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Calsequestrin-1 (CASQ1) is a calcium-binding protein with high expression in fast twitch fibers of skeletal muscle. Due to its high binding capacity and low affinity for calcium, CASQ1 is the most crucial Ca2+ buffering protein in the sarcoplasmic reticulum, facilitating rapid Ca2+ release to initiate muscle contraction. CASQ1 polymerizes upon Ca2+ binding, which in turn increases its ability to bind Ca2+. The C-terminal CAS domain mediates polymerization as well as interactions with other sarcoplasmic reticulum proteins. Variants in CASQ1 have been associated with vacuolar myopathy with CASQ1 aggregates. Six pathogenic variants have been published to date. The disease is an adult-onset, often slowly progressive muscle disorder, the typical symptoms being proximal and/or distal muscle weakness, myalgia, exercise intolerance, cramps, and fatigue. We have identified three novel dominant CASQ1 variants in seven patients with mild adult-onset myopathy: p.Glu89Lys, p.Ser356Gly, and p.Gly383Alafs*39 (RefSeq NP_001222.3). All variants are predicted to damage the protein. The variant p.Gly383Alafs*39 deletes the aspartic acid residues of the CAS domain and the protein is extended by 24 amino acids. Transfection of CASQ1 -wt and CASQ1 -Ser356Gly to HeLa cells resulted in polymeric CASQ1 networks, whereas CASQ1 -Gly383Alafs*39 ended up in aggregates and CASQ1 -Glu89Lys appeared monomeric. Treatment with the sarcoplasmic/endoplasmic reticulum calcium ATPase (SERCA) inhibitor thapsigargin revealed that CASQ1-Gly383Alafs*39 and CASQ1 -Glu89Lys remain aggregated or monomeric, respectively, also when the ER is depleted of Ca2+. Experiments measuring the Ca2+ binding ability of the variants are underway. In conclusion, this study adds to our understanding of CASQ1-related myopathies. [ABSTRACT FROM AUTHOR]

Distal myopathies are a group of rare progressive genetic muscle disorders typically causing weakness and atrophy in the feet and/or hand muscles. The family under study is affected by an adult-onset dominant distal myopathy and the objective was to identify the pathogenic variant causing the disease. After negative gene panel and exome data, linkage analysis was conducted with genotyped single nucleotide polymorphisms to identify loci that co-segregated with the symptoms. Rare small-scale variants were analyzed with short-read whole exome and genome sequencing. Structural variants (SV) and repeat expansions were detected from whole genome sequenced samples with bioinformatic tools. SVs were further revealed with optical genome mapping. A muscle biopsy of a patient was used for untargeted proteomic profiling to recognize dysregulated proteins, and RNA sequencing was performed to identify splicing alterations and altered transcriptional regulation. Genetic regions identified by linkage analysis did not contain rare segregating variants likely to be pathogenic. Linkage analysis was then repeated several times with different parameters, to account for possible presymptomatic carriers, and a patient with uncertain disease status. But none of the potential new linked regions did contain any potentially promising variants. Proteomic profiling identified over 200 dysregulated proteins, many of which were encoded by muscle-associated genes but were not located in any of the linked regions and did not contain rare segregating variants. RNA sequencing detected splicing alterations in five genes and expression changes in two genes, which were all seemingly unrelated to muscle disorders. To date we haven't been able to identify the causative variant, but further analyses are ongoing. The cause may turn out to be a complex mechanisms such as digenic/oligenic inheritance or limitations of the current next-generation sequencing. [ABSTRACT FROM AUTHOR]

X-linked myotubular myopathy (XL-MTM) is a rare and often lethal congenital myopathy, clinically characterized by muscle weakness. Its underlying molecular mechanisms remain incompletely understood. As myosin has been implicated in the pathophysiology of other forms of congenital myopathies, in the present study, we aimed to define whether muscle myosin is dysfunctional in the context of XL-MTM. For that, we used muscle tissue from human patients and a canine model. We isolated individual muscle fibres from these two species and performed loaded Mant-ATP chase experiments. Our preliminary results indicate a shift of myosin metabolic/biochemical states towards highly energy-consuming conformations in human and canine XL-MTM. Subsequently, to reverse this alteration potentially contributing to muscle fatigue, we investigated the effects of an ATP-conserving drug targeting myosin, mavacamten, using a well-defined mouse model of XL-MTM lacking myotubularin. The results are currently being analysed and include histology as well as untargeted global proteomics. Potentially, taken together, these data will lead to a better understanding of XL-MTM and the potency of myosin as a drug target. [ABSTRACT FROM AUTHOR]

Comprehensive research on the possible difficulties in eating or the nutritional status of persons with nemaline myopathy (NM) and related disorders (NMR) has not been done despite the fact that the muscle weakness in these disorders often affects the muscles used for eating and dining, and although some scientific evidence of poorer nutritional status of patients with myopathy exists. We conducted a pilot study among adult Finnish NM or NMR patients to investigate their food consumption, nutrient intakes, selected nutrient-related laboratory parameters in blood, and self-assessed functioning at dining and eating and of the gastrointestinal tract. The methods included a survey including eating and dining related questions, a food frequency questionnaire (FFQ), food diaries, and laboratory analyses from blood samples. We invited 32 patients, and 20 (15 females. 5 males; 16 ambulatory, 4 non-ambulatory) returned the survey and FFQ. Food diaries were returned by 17, and blood samples were obtained from 16. The inter-individual differences were large in food consumption as well as nutrient and energy intake. Energy intakes, and intakes of vitamin D, calcium, dietary fiber, vitamin C, folate, and iron as well as consumption of healthy foods (such as fruits, vegetables, and whole grains) were low, especially in non-ambulatory participants, who also reported challenges in eating and dining-related functioning. The ability to walk did not, however, determine the quality of diet, and by selecting healthy foods easy to eat (e.g. porridge and smoothies), a good diet was achievable. The laboratory parameters failed to indicate severe undernourishment in any of the participants. Thus, evaluation of food consumption and nutrient intakes were needed to find patients at risk of undernourishment. The results underline the importance of monitoring adequate intake of calcium and vitamin D in this group of patients, especially considering the immobility-induced risk of osteoporosis. [ABSTRACT FROM AUTHOR]

Structural variants of the nebulin gene (NEB), including duplications and deletions of up to 24 exons, and copy number variation of the triplicate region, are an established cause of recessive nemaline myopathies and related neuromuscular disorders. Large in-frame deletions have been shown to cause dominantly inherited distal myopathies. Here, we provide an overview of 32 families with muscle disorders caused by such intragenic structural variants in NEB. The sub-cohorts with recessively inherited phenotypes consist of 12 families with pathogenic gains of the triplicate region, and 11 families with ten unique intragenic deletions or duplications. In these 23 families, the structural variants cause disease through compound heterozygosity with small variants in trans, except in two families, in which the structural variant is homozygous. Eight families have not been previously described. The third sub-cohort consists of 12 families, of which 10 have not been previously described, with distal myopathy phenotypes caused by altogether eight unique deletions, encompassing 51 to 97 exons in size, in either heterozygous (n = 10) or mosaic (n = 2) state. The structural variants were identified using whole exome sequencing, whole genome sequencing, custom Droplet Digital PCR, or custom Comparative Genomic Hybridization arrays. Sanger sequencing was used to validate breakpoints in cases where applicable. In the families where inheritance was recessive, no correlation could be made between the types of variants and the severity of the disease. In contrast, all patients carrying a large dominant deletion in NEB were affected by milder, predominantly distal muscle weakness. Thus, for the first time, we establish a clear and statistically significant correlation (T-test, p < 0.0001) between large NEB deletions and distal myopathy. In addition, we provide the hitherto largest overview of the spectrum of structural variants in NEB. [ABSTRACT FROM AUTHOR]

Background: During the COVID-19 pandemic, intensive care units (ICUs) were under heavy pressure, with a significantly increased number of severely ill patients. Hospitals introduced restrictions, and families could not visit their ill and dying family members. Patients were cared for without privacy, and several died in shared patient rooms, leaving the intensive care nurse to protect the patient's need for loving care in a vulnerable situation at the end of life., Aim: This study aimed to investigate how piloting and watch over were revealed in end-of-life care for patients with COVID-19 in intensive care COVID-19., Study Design: A qualitative study was conducted with an abductive approach was conducted. Data were collected via semi-structured interviews to cover the research area while allowing the informant to talk freely about the topic; 11 informants were interviewed., Results: The findings are presented based on four categories: The road to the decision, End-of-life care, Farewell of close family members and Closure. Each category and subcategory reveal how piloting and watch over were addressed in the end-of-life care of patients with COVID-19 in the ICU during the pandemic. Overall findings indicated that workload and organization of care directly affect the quality of care given, the acceptance of privacy and the possibility of dignified end-of-life care., Conclusions: Workload directly affects the quality of care, risking dehumanization of the patient. Visiting restrictions hindered supporting family members through the various piloting phases. Visiting restrictions also forced the ICU nurses to take on the role of the relative in watching over the patient., Relevance to Clinical Practice: Collaboration with family members is essential for the intensive care nurse to be able to provide a person-centred and dignified end-of-life care., (© 2024 The Author(s). Nursing in Critical Care published by John Wiley & Sons Ltd on behalf of British Association of Critical Care Nurses.)

The entitlement to access assistive technology (AT) is fundamental for all individuals. However, challenges encountered during societal crises can significantly impact opportunities for participation and engagement among AT users. Understanding the implications of crises and disasters on AT provision along with their repercussions for end users is crucial. This research endeavors to investigate the experiences of managers overseeing AT provision during crises, using the first wave of the COVID-19 pandemic as a case study. An open-ended questionnaire was distributed to health care managers ( n  = 18) within AT organizations in Sweden. The responses were analyzed using qualitative content analysis, and four categories derived: Embracing change and navigating new realities, Optimizing strategies due to decreased and limiting prescriptions, Unlocking access and addressing challenges in AT provision, The impact on the staff and their well-being vs effectiveness in the AT organization . The findings indicate that AT organizations have demonstrated remarkable resilience and adaptability in the face of reduced consultations and growing care burden. Despite these challenges, managers have gained valuable insights into developing AT provision more efficiently and sustainably, particularly in digitization. The lessons learned will be critical in ensuring AT provision remains responsive to the needs of patients and society in the future.

Urinary tract infection (UTI) is one of the most common bacterial infections worldwide and the most common cause is uropathogenic Escherichia coli (UPEC). Current research is mostly focused on how UPEC affects host factors, whereas the effect of host factors on UPEC is less studied. Our previous studies have shown that estrogen alters UPEC virulence. However, the effect of this altered UPEC virulence on neutrophils is unknown. The aim of the present study was to investigate how the altered UPEC virulence mediated by estrogen modulates neutrophil responses. We found that estradiol-stimulated CFT073 increased neutrophil phagocytosis, NETs formation and intracellular ROS production. We observed that the total ROS production from neutrophils was reduced by estradiol-stimulated CFT073. We also found that estradiol-stimulated CFT073 induced less cytotoxicity in neutrophils. Additionally, we found that several cytokines and chemokines like IL-8, IL-1β, CXCL6, MCP-1 and MCP-4 were increased upon estradiol-stimulated CFT073 infection. In conclusion, this study demonstrates that the estrogen-mediated alterations to UPEC virulence modulates neutrophil responses, most likely in a host-beneficial manner., (© 2024. The Author(s).)

Nemaline myopathy (NM) is a genetic muscle disease, primarily caused by mutations in the NEB gene (NEB-NM) and with muscle myosin dysfunction as a major molecular pathogenic mechanism. Recently, we have observed that the myosin biochemical super-relaxed state was significantly impaired in NEB-NM, inducing an aberrant increase in ATP consumption and remodelling of the energy proteome in diseased muscle fibres. Because the small-molecule Mavacamten is known to promote the myosin super-relaxed state and reduce the ATP demand, we tested its potency in the context of NEB-NM. We first conducted in vitro experiments in isolated single myofibres from patients and found that Mavacamten successfully reversed the myosin ATP overconsumption. Following this, we assessed its short-term in vivo effects using the conditional nebulin knockout (cNeb KO) mouse model and subsequently performing global proteomics profiling in dissected soleus myofibres. After a 4 week treatment period, we observed a remodelling of a large number of proteins in both cNeb KO mice and their wild-type siblings. Nevertheless, these changes were not related to the energy proteome, indicating that short-term Mavacamten treatment is not sufficient to properly counterbalance the metabolically dysregulated proteome of cNeb KO mice. Taken together, our findings emphasize Mavacamten potency in vitro but challenge its short-term efficacy in vivo. KEY POINTS: No cure exists for nemaline myopathy, a type of genetic skeletal muscle disease mainly derived from mutations in genes encoding myofilament proteins. Applying Mavacamten, a small molecule directly targeting the myofilaments, to isolated membrane-permeabilized muscle fibres from human patients restored myosin energetic disturbances. Treating a mouse model of nemaline myopathy in vivo with Mavacamten for 4 weeks, remodelled the skeletal muscle fibre proteome without any noticeable effects on energetic proteins. Short-term Mavacamten treatment may not be sufficient to reverse the muscle phenotype in nemaline myopathy., (© 2024 The Author(s). The Journal of Physiology published by John Wiley & Sons Ltd on behalf of The Physiological Society.)

Nemaline myopathy (NM) is a congenital myopathy with generalised muscle weakness, most pronounced in neck flexor, bulbar and respiratory muscles. The aim of this cross-sectional study was to assess the Dutch NM patient cohort. We assessed medical history, physical examination, quality of life (QoL), fatigue severity, motor function (MFM), and respiratory muscle function. We included 18 of the 28 identified patients (13 females (11-67 years old); five males (31-74 years old)) with typical or mild NM and eight different genotypes. Nine patients (50 %) used a wheelchair, eight patients (44 %) used mechanical ventilation, and four patients (22 %) were on tube feeding. Spinal deformities were found in 14 patients (78 %). The median Medical Research Council (MRC) sum score was 38/60 [interquartile range 32-51] in typical and 48/60 [44-50] in mild NM. The experienced QoL was lower and fatigue severity was higher than reference values of the healthy population. The total MFM score was 55 % [49-94] in typical and 88 % [72-93] in mild NM. Most of the patients who performed spirometry had a restrictive lung function pattern (11/15). This identification and characterisation of the Dutch NM patient cohort is important for international collaboration and can guide the design of future clinical trials., Competing Interests: Declaration of competing interest None., (Copyright © 2024. Published by Elsevier B.V.)

Background: We sought to analyze, in well-defined clinical setting, the first 100 patients treated at the intraoperative MRI (iMRI) hybrid surgical theatre at our facility in a population-based setting to evaluate which pathologies are best approached with iMRI assisted surgeries, as this is not yet clearly defined., Methods: Patients undergoing surgery in the 3T iMRI hybrid surgical theatre at our neurosurgical department between December 2017 to May 2021 were included after informed consent. Demographic, clinical, surgical, histological, radiological and outcome parameters, as well as variables related to iMRI, were retrospectively collected and analyzed. Patients were subdivided into adult and pediatric cohorts., Results: Various neurosurgical procedures were performed; resection of tumors and epileptic foci, endoscopic skull base procedures including pituitary lesions, deep brain stimulation (DBS) and laser interstitial thermal therapy (LITT). In total, 41 patients were pediatric. An iMRI scan was carried out in 96% of cases and led to continuation of surgery in 50% of cases, mainly due to visualized remaining pathological tissue (95.2%). Median time to iMRI from intubation was 280 min and median total duration of surgery was 445 min. The majority of patients experienced no postoperative complications (70%), 13 patients suffered permanent postoperative deficits, predominantly visual., Conclusion: Herein, we demonstrate the first 100 patients undergoing neurosurgery aided by iMRI at our facility since introduction. Indications for surgery differed between pediatric and adult patients. The iMRI was utilized for tumor surgeries, particularly adult low-grade gliomas and pediatric tumors, as well as for epilepsy surgery and DBS. In this heterogenous population, iMRI led to continuation of surgery in 50%. To establish the benefit in maximizing the extent of resection in these brain pathologies future studies are recommended., Clinical Trial Number: Not applicable., (© 2024. The Author(s).)

Background: Lipids such as phosphatidic acids (PAs) and cardiolipins (CLs) present strongly tailing peaks in reversed phase liquid chromatography, which entails low detectability. They are usually analyzed by hydrophilic interaction liquid chromatography (HILIC), which hampers high-throughput lipidomics. Thus, there is a great need for improved analytical methods in order to obtain a broader coverage of the lipidome in a single chromatographic method. We investigated the effect of ammonium bicarbonate (ABC) on peak asymmetry and detectability, in comparison with ammonium formate (AFO) on both a conventional BEH C18 column and an HST-CSH C18 column., Results: The combination of 2.5 mM ABC buffer pH 8 with an HST-CSH C18 column produced significantly improved results, reducing the asymmetry factor at 10 % peak height of PA 16:0/18:1 from 8.4 to 1.6. Furthermore, on average, there was up to a 54-fold enhancement in the peak height of its [M - H] - ion compared to AFO and the BEH C18 column. We confirmed this beneficial effect on other strongly tailing lipids, with accessible phosphate moieties e.g., cardiolipins, phosphatidylinositol phosphate, phosphatidylinositol bisphosphate, phosphorylated ceramide and phosphorylated sphingosine. Furthermore, we found an increased detectability of phospho- and sphingolipids up to 28 times in negative mode when using an HST-CSH C18 column. The method was successfully applied to mouse liver samples, where previously undetected endogenous phospholipids could be analyzed with improved chromatographic separation., Significance: In conclusion, the use of 2.5 mM ABC substantially improved the peak shape of PAs and enhanced the detectability of the lipidome in negative mode on an RPLC-ESI-Q-TOF-MS system on both BEH C18 and HST-CSH C18 columns. This method provides a wider coverage of the lipidome with one single injection for future lipidomic applications in negative mode., Competing Interests: Declaration of competing interest There is no conflict of interest from the authors of this article., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: A prerequisite before introducing a screening program is that the screening examinations are acceptable to participants., Objective: To evaluate the acceptance and bother of prostate cancer screening examinations., Design, Setting, and Participants: The randomized population-based GÖTEBORG-2 prostate cancer screening trial invited >37 000 men for prostate-specific antigen (PSA) testing followed by magnetic resonance imaging (MRI) in case of elevated PSA and prostate biopsy (targeted and/or systematic) if indicated., Outcome Measurements and Statistical Analysis: Participants were asked to fill out a questionnaire and rate the level of bother associated with each examination (PSA, MRI, and prostate biopsy) on a categorical scale ranging from 1 to 5 (1 = "not at all bothersome" and 5 = "very bothersome"), and to rate their willingness to repeat the examinations, by marking an X on a continuous scale ranging from 0 to 10 (0 = "yes, without any hesitation" and 10 = "no, absolutely not"). Wilcoxon signed rank test was used., Results and Limitations: Compliance with MRI was 96% (1790/1872), compliance with biopsy was 89% (810/907), and the response rate to the questionnaire was 75% (608/810). Men who underwent all examinations (n = 577) responded that biopsy was more bothersome than PSA test (p < 0.001) and MRI (p < 0.001). High levels of bother (≥4 out of 5) were reported by 2% (12/577) for PSA test, 8% (46/577) for MRI, and 43% (247/577) for biopsy. Men were more willing to repeat MRI than biopsy (p < 0.001), but the difference was small (median 0.2 [interquartile range 0.1-0.6] vs 0.5 [0.1-2.0])., Conclusions: Biopsies are more bothersome than MRI, but a large majority of men accept to repeat both examinations if necessary. Omitting biopsy for MRI-negative men and shifting to targeted biopsies only will reduce bother for men participating in prostate cancer screening., Patient Summary: We asked men how bothersome they found the prostate-specific antigen (PSA) test, magnetic resonance imaging (MRI), and prostate biopsies. Biopsies were more bothersome than PSA and MRI, but most men were willing to repeat all procedures if necessary., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Uropathogenic Escherichia coli (UPEC) is the most common cause of urinary tract infections (UTIs) in humans. Testosterone negatively impacts UTIs by affecting the immune response, leading to higher susceptibility of chronic cystitis in individuals with elevated testosterone levels, regardless of gender. Current research is mostly focused on how testosterone affects the host response to UPEC, but not so much is known about how testosterone directly affect UPEC virulence. The aim of the present study was to investigate the impact of testosterone exposure on the virulence of UPEC. We found that testosterone directly increases UPEC growth, endotoxin release and biofilm formation. We also found that testosterone-stimulated CFT073 increased colonization and invasion of bladder epithelial cells. Testosterone-stimulated CFT073 also increased the release of IL-1β and LDH from bladder epithelial cells. Additionally, by using a Caenorhabditis elegans survival assay we also showed that testosterone decreased the survival of CFT073 infected C. elegans worms. Taken together, our findings show that testosterone directly increases the virulence traits of UPEC., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Wu, Pettersson and Demirel.)

Purpose: The overall aim of this study was to describe experiences of discrimination due to inaccessibility among people using mobility devices., Material and Methods: We conducted a thematic qualitative analysis of 88 complaints about wheeled mobility device use, inaccessibility, and discrimination submitted to the Swedish Equality Ombudsman (DO) during 2015 and 2016., Results: The analysis resulted in three themes: instigating change by invoking laws and regulations and highlighting lack of compliance; demanding to be recognised, understood, and listened to; and struggling for equal access and social participation. Regulations and treaties were invoked as the basis for complaints by people using mobility devices regarding their lack of access to physical environments and impediments to their enjoyment of their full right to participate in and contribute to society. The complaints described feelings of discrimination, the disadvantages and exclusion due to physical inaccessibility, and experiences of being prevented from living one's life as others do., Conclusions: Complaints filed by people using mobility devices showed that they were denied access to a wide range of contexts, including offices, theatres, restaurants, schools, and public transportation, though they desired to live an active and social life outside their homes. Filing a complaint was a way to take action, highlight present inaccessibility, and express a hope for change.IMPLICATIONS FOR REHABILITATIONDifficulties experienced by people using wheeled mobility devices can reveal knowledge important for revising existing design and renovation standards for housing and public buildings.Documenting facilitators and barriers in different environments is important for giving voice to the needs of wheeled mobility device users and revealing standards that need to be strongly enforced or revised.People using wheeled mobility devices should be supported in finding solutions in inaccessible environments, both to fulfil their wishes and to enable their participation in society.

Rare or novel missense variants in large genes such as TTN and NEB are frequent in the general population, which hampers the interpretation of putative disease-causing biallelic variants in patients with sporadic neuromuscular disorders. Often, when the first initial genetic analysis is performed, the reconstructed haplotype, i.e. phasing information of the variants is missing. Segregation analysis increases the diagnostic turnaround time and is not always possible if samples from family members are lacking. To overcome this difficulty, we investigated how well the linked-read technology succeeded to phase variants in these large genes, and whether it improved the identification of structural variants. Linked-read sequencing data of nemaline myopathy, distal myopathy, and proximal myopathy patients were analyzed for phasing, single nucleotide variants, and structural variants. Variant phasing was successful in the large muscle genes studied. The longest continuous phase blocks were gained using high-quality DNA samples with long DNA fragments. Homozygosity increased the number of phase blocks, especially in exome sequencing samples lacking intronic variation. In our cohort, linked-read sequencing added more information about the structural variation but did not lead to a molecular genetic diagnosis. The linked-read technology can support the clinical diagnosis of neuromuscular and other genetic disorders., (© 2024. The Author(s).)

We describe three patients with asymmetric congenital myopathy without definite nemaline bodies and one patient with severe nemaline myopathy. In all four patients, the phenotype had been caused by pathogenic missense variants in ACTA1 leading to the same amino acid change, p.(Gly247Arg). The three patients with milder myopathy were mosaic for their variants. In contrast, in the severely affected patient, the missense variant was present in a de novo, constitutional form. The grade of mosaicism in the three mosaic patients ranged between 20 % and 40 %. We speculate that the milder clinical and histological manifestations of the same ACTA1 variant in the patients with mosaicism reflect the lower abundance of mutant actin in their muscle tissue. Similarly, the asymmetry of body growth and muscle weakness may be a consequence of the affected cells being unevenly distributed. The partial improvement in muscle strength with age in patients with mosaicism might be due to an increased proportion over time of nuclei carrying and expressing two normal alleles., Competing Interests: Declaration of Competing Interest The authors declare no competing interests. Author MV is and stockholder of Invitae, and TLW is a former employee of Invitae., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)