When treating patients with orthognathic surgery, there might be a risk of obstructive sleep apnoea (OSA) due to soft tissue changes in the upper airways, especially in patients treated with isolated mandibular setback or mandibular setback in combination with maxillary advancement. In the present study, we assessed respiratory function during sleep with home cardiorespiratory polygraphy in 62 patients who had not been previously been diagnosed with OSA at three times: prior to orthognathic surgery for aesthetic and functional indications, and then 3 months and 1 year after surgery. We evaluated surgical displacement based on measurements in three dimensions using pre- and post-operative computed tomography. There were only minor changes in the respiratory parameters such as the apnoea-hypopnoea index (AHI), the apnoea-hypopnoea index in the supine position (AHIsup), the oxygen saturation index (ODI) and the snore index. There was no significant correlation between surgical displacement and the AHI, AHIsup and ODI. There was a weak but significant correlation between vertical displacement of the anterior mandible and the snore index. Within the limitations of the present study, the risk for iatrogenic obstruction of the upper airways seems to be low in patients without OSA treated with orthognathic surgery.

This study has followed a birth cohort for over 20 years to find factors associated with neurodevelopmental disorder (ND) diagnosis. Detailed, early -life longitudinal questionnaires captured infection and antibiotic events, stress, prenatal factors, family history, and more. Biomarkers including cord serum metabolome and lipidome, human leukocyte antigen (HLA) genotype, infant microbiota, and stool metabolome were assessed. Among the 16,440 Swedish children followed across time, 1,197 developed an ND. Significant associations emerged for future ND diagnosis in general and for specific ND subtypes, spanning intellectual disability, speech disorder, attention-deficit/hyperactivity disorder, and autism. This investigation revealed microbiome connections to future diagnosis as well as early emerging mood and gastrointestinal problems. The findings suggest links to immunodysregulation and metabolism, compounded by stress, early -life infection, and antibiotics. The convergence of infant biomarkers and risk factors in this prospective, longitudinal study on a large-scale population establishes a foundation for early -life prediction and intervention in neurodevelopment.

Objective: The objective of this study was to evaluate the diagnostic yield and clinical utility of an automated AI video-based seizure detection device, Nelli (R), (SDD) in pharmacoresistant epilepsy patients. The SDD captures and automatically classifies nocturnal motor behavior suggestive of epileptic seizures or non-epileptic motor behavior of potential clinical value. Methods: Patients with focal epilepsy and pharmacoresistance referred for inpatient long-term video-EEG monitoring were prospectively recruited. Participants were monitored in their home at night with the SDD for a median of 15.5 nights. Captured video recordings were analyzed by clinical experts and each SDD-registration session was classified as diagnostic or not. Clinical utility for each participant was assessed from pre-specified utility measures. The outcome measures were compared between major focal motor and subtle focal motor seizures. Results: One SDD-registration session in each of the 20 participants was performed and analyzed. Video recordings were captured in 18 sessions. Diagnostic yield was found in 11 registration sessions (55.0 %) and clinical utility in 8 registration sessions (40.0 %). No significant difference was found between the AI-algorithm classification and clinical experts' consensus assessment of captured video recordings as epileptic or not. Positive predictive value was 81.8 % for registration sessions containing video recordings classified as epileptic seizures. The diagnostic yield and clinical utility were significantly higher among major focal motor seizures (81.8 % and 63.6 %) compared to subtle focal motor seizures. Significance: The SDD is useful to evaluate patients with pharmacoresistant epilepsy and major focal motor seizures (hyperkinetic, tonic, clonic, focal to bilateral tonic-clonic seizures); it may facilitate the diagnostic process in patients referred for long-term inpatient video-EEG evaluation and beneficially change anti-seizure treatments. The SDD provided accurate classification of major focal motor seizures as epileptic, or non- epileptic, and may serve as a useful diagnostic tool to distinguish epileptic and non-epileptic episodic events with a prominent motor component.

The biological response to nanomaterials exposure depends on their properties, route of exposure, or model organism. Titanium dioxide nanoparticles (TiO2 NPs) are among the most used nanomaterials; however, concerns related to oxidative stress and metabolic effects resulting from their ingestion are rising. Therefore, in the present work, we addressed the metabolic effects of citrate-coated 45 nm TiO2 NPs combining bioaccumulation, tissue ultrastructure, and proteomics approaches on gilthead seabream, Sparus aurata and Japanese carpet shell, Ruditapes philippinarum. Sparus aurata was exposed through artificially contaminated feeds, while R. philippinarum was exposed using TiO2 NPs-doped microalgae solutions. The accumulation of titanium and TiO2 NPs in fish liver is associated with alterations in hepatic tissue structure, and alteration to the expression of proteins related to lipid and fatty acid metabolism, lipid breakdown for energy, lipid transport, and homeostasis. While cellular structure alterations and the expression of proteins were less affected than in gilthead seabream, atypical gill cilia and microvilli and alterations in metabolic-related proteins were also observed in the bivalve. Overall, the effects of TiO2 NPs exposure through feeding appear to stem from various interactions with cells, involving alterations in key metabolic proteins, and changes in cell membranes, their structures, and organelles. The possible appearance of metabolic disorders and the environmental risks to aquatic organisms posed by TiO2 NPs deserve further study.

Chronic wasting disease (CWD) is a prion disease affecting deer, elk and moose in North America and reindeer, moose and red deer in Northern Europe. Pathogenesis is driven by the accumulation of PrPSc, a pathological form of the host's cellular prion protein (PrPC), in the brain. CWD is contagious among North American cervids and Norwegian reindeer, with prions commonly found in lymphatic tissue. In Nordic moose and red deer CWD appears exclusively in older animals, and prions are confined to the CNS and undetectable in lymphatic tissues, indicating a sporadic origin.We aimed to determine transmissibility, neuroinvasion and lymphotropism of Nordic CWD isolates using gene-targeted mice expressing either wild-type (138SS/226QQ) or S138N (138NN/226QQ) deer PrP. When challenged with North American CWD strains, mice expressing S138N PrP did not develop clinical disease but harbored prion seeding activity in brain and spleen. Here, we infected these models intracerebrally or intraperitoneally with Norwegian moose, red deer and reindeer CWD isolates. The moose isolate was the first CWD type to cause full-blown disease in the 138NN/226QQ model in the first passage, with 100% attack rate and shortened survival times upon second passage. Furthermore, we detected prion seeding activity or PrPSc in brains and spinal cords, but not spleens, of 138NN/226QQ mice inoculated intraperitoneally with the moose isolate, providing evidence of prion neuroinvasion. We also demonstrate, for the first time, that transmissibility of the red deer CWD isolate was restricted to transgenic mice overexpressing elk PrPC (138SS/226EE), identical to the PrP primary structure of the inoculum.Our findings highlight that susceptibility to clinical disease is determined by the conformational compatibility between prion inoculum and host PrP primary structure. Our study indicates that neuroinvasion of Norwegian moose prions can occur without, or only very limited, replication in the spleen, an unprecedented finding for CWD. Chronic wasting disease (CWD) is a prion disease of cervids that is expanding its global footprint. The pathogenesis of prion disease is driven by the accumulation of PrPSc, a misfolded isoform of the cellular prion protein (PrPC). CWD prion strains from North America are lymphotropic, while Norwegian moose and red deer prions are not, and therefore considered non-contagious, sporadic CWD forms.We studied the propagation of Norwegian CWD prions in gene-targeted mice carrying cervid PrPC variants. We reveal that the Norwegian moose isolate induces clinical disease in mice expressing a PrPC variant previously shown to only display subclinical infection upon challenge with North American CWD. We report the first instance of red deer CWD transmission exclusively to mice overexpressing elk PrPC.Notably, our findings suggest a neuroinvasion route for Norwegian moose CWD prions that potentially bypasses spleen replication, underscoring the complexity of prion disease transmission, and the need for continued research into the behavior of prions across different species and protein variants.

Background: Aneurysmal subarachnoid haemorrhage (aSAH) is a life-threatening disease with high mortality and morbidity. Patients with aSAH in Sweden are cared for at one of six neuro intensive care units (NICU) or at a general intensive care unit (ICU).This study aimed to describe the incidence, length of stay, time in ventilator and mortality for these patients. Methods: This is a retrospective, descriptive study of patients with aSAH, registered in the Swedish Intensive care Registry between 2017 and 2019. The cohort was divided in sub-cohorts (NICU and general ICU) and regions. Mortality was analysed with logistic regression. Results: A total of 1520 patients with aSAH from five regions were included in the study. Mean age of the patients were 60.6 years and 58% were female. Mortality within 180 days of admission was 30% (n = 456) of which 17% (n = 258) died during intensive care. A majority of the patients were treated at one hospital and in one ICU (70%, n = 1062). More than half of the patients (59%, n = 897) had their first intensive care admission at a hospital with a NICU. Patients in the North region had the lowest median GCS (10) and the highest SAPS3 score (60) when admitted to NICU. Treatment with invasive mechanical ventilation differed significantly between regions; 91% (n = 80) in the region with highest proportion versus 56% (n = 94) in the region with the lowest proportion, as did mortality; 16% (n = 44) versus 8% (n = 23). No differences between regions were found regarding age, sex and length of stay. Conclusions: Patients with aSAH treated in a NICU or in an ICU in Sweden differs in characteristics. The study further showed some differences between regions which might be reduced if there were national consensus and treatment guidelines implemented.

Psoriasis is a common immune-mediated inflammatory disease, caused by a combination of genetic predisposition and environmental trigger factors. In spite of extensive research regarding the pathogenesis of psoriasis, many specific immunopathogenic mechanisms still remain unclear.

The aim of this thesis is to gain further insight into the pathogenesis of psoriasis through studying the bacterial microbiome of the skin and pharynx, the correlation between the microbiome and genetic susceptibility, and through studying pathway and cluster analysis of gene expression in patients with psoriasis.

Study I compared the bacterial microbiome of lesional and non-lesional skin of patients with psoriasis before and after treatment with narrowband ultraviolet B. Study II compared the bacterial microbiome of the pharynx and lesional and non-lesional skin of patients with psoriasis to healthy controls. Study III explored the association between the severity of psoriasis and abundance of the Streptococcus genus on the skin in individuals with specific psoriasis-associated gene variants. Study IV aimed to find novel pathways associated with psoriasis and to identify possible future therapeutic targets by conducting pathway and cluster analysis of differentially expressed genes of lesional and non-lesional skin of patients with psoriasis compared to healthy controls.

We found significant differences in the relative abundance of the bacterial genera Prevotella and Cutibacterium in both pharynx and skin and Finegoldia and Anaerococcus in the skin, which could be related to differences in regulated pathways relating to mitochondria and neutrophil function. We also found that the abundance of Streptococcus on the skin associated with severity of psoriasis in patients with certain genetic variants, suggesting that the microbiome’s effect on the manifestation of psoriasis is dependent on genetic predisposition.

Psoriasis results from both genetic predisposition and environmental triggers, such as Streptococcal infections. This study aimed to explore the correlation between the abundance of the Streptococcus genus on the skin and psoriasis severity in individuals carrying specific psoriasis -associated genetic variants. Studying 39 chronic plaque psoriasis patients, the elbow skin microbiome and 49 psoriasis -related single nucleotide polymorphisms (SNPs) were analysed using a MiSeq instrument for 16S rDNA sequencing, and CLC Genomic Workbench for processing and analysis. Through multivariate linear regression analysis, a positive correlation was found between Streptococcus genus abundance and psoriasis severity in patients with certain FBXL19 gene -related heterozygous SNPs (rs12924903, rs10782001, rs12445568). Conversely, a negative association was observed in patients with homozygous genotypes. Moreover, we identified an association between Streptococcus abundance and psoriasis severity in patients with genetic variants related to IL -22, ERAP1, NOS2, and ILF3. This is the first study highlighting a positive association between Streptococcus skin colonization and psoriasis severity in patients with heterozygous genotypes within the FBXL19 gene region. FXBL19 targets the IL-33/IL1RL1 axis, crucial in infectious diseases and innate immunity promotion. These novel results suggests an intricate interaction among host genetics, Streptococcus skin colonization, and psoriasis inflammation, offering potential avenues for novel treatment approaches.

AIMS: We have evaluated long-term weighted mean HbA_1c (wHbA_1c), HbA_1c variability, diabetes duration, and lipid profiles in relation to the development of diabetic peripheral neuropathy (DPN), nephropathy, and retinopathy in childhood-onset type 1 diabetes.

MATERIALS AND METHODS: In a longitudinal cohort study, 49 patients (21 women) with childhood-onset type 1 diabetes were investigated with neurophysiological measurements, blood tests, and clinical examinations after a diabetes duration of 7.7 (±3.3) years (baseline) and followed with repeated examinations for 30.6 (±5.2) years. We calculated wHbA_1c by integrating the area under all HbA_1c values since the diabetes diagnosis. Lipid profiles were analysed in relation to the presence of DPN. Long-term fluctuations of HbA_1c variability were computed as the standard deviation of all HbA_1c measurements. Data regarding the presence of other diabetes complications were retrieved from medical records.

RESULTS: In this follow-up study, 51% (25/49) of the patients fulfilled electrophysiological criteria for DPN. In nerve conduction studies, there was a deterioration in the amplitudes and conduction velocities for the median, peroneal, and sural nerves over time. Patients with DPN had a longer duration of diabetes, higher wHbA_1c, and increased HbA_1c variability. The lowest wHbA_1c value associated with the development of DPN was 62 mmol/mol (7.8%). The presence of albuminuria and retinopathy was positively correlated with the presence of neuropathy.

CONCLUSIONS: More than half of the patients had developed DPN after 30 years. None of the patients who developed DPN had a wHbA_1c of less than 62 mmol/mol (7.8%).

Autism spectrum conditions (ASC) encompass a range of sensory, motor, and social-communicative differences, reflecting the considerable heterogeneity within the autism spectrum. This diversity underscores the limitations of categorical diagnostic approaches, which often fail to capture the individualized manifestations of autism. Advances in genetics and neuroscience have driven a shift towards dimensional frameworks that emphasize the spectrum nature of autism and the broad autism phenotype (BAP). BAP encapsulates subclinical traits that mirror those of autism in the general population, challenging the conventional boundaries between clinical and non-clinical populations. Furthermore, sensorimotor differences, which are particularly prevalent in individuals with ASC, follow a spectrum-like pattern similar to the BAP and are predictive of developmental outcomes related to social participation, communication, and overall quality of life in people with and without ASC. However, specific descriptions of these relationships are lacking.

This dissertation investigated the complex relationships between sensorimotor differences and autistic traits (ATs). Through a series of five interconnected studies, we examined broad sensory processing patterns and specific sensory modalities, namely auditory processing and motor/proprioception, to explore their roles in autistic phenotypes.

Study 1 of the dissertation validated a Swedish translation of the Broad Autism Phenotype Questionnaire (BAPQ). By examining parents' BAP traits, the study highlighted significant associations between higher ATs and having a child with ASC. Furthermore, it confirmed the presence of all three AT domains—social interaction (AT_SOC), communication (AT_COM), and cognitive rigidity (AT_RIG), reinforcing the genetic and phenotypic continuity between clinical and subclinical ATs.

Study 2 and Study 3 served as broader investigations into all seven sensory modalities and their associations with ATs. Study 2 explored these modality-specific associations, using Bayesian stochastic search variable selection (SSVS) and dominance analysis. This study highlighted auditory processing difficulties as the most consistent predictor of all three AT domains. Additionally, proprioceptive and tactile processing difficulties were specifically associated with AT_COM and AT_SOC, respectively.

Study 3 extended this analysis to a developing population, focusing on the relationship between sensorimotor processing, ATs, and anxiety in children aged 6-11 years. Identical to Study 2, we found tactile symptoms as a predictor of AT_SOC, proprioceptive symptoms for AT_COM, and auditory symptoms for AT_RIG. In addition, olfactory symptoms were selected as a predictor of AT_COM, and motor coordination was a consistent predictor of all AT domains. Using SSVS, this study also identified that auditory and olfactory processing difficulties were strong predictors of anxiety symptoms.

Building on the previous studies, Study 4 narrowed the focus to auditory processing differences, investigating specific auditory problems and their associations with the AT domains. All AT domains significantly predicted affective reactions to sounds, while difficulties with speech perception, spatial perception, and auditory stream segregation were most strongly predicted by AT_COM.

Study 5 focused on the previously found links between motor coordination and proprioceptive processing and AT_COM. Using causal mediation analysis within a counterfactual framework, this study found that cerebellar error correction deficits, measured through a finger tapping task, significantly impacted AT_COM through motor skills in childhood.

Together, this dissertation provides a comprehensive overview of the sensory processing dimensions related to the core AT domains. Specifically, the studies underscored the clinical significance of monitoring auditory and olfactory complaints in children, as these were predictive of anxiety, and emphasized that early motor deficits impact social communication development. The findings advocate for the inclusion of detailed sensory and motor assessments in neurodevelopmental evaluations to identify children at risk for poor mental health outcomes. Future research should continue to explore the mechanisms underlying sensory processing differences. Particular focus should be placed on auditory and motor/proprioceptive functions and their contributions to ATs and clinical outcomes, such as anxiety. Emphasis should also be given to longitudinal studies that track these relationships over time.

BackgroundAutism spectrum conditions (ASC) and quantitative autistic traits (QATs) are associated with sensory symptoms, which may contribute to anxiety and adversely affect social and cognitive development. Although sensory symptoms can occur across all senses, the relative roles of specific sensory modalities as contributors to the autistic phenotype and to anxiety are not well understood. The objective of this study was to examine which sensory symptoms were most predictive of high anxiety.MethodsWe recruited 257 female primary caregivers of children aged 6 to 11 years (49% girls) to a questionnaire study comprising parent-report measures for classical QATs (social, communicative, and rigid), autism-related sensorimotor symptoms (visual, auditory, tactile, olfactory, gustatory, vestibular, proprioceptive, and motor), and anxiety symptoms. First, Bayesian stochastic search variable selection (SSVS) was used to identify the most probable sensorimotor predictors of specific QATs as well as diagnosed ASC. Then, the selected predictors were used in another SSVS, using anxiety symptoms as a dependent variable, to identify which of the autism-relevant sensorimotor symptoms were most robustly predictive of anxiety. Finally, the effect sizes of anxiety-related sensory symptoms were estimated with linear regressions.ResultsWe found that auditory symptoms and motor difficulties were most predictive of ASC diagnosis. Developmental motor difficulties were also strongly related to all individual QATs, whereas auditory symptoms were more selectively predictive of rigid traits. Tactile symptoms robustly predicted social interaction QATs, and proprioceptive symptoms predicted communicative QATs. Anxiety outcomes were most strongly predicted by difficulties with auditory and olfactory processing.ConclusionsThe results support the clinical importance of being alert to complaints about sounds and hearing in neurodevelopmental populations, and that auditory processing difficulties may be evaluated as an early marker of poor mental health in children with and without diagnosed autism. Olfactory processing differences appeared to be an anxiety marker less strongly associated with ASC or QATs, while motor difficulties were highly autism-relevant but not equally strongly associated with anxiety outcomes. We suggest that future studies may focus on the mechanisms and consequences of neurodevelopmental central auditory processing dysfunction and its potential relationship to anxiety disorders.

Background: Decreased sound tolerance (DST) is a disabling transdiagnostic phenomenon with high clinical relevance in autism. Neurodevelopmental DST is often studied as part of a general multisensory construct that includes both hyper- and hyposensitivity. Therefore, knowledge about the potential relevance of individual differences in the auditory modality is lacking. The purpose of the study was to begin to differentiate between commonly pooled auditory functions, by incorporating psychometric tools from the field of audiology.Methods: In a pilot sample (N = 520 adults, 23% autistic), we used Bayesian correlations to quantify the contribution of individual auditory items from the Glasgow Sensory Questionnaire to the degree of social, communicative, and rigid autistic traits measured with the Broad Autism Phenotype Questionnaire (BAPQ) subscales. Then, we recruited an independent sample (N = 175 adults, 18% autistic) to measure, more specifically, (1) emotional reactions to sounds (affective DST), (2) speech understanding difficulties, and (3) nonsocial auditory processing (spatial perception and stream segregation), using self-report questionnaires. We used multiple regression to test for associations with the autistic trait domains.Results: We found that all autistic traits measured by the BAPQ (social, communicative, and rigid) linearly predicted affective DST, and these associations remained when autistic participants were excluded. Difficulties with speech perception, as well as spatial perception and auditory stream segregation, were most strongly predicted by communication differences.Conclusion: The robust relationship between autistic traits and emotional sound reactivity suggests that affective DST falls on a spectrum just like autism. This argues against strict dichotomization and encourages the use of continuous measures. The results support a dominant role for emotional and stress systems in autism-related DST and may suggest that detailed audiological tests are clinically useful, in particular, in the context of pragmatic language difficulties. Community Brief Why is this an important issue? Differences in sensory perception go hand in hand with an autistic neurotype. Many individuals find everyday noises intolerable or difficult to understand, but there is little knowledge of how these experiences vary among individuals. What was the purpose of this study? To understand which aspects of auditory processing are related to autistic characteristics, using assessment tools from the audiology field. What did the researchers do? They studied 175 adults with different levels of autistic-like social function, communication style, and rigid thinking (18% with an autism diagnosis). They used clinical questionnaires to measure emotional reactions to sounds, speech understanding, auditory space perception, and the ability to distinguish overlapping sounds. What were the results of the study? All autistic characteristics were linked to strong emotional reactions to sounds. Those with an autistic communication style also reported difficulties with speech perception, auditory space perception, and separating overlapping sounds. What do these findings add to what was already known? The results highlight a role of emotional and stress responses in autistic sound sensitivity. In addition, they point to the existence of broader auditory difficulties in adults with an autistic communication style. What are potential weaknesses in the study? It did not include laboratory measurements of auditory function. How will these findings help autistic adults now or in the future? This study helps by breaking down specific hearing challenges in autism, such as trouble understanding speech or reacting strongly to everyday sounds, rather than looking at sensory issues as a whole. This can lead to better support and solutions for these specific difficulties.

Background. Decreased sound intolerance (DST) is a disabling transdiagnostic phenomenon with high clinical relevance in autism. Neurodevelopmental DST is often studied as part of a general multisensory construct that includes both hyper- and hyposensitivity. Therefore, knowledge about the potential relevance of individual differences in the auditory modality is lacking. The purpose of the study was to begin to differentiate between commonly pooled auditory functions, by incorporating psychometric tools from the field of audiology. 

Methods. In a pilot sample (N = 520 adults, 23% autistic), we used Bayesian correlations to quantify the contribution of individual auditory items from the Glasgow Sensory Questionnaire to the degree of social, communicative and rigid autistic traits measured with the Broad Autism Phenotype Questionnaire (BAPQ) subscales. Then, we recruited an independent sample (N = 175 adults, 18% autistic) to measure, more specifically, 1) emotional reactions to sounds (affective DST), 2) speech understanding difficulties, and 3) non-social auditory processing (spatial perception and stream segregation), using self-report questionnaires. We used multiple regressions to test for associations with the autistic trait domains. 

Results. We found that all autistic traits measured by the BAPQ (social, communicative and rigid) linearly predicted affective DST, and these associations remained when autistic participants were excluded. Difficulties with speech perception, as well as spatial perception and auditory stream segregation, were most strongly predicted by communication differences. 

Conclusion. The robust relationship between autistic traits and emotional sound reactivity suggest that affective DST falls on a spectrum just like autism. This argues against strict dichotomization and encourages the use of continuous measures. The results support a dominant role for emotional and stress systems in autism-related DST, and may suggest that detailed audiological tests are clinically useful, in particular in the context of pragmatic language difficulties. 

Early motor function is important for emerging social pragmatic communication (SPC) skills in both typical and atypical development. However, the nature of motor impairments relevant for higher-level communication is not well understood. Inefficient cerebellar error correction might directly cause both developmental coordination disorder (DCD) symptoms and SPC difficulties, through the extensive communication between cerebellar zones and brain-wide sensorimotor and higher-order networks. DCD symptoms related to cerebellar deficits could also impact SPC through affecting the developmental trajectory of social development, which requires motor skills. This study aimed to test the hypothesis that error correction deficits affect SPC outcomes through childhood DCD symptoms, by using contemporary causal inference methodology. We used a finger tapping task and computational modeling to measure cerebellar error correction in adult participants (n = 138), and quantified childhood DCD symptoms and SPC skills using psychometric measures. The results confirmed that error correction ability likely affects SPC skills, and indicated that childhood motor skills significantly mediated this. These results argue against a direct effect of domain-general error correction deficits on SPC, and instead suggest that cerebellum-related DCD symptoms affect sociocommunicative development more directly through motor deficits during development. Further research is required to test whether cerebellar error correction could be used as an early marker to identify children in need for early SPC interventions.    

New antibacterial compounds are urgently needed, especially for infections caused by the top-priority Gramnegative bacteria that are increasingly difficult to treat. Lipid A is a key component of the Gram-negative outer membrane and the LpxH enzyme plays an important role in its biosynthesis, making it a promising antibacterial target. Inspired by previously reported ortho-N-methyl-sulfonamidobenzamide-based LpxH inhibitors, novel benzamide substitutions were explored in this work to assess their in vitro activity. Our findings reveal that maintaining wild-type antibacterial activity necessitates removal of the N-methyl group when shifting the orthoN-methyl-sulfonamide to the meta-position. This discovery led to the synthesis of meta-sulfonamidobenzamide analogs with potent antibacterial activity and enzyme inhibition. Moreover, we demonstrate that modifying the benzamide scaffold can alter blocking of the cardiac voltage-gated potassium ion channel hERG. Furthermore, two LpxH-bound X-ray structures show how the enzyme-ligand interactions of the meta-sulfonamidobenzamide analogs differ from those of the previously reported ortho analogs. Overall, our study has identified meta-sulfonamidobenzamide derivatives as promising LpxH inhibitors with the potential for optimization in future antibacterial hit-to-lead programs.

Background: It has been suggested that carbidopa at high blood concentrations may counter the therapeutic effect of levodopa in Parkinson's disease by entering the brain and blocking central levodopa conversion to dopamine. We previously demonstrated equivalent plasma levodopa concentration in patients with Parkinson's disease during 16 h of (1) intravenous carbidopa/levodopa (DIZ101) infusion, (2) subcutaneous carbidopa/levodopa (DIZ102) infusion or (3) intestinal carbidopa/levodopa gel infusion. Plasma levels of carbidopa were however approximately four times higher with DIZ101 and DIZ102 than with LCIG, and higher than those usually observed with oral levodopa/carbidopa. Objectives: To investigate if high carbidopa blood concentrations obtained with parenteral levodopa/carbidopa (ratio 8:1) counter the effect of levodopa on motor symptoms. Methods: Eighteen patients with advanced Parkinson's disease were administered DIZ101, DIZ102, and intestinal levodopa/carbidopa gel for 16 h on different days in randomized order. Video recordings of a subset of the motor examination in the Unified Parkinson's Disease Rating Scale (UPDRS) were evaluated by raters blinded for treatment and time. Motor function was also measured using a wrist-worn device monitoring bradykinesia, dyskinesia, and tremor (Parkinson KinetiGraph). Results: There was no tendency for poorer levodopa effect with DIZ101 or DIZ102 as compared to LCIG. Conclusion: Although DIZ101 or DIZ102 causes approximately four times higher plasma carbidopa levels than LCIG, patients responded equally well to all treatments. The results do not indicate that high plasma carbidopa levels hamper the motor efficacy of levodopa.

Background: The role of environmental factors in neurological disorders constitutes a topic of increasing importance. Teaching in European universities should expand and update this field gaining future health professionals including adjacent disciplines. Aim: To describe recent efforts to create courses that cover crucial interdisciplinary content that we believe should be included in modern education, and to adapt modern pedagogic strategies. Methods: In collaboration with RISE (Rencontres Internationales Sante<acute accent> Environnement), elective courses focused on Environmental Health and Medicine (EHM) were developed, in France, Sweden, and Turkey. The courses combined classic teaching methods and new pedagogic and digital solutions to create environment-related health awareness and facilitate future interprofessional collaboration in this field. Results: UPRISE is an innovative elective course introduced in 2020 in Sweden's Uppsala University with the participation of lecturers from several countries and aim to recruit students from different universities. A total of 45, mainly female students (68%), participated in the course. In Strasbourg, France, a novel course on environmental medicine was held in 2019-2023 and examined 90 students, of which more than half were female. Nine graduate nurse students in Turkey attended ten seminar series focused on EHM. Overall, students expressed satisfaction with the courses. Conclusions: This European project for courses in higher education arising from RISE was met with appreciation and challenges from academic institutions. However, due to considerable efforts to introduce the EHM concept, a unique compulsory course for all medical students in the second year of training started in 2023 in all French medical faculties. In 2023, UPRISE was integrated into ENLIGHT, the European University Network to promote equitable quality of Life, sustainability, and Global engagement through Higher education Transformation.

Restless legs syndrome (RLS) is a common neurological disorder characterised by an urge to move arms and legs, usually associated with discomfort, pain, motor restlessness, and sleep disturbance. An individually adapted treatment is needed but difficult to optimise, which makes shared decision-making (SDM) important. However, brief validated instruments on how patients with RLS perceive their involvement in treatment decisions are lacking. Therefore, the aim was to validate two instruments, SURE (Sure of myself, Understand information, Risk-benefit ratio, Encouragement, i.e., to assess decisional conflict) and CollaboRATE (brief patient survey focused on SDM, i.e., to assess SDM), in patients with RLS. A cross-sectional design, including 788 participants with RLS (65% females, mean [SD] age 70.8 [11.4] years) from a national patient organisation for RLS, was used. A postal survey was sent out to collect data regarding weight, height, comorbidities, demographics, and RLS-related treatment data. The following instruments were included: the SURE, CollaboRATE, Restless Legs Syndrome-6 Scale, and eHealth Literacy Scale. Confirmatory factor analysis and Rasch models were used to assess the validity and reliability of the SURE and CollaboRATE. Measurement invariance, unidimensionality, and differential item functioning (DIF) across age, gender, and medication groups were assessed. The SURE and CollaboRATE were both identified as unidimensional instruments with satisfactory internal consistency. No DIF across age and gender was identified, while significant DIF was observed for both the SURE and CollaboRATE regarding medication use categories. However, both the SURE and CollaboRATE are potential instruments to be used in research, but also as reflection tools by healthcare professionals, patients, and students to explore and assess SDM, and support its development in clinical care.

Objective: The primary aim was to estimate the pooled mean score of quality of life (QoL) (total, mental and physical health components) among patients with Restless Legs Syndrome (RLS). Secondary aims were to assess: (I) QoL differences for RLS vs. control groups, (II) heterogeneity and possible sources; and (III) moderating variables. Methods: Studies identified in PubMed, Scopus, Web of Science, and ProQuest between January 2000 and December 2022 were included. Methodological quality was assessed with Newcastle Ottawa Scale. The protocol was pre -registered (PROSPERO, CRD42023387318). Results: Twenty-seven studies (20121 participants, 12 countries) were included. The corrected pooled estimated mean score of QoL was 47.92 (27 studies, CI 95 %: 43.11 to 52.72, range 0 -100, i.e., low -high QoL) and was marginally affected by publication year (increased 0.89 by each year, p = 0.12). The corrected pooled estimated mean score of the mental health component was 47.32 (17 studies, 95 % CI: 43.12 to 51.51, range 0 -100) and influenced by RLS instrument (decreased with recent versions, p = 0.05). The corrected pooled estimated mean score of the physical health component was 39.08 (17 studies, 95 % CI: 33.05 to 45.10, range 0 -100), with no statistically significant moderator. The pooled estimated QoL scores were statistically significantly lower in RLS patients compared to control groups with standardized mean difference (SMD) of -0.78, -0.57 and -0.50 respectively for overall QoL (24 studies), physical and mental health components (14 studies). Total QoL SMD was affected by proportion of women. Conclusion: Low QoL was revealed among RLS patients, which was statistically significantly reduced compared to control groups.

Background Glioblastoma is the most common malignant brain tumor in adults. Non-invasive clinical parameters could play a crucial role in treatment planning and serve as predictors of patient survival. Our register-based real-life study aimed to investigate the prognostic value of presenting symptoms. Methods Data on presenting symptoms and survival, as well as known prognostic factors, were retrieved for all glioblastoma patients in Sweden registered in the Swedish Brain Tumor Registry between 2018 and 2021. The prognostic impact of different presenting symptoms was calculated using the Cox proportional hazard model. Results Data from 1458 adults with pathologically verified IDH wild-type glioblastoma were analyzed. Median survival time was 345 days. The 2-year survival rate was 21.5%. Registered presenting symptoms were focal neurological deficits, cognitive dysfunction, headache, epilepsy, signs of raised intracranial pressure, and cranial nerve symptoms, with some patients having multiple symptoms. Patients with initial cognitive dysfunction had significantly shorter survival than patients without; 265 days (245-285) vs. 409 days (365-453; P < .001). The reduced survival remained after Cox regression adjusting for known prognostic factors. Patients presenting with seizures and patients with headaches had significantly longer overall survival compared to patients without these symptoms, but the difference was not retained in multivariate analysis. Patients with cognitive deficits were less likely to have radical surgery and to receive extensive anti-neoplastic nonsurgical treatment. Conclusions This extensive real-life study reveals that initial cognitive impairment acts as an independent negative predictive factor for treatment decisions and adversely affects survival outcomes in glioblastoma patients.

Low-frequency hearing is critically important for speech and music perception. However, technical and anatomical limitations previously made it difficult to study the mechanics of the low-frequency parts of the cochlea, but this changed with the introduction of optical coherence tomography vibrometry. With this technique, sound-evoked vibration can be measured from the apex of a fully intact cochlea. Results of such measurements generated controversy because conventional traveling waves, the hallmark of which is longer group delay closer to the helicotrema, were absent within the apical 20% of the guinea pig cochlea (Burwood et al, Science Advances 8:eabq2773, 2022). The validity of this result was questioned, primarily because group delays were calculated from phase values averaged across many points within the organ of Corti. Here we show that variations in phase across the organ of Corti are minor and does not affect the group delay significantly. We also assess the precision of phase measurements with optical coherence tomography. An artificial target with reflectivity similar to the organ of Corti was used. These measurements revealed that a commonly used commercial optical coherence tomography system produces half-cycle errors in 1-5 % of pixels, leading to a bimodal distribution of phase values. This problem can be easily addressed by using medians when computing averages, as was done by Burwood et al (2022). Hence, neither averaging across pixels nor technical factors can explain the apparent lack of conventional traveling waves at the apex of the guinea pig cochlea at low stimulus levels. The physiological mechanisms that operate at the apex apparently differ from other cochlear regions.

The ability to detect and respond to sickness in others promotes survival. Here we show that mouse dams respond to immune challenged pups by mirroring their inflammatory response. Dams with pups subjected to immune challenge displayed a marked induction of inflammatory mediators in both the brain and the periphery, accompanied by an increase in maternal behaviors and corticosterone levels. This social transmission of inflammation did not require physical contact, and it contributed to the stress hormone response in the dams. In adult dyads, interaction with an immune challenged cagemate did not elicit robust inflammatory signaling but induced an increased responsiveness to a subsequent immune challenge. The identification of social transmission of inflammation, or inflammatory responsiveness, may open new avenues for research on social behavior, just like the description of similar phenomena such as observational fear and transmitted pain has done.

Inflammation, corticosteroids, and loading all affect tendon healing, with an interaction between them. However, underlying mechanisms behind the effect of corticosteroids and the interaction with loading remain unclear. The aim of this study was to investigate the role of dexamethasone during tendon healing, including specific effects on tendon cells. Rats (n = 36) were randomized to heavy loading or mild loading, the Achilles tendon was transected, and animals were treated with dexamethasone or saline. Gene and protein analyses of the healing tendon were performed for extracellular matrix-, inflammation-, and tendon cell markers. We further tested specific effects of dexamethasone on tendon cells in vitro. Dexamethasone increased mRNA levels of S100A4 and decreased levels of ACTA2/alpha-SMA, irrespective of load level. Heavy loading + dexamethasone reduced mRNA levels of FN1 and TenC (p < 0.05), while resolution-related genes were unaltered (p > 0.05). In contrast, mild loading + dexamethasone increased mRNA levels of resolution-related genes ANXA1, MRC1, PDPN, and PTGES (p < 0.03). Altered protein levels were confirmed in tendons with mild loading. Dexamethasone treatment in vitro prevented tendon construct formation, increased mRNA levels of S100A4 and decreased levels of SCX and collagens. Dexamethasone during tendon healing appears to act through immunomodulation by promoting resolution, but also through an effect on tendon cells.

A major driver of neuronal hyperexcitability is dysfunction of K+ channels, including voltage-gated KCNQ2/3 channels. Their hyperpolarized midpoint of activation and slow activation and deactivation kinetics produce a current that regulates membrane potential and impedes repetitive firing. Inherited mutations in KCNQ2 and KCNQ3 are linked to a wide spectrum of neurodevelopmental disorders (NDDs), ranging from benign familial neonatal seizures to severe epileptic encephalopathies and autism spectrum disorders. However, the impact of these variants on the molecular mechanisms underlying KCNQ3 channel function remains poorly understood and existing treatments have significant side effects. Here, we use voltage clamp fluorometry, molecular dynamic simulations, and electrophysiology to investigate NDD-associated variants in KCNQ3 channels. We identified two distinctive mechanisms by which loss- and gain-of function NDD-associated mutations in KCNQ3 affect channel gating: one directly affects S4 movement while the other changes S4-to-pore coupling. MD simulations and electrophysiology revealed that polyunsaturated fatty acids (PUFAs) primarily target the voltage-sensing domain in its activated conformation and form a weaker interaction with the channel's pore. Consistently, two such compounds yielded partial and complete functional restoration in R227Q- and R236C-containing channels, respectively. Our results reveal the potential of PUFAs to be developed into therapies for diverse KCNQ3-based channelopathies. Fatty acids restore the function of neurodevelopmental-associated KCNQ3 mutations that either disrupt channel function by directly affecting voltage sensor movement or by changing voltage sensor-to-pore coupling.

The superfamily of voltage-gated potassium (KV) channels is crucial for the normal function of several tissues and represents an attractive pharmacological target for treating disorders such as epilepsy and cardiac arrhythmias. However, any drug designed to target a KV channel must be capable of discriminating between different members within the superfamily, lest they plague the user with deleterious side effects. Such rational design requires structural and functional insight into how the selectivity of a molecule can be tailored to suit the intended target.  

This thesis combines the use of electrophysiological and computational techniques to investigate the molecular basis for how the function of hKV7 and hERG channels can be modulated by different lipophilic compounds with known or suspected effects on ion channels. These include polyunsaturated fatty acids (PUFAs), cannabidiol (CBD), and synthetic cannabinoid receptor agonists (SCRAs).   

Using the two-electrode voltage clamp technique on Xenopus oocytes, we find that both PUFAs and CBD modulate the function of hKV7 channels in subtype-specific manners. PUFAs facilitated the activation of hKV7 channels, except for hKV7.4 channels which were instead inhibited. Molecular dynamics simulations revealed that structural differences in the voltage-sensing domain of hKV7.4 conferred a unique, inhibitory PUFA interaction site absent in the other hKV7 subtypes. Once this site was neutralised by mutagenesis, PUFAs facilitated hKV7.4 activation. In the case of CBD, we observed three different responses: inhibition of channels with hKV7.1 subunits, potentiated voltage-sensitivity of channels with hKV7.2 or hKV7.3 subunits and enhanced maximum conductance of channels with hKV7.4 or hKV7.5 subunits. However, these responses were evoked from the same interaction site in the pore domain, indicating a more complex subtype-specific mechanism of action. Finally, using an automated patch-clamp system we screened 36 different SCRAs on the cardiac channels responsible for repolarisation: hERG and hKV7.1/KCNE1. We find 28 of the SCRAs to be inhibitors of hERG and 22 to be inhibitors of hKV7.1/KCNE1. Molecular dynamics simulations suggest the increased susceptibility of hERG to SCRA-mediated inhibition may be due to a unique central cavity site that is absent from the pore domain of hKV7.1/KCNE1.   

In conclusion, structurally diverse lipophilic molecules of endogenous and exogenous origins can interact with KV channels and influence their function by enhancing or interfering with functional domains. In some instances, structural differences in the channel protein can explain the discrepancies in pharmacology. These findings have implications for both pharmacology (informing rational drug design) and toxicology (identifying targets through which adverse effects may occur).   

Objectives: To determine prevalence and clinical associations of anti-Four-and-a-half-LIM-domain 1 (FHL1) autoantibodies in patients with idiopathic inflammatory myopathies (IIM) and to evaluate autoantibody levels over time. Methods: Sera at the time of diagnosis from patients with IIM (n = 449), autoimmune disease controls (DC, n = 130), neuromuscular diseases (NMDs, n = 16) and healthy controls (HC, n = 100) were analysed for anti-FHL1 autoantibodies by enzyme-linked immunosorbent assay (ELISA). Patients with IIM FHL1+ and FHL1- were included in a longitudinal analysis. Serum levels were correlated to disease activity. Results: Autoantibodies to FHL1 were more frequent in patients with IIM (122/449, 27%) compared with DC (autoimmune DC and NMD, 13/146, 9%, P < 0.001) and HC (3/100.3%, P < 0.001). Anti-FHL1 levels were higher in IIM [median (IQR)=0.62 (0.15-1.04)] in comparison with DC [0.22 (0.08-0.58)], HC [0.35 (0.23-0.47)] and NMD [0.48 (0.36-0.80)] P < 0.001. Anti-FHL1+ patients with IIM were younger at the time of diagnosis compared with the anti-FHL1- group (P = 0.05) and were seronegative for other autoantibodies in 25%.<br /> In the first follow-up, anti-FHL1+ sample 20/33 (60%) positive at baseline had turned negative for anti-FHL1 autoantibodies. Anti-FHL1 autoantibodies rarely appeared after initiating treatment. Anti-FHL1 autoantibody levels correlated with CK (r = 0.62, P= 0.01), disease activity measured using the Myositis Disease Activity Assessment Tool (MYOACT) (n = 14, P = 0.004) and inversely with Manual Muscle Test-8 (r = -0.59, P = 0.02) at baseline. Conclusion: Anti-FHL1 autoantibodies were present in 27% of patients with IIM; of these, 25% were negative for other autoantibodies. Other autoimmune diseases had lower frequencies and levels. Anti-FHL1 levels often decreased with immunosuppressive treatment, correlated with disease activity measures at diagnosis and rarely appeared after start of treatment.

Meiotic recombination between homologous chromosomes is vital for maximizing genetic variation among offspring. However, sex-determining regions are often rearranged and blocked from recombination. It remains unclear whether rearrangements or other mechanisms might be responsible for recombination suppression. Here, we uncover that the deficiency of the DNA cytosine methyltransferase DNMT1 in the green alga Chlamydomonas reinhardtii causes anomalous meiotic recombination at the mating-type locus (MT), generating haploid progeny containing both plus and minus mating-type markers due to crossovers within MT. The deficiency of a histone methyltransferase for H3K9 methylation does not lead to anomalous recombination. These findings suggest that DNA methylation, rather than rearrangements or histone methylation, suppresses meiotic recombination, revealing an unappreciated biological function for DNA methylation in eukaryotes.

Introduction: The Swedish Sleep Apnea Registry (SESAR) collects clinical data from individual obstructive sleep apnea (OSA) patients since 2010. SESAR has recently been integrated with additional national healthcare data. The current analysis presents the SESAR structure and representative clinical data of a national sleep apnea cohort. Methods: Clinical data from unselected patients with a diagnosis of OSA are submitted to the SESAR registry. 48 sleep centers report data from diagnosis, treatment starts with Continuous Positive Airway Pressure (CPAP), oral devices (OD), and Upper Airway Surgery (UAS). Data from follow-up are included. SESAR is linked to mandatory national healthcare data (mortality, comorbidities, procedures, prescriptions) and diagnosis-specific quality registries (e.g. stroke, heart failure, diabetes) within the DISCOVERY project. Results: 83,404 OSA patients have been reported during the diagnostic workup (age 55.4 +/- 14.1 years, BMI 30.8 +/- 6.5 kg/m2, 2 , AHI 25.8 +/- 21.6n/h, respectively). At least one cardiometabolic and respiratory comorbidity is recognized in 57 % of female and 53 % of male OSA patients with a linear increase across OSA severity. In 54,468, 7,797, and 390 patients, start of CPAP, OD or UAS treatment is reported, respectively. OD patients have 4 units lower BMI and 10 units lower AHI compared to patients started on CPAP. UAS patients are characterized by 10 years lower age. The degree of daytime sleepiness is comparable between treatment groups with mean Epworth Sleepiness Scale Scores between 9 and 10. Conclusion: SESAR is introduced as a large national registry of OSA patients. SESAR provides a useful tool to highlight OSA management and to perform relevant outcome research.

Radiotherapy treatment planning takes substantial time, several hours per patient, as it involves manual segmentation of tumor and risk organs. Segmentation networks can be trained to automatically perform the segmentations, but typically require large annotated datasets for training. Sharing of sensitive data between hospitals, to create a larger dataset, is often difficult due to ethics and GDPR. Here we therefore demonstrate that federated learning is a solution to this problem, as then only the segmentation model is sent between each hospital and a global server. We export and preprocess brain tumor images from the oncology departments in Linköping and Lund, and use federated learning to train a global segmentation model using two different frameworks.

Tendon healing involves mechanosensitive cells that adapt to mechanical stimuli through mechanotransduction, resulting in increased tissue strength. However, detailed insights into this process in response to different loads remain limited. We aimed to investigate how different loading regimes impact the spatial composition of elastin and collagens during Achilles tendon healing. Histological analysis was conducted on healing rat Achilles tendons exposed to (1) full loading, (2) reduced loading, or (3) minimal loading. Histological analysis included Hematoxylin & Eosin and immunohistochemical staining targeting elastin, Collagen 1, Collagen 3, and CD31. Our results showed that the impact of mechanical stimuli on healing tendons varied with the degree of loading. Unexpectedly, minimal loading led to higher staining intensity for collagens and elastin. However, tendons exposed to minimal loading appeared thinner and exhibited a less organized matrix structure, with fewer, less aligned, and more rounded cells. Additionally, our findings indicated an inverse correlation between angiogenesis and load level, with more blood vessels in tendons subjected to less loading. Tissue integrity improved by 12 weeks post-injury, but the healing process continued and did not regain the structure seen in intact tendons even after 20 weeks. This study reveals a load-dependent effect on matrix alignment, cell density, and cell alignment.

Background The treatment of elderly/ frail patients with glioblastoma is a balance between avoiding undue toxicity, while not withholding effective treatment. It remains debated, whether these patients should receive combined chemo-radiotherapy with temozolomide (RT/TMZ -> TMZ) regardless of the O6-methylguanine DNA methyltransferase gene promoter (MGMTp) methylation status. MGMT is a well-known resistance factor blunting the treatment effect of TMZ, by repairing the most genotoxic lesion. Epigenetic silencing of the MGMTp sensitizes glioblastoma to TMZ. For risk-adapted treatment, it is of utmost importance to accurately identify patients, who will not benefit from TMZ treatment.Methods Here, we present a reanalysis of the clinical trials CE.6 and the pooled NOA-08 and Nordic trials in elderly glioblastoma patients that compared RT to RT/TMZ -> TMZ, or RT to TMZ, respectively. For 687 patients with available MGMTp methylation data, we applied a cutoff discerning truly unmethylated glioblastoma, established in a pooled analysis of 4 clinical trials for glioblastoma, with RT/TMZ -> TMZ treatment, using the same quantitative methylation-specific MGMTp PCR assay.Results When applying this restricted cutoff to the elderly patient population, we confirmed that glioblastoma with truly unmethylated MGMTp derived no benefit from TMZ treatment. In the Nordic/NOA-08 trials, RT was better than TMZ, suggesting little or no benefit from TMZ.Conclusions For evidence-based treatment of glioblastoma patients validated MGMTp methylation assays should be used that accurately identify truly unmethylated patients. Respective stratified management of patients will reduce toxicity without compromising outcomes and allow testing of more promising treatment options. For the podcast associated with this article, please visit 'https://soc-neuro-onc.libsyn.com/tmz-and-mgmt-in-elderly-gbm-patients'

Introduction Restless Legs Syndrome (RLS) is a neurological disorder characterized by an urge to move arms and legs, commonly combined with distress, pain and motor restlessness. It can cause fragmented sleep, daytime symptoms, and decreased quality of life. Pharmacological treatment can suppress symptoms, but not cure. When challenged with illness, people may turn to their inner psychological resources such as self-esteem and mastery. The Pearlin Mastery scale was developed to study stress and coping, is commonly used in people with chronic illnesses, however, not yet validated in people with RLS.Aim The aim was to test reliability and construct validity of the Pearlin Mastery Scale in persons with RLS.Methods A cross-sectional postal survey including the Pearlin Mastery Scale, Restless Legs Syndrome-6 Scale, Pittsburgh Sleep Quality Index and Patient Health Questionnaire was sent out to members (n = 1500) of the national RLS association and 788 (52.5%) agreed to participate. Data were analyzed using classical test theory, Confirmatory factor analysis and Rasch measurement theory analysis. Hypothesis testing for construct validity was done by bivariate correlation analyses.Results Most respondents were women (65%), retired (71%) and had a mean-age of 70.8 years (SD 11.4). The 7-item version of the Pearlin Mastery Scale showed poor fit to the one factor model. After omitting the two positively worded items (i.e., item 4 and 6), the 5-item version was found to be unidimensional, with satisfactory internal consistency. However, all items showed considerable ceiling effects. No measurement variance was seen regarding age-groups or sex. Higher level of mastery was moderately correlated with less depressive symptoms but only weakly correlated to RLS-related sleep problems.Conclusion The 5-item version of the Pearlin Mastery Scale is suggested to be used in persons with RLS due to its acceptable psychometric properties. The instrument could be applied as an outcome measure for behavioral change interventions aiming to support mastery in RLS.

The endogenous endocannabinoid-like compound N-arachidonoyl-L-serine (ARA-S) facilitates activation of the human Kv7.1/KCNE1 channel and shortens a prolonged action potential duration and QT interval in guinea pig hearts. Hence, ARA-S is interesting to study further in cardiac models to explore the functional impact of such Kv7.1/KCNE1-mediated effects. To guide which animal models would be suitable for assessing ARA-S effects, and to aid interpretation of findings in different experimental models, it is useful to know whether Kv7.1/KCNE1 channels from relevant species respond similarly to ARA-S. To this end, we used the two-electrode voltage clamp technique to compare the effects of ARA-S on Kv7.1/KCNE1 channels from guinea pig, rabbit, and human Kv7.1/KCNE1, when expressed in Xenopus laevis oocytes. We found that the activation of Kv7.1/KCNE1 channels from all tested species was facilitated by ARA-S, seen as a concentration-dependent shift in the voltage-dependence of channel opening and increase in current amplitude and conductance over a broad voltage range. The rabbit channel displayed quantitatively similar effects as the human channel, whereas the guinea pig channel responded with more prominent increase in current amplitude and maximal conductance. This study suggests that rabbit and guinea pig models are both suitable for studying ARA-S effects mediated via Kv7.1/KCNE1.

The aim of the article is to describe our work on peptides discovered in Dr. Mutt's laboratory, particularly galanin. Some personal recollections of meetings with Viktor Mutt and a brief overview of early neuropeptide research at Karolinska Institutet are provided. General aspects on neuropeptide signalling and neuropeptide-neurotransmitter coexistence are followed by the presentation of a possible involvement of the galanin system in pain and depression. Special emphasis is on the role of galanin in the rat and human locus coeruleus. Additional analyses of the human postmortem brains have given results on galanin and other peptides both in the normal prefrontal cortex as well as in different brain regions of depressed patients who have committed suicide and in control subjects. Possible options for developing treatment strategies for pain and depression based on galaninergic mechanisms are discussed. Finally, some recent drugs approved by the FDA for the treatment of conditions such as migraine, which target the signalling of other peptides, are highlighted. In conclusion, the aim of the article is to highlight the potential of the large group of neuropeptides as targets for the development of drugs that may further help patients with illnesses afflicting the nervous system.

Primary cutaneous lymphomas (PCLs), especially mycosis fungoides (MF), pose significant diagnostic and therapeutic challenges. This study aims to correlate initial histological features with the disease course and survival in MF patients. A retrospective-prospective cohort study was conducted on 83 patients diagnosed with early-stage MF at the Departments of Dermatovenerology and Pathology, UHC Zagreb, from January 2003 to December 2012. The analyzed histopathological parameters included lichenoid dermal lymphocyte infiltrate, Pautrier microabscesses, and lymphocyte atypia. Patients with more than 30 guardian lymphocytes per 100 keratinocytes exhibited worse overall and progression-free survival. Furthermore, those with over 50% atypical lymphocytes demonstrated a faster progression rate. A dense lichenoid dermal infiltrate and a high count of lymphocyte "keepers" significantly increased the mortality risk within five years of diagnosis. This study did not fully confirm the hypothesis regarding the prognostic value of large Pautrier microabscesses but highlighted the importance of dense lichenoid infiltrates. The study identified new potential histopathological prognostic factors in early-stage MF, suggesting the need for larger studies to confirm these findings. The identification of such predictors could enhance the prognostic stratification and guide more tailored therapeutic approaches for MF patients.

Background: Data on the incidence and comorbidity of Lichen sclerosus (LS), based on validated nationwide population-based registries, remains scarce. Objective: To explore the incidence and association of comorbidities with LS in Sweden, emphasizing its potential links to malignancies and autoimmune disorders. Methods: A population-based retrospective open cohort study was conducted using the National Patient Register to identify all individuals diagnosed with LS (ICD-10 code L90.0) from 1 January 2001 to 1 January 2021. The study included 154,424 LS patients and a sex and age matched control group of 463,273 individuals to assess the incidence and odds ratios for various cancers and premalignant conditions. Results: The incidence of LS in Sweden was 80.9 per 100,000 person per year, with higher incidence in females (114.4) than in males (47.2). LS patients showed an increased odds ratio for vulvar cancer (OR = 8.3; 95% CI = 7.5-9.0), penile cancer (OR = 8.9; 95% CI = 7.3-11.0), prostate cancer (OR = 1.2; 95% CI = 1.1-1.2), testicular cancer (OR = 1.4; 95% CI = 1.1-1.7), bladder cancer (OR = 1.1; 95% CI = 1.1-1.2), breast cancer (OR = 1.4; 95% CI = 1.3-1.4), leukoplakia of the vulva (OR = 253.5; 95% CI = 221.9-289.6), and leukoplakia of the penis (OR = 5.1; 95% CI = 4.9-5.4). Conclusions: This study underscores the significantly increased association of various cancers and premalignant conditions in LS patients, highlighting the critical need for efficacious treatment and diligent follow-up. The association between LS and autoimmune diseases further necessitates comprehensive investigation to understand the underlying mechanisms and clinical management implications. Future research is essential to confirm these findings and elucidate the role of LS in cancer development.

Alzheimer’s disease (AD) is the leading cause of dementia, and clinical symptoms develop due to neuronal death. This neurodegeneration is believed to be primarily initiated by the abnormal accumulation of Amyloid β (Aβ) peptide and tau protein aggregates. Among these, Aβ accumulation is considered the primary driver of AD and forms the main focus of this thesis. Different alloforms of Aβ (e.g. Aβ1-40 and Aβ1-42) are produced by differential cleavage of the Amyloid precursor protein (APP). The lifecycles of APP and Aβ depend on their localization to different cellular compartments such as endosomes, but they can also be trafficked out of the cells in exosomes, a type of extracellular vesicle (EV). It is hypothesized that halting the spread and accumulation of Aβ via EVs could slow neurodegeneration and disease progression. However, the mechanisms underlying Aβ packaging into these vesicles and how other AD-related cellular dysfunctions influence EV propagation, remain largely unclear. Understanding these mechanisms could be crucial for identifying new treatment targets. This thesis aims to elucidate some of these gaps.   

Differences in Aβ fibril conformations have been hypothesized to result in diverse clinical phenotypes of AD. Therefore, Paper I examined how different ratios of Aβ1-40 to Aβ1-42 affect fibril conformation. Given that there is no standard timeframe for fibril formation, we further investigated the effect of maturation time. Using electron microscopy, different dyes, and antibodies, we found that both Aβ1-40:42 ratio and maturation significantly influenced fibril conformation. Fibrils with higher Aβ1-42 ratio showed increased cellular association, although this did not substantially impact cytotoxicity and autophagy.

Mutations or dysfunctions related to endosomal trafficking (e.g. dysfunction of the retromer subunit Vps35), can cause Aβ accumulation but the impact on exosome biogenesis and EV release is lacking. Therefore, Paper II investigated the effect of Vps35 knockout (KO) on exosome biogenesis and EV cargo during Aβ challenge. We found that lack of Vps35 resulted in lower EV abundance, caused by a decrease in exosome biogenesis-related proteins, leading to increased Aβ cargo. Aβ challenge also led to increased EV release and affected EV cargo, revealing new mechanisms by which Vps35 dysfunction contributes to Aβ accumulation.  

Aβ is hypothesized to be released in exosomes via a neutral sphingomyelinase 2 (nSMase2)-dependent pathway. However, knowledge regarding how Aβ is packaged is still lacking. Therefore, Paper III aimed to confirm the nSmase2-dependency and to investigate whether the Aβ-interacting cellular prion protein (PrPC) could influence Aβ packaging into exosomes. Our findings confirmed that Aβ release occurs via an nSMase2-dependent pathway, independent of PrPC. Surprisingly, lack of PrPC caused cellular Aβ accumulation, changes to exosome biogenesis-related proteins, and an increase in EV production.   

Autophagy is heavily involved in both the accumulation and degradation of Aβ and has been connected to EV abundance. Therefore, Paper IV examined the role of autophagy in Aβ accumulation and EV dynamics. Using a cross between an Atg7 conditional KO mouse and a knock in APP mouse (with Swedish and Iberian mutation, APP^NL-F) we could demonstrate that plaques and intracellular Aβ was decreased while EV abundance was increased upon autophagy deficiency. However, the Aβ content of EVs was not altered. With additional mutations (Arctic mutation, APP^NL-G-F), leading to excessive Aβ production in autophagy deficient mice, similar decreases in Aβ plaque were seen but intracellular Aβ remained unchanged. This resulted in different cell death mechanisms, gamma oscillations, and behavioral outcomes between the autophagy deficient APP^NL-F and APP^NL-G-F mice.   In conclusion, this thesis expands our understanding of the impact of amyloid conformation, endosomal trafficking dysfunction, autophagy dysfunction, exosome biogenesis and EVs dynamics in the progression of AD. 

Alzheimer's disease (AD) progression is closely linked to the propagation of pathological Amyloid beta (A beta), a process increasingly understood to involve extracellular vesicles (EVs), namely exosomes. The specifics of A beta packaging into exosomes remain elusive, although evidence suggests an ESCRT (Endosomal Sorting Complex Required for Transport)-independent origin to be responsible in spreading of AD pathogenesis. Intriguingly, PrPC, known to influence exosome abundance and bind oligomeric A beta (oA beta), can be released in exosomes via both ESCRT-dependent and ESCRT-independent pathways, raising questions about its role in oA beta trafficking. Thus, we quantified A beta levels within EVs, cell medium, and intracellularly, alongside exosome biogenesis-related proteins, following deletion or overexpression of PrPC. The same parameters were also evaluated in the presence of specific exosome inhibitors, namely Manumycin A and GW4869. Our results revealed that deletion of PrPC increases intracellular A beta accumulation and amplifies EV abundance, alongside significant changes in cellular levels of exosome biogenesis-related proteins Vps25, Chmp2a, and Rab31. In contrast, cellular expression of PrPC did not alter exosomal A beta levels. This highlights PrPC's influence on exosome biogenesis, albeit not in direct A beta packaging. Additionally, our data confirm the ESCRT-independent exosome release of A beta and we show a direct reduction in Chmp2a levels upon oA beta challenge. Furthermore, inhibition of opposite exosome biogenesis pathway resulted in opposite cellular PrPC levels. In conclusion, our findings highlight the intricate relationship between PrPC, exosome biogenesis, and A beta release. Specifically, they underscore PrPC's critical role in modulating exosome-associated proteins, EV abundance, and cellular A beta levels, thereby reinforcing its involvement in AD pathogenesis.Graphical abstractThere are two main exosome biogenesis pathways: ESCRT dependent and ESCRT independent. In this study, we explored the effect of the cellular prion protein (PrPC) on the release of Amyloid beta via exosomes. Our findings demonstrate that Amyloid beta mainly is released via an ESCRT-independent pathway, independent of PrPC. However, lack of PrPC resulted in upregulation of the ESCRT-dependent proteins Tsg101 and VPS25, a decrease in Chmp2a, and an overall increase in extracellular vesicles. Lack of PrPC also caused an accumulation of cellular, but not exosomal, Amyloid beta.

The amyloid beta (A beta) peptide has a central role in Alzheimer's disease (AD) pathology. The peptide length can vary between 37 and 49 amino acids, with A beta 1-42 being considered the most disease-related length. However, A beta 1-40 is also found in A beta plaques and has shown to form intertwined fibrils with A beta 1-42. The peptides have previously also shown to form different fibril conformations, proposed to be related to disease phenotype. To conduct more representative in vitro experiments, it is vital to uncover the impact of different fibril conformations on neurons. Hence, we fibrillized different A beta 1-40:42 ratios in concentrations of 100:0, 90:10, 75:25, 50:50, 25:75, 10:90 and 0:100 for either 24 h (early fibrils) or 7 days (aged fibrils). These were then characterized based on fibril width, LCO-staining and antibody-staining. We further challenged differentiated neuronal-like SH-SY5Y human cells with the different fibrils and measured A beta content, cytotoxicity and autophagy function at three different time-points: 3, 24, and 72 h. Our results revealed that both A beta 1-40:42 ratio and fibril maturation affect conformation of fibrils. We further show the impact of these conformation changes on the affinity to commonly used A beta antibodies, primarily affecting A beta 1-40 rich aggregates. In addition, we demonstrate uptake of the aggregates by neuronally differentiated human cells, where aggregates with higher A beta 1-42 ratios generally caused higher cellular levels of A beta. These differences in A beta abundance did not cause changes in cytotoxicity nor in autophagy activation. Our results show the importance to consider conformational differences of A beta fibrils, as this can have fundamental impact on A beta antibody detection. Overall, these insights underline the need for further exploration of the impact of conformationally different fibrils and the need to reliably produce disease relevant A beta aggregates.image

Background. TTFields is recommended internationally for the treatment of glioblastoma. In Sweden, TTFields requires a possibly challenging collaboration between the patient, next-of-kin, healthcare, and the private company providing the device, both from an ethical and practical perspective. Little is known about glioblastoma patients' own experiences of TTFields treatment. Methods. Semi-structured individual interviews were conducted with 31 patients with glioblastoma who had been offered TTFields by the healthcare. These were analyzed by qualitative content analysis. Results. Participants described there being multiple actors around them as TTFields users; (1) device prescription from physicians, sometimes providing insufficient information, (2) practical assistance from next-of-kin, necessary to access treatment, (3) home visits from the private company staff for device control, where close bonds between patients and TTFields staff occurred. TTFields treatment created hope and a feeling of control in an otherwise hopeless situation, sometimes evoking worries at the time of planned treatment stop. Some refrained from TTFields or discontinued early due to fear or experience of negative effects on quality of life. Others described finding practical and mental solutions for coping with the treatment in everyday life. Conclusions. Our study identified a need for better support and information from healthcare providers for TTFields. A solution is necessary for assistance with TTFields for those without support from next-of-kin. The study raises the question of possible advantages of healthcare handling the technical support of the device instead of a private company, thereby avoiding a true or perceived influence on the patient's decision to continue or stop treatment.

Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels1 are essential for pacemaking activity and neural signalling2,3. Drugs inhibiting HCN1 are promising candidates for management of neuropathic pain4 and epileptic seizures5. The general anaesthetic propofol (2,6-di-iso-propylphenol) is a known HCN1 allosteric inhibitor6 with unknown structural basis. Here, using single-particle cryo-electron microscopy and electrophysiology, we show that propofol inhibits HCN1 by binding to a mechanistic hotspot in a groove between the S5 and S6 transmembrane helices. We found that propofol restored voltage-dependent closing in two HCN1 epilepsy-associated polymorphisms that act by destabilizing the channel closed state: M305L, located in the propofol-binding site in S5, and D401H in S6 (refs. 7,8). To understand the mechanism of propofol inhibition and restoration of voltage-gating, we tracked voltage-sensor movement in spHCN channels and found that propofol inhibition is independent of voltage-sensor conformational changes. Mutations at the homologous methionine in spHCN and an adjacent conserved phenylalanine in S6 similarly destabilize closing without disrupting voltage-sensor movements, indicating that voltage-dependent closure requires this interface intact. We propose a model for voltage-dependent gating in which propofol stabilizes coupling between the voltage sensor and pore at this conserved methionine-phenylalanine interface in HCN channels. These findings unlock potential exploitation of this site to design specific drugs targeting HCN channelopathies. Propofol repairs malfunctioning mutant HCN1 channels associated with epilepsy, and its unusual mechanism of action on these ion channels can potentially be exploited to design precision drugs targeting HCN channelopathies.

Background & aim: Clinical trials supplementing the long-chain polyunsaturated fatty acids (LCPUFAs) docosahexaenoic acid (DHA) and arachidonic acid (AA) to preterm infants have shown positive effects on in flammation-related morbidities, but the molecular mechanisms underlying these effects are not fully elucidated. This study aimed to determine associations between DHA, AA, and in flammation-related proteins during the neonatal period in extremely preterm infants. Methods: A retrospective exploratory study of infants (n 1 / 4 183) born below 28 weeks gestation from the Mega Donna Mega trial, a randomized multicenter trial designed to study the effect of DHA and AA on retinopathy of prematurity. Serial serum samples were collected after birth until postnatal day 100 (median 7 samples per infant) and analyzed for phospholipid fatty acids and proteins using targeted proteomics covering 538 proteins. Associations over time between LCPUFAs and proteins were explored using mixed effect modeling with splines, including an interaction term for time, and adjusted for gestational age, sex, and center. Results: On postnatal day one, 55 proteins correlated with DHA levels and 10 proteins with AA levels. Five proteins were related to both fatty acids, all with a positive correlation. Over the first 100 days after birth, we identi fied 57 proteins to be associated with DHA and/or AA. Of these proteins, 41 (72%) related to in flammation. Thirty-eight proteins were associated with both fatty acids and the overall direction of association did not differ between DHA and AA, indicating that both LCPUFAs similarly contribute to upand down-regulation of the preterm neonate in flammatory proteome. Primary examples of this were the in flammation-modulating cytokines IL -6 and CCL7, both being negatively related to levels of DHA and AA in the postnatal period. Conclusions: This study supports postnatal non -antagonistic and potentially synergistic effects of DHA and AA on the inflammation proteome in preterm infants, indicating that supplementation with both fatty acids may contribute to limiting the disease burden in this vulnerable population. Clinical registration number: ClinicalTrials.gov (NCT03201588). (c) 2024 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Purpose The aim of this study was to validate the Ethos Brief Index (EBI) in patients with Restless Legs Syndrome (RLS).Methods A cross-sectional design, including 788 subjects with RLS (65% women, 70.8 years, SD 11.3) from the Swedish RLS Association, was used. A postal survey was sent out to collect data regarding socio demographics, comorbidities, and RLS-related treatment data. Questionnaires included were EBI, the Restless Legs Syndrome-6 Scale (RLS-6), Restless Legs Syndrome-Quality of Life questionnaire (RLSQoL), the Insomnia Severity Index (ISI), and the Epworth Sleepiness Scale (ESS). The validity and reliability of the EBI were investigated using Rasch and confirmatory factor analysis (CFA) models. Measurement invariance, unidimensionality, and differential item functioning (DIF) across age and gender groups, as well as insomnia, daytime sleepiness, RLS-related QoL and RLS severity were assessed.Results The results supported the unidimensionality of the EBI in the CFA (i.e., explaining 61.5% of the variance) and the Rasch model. The reliability of the EBI was confirmed using composite reliability and Cronbach's alpha. No DIF was identified for gender, age, insomnia, daytime sleepiness, RLS severity or RLS-related QoL.Conclusion The EBI showed good validity and reliability and operated equivalently for male and female patients with RLS. Accordingly, healthcare professionals can use the EBI as a psychometrically sound tool to explore and identify patient-centered problems related to the whole life situation.

Psoriasis is an IL-23/Th17-mediated skin disorder with a strong genetic predisposition. The impact of its susceptibility gene nitric oxide synthase 2 (NOS2) remains unknown. Here, we demonstrate strong NOS2 mRNA expression in psoriatic epidermis, an effect that is IL-17 dependent. However, its complete translation to protein is prevented by the IL-17-induced miR-31 implying marginally upregulated NO levels in psoriatic skin. We demonstrate that lower levels of NO, as opposed to higher levels, increase keratinocyte proliferation and mediate IL-17 downstream effects. We hypothesized that the psoriatic phenotype may be alleviated by either eliminating or increasing cellular NO levels. In fact, using the imiquimod psoriasis mouse model, we found a profound impact on the psoriatic inflammation in both IMQ-treated NOS2 KO mice and wild-type mice treated with IMQ and the NO-releasing berdazimer gel. In conclusion, we demonstrate that IL-17 induces NOS2 and fine-tunes its translation towards a window of proinflammatory and hyperproliferative effects and identify NO donor therapy as a new treatment modality for psoriasis.

Pain and social behavior are subject to reciprocal modulation. Social animals show attenuated pain behavior in the presence of conspecifics, while observers are in turn affected by exposure to another individual in pain. Both phenomena have been established in rodents, which in addition to experiencing emotional contagion from afflicted conspecifics may act to relieve their afflicted state. Little has been done to investigate the motivation of such prosocial behavior in naturalistic conditions. Here, using a novel formalin test targeting the nape of the neck, a socially relevant area, we investigated nocifensive behaviors and social interactions in mice group-housed in a seminatural environment (SNE). In the SNE, formalin-injected mice displayed fewer back-scratching occurrences than when housed alone, which was inversely correlated to the social behavior received. These mice also emitted and received fewer social interactions, attesting of social withdrawal. With respect to dyadic exchanges, saline-treated mice initiated fewer allosniffing and anogenital sniffing episodes towards formalin-treated mice than towards other saline-treated ones. These findings are counter to those showing empathy and prosocial approach in mouse pain models. It is possible that in naturalistic conditions that allow the mice to express a wide range of their behavioral repertoire, healthy mice simply avoid individuals in pain and the cost associated with emotional contagion. Interestingly, behaviors involving direct body contact, namely allogrooming and pair-resting, were not different between saline- and formalin-treated mice and thus may carry a prosocial, altruistic component. These findings unveil new patterns of social modulation by pain in a naturalistic laboratory setting holding high translational value.

The CorA/Mrs2 family of pentameric proteins are cardinal for the influx of Mg2+ across cellular membranes, importing the cation to mitochondria in eukaryotes. Yet, the conducting and regulation mechanisms of permeation remain elusive, particularly for the eukaryotic Mrs2 members. Here, we report closed and open Mrs2 cryo-electron microscopy structures, accompanied by functional characterization. Mg2+ flux is permitted by a narrow pore, gated by methionine and arginine residues in the closed state. Transition between the conformations is orchestrated by two pairs of conserved sensor-serving Mg2+-binding sites in the mitochondrial matrix lumen, located in between monomers. At lower levels of Mg2+, these ions are stripped, permitting an alternative, symmetrical shape, maintained by the RDLR motif that replaces one of the sensor site pairs in the open conformation. Thus, our findings collectively establish the molecular basis for selective Mg2+ influx of Mrs2 and an auto-ligand-gating regulation mechanism. Here, the authors demonstrate how Mrs2, critical for import of Mg2+ into the mitochondria, transitions from open and closed five-fold symmetric states to control the influx of Mg2+ using an auto-ligand-gating regulation mechanism.

OBJECTIVE MRI volumetry could be used as an alternative to invasive tests of shunt function. In this study, the authors aimed to assess the difference in ventricular volume (VV) before and after surgery and at different performance levels (PLs) of the shunt. METHODS This study was a randomized, double-blind trial with a crossover design. The study sample consisted of 36 patients (25 men, 11 women) with a median age of 76 years. All patients had idiopathic normal pressure hydrocephalus (iNPH) and received a Strata shunt at the regional hospital in & Ouml;stersund, Sweden, with an initial PL of 1.5. Participants underwent MRI with volumetric sequences before surgery and four times postoperatively: at 1 month before randomization to either PL 1.0 (n = 15) or 2.5 (n = 17); at 2 months before crossover to PL 2.5 or 1.0; at 3 months before lowering the PL to 0.5; and finally, at 3 months and 1 day after surgery before resetting the PL to 1.5. VV was measured semiautomatically using quantitative MRI. Both the patient and the examiner of clinical tests and volumetry were blinded to the PL. RESULTS VV changed significantly between the presurgical level (median 129 ml) and the different shunt settings, i.e., PL 1.0 (median 115 ml), 1.5 (median 120 ml), and 2.5 (median 128 ml; p < 0.001). A unidirectional change in VV was observed for all participants between PL 1.0 and PL 2.5 (median 12 ml, range 2.1-40.7 ml, p < 0.001). No significant change was noted in VV after 24 hours at PL 0.5. Eight participants had asymptomatic subdural effusions at PL 1.0. CONCLUSIONS The consistent decrease in VV after shunt surgery and between PL 2.5 and 1.0 supports the idea that MRI volumetry could be a noninvasive method for evaluating shunt function in iNPH, preventing unnecessary shunt revisions. However, further studies on retest variability of VV as well as verification against advanced testing of shunt function are needed before a clinical implementation of this method can be performed.

Sleepiness and apathy are often reported in patients with normal pressure hydrocephalus. However, research on outcomes after shunt surgery has mainly focused on the classical triad symptoms, that is, gait, cognition, and bladder dysfunction. This study aimed to describe the effects of shunt treatment on excessive daytime sleepiness and whether there was a relation to changes in ventricular volume. Pre- and postsurgical excessive daytime sleepiness was investigated using the Epworth sleepiness scale in a sample of 32 patients with normal pressure hydrocephalus who underwent shunt surgery. Data were gathered before surgery and at 1, 2, and 3 months after surgery and with different settings of the shunt. In the total sample, the Epworth sleepiness scale improved by a median of 1.5 points at 1 month after surgery, p = 0.026. The improvement was predominately found in the group (n = 6) with high presurgical daytime sleepiness (Epworth sleepiness scale >12) (median = 12 points, p = 0.035) compared with a median change of 0 points (p = 0.47) in the group with Epworth sleepiness scale <= 12 (n = 26). Between the postsurgical follow-ups, no further change in the Epworth sleepiness scale score was observed. The Epworth sleepiness scale score did not correlate with clinical tests nor with ventricular volume. Daytime sleepiness seems to be another domain of normal pressure hydrocephalus symptomatology in addition to the classical triad that is responsive to treatment, at least when pronounced. The Epworth sleepiness scale is a quick test to administer and could be a valuable addition to pre-surgical screening for treatable symptoms.

Tubulin beta-3 staining pattern and staining intensity of 5-hydroxymethyl cytosine (5-hmC) are potential diagnostic and prognostic markers in melanocytic lesions that need further evaluation. Melanocytic nevi and primary cutaneous melanomas were immunohistochemically stained for tubulin-beta-3 and 5-hmC. Immunoreactivity and staining patterns were correlated with Breslow-thickness, clinical and pathological characteristics, and progression-free survival. Melanocytes showed positive tubulin beta-3 staining. However, in most nevi, tubulin beta-3 staining appeared as a gradient with intense cytoplasmic staining in cells of the superficial part of the lesion that faded to weak staining in the deep dermal part, while no gradient was found in deep penetrating nevi and melanomas. In 53 % of the melanomas, areas with loss of tubulin beta-3 staining were found. 5-hmC staining intensity was significantly higher in melanocytic nevi compared to melanomas. Breslow thickness in combination with low 5-hmC score and loss of tubulin-beta-3 staining was predictive for poor prognosis. As single markers, tubulin-beta-3 and 5-hmC can be useful to distinguish between melanocytic nevi and melanoma, but staining variability limits the use of 5-hmC. In melanomas measuring >1.5 mm, combination of low 5-hmC score and loss of tubulin-beta-3 staining may have prognostic value.