Rituximab abrogates aquaporin-4-specific germinal center activity in patients with neuromyelitis optica spectrum disorders.

Neuromyelitis optica spectrum disorders (NMOSDs) are caused by immunoglobulin G (IgG) autoantibodies directed against the water channel aquaporin-4 (AQP4). In NMOSDs, discrete clinical relapses lead to disability and are robustly prevented by the anti-CD20 therapeutic rituximab; however, its mechanism of action in autoantibody-mediated disorders remains poorly understood. We hypothesized that AQP4-IgG production in germinal centers (GCs) was a core feature of NMOSDs and could be terminated by rituximab. To investigate this directly, deep cervical lymph node (dCLN) aspirates (n = 36) and blood (n = 406) were studied in a total of 63 NMOSD patients. Clinical relapses were associated with AQP4-IgM generation or shifts in AQP4-IgG subclasses (odds ratio = 6.0; range of 3.3 to 10.8; P < 0.0001), features consistent with GC activity. From seven dCLN aspirates of patients not administered rituximab, AQP4-IgGs were detected alongside specific intranodal synthesis of AQP4-IgG. AQP4-reactive B cells were isolated from unmutated naive and mutated memory populations in both blood and dCLNs. After rituximab administration, fewer clinical relapses (annual relapse rate of 0.79 to 0; P < 0.001) were accompanied by marked reductions in both AQP4-IgG (fourfold; P = 0.004) and intranodal B cells (430-fold; P < 0.0001) from 11 dCLNs. Our findings implicate ongoing GC activity as a rituximab-sensitive driver of AQP4 antibody production. They may explain rituximab's clinical efficacy in several autoantibody-mediated diseases and highlight the potential value of direct GC measurements across autoimmune conditions.

Do Early Relapses Predict the Risk of Long-Term Relapsing Disease in an Adult and Paediatric Cohort with MOGAD?

OBJECTIVE: Myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD) can be monophasic or relapsing, with early relapse being a feature. However, the relevance of early relapse on longer-term relapse risk is unknown. Here, we investigate whether early relapses increase longer-term relapse risk in patients with MOGAD. METHODS: A retrospective analysis of 289 adult- and pediatric-onset patients with MOGAD followed for at least 2 years in 6 specialized referral centers. "Early relapses" were defined as attacks within the first 12 months from onset, with "very early relapses" defined within 30 to 90 days from onset and "delayed early relapses" defined within 90 to 365 days. "Long-term relapses" were defined as relapses beyond 12 months. Cox regression modeling and Kaplan-Meier survival analysis were used to estimate the long-term relapse risk and rate. RESULTS: Sixty-seven patients (23.2%) had early relapses with a median number of 1 event. Univariate analysis revealed an elevated risk for long-term relapses if any "early relapses" were present (hazard ratio [HR] = 2.11, p < 0.001), whether occurring during the first 3 months (HR = 2.70, p < 0.001) or the remaining 9 months (HR = 1.88, p = 0.001), with similar results yielded in the multivariate analysis. In children with onset below aged 12 years, only delayed early relapses were associated with an increased risk of long-term relapses (HR = 2.64, p = 0.026). INTERPRETATION: The presence of very early relapses and delayed early relapses within 12 months of onset in patients with MOGAD increases the risk of long-term relapsing disease, whereas a relapse within 90 days appears not to indicate a chronic inflammatory process in young pediatric-onset disease. ANN NEUROL 2023;94:508-517.

Chitosan-Calcium Aluminate as a Cell-homing Scaffold: Its Bioactivity Testing in a Microphysiological Dental Pulp Platform.

In vitro models of the dental pulp microenvironment have been proposed for the assessment of biomaterials, to minimise animal use in operative dentistry. In this study, a scaffold/3-D dental pulp cell culture interface was created in a microchip, under simulated dental pulp pressure, to evaluate the cell-homing potential of a chitosan (CH) scaffold functionalised with calcium aluminate (the 'CHAlCa scaffold'). This microphysiological platform was cultured at a pressure of 15 cm H2O for up to 14 days; cell viability, migration and odontoblastic differentiation were then assessed. The CHAlCa scaffold exhibited intense chemotactic potential, causing cells to migrate from the 3-D culture to its surface, followed by infiltration into the macroporous structure of the scaffold. By contrast, the cells in the presence of the non-functionalised chitosan scaffold showed low cell migration and no cell infiltration. CHAlCa scaffold bioactivity was confirmed in dentin sialophosphoprotein-positive migrating cells, and odontoblastic markers were upregulated in 3-D culture. Finally, in situ mineralised matrix deposition by the cells was confirmed in an Alizarin Red-based assay, in which the CHAlCa and CH scaffolds were adapted to fit within dentin discs. More intense deposition of matrix was observed with the CHAlCa scaffold, as compared to the CH scaffold. In summary, we present an in vitro platform that provides a simple and reproducible model for selecting and developing innovative biomaterials through the assessment of their cell-homing potential. By using this platform, it was shown that the combination of calcium aluminate and chitosan has potential as an inductive biomaterial that can mediate dentin tissue regeneration during cell-homing therapies.

Diagnostic value of intereye difference metrics for optic neuritis in aquaporin-4 antibody seropositive neuromyelitis optica spectrum disorders.

BACKGROUND: The novel optic neuritis (ON) diagnostic criteria include intereye differences (IED) of optical coherence tomography (OCT) parameters. IED has proven valuable for ON diagnosis in multiple sclerosis but has not been evaluated in aquaporin-4 antibody seropositive neuromyelitis optica spectrum disorders (AQP4+NMOSD). We evaluated the diagnostic accuracy of intereye absolute (IEAD) and percentage difference (IEPD) in AQP4+NMOSD after unilateral ON >6 months before OCT as compared with healthy controls (HC). METHODS: Twenty-eight AQP4+NMOSD after unilateral ON (NMOSD-ON), 62 HC and 45 AQP4+NMOSD without ON history (NMOSD-NON) were recruited by 13 centres as part of the international Collaborative Retrospective Study on retinal OCT in Neuromyelitis Optica study. Mean thickness of peripapillary retinal nerve fibre layer (pRNFL) and macular ganglion cell and inner plexiform layer (GCIPL) were quantified by Spectralis spectral domain OCT. Threshold values of the ON diagnostic criteria (pRNFL: IEAD 5 µm, IEPD 5%; GCIPL: IEAD: 4 µm, IEPD: 4%) were evaluated using receiver operating characteristics and area under the curve (AUC) metrics. RESULTS: The discriminative power was high for NMOSD-ON versus HC for IEAD (pRNFL: AUC 0.95, specificity 82%, sensitivity 86%; GCIPL: AUC 0.93, specificity 98%, sensitivity 75%) and IEPD (pRNFL: AUC 0.96, specificity 87%, sensitivity 89%; GCIPL: AUC 0.94, specificity 96%, sensitivity 82%). The discriminative power was high/moderate for NMOSD-ON versus NMOSD-NON for IEAD (pRNFL: AUC 0.92, specificity 77%, sensitivity 86%; GCIP: AUC 0.87, specificity 85%, sensitivity 75%) and for IEPD (pRNFL: AUC 0.94, specificity 82%, sensitivity 89%; GCIP: AUC 0.88, specificity 82%, sensitivity 82%). CONCLUSIONS: Results support the validation of the IED metrics as OCT parameters of the novel diagnostic ON criteria in AQP4+NMOSD.

The effect of smoking on MRI lesion resolution in NMOSD-AQP4 and MOGAD.

BACKGROUND: The effect of smoking on the resolution of magnetic resonance imaging (MRI) lesions in patients with neuromyelitis optica spectrum disorders with aquaporin-4 positive antibody (NMOSD-AQP4) and myelin oligodendrocyte glycoprotein antibody-associated disease (MOGAD) has not been studied before. OBJECTIVE: We aimed to determine the effect of smoking on lesion resolution in MRI and assess its correlation with clinical recovery after a relapse. METHODS: We conducted a cohort study including NMOSD-AQP4 and MOGAD patients with acute and follow-up MRI scans. We collected demographic, clinical, imaging and smoking data. Logistic regression models were fitted to predict the effect of smoking on lesion resolution and to assess whether clinical recovery was associated with MRI lesion resolution. RESULTS: A total of 105 patients were included (57 with NMOSD-AQP4 and 48 with MOGAD). Current and past smoking was associated with a higher risk of persistent lesions in NMOSD-AQP4 and MOGAD (risk ratio (RR) = 3.4, 95% confidence interval (CI) = 2.5-4.7, p < 0.001). Additionally, the presence of lesion resolution was associated with better clinical recovery (RR = 1.9, 95% CI = 1.7-2.2, p < 0.001). CONCLUSION: Smoking is associated with worse MRI lesion resolution in patients with NMOSD-AQP4 and MOGAD, and lesion resolution correlates with clinical recovery. Our findings suggest a detrimental effect of smoking in inflammatory central nervous system (CNS) diseases.

Role of autoantibody levels as biomarkers in the management of patients with myasthenia gravis: A systematic review and expert appraisal.

BACKGROUND AND PURPOSE: Although myasthenia gravis (MG) is recognized as an immunoglobulin G autoantibody-mediated disease, the relationship between autoantibody levels and disease activity in MG is unclear. We sought to evaluate this landscape through systematically assessing the evidence, testing the impact of predefined variables on any relationship, and augmenting with expert opinion. METHODS: In October 2020, a forum of leading clinicians and researchers in neurology from across Europe (Expert Forum for Rare Autoantibodies in Neurology in Myasthenia Gravis) participated in a series of virtual meetings that took place alongside the conduct of a systematic literature review (SLR). RESULTS: Forty-two studies were identified meeting inclusion criteria. Of these, 10 reported some correlation between a patient's autoantibody level and disease severity. Generally, decreased autoantibody levels (acetylcholine receptor, muscle-specific kinase, and titin) were positively and significantly correlated with improvements in disease severity (Quantitative Myasthenia Gravis score, Myasthenia Gravis Composite score, Myasthenia Gravis Activities of Daily Living score, Myasthenia Gravis Foundation of America classification). Given the limited evidence, testing the impact of predefined variables was not feasible. CONCLUSIONS: This first SLR to assess whether a correlation exists between autoantibody levels and disease activity in patients with MG has indicated a potential positive correlation, which could have clinical implications in guiding treatment decisions. However, in light of the limited and variable evidence, we cannot currently recommend routine clinical use of autoantibody level testing in this context. For now, patient's characteristics, clinical disease course, and laboratory data (e.g., autoantibody status, thymus histology) should inform management, alongside patient-reported outcomes. We highlight the need for future studies to reach more definitive conclusions on this relationship.

Relationship between polyvictimization and overweight among adolescents from São Paulo city, Brazil.

This study is to identify victimization patterns and analyze the association between the experience of polyvictimization and overweight or obesity among adolescent girls and boys. The sample consisted of 2680 Brazilian ninth-graders enrolled in public and private schools, taken from the São Paulo Project for the social development of children and adolescents (SP-PROSO). Victimization was explored in two ways: (i) as per Finkelhor and (ii) by latent class analysis (LCA). The interest outcomes were overweight and obesity. Multinomial logistic regression models were used to study the relationship between victimization and overweight or obesity, controlling for confounders. LCA grouped boys and girls adolescents into three classes. These classes received the same labels, but the patterns differed between sexes. Class 1 was characterized by fewer types of victimization suffered and lower endorsement values in the analyzed items and was named bullying and indirect victimization (♀: 42.7%, n = 546; ♂: 21.6%, n = 293). Class 2 included more victimization types than Class 1 and less than Class 3. This class was labeled family violence and peer victimization (♀: 29.1%, n = 356; ♂: 47.9%, n = 652). Class 3 was named high polyvictimization (♀: 28.2%, n = 345; ♂: 30.5%, n = 418). According to Finkelhor, polyvictimization was not associated with overweight or obesity in both sexes. Only the class of high polyvictimization was associated with being overweight (ORadj: 1.60, 95%CI: 1.01-2.54) in girls. In this study, polyvictimization was associated with being overweight only among adolescent girls. Longitudinal studies in different contexts and populations are needed to understand this relationship.

Omphalocele: clinical and epidemiological profile of patients born in a tertiary care center in Rio de Janeiro.

BACKGROUND: The objective of this paper is to describe the clinical and epidemiological profile and the early outcomes of patients with omphalocele born in a fetal medicine, pediatric surgery, and genetics reference hospital in Rio de Janeiro - Brazil. To determine its prevalence, describe the presence of genetic syndromes, and congenital malformations, emphasizing the characteristics of congenital heart diseases and their most common types. METHODS: Using Latin-American Collaborative Study of Congenital Malformations (ECLAMC) database and records review, a retrospective cross-sectional study was performed, including all patients born with omphalocele between January 1st, 2016, and December 31st, 2019. RESULTS: During the period of the study, our unity registered 4,260 births, 4,064 were live births and 196 stillbirths. There were 737 diagnoses of any congenital malformation, among them 38 cases of omphalocele, 27 were live born, but one was excluded for missing data. 62.2% were male, 62.2% of the women were multiparous and 51.3% of the babies were preterm. There was an associated malformation in 89.1% of the cases. Heart disease was the most common (45.9%) of which tetralogy of Fallot was the most frequent (23.5%). Mortality rate was 61.5%. CONCLUSIONS: Our data showed a good correspondence with the existing literature. Most patients with omphalocele had other malformations, especially congenital heart disease. No pregnancy was interrupted. The presence of concurrent defects showed a huge impact on prognosis, since, even if most survived birth, few remained alive and received hospital discharge. Based on these data, fetal medicine and neonatal teams must be able to adjust parents counseling about fetal and neonatal risks, especially when other congenital diseases are present.

Nova diet quality scores and risk of weight gain in the NutriNet-Brasil cohort study.

OBJECTIVE: To assess the prospective association of two diet quality scores based on the Nova food classification with BMI gain. DESIGN: The NutriNet-Brasil cohort is an ongoing web-based prospective study with continuous recruitment of participants aged ≥ 18 years since January 2020. A short 24-h dietary recall screener including 'yes/no' questions about the consumption of whole plant foods (WPF) and ultra-processed foods (UPF) was completed by participants at baseline. The Nova-WPF and the Nova-UPF scores were computed by adding up positive responses regarding the consumption of thirty-three varieties of WPF and twenty-three varieties of UPF, respectively. Participants reported their height at baseline and their weight at both baseline and after approximately 15 months of follow-up. A 15-month BMI (kg/m2) increase of ≥5 % was coded as BMI gain. SETTING: Brazil. PARTICIPANTS: 9551 participants from the NutriNet-Brasil cohort. RESULTS: Increasing quintiles of the Nova-UPF score were linearly associated with higher risk of BMI gain (relative risk Q5/Q1 = 1·34; 95 % CI 1·15, 1·56), whereas increasing quintiles of the Nova-WPF score were linearly associated with lower risk (relative risk Q5/Q1 = 0·80; 95 % CI 0·69, 0·94). We identified a moderate inverse correlation between the two scores (-0·33) and a partial mediating effect of the alternative score: 15 % for the total effect of the Nova-UPF score and 25 % for the total effect of the Nova-WPF score. CONCLUSIONS: The Nova-UPF and Nova-WPF scores are independently associated with mid-term BMI gain further justifying their use in diet quality monitoring systems.

A new approach for professional dental bleaching using a polymeric catalyst primer.

OBJECTIVE: Evaluate the influence of a polymeric catalyst primer (PCP) on esthetic efficacy (EE), degradation kinetics of hydrogen peroxide (H2 O2 ), and trans-amelodentinal cytotoxicity (TC) of bleaching gels. MATERIALS AND METHODS: The following groups were established: G1: No treatment (NC, negative control); G2: PCP; G3: 10% H2 O2 ; G4: PCP + 10% H2 O2 ; G5: 20% H2 O2 ; G6: PCP + 20% H2 O2 ; G7: 35% H2 O2 (positive control); G8: PCP + 35% H2 O2 . To determine EE, enamel/dentin discs (E/DDs) were stained and subjected or not to bleaching protocols for 45 min. To assess TC, the E/DDs were adapted to artificial pulp chambers. The extracts (culture medium + gel components diffused through E/DDs) were applied to odontoblast-like MDPC-23 cells. The viability (VB), oxidative stress (OxS), morphology (SEM), amount of H2 O2 diffused and the production of hydroxyl radical (OH• ) were assessed (two-way ANOVA/Tukey/paired Student t-test; p < 0.05). RESULTS: The highest EE was found in G8 (p < 0.05), and G4, G6, and G7 did not differ statistically (p > 0.05). In G4, the limited H2 O2 diffusion reduced OxS and increased cell VB (p < 0.05). CONCLUSIONS: Coating the enamel with PCP containing 10 mg/ml of manganese oxide before applying the 10% H2 O2 bleaching gel maintains the EE of conventional in-office bleaching and minimizes the toxic effects of this esthetic therapy. CLINICAL SIGNIFICANCE: Coating the enamel with a PCP before applying the bleaching gel may potentiate the EE of the conventional in-office tooth bleaching and reduce the toxicity of this professional therapy to the dental pulp.

Transdentinal cytotoxicity of resin luting cements using the bovine and human dentin barrier.

STATEMENT OF PROBLEM: Based upon ethical questions and because of the difficulty of obtaining intact human teeth, researchers have used bovine teeth to assess the physical and mechanical properties of different dental materials. However, data from transdentinal cytotoxicity tests showing that the bovine dentin barrier is similar to the human dentin barrier is lacking. PURPOSE: The purpose of this in vitro study was to evaluate whether the bovine dentin barrier produces similar results to those obtained when the human dentin barrier is used to assess the transdentinal cytotoxicity of resin luting cements. MATERIAL AND METHODS: The number and diameter of dentinal tubules present in the human dentin barrier and bovine dentin barrier were evaluated and assessed with a t test (α=.05). After inserting the standardized dentin barriers into artificial pulp chambers, murine dental papilla-derived cells (MDPC-23) were seeded on the pulpal surface of the specimens, and the luting cements were applied to their occlusal surfaces. Then, the following groups were established for both human and bovine dentin barriers: no treatment (negative control); Single Bond Universal; RelyX Luting 2; RelyX U200; and RelyX Ultimate. After 24 hours, the viability (alamarBlue) and morphology (scanning electron microscopy) of the cells were evaluated with a 2-way analysis of variance and the Tukey honest significance test (α=.05). RESULTS: Dentinal tubules with larger diameters were observed in bovine dentin (P<.05), but the number of tubules was similar (P>.05). A reduction in viability and notable changes in the morphology of MDPC-23 cells occurred in the Single Bond Universal and RelyX Luting 2 groups in comparison with the negative control (P<.05). The RelyX U200 and RelyX Ultimate groups were statistically similar to the negative control (P>.05). No difference was found in cytotoxicity when the same luting cement was applied on human or bovine dentin barriers (P>.05). CONCLUSIONS: For transdentinal cytotoxicity tests of resin luting cements, the bovine dentin barrier proved similar results to the human dentin barrier.

Early predictors of disability of paediatric-onset AQP4-IgG-seropositive neuromyelitis optica spectrum disorders.

OBJECTIVE: To describe onset clinical features predicting time to first relapse and time to long-term visual, motor and cognitive disabilities in paediatric-onset aquaporin-4 antibody (AQP4-IgG) neuromyelitis optica spectrum disorders (NMOSDs). METHODS: In this retrospective UK multicentre cohort study, we recorded clinical data of paediatric-onset AQP4-IgG NMOSD. Univariate and exploratory multivariable Cox proportional hazard models were used to identify long-term predictors of permanent visual disability, Expanded Disability Status Scale (EDSS) score of 4 and cognitive impairment. RESULTS: We included 49 paediatric-onset AQP4-IgG patients (38.8% white, 34.7% black, 20.4% Asians and 6.1% mixed), mean onset age of 12±4.1 years, and 87.7% were female. Multifocal onset presentation occurred in 26.5% of patients, and optic nerve (47%), area postrema/brainstem (48.9%) and encephalon (28.6%) were the most involved areas. Overall, 52.3% of children had their first relapse within 1 year from disease onset. Children with onset age <12 years were more likely to have an earlier first relapse (p=0.030), despite showing no difference in time to immunosuppression compared with those aged 12-18 years at onset. At the cohort median disease duration of 79 months, 34.3% had developed permanent visual disability, 20.7% EDSS score 4 and 25.8% cognitive impairment. Visual disability was associated with white race (p=0.032) and optic neuritis presentations (p=0.002). Cognitive impairment was predicted by cerebral syndrome presentations (p=0.048), particularly if resistant to steroids (p=0.034). CONCLUSIONS: Age at onset, race, onset symptoms and resistance to acute therapy at onset attack predict first relapse and long-term disabilities. The recognition of these predictors may help to power future paediatric clinical trials and to direct early therapeutic decisions in AQP4-IgG NMOSD.

Astrocytic outer retinal layer thinning is not a feature in AQP4-IgG seropositive neuromyelitis optica spectrum disorders.

BACKGROUND: Patients with anti-aquaporin-4 antibody seropositive (AQP4-IgG+) neuromyelitis optica spectrum disorders (NMOSDs) frequently suffer from optic neuritis (ON) leading to severe retinal neuroaxonal damage. Further, the relationship of this retinal damage to a primary astrocytopathy in NMOSD is uncertain. Primary astrocytopathy has been suggested to cause ON-independent retinal damage and contribute to changes particularly in the outer plexiform layer (OPL) and outer nuclear layer (ONL), as reported in some earlier studies. However, these were limited in their sample size and contradictory as to the localisation. This study assesses outer retinal layer changes using optical coherence tomography (OCT) in a multicentre cross-sectional cohort. METHOD: 197 patients who were AQP4-IgG+ and 32 myelin-oligodendrocyte-glycoprotein antibody seropositive (MOG-IgG+) patients were enrolled in this study along with 75 healthy controls. Participants underwent neurological examination and OCT with central postprocessing conducted at a single site. RESULTS: No significant thinning of OPL (25.02±2.03 µm) or ONL (61.63±7.04 µm) were observed in patients who were AQP4-IgG+ compared with patients who were MOG-IgG+ with comparable neuroaxonal damage (OPL: 25.10±2.00 µm; ONL: 64.71±7.87 µm) or healthy controls (OPL: 24.58±1.64 µm; ONL: 63.59±5.78 µm). Eyes of patients who were AQP4-IgG+ (19.84±5.09 µm, p=0.027) and MOG-IgG+ (19.82±4.78 µm, p=0.004) with a history of ON showed parafoveal OPL thinning compared with healthy controls (20.99±5.14 µm); this was not observed elsewhere. CONCLUSION: The results suggest that outer retinal layer loss is not a consistent component of retinal astrocytic damage in AQP4-IgG+ NMOSD. Longitudinal studies are necessary to determine if OPL and ONL are damaged in late disease due to retrograde trans-synaptic axonal degeneration and whether outer retinal dysfunction occurs despite any measurable structural correlates.

Contrasting the brain imaging features of MOG-antibody disease, with AQP4-antibody NMOSD and multiple sclerosis.

BACKGROUND: Identifying magnetic resonance imaging (MRI) markers in myelin-oligodendrocytes-glycoprotein antibody-associated disease (MOGAD), neuromyelitis optica spectrum disorder-aquaporin-4 positive (NMOSD-AQP4) and multiple sclerosis (MS) is essential for establishing objective outcome measures. OBJECTIVES: To quantify imaging patterns of central nervous system (CNS) damage in MOGAD during the remission stage, and to compare it with NMOSD-AQP4 and MS. METHODS: 20 MOGAD, 19 NMOSD-AQP4, 18 MS in remission with brain or spinal cord involvement and 18 healthy controls (HC) were recruited. Volumetrics, lesions and cortical lesions, diffusion-imaging measures, were analysed. RESULTS: Deep grey matter volumes were lower in MOGAD (p = 0.02) and MS (p = 0.0001), compared to HC and were strongly correlated with current lesion volume (MOGAD R = -0.93, p < 0.001, MS R = -0.65, p = 0.0034). Cortical/juxtacortical lesions were seen in a minority of MOGAD, in a majority of MS and in none of NMOSD-AQP4. Non-lesional tissue fractional anisotropy (FA) was only reduced in MS (p = 0.01), although focal reductions were noted in NMOSD-AQP4, reflecting mainly optic nerve and corticospinal tract pathways. CONCLUSION: MOGAD patients are left with grey matter damage, and this may be related to persistent white matter lesions. NMOSD-AQP4 patients showed a relative sparing of deep grey matter volumes, but reduced non-lesional tissue FA. Observations from our study can be used to identify new markers of damage for future multicentre studies.

Comorbidities in older patients with myasthenia gravis - Comparison between early- and late-onset disease.

BACKGROUND: Multimorbidity is an emerging challenge in older myasthenia gravis patients, which can have even greater impact on quality of life and outcome than symptoms of myasthenia. AIMS OF THE STUDY: We aimed to investigate comorbidities in older population and compare early-onset (EOMG) and late-onset (LOMG) myasthenia patients. METHODS: We investigated clinical information of patients from Oxford Myasthenia Centre age 50 or older. Data on 60 chronic disorders were extracted. RESULTS: We included 327 myasthenia patients (30.9% EOMG and 69.1% LOMG) with a median age of 70 years. Comorbidities were present in 94.5% of patients and accumulated with age. Hypertension (58.4% vs. 31.7%), hypercholesterolemia (41.2% vs. 23.8%), diabetes (24.8% vs. 11.9%), cataract (15.5% vs. 5.0%) and prostate disorders (15.0% vs. 2.0%) were more common in LOMG than EOMG, but there were no differences between 70 EOMG and 70 LOMG patients matched according to age and sex. CONCLUSIONS: Comorbidities in older patients with myasthenia are very common, increase with age, and do not differ between early- and late-onset disease.

Quantitative spinal cord MRI in MOG-antibody disease, neuromyelitis optica and multiple sclerosis.

Spinal cord involvement is a hallmark feature of multiple sclerosis, neuromyelitis optica with AQP4 antibodies and MOG-antibody disease. In this cross-sectional study we use quantitative spinal cord MRI to better understand these conditions, differentiate them and associate with relevant clinical outcomes. Eighty participants (20 in each disease group and 20 matched healthy volunteers) underwent spinal cord MRI (cervical cord: 3D T1, 3D T2, diffusion tensor imaging and magnetization transfer ratio; thoracic cord: 3D T2), together with disability, pain and fatigue scoring. All participants had documented spinal cord involvement and were at least 6 months post an acute event. MRI scans were analysed using publicly available software. Those with AQP4-antibody disease showed a significant reduction in cervical cord cross-sectional area (P = 0.038), thoracic cord cross-sectional area (P = 0.043), cervical cord grey matter (P = 0.011), magnetization transfer ratio (P ≤ 0.001), fractional anisotropy (P = 0.004) and increased mean diffusivity (P = 0.008). Those with multiple sclerosis showed significantly increased mean diffusivity (P = 0.001) and reduced fractional anisotropy (P = 0.013), grey matter volume (P = 0.002) and magnetization transfer ratio (P = 0.011). In AQP4-antibody disease the damage was localized to areas of the cord involved in the acute attack. In multiple sclerosis this relationship with lesions was absent. MOG-antibody disease did not show significant differences to healthy volunteers in any modality. However, when considering only areas involved at the time of the acute attack, a reduction in grey matter volume was found (P = 0.023). This suggests a predominant central grey matter component to MOG-antibody myelitis, which we hypothesize could be partially responsible for the significant residual sphincter dysfunction. Those with relapsing MOG-antibody disease showed a reduction in cord cross-sectional area compared to those with monophasic disease, even when relapses occurred elsewhere (P = 0.012). This suggests that relapsing MOG-antibody disease is a more severe phenotype. We then applied a principal component analysis, followed by an orthogonal partial least squares analysis. MOG-antibody disease was discriminated from both AQP4-antibody disease and multiple sclerosis with moderate predictive values. Finally, we assessed the clinical relevance of these metrics using a multiple regression model. Cervical cord cross-sectional area associated with disability scores (B = -0.07, P = 0.0440, R2 = 0.20) and cervical cord spinothalamic tract fractional anisotropy associated with pain scores (B = -19.57, P = 0.016, R2 = 0.55). No spinal cord metric captured fatigue. This work contributes to our understanding of myelitis in these conditions and highlights the clinical relevance of quantitative spinal cord MRI.

The relationship between ultra-processed food consumption and internalising symptoms among adolescents from São Paulo city, Southeast Brazil.

OBJECTIVE: To investigate the association between ultra-processed food (UPF) consumption and internalising symptoms (IS) among adolescents. DESIGN: It is a cross-sectional study. Paper-pencil survey was completed in classroom with information on UPF consumption, IS and selected covariates. IS were assessed with the Internalizing Symptoms sub-scale from the Social Behaviour Questionnaire (IS-SBQ). UPF was evaluated with a FFQ extracted from the Brazilian National School Health Survey. Crude and adjusted association between UPF and IS was investigated with structural equation models. SETTING: São Paulo, SP, Brazil. PARTICIPANTS: A total of 2680 students, Mage = 14·85; (95 % CI 14·81, 14·88). RESULTS: UPF consumption was associated with higher scores in IS in the crude (ß = 0·14; P < 0·001) and adjusted (ß = 0·12; P < 0·001) models. The higher the consumption of UPF, the higher is the IS score. The following variables were associated with a lower risk of UPF consumption: male sex, public school and having more meals with parents. The change in the magnitude of the standardised score was almost negligible, but the model was significantly improved with the inclusion of covariates. CONCLUSIONS: Our results provide evidence about the positive association between UPF consumption and IS among adolescents. The association, despite its low magnitude, remained significant after adjusting for potential confounders. These results are relevant considering the increase in UPF consumption worldwide and in low- and middle-income countries. Also, our study emphasises the importance of a healthy diet with a reduction in UPF consumption among adolescents.

Poly(caprolactone)-aligned nanofibers associated with fibronectin-loaded collagen hydrogel as a potent bioactive scaffold for cell-free regenerative endodontics.

AIM: Guided tissue regeneration has been considered a promising strategy to replace conventional endodontic therapy of teeth with incomplete root formation. Therefore, the objective of this study was to develop a tubular scaffold (TB-SC) with poly (caprolactone)-aligned nanofibres associated with a fibronectin (FN)-loaded collagen hydrogel and assess the pulp regeneration potential mediated by human apical papilla cells (hAPCs) using an in vitro model of teeth with incomplete root formation. METHODOLOGY: Aligned nanofibre strips based on 10% poly(caprolactone) (PCL) were synthesized with the electrospinning technique to produce the TB-SCs. These were submitted to different treatments, according to the following groups: TB-SC (negative control): TB-SC without treatment; TB-SC + FN (positive control): TB-SC coated with 10 µg/ml of FN; TB-SC + H: TB-SC associated with collagen hydrogel; TB-SC + HFN: TB-SC associated with FN-loaded collagen hydrogel. Then, the biomaterials were inserted into cylindrical devices to mimic the regenerative therapy of teeth with incomplete root formation. The hAPCs were seeded on the upper surface of the TB-SCs associated or not with any treatment, and cell migration/proliferation and the gene expression of markers related to pulp regeneration (ITGA5, ITGAV, COL1A1 and COL1A3) were evaluated. The data were submitted to anova/Tukey's tests (α = 5%). RESULTS: Higher values of cell migration/proliferation and gene expression of all markers tested were observed in groups TB-SC + FN, TB-SC + H, and TB-SC + HFN compared with the TB-SC group (p < .05). The hAPCs in the TB-SC + HFN group showed the highest values of cell proliferation and gene expression of COL1A1 and COL3A1 (p < .05), as well as superior cell migration results to groups TB-SC and TB-SC + H (p < .05). CONCLUSION: Aligned nanofibre scaffolds associated with the FN-loaded collagen hydrogel enhanced the migration and proliferation of hAPCs and gene expression of pulp regeneration markers. Therefore, the use of these biomaterials may be considered an interesting strategy for regenerative pulp therapy of teeth with incomplete root formation.

Cytocompatibility and bioactivity of calcium hydroxide-containing nanofiber scaffolds loaded with fibronectin for dentin tissue engineering.

OBJECTIVES: The aim of this study was to characterize polycaprolactone-based nanofiber scaffolds (PCL) incorporated with calcium hydroxide (CH) and evaluate their bioactivity on human dental pulp cells (HDPCs) when loaded with fibronectin (FN). MATERIALS AND METHODS: CH (0.1%; 0.2%; 0.4% w/v; or 0%) was incorporated into PCL (10% w/v) scaffolds prepared by electrospinning. Morphology and composition were characterized using SEM/EDS. HDPCs were seeded on the scaffolds and evaluated for viability (alamarBlue; Live/Dead), and adhesion/spreading (F-actin). Next, scaffolds containing 0.4% CH were loaded with FN (20 µg/mL). HDPCs were evaluated for viability, adhesion/spreading, migration (Trans-well), gene expression (RT-qPCR), alkaline phosphatase activity (ALP), and mineralization nodules (Alizarin Red). Data were submitted to ANOVA and post-hoc tests (α = 5%). RESULTS: Nanofibers with larger diameter were seen as CH concentration increased, while there was no effect on interfibrillar spaces. An increase in cell viability was seen for 0.4% CH, in all periods. Incorporation of CH and FN into the scaffolds increased cellular migration, spread, and viability, all intensified when CH and FN were combined. ALPL and DSPP expression, and ALP activity were not affected by CH and FN. COL1A1 was downregulated in all groups, while DMP1 was upregulated in the presence of CH, with no differences for the groups loaded with FN. CH increased the formation of mineralized matrix, which was not influenced by FN. CONCLUSIONS: In conclusion, the incorporation of CH enhanced the odontogenic potential of HDPCs, irrespective of the presence of FN. The PCL + 0.4% CH formulation may be a useful strategy for use in dentin tissue engineering. CLINICAL RELEVANCE: A change in the form of presentation of calcium hydroxide-based materials used for direct pulp capping can increase biocompatibility and prolong the vitality of dental pulp.

Strategy for reducing cytotoxicity and obtaining esthetic efficacy with 15 min of in-office dental bleaching.

OBJECTIVES: Evaluate in vitro the esthetic efficacy and cytotoxicity of a bleaching gel containing 35% hydrogen peroxide (BG-35%H2O2), applied for different time intervals, on enamel coated or not with polymeric biomaterials. MATERIALS AND METHODS: Nanofiber scaffolds (NSc) and a primer catalyst (PrCa) were used to coat the bovine enamel/dentin discs before the application of BG-35%H2O2, according to the following groups: G1-negative control (NC, without treatment); G2, G3, and G4-BG-35%H2O2 applied for 3 × 15, 2 × 15, and 15 min; G5, G6, and G7-BG-35%H2O2 applied on enamel coated with NSc and PrCa for 3 × 15; 2 × 15, and 15 min, respectively. The culture medium with components of gel diffused through the discs was applied on MDPC-23 cells, which were evaluated regarding to viability (VB), integrity of the membrane (IM), and oxidative stress (OxS). The quantity of H2O2 diffused and esthetic efficacy (ΔE/ΔWI) of the dental tissues were also analyzed (ANOVA/Tukey; p < 0.05). RESULTS: Only G7 was similar to G1 regarding VB (p > 0.05). The lowest value of H2O2 diffusion occurred in G4 and G7, where the cells exhibited the lowest OxS than G2 (p < 0.05). Despite G5 showing the greatest ΔE regarding other groups (p < 0.05), the esthetic efficacy observed in G7 was similar to G2 (p > 0.05). ΔWI indicated a greater bleaching effect for groups G5, G6, and G7 (p < 0.05). CONCLUSION: Coating the dental enamel with polymeric biomaterials reduced the time and the cytotoxicity of BG-35%H2O2. CLINICAL SIGNIFICANCE: Coating the dental enamel with polymeric biomaterials allows safer and faster BG-35%H2O2 application.