobe epilepsy with hippocampal sclerosis (TLE-HS), (3) nonlesional temporal lobe epilepsy, (4) genetic generalized epilepsy, and (5) extratemporal focal epilepsy (ETLE). Relationships were examined for cerebellar volume versus age at seizure onset, duration of epilepsy, phenytoin treatment, and cerebral cortical thickness. RESULTS:Across all epilepsies, reduced total cerebellar volume was observed (d = .42). Maximum volume loss was observed in the corpus medullare (dmax = .49) and posterior lobe gray matter regions, including bilateral lobules VIIB (dmax = .47), crus I/II (dmax = .39), VIIIA (dmax = .45), and VIIIB (dmax = .40). Earlier age at seizure onset ( η ρ max 2  = .05) and longer epilepsy duration ( η ρ max 2  = .06) correlated with reduced volume in these regions. Findings were most pronounced in TLE-HS and ETLE, with distinct neuroanatomical profiles observed in the posterior lobe. Phenytoin treatment was associated with reduced posterior lobe volume. Cerebellum volume correlated with cerebral cortical thinning more strongly in the epilepsy cohort than in controls. SIGNIFICANCE:We provide robust evidence of deep cerebellar and posterior lobe subregional gray matter volume loss in patients with chronic epilepsy. Volume loss was maximal for posterior subregions implicated in nonmotor functions, relative to motor regions of both the anterior and posterior lobe. Associations between cerebral and cerebellar changes, and variability of neuroanatomical profiles across epilepsy syndromes argue for more precise incorporation of cerebellar subregional damage into neurobiological models of epilepsy.

SUMMARYEndoreduplication, during which cells increase their DNA content through successive rounds of full genome replication without cell division, is the major source of endopolyploidy in higher plants. Endoreduplication plays pivotal roles in plant growth and development and is associated with the activation of specific transcriptional programmes that are characteristic of each cell type, thereby defining their identity. In plants, endoreduplication is found in numerous organs and cell types, especially in agronomically valuable ones, such as the fleshy fruit (pericarp) of tomato presenting high ploidy levels. We used the tomato pericarp tissue as a model system to explore the transcriptomes associated with endoreduplication progression during fruit growth. We confirmed that expression globally scales with ploidy level and identified sets of differentially expressed genes presenting only developmental‐specific, only ploidy‐specific expression patterns or profiles resulting from an additive effect of ploidy and development. When comparing ploidy levels at a specific developmental stage, we found that non‐endoreduplicated cells are defined by cell division state and cuticle synthesis while endoreduplicated cells are mainly defined by their metabolic activity changing rapidly over time. By combining this dataset with publicly available spatiotemporal pericarp expression data, we proposed a map describing the distribution of ploidy levels within the pericarp. These transcriptome‐based predictions were validated by quantifying ploidy levels within the pericarp tissue. This in situ ploidy quantification revealed the dynamic progression of endoreduplication and its cell layer specificity during early fruit development. In summary, the study sheds light on the complex relationship between endoreduplication, cell differentiation and gene expression patterns in the tomato pericarp.

ABSTRACT In tomato ( Solanum lycopersicum L .) fruit, the locular tissue (LT) is a unique jelly-like tissue that differentiates from the central axis of the fruit after ovule fertilization. LT is essential for seed development and dispersal by preventing early germination and initiating fruit ripening. In this work, we studied a “ gel-less ” mutant and identified the underlying mutation in the coding sequence of the C2H2 zinc finger transcription factor (TF) Sl ZFP2. Histological, cytological and molecular characterization from knockout-CRISPR/Cas9 lines for this gene revealed the strong and early impact of zfp2 mutation on cell cycle and endocycle in LT. Additionally, model-based analysis of cellular data revealed that cell cycle was the main altered process, explaining the zfp2 mutant phenotype. Further laser capture microdissection coupled with RNA-Seq analysis of young LT highlighted global expression changes between WT and zfp2 mutant and led to a preliminary list of potential direct targets of the Sl ZFP2 TF. This multifaceted approach not only uncovered a new role for Sl ZFP2 TF as an essential regulator of LT morphogenesis, but also provides a foundation for future works aimed at deciphering the intricate regulatory networks governing fruit tissue development in tomato. One sentence summary Alteration of cell division and endoreduplication in a gel-less mutant reveals the role of the transcription factor Sl ZFP2 in tomato locular tissue morphogenesis

The assessment of MRI as negative in epilepsy patients depends on the MRI protocol used, the magnetic field strength of the MRI, the expertise of the examiner, the background information on the presumed location of the epileptogenic zone and if necessary, the automated image analysis methods available at the center. Indeed, in the majority of surgically treated MRI negative patients an epileptogenic lesion can be demonstrated histopathologically, which then usually corresponds to subtle focal cortical dysplasia (FCD), hippocampus sclerosis (HS) or gliosis; however, preoperative visualization of the lesion is crucial for adequate resection of the epileptogenic zone and thus for a favorable postoperative outcome. This is why optimized MRI methods for epilepsy detection should be exploited in patients with focal epilepsy refractory to medication. These include the HARNESS MRI protocol with volumetric T1 and FLAIR sequences as well as a high-resolution T2 sequence at 3 T, which already enables a significantly higher lesion detection visually than with conventional sequences. The human eye should be supported by automated postprocessing methods, which have now achieved a high sensitivity in FCD and HS detection through continuous technical developments and the application of machine learning, but which require interpretation by experienced users due to the limited specificity. In the medium term, the superiority of high-field MRI with 7 T and more in selected patients, which has already been described in the experimental environment, will also bring further advantages in preoperative visualization in the clinical context.

Image templates are a common tool for neuroscience research. Often, they are used for spatial normalization of magnetic resonance imaging (MRI) data, which is a necessary procedure for analyzing brain morphology and function via voxel-based analysis. This allows the researcher to reduce individual shape differences across images and make inferences across multiple subjects. Many templates have a small field-of-view typically focussed on the brain, limiting the use for applications requiring detailed information about other extra-cranial structures in the head and neck area. However, there are several applications where such information is important, for example source reconstruction of electroencephalography (EEG) and/or magnetoencephalography (MEG). We have constructed a new template based on 225 T1w and FLAIR images with a big field-of-view that can serve both as target for across subject spatial normalization as well as a basis to build high-resolution head models. This template is based on and iteratively re-registered to the MNI152 space to provide maximal compatibility with the most commonly used brain MRI template.