Global DNA methylation is not elevated in blood samples from Machado-Joseph disease mutation carriers.

Machado-Joseph disease (MJD) is an autosomal dominant spinocerebellar ataxia (SCA) caused by a polyglutamine expansion in the ataxin-3 protein, which initiates a cascade of pathogenic events, including transcriptional dysregulation. Genotype-phenotype correlations in MJD are incomplete, suggesting an influence of additional factors, such as epigenetic modifications, underlying the MJD pathogenesis. DNA methylation is known to impact the pathophysiology of neurodegenerative disorders through gene expression regulation and increased methylation has been reported for other SCAs. In this work we aimed to analyse global methylation in MJD carriers. Global 5-mC levels were quantified in blood samples of 33 MJD mutation carriers (patients and preclinical subjects) and 33 healthy controls, matched by age, sex, and smoking status. For a subset of 16 MJD subjects, a pilot follow-up analysis with two time points was also conducted. No differences were found in median global 5-mC levels between MJD mutation carriers and controls and no correlations between methylation levels and clinical or genetic variables were detected. Also, no alterations in global 5-mC levels were observed over time. Our findings do not support an increase in global blood methylation levels associated with MJD.

Hereditary transthyretin amyloidosis: a myriad of factors that influence phenotypic variability.

Hereditary transthyretin-related amyloidosis (ATTRv amyloidosis) is a rare and progressively debilitating disease characterized by the deposition of transthyretin (TTR) amyloid fibrils in various organs and tissues, most commonly in the heart and peripheral nerves. This pathological deposition can lead to significant organ dysfunction and, ultimately, organ failure. ATTRv amyloidosis exhibits a broad range of clinical presentations, from purely neurological symptoms to purely cardiac manifestations, as well as mixed phenotypes which result from both neurological and cardiac implications. This wide phenotypical spectrum realistically challenges disease diagnosis and prognosis, especially in individuals without or with an unknown family history. Multiple factors are thought to contribute to this variability, including genetic, epigenetic, and even environmental influences. Understanding these factors is crucial, as they can significantly affect disease expression and progression. This review aims to summarize each of these contributing factors, to help elucidate the current knowledge on the phenotypical variability of ATTRv amyloidosis.

Sit4 Genetically Interacts with Vps27 to Regulate Mitochondrial Function and Lifespan in Saccharomyces cerevisiae.

The Sit4 protein phosphatase plays a key role in orchestrating various cellular processes essential for maintaining cell viability during aging. We have previously shown that SIT4 deletion promotes vacuolar acidification, mitochondrial derepression, and oxidative stress resistance, increasing yeast chronological lifespan. In this study, we performed a proteomic analysis of isolated vacuoles and yeast genetic interaction analysis to unravel how Sit4 influences vacuolar and mitochondrial function. By employing high-resolution mass spectrometry, we show that sit4Δ vacuolar membranes were enriched in Vps27 and Hse1, two proteins that are part of the endosomal sorting complex required for transport-0. In addition, SIT4 exhibited a negative genetic interaction with VPS27, as sit4∆vps27∆ double mutants had a shortened lifespan compared to sit4∆ and vps27∆ single mutants. Our results also show that Vps27 did not increase sit4∆ lifespan by improving protein trafficking or vacuolar sorting pathways. However, Vps27 was critical for iron homeostasis and mitochondrial function in sit4∆ cells, as sit4∆vps27∆ double mutants exhibited high iron levels and impaired mitochondrial respiration. These findings show, for the first time, cross-talk between Sit4 and Vps27, providing new insights into the mechanisms governing chronological lifespan.

The socioeconomic epidemiology of inherited retinal diseases in Portugal.

BACKGROUND: Inherited retinal diseases (IRDs) are a group of rare degenerative disorders of the retina that can lead to blindness from birth to late middle age. Knowing the target population and its resources is essential to better plan support measures. The aim of this study was to evaluate the socioeconomic characteristics of regions in Portugal where IRD patients reside to inform the planning of vision aid and rehabilitation intervention measures. RESULTS: This study included 1082 patients from 973 families, aged 3 to 92 years, with a mean age of 44.8 ± 18.1 years. Patients living with an IRD were identified in 190 of the 308 municipalities. According to this study, the estimated IRD prevalence in Portugal was 10.4 per 100,000 inhabitants, and by municipalities, it ranged from 0 to 131.2 per 100,000 inhabitants. Overall, regions with a higher prevalence of IRD have a lower population density (r=-0.371, p < 0.001), a higher illiteracy rate (r = 0.404, p < 0.001) and an overall older population (r = 0.475, p < 0.001). Additionally, there is a lower proportion of doctor per capita (r = 0.350, p < 0.001), higher social security pensions beneficiaries (r = 0.439, p < 0.001), worse water quality for human consumption (r=-0.194, p = 0.008), fewer audiences at the cinema (r=-0.315, p < 0.001) and lower proportion of foreign guests in tourist accommodations (r=-0.287, p < 0.001). CONCLUSION: The number of identified patients with IRD varied between regions. Using data from national statistics (PORDATA), we observed differences in socioeconomic characteristics between regions. Multiple targeted aid strategies can be developed to ensure that all IRD patients are granted full clinical and socioeconomic support.

Vertical Dentofacial Skeletal Divergency Is Not Linked with Oral Health-Related Quality of Life.

The aim of this study is to assess how vertical skeletal malocclusion affects oral health-related quality of life (OHRQoL) among a sample of individuals comprising adolescents, young adults, and adults seeking orthodontic treatment. From January 2019 to March 2020, participants were consecutively enrolled. The assessment of OHRQoL involved measurement using the oral health impact profile (OHIP-14). Lateral cephalograms were performed to measure the vertical skeletal divergency with four cephalometric measurements. Descriptive and inferential statistical analyses were performed. The Mann-Whitney test was applied to compare OHRQoL scores according to the vertical dimension category. The mean age of the participants ranged between 30.3 ± 14.9 and 29.9 ± 14.4 and there was a majority of female participants, between 64.1% and 65.9%. There were no statistically significant differences observed between hyperdivergent and normodivergent groups in either the total score or any domain of the OHIP-14 questionnaire. Individuals with hyperdivergent facial morphology did not show a reduced OHRQoL compared with a normodivergent facial type.

The Visual Impairment of Inherited Retinal Diseases in Portugal as per the National Table of Disabilities.

Purpose: To evaluate the visual impairment of patients with inherited retinal diseases (IRDs), as per the national table of disabilities (TNI). Design: Retrospective, single-center cohort study. Participants: Patients with a clinical diagnosis of IRD were recruited at a referral center in Portugal. Methods: Demographics and clinical data were collected from each individual patient file. The estimated visual disability coefficient was calculated through the evaluation of 7 graduated categories: orbital or eyelid deformities, low vision, visual field change, loss of bi-foveolar fixation, oculomotor palsy, photophobia, and chronic conjunctivitis. The TNI provides minimum and maximum disability values for numerous conditions within each category, which were summed to calculate an overall summary disability coefficient for each patient. Main Outcome Measures: Demographic/clinical and estimated minimum and maximum visual disability coefficient according to the TNI for each patient. Results: This study included 253 patients from 214 families, aged 3 to 80 years, with a mean age of 39.8 ± 20.0 years. The mean estimated minimum and maximum visual disability coefficients as per the TNI were 0.6 ± 0.4 and 0.7 ± 0.4, respectively. The low vision was the single most frequent contributor category (21.7%) present in the calculation of visual impairment. Low vision and visual field changes were the most frequent double combination (18.2%), and the addition of loss of bi-foveolar fixation was the most frequent triple combination (8.3%). Conclusions: This study found that IRD patients had a significant visual disability, with the majority having a disability coefficient ≥0.6, which would qualify them for a "multipurpose disability medical certificate." Financial Disclosures: The authors have no proprietary or commercial interest in any materials discussed in this article.

Integrating functional scoring and regulatory data to predict the effect of non-coding SNPs in a complex neurological disease.

Most SNPs associated with complex diseases seem to lie in non-coding regions of the genome; however, their contribution to gene expression and disease phenotype remains poorly understood. Here, we established a workflow to provide assistance in prioritising the functional relevance of non-coding SNPs of candidate genes as susceptibility loci in polygenic neurological disorders. To illustrate the applicability of our workflow, we considered the multifactorial disorder migraine as a model to follow our step-by-step approach. We annotated the overlap of selected SNPs with regulatory elements and assessed their potential impact on gene expression based on publicly available prediction algorithms and functional genomics information. Some migraine risk loci have been hypothesised to reside in non-coding regions and to be implicated in the neurotransmission pathway. In this study, we used a set of 22 non-coding SNPs from neurotransmission and synaptic machinery-related genes previously suggested to be involved in migraine susceptibility based on our candidate gene association studies. After prioritising these SNPs, we focused on non-reported ones that demonstrated high regulatory potential: (1) VAMP2_rs1150 (3' UTR) was predicted as a target of hsa-mir-5010-3p miRNA, possibly disrupting its own gene expression; (2) STX1A_rs6951030 (proximal enhancer) may affect the binding affinity of zinc-finger transcription factors (namely ZNF423) and disturb TBL2 gene expression; and (3) SNAP25_rs2327264 (distal enhancer) expected to be in a binding site of ONECUT2 transcription factor. This study demonstrated the applicability of our practical workflow to facilitate the prioritisation of potentially relevant non-coding SNPs and predict their functional impact in multifactorial neurological diseases.

In vitro micropropagation and tiliroside production in Paratecoma peroba (Record) Kuhlm, an endemic and endangered Brazilian tree.

In vitro tissue culture can be an alternative method for endangered species propagation, biodiversity conservation and secondary metabolite studies. Paratecoma peroba (Record) Kuhlm. (Bignoniaceae) is an endemic and endangered Brazilian species. This work aimed to establish in vitro morphogenesis and callus induction and to perform a phytochemical analysis of P. peroba callus extract. Higher seed germination (43%) was obtained in Wood Plant Medium culture without activated charcoal (AC). Combination of 5 µM benzyladenine + 10 µM gibberellic acid, without AC, resulted in a higher number of shoots (2 shoots/explant). A callus culture was stabilised from zygotic embryos using 2,4-dichlorophenoxyacetic acid. A callus methanolic extract was used for phytochemical analysis. The isolated substance was identified as tiliroside (kaempferol 3-O-ß-D-(6''-O-E-p-coumaroyl)-glucopyranoside) by NMR and quantified in callus and leaf extracts by HPLC. This study adds to the chemical knowledge of this species and it is the first report of a flavonol in Paratecoma.

The Role of Extracorporeal Shock Wave Therapy in the Treatment of Muscle Injuries: A Systematic Review.

Muscle injuries commonly occur in sports and can be classified as indirect and direct, according to the 2013 Munich Consensus Statement (MCS). Since recent evidence suggests that extracorporeal shock wave therapy (ESWT) improves muscular microcirculation and may increase regeneration after acute muscle injury, we performed a systematic review following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement guidelines to access the efficacy and safety of ESWT in the treatment of patients with muscle injuries. PubMed and Cochrane were searched to screen for potentially relevant articles and the literature search was last updated in June 2023. The inclusion criteria were randomized controlled trials, observational studies, or case controls published in English, Portuguese, or Spanish that studied the effect of ESWT on indirect and direct muscle injuries in individuals aged ≥18, with at least one of the following reported outcomes: pain on the visual analog scale (VAS), functionality assessed either with disability scales or subjectively, time for return to play (RTP), re-injury rate, and ultrasonographic evaluation. The exclusion criteria were literature reviews, systematic reviews, studies in animals, studies in other languages, studies that failed to meet the targeted population or intervention and studies that didn't report any of the outcomes of interest. The quality of the studies was analyzed using the Cochrane Assessment Tool, the Newcastle-Ottawa Quality Assessment Scale, and the JBI Critical Appraisal Checklist. Eight studies were included in the systematic review (two randomized controlled trials, one prospective observational study, two retrospective observational studies, and three case reports), with a total of 143 adult participants. ESWT was associated with less pain on VAS, better function, reduction of size of lesion on ultrasound evaluation, faster RTP and/or lower re-injury rate in patients with indirect and direct muscle injuries and muscular hematomas, a frequent secondary complication of muscle injuries. The evidence regarding the use of ESWT for these types of injuries is therefore promising. Nevertheless, higher-quality studies are needed in the future to prove its efficacy, better comprehend its mechanisms of action and define treatment protocols (timing, type and parameters of ESWT).

The Role of Systemic Steroids in Sciatica Due to Herniated Lumbar Disc: A Systematic Review and Meta-analysis.

STUDY DESIGN: Systematic review and meta-analysis. OBJECTIVE: Evaluate the role of systemic steroids in treating patients with sciatica due to lumbar disk herniation (LDH). SUMMARY OF BACKGROUND DATA: The association between LDH and sciatica has been well described. The use of steroids seems logical in this context; however, their efficacy is not well described, and their use remains controversial. METHODS: A comprehensive search on PubMed, EMBASE, and Scopus databases (up to February 15, 2022) was performed to identify randomized clinical trials that included patients with symptoms of sciatica due to LDH that were treated with systemic steroids. The risk of bias was judged using the Cochrane risk-of-Bias2 tool. Meta-analysis was conducted using a random-effects model to estimate the between-group effect size for pain and functional outcomes. The risk of developing adverse events (AE) was computed using relative risks. All pooled results are reported with their 95% confidence interval (CI) and certainty of evidence analyzed using the Grading of Recommendations Assessment, Development, and Evaluation framework. RESULTS: Ten studies met inclusion criteria, comprising a total of 1017 participants: 540 in the treatment group and 477 in the control group. Steroid treatment was associated with a significant superior reduction of pain (SMD = -0.42, 95% CI -0.76 to -0.08, weak effect, very-low certainty) and reduction in disability (SMD = -0.30, 95% CI -0.51 to -0.10, weak effect, very-low certainty). Corticosteroid administration was associated with a significant increased risk of developing an AE (relative risks = 2.00, 95% CI 1.40 to 2.85, low certainty). CONCLUSION: The use of systemic steroids in the treatment of sciatica due to LDH seems reasonable despite a 2-fold higher risk of developing mild AEs. However, the effect size is small for reducing pain in the short term and improving functional outcomes at long-term follow-up.

Non-coding variants in VAMP2 and SNAP25 affect gene expression: potential implications in migraine susceptibility.

Migraine is a common and complex neurological disease potentially caused by a polygenic interaction of multiple gene variants. Many genes associated with migraine are involved in pathways controlling the synaptic function and neurotransmitters release. However, the molecular mechanisms underpinning migraine need to be further explored.Recent studies raised the possibility that migraine may arise from the effect of regulatory non-coding variants. In this study, we explored the effect of candidate non-coding variants potentially associated with migraine and predicted to lie within regulatory elements: VAMP2_rs1150, SNAP25_rs2327264, and STX1A_rs6951030. The involvement of these genes, which are constituents of the SNARE complex involved in membrane fusion and neurotransmitter release, underscores their significance in migraine pathogenesis. Our reporter gene assays confirmed the impact of at least two of these non-coding variants. VAMP2 and SNAP25 risk alleles were associated with a decrease and increase in gene expression, respectively, while STX1A risk allele showed a tendency to reduce luciferase activity in neuronal-like cells. Therefore, the VAMP2_rs1150 and SNAP25_rs2327264 non-coding variants affect gene expression, which may have implications in migraine susceptibility. Based on previous in silico analysis, it is plausible that these variants influence the binding of regulators, such as transcription factors and micro-RNAs. Still, further studies exploring these mechanisms would be important to shed light on the association between SNAREs dysregulation and migraine susceptibility.

Intraoperative fluoroscopic radiation in orthopedic trauma: correlation with surgery type and surgeon experience.

Background: While fluoroscopy is widely used in orthopedic trauma surgeries, it is associated with harmful effects and should, therefore, be minimized. However, reference values for these surgeries have not been defined, and it is not known how surgeon experience affects these factors. The aims of this study were to analyze the radiation emitted and exposure time for common orthopedic trauma surgeries and to assess whether they are affected by surgeon experience. Methods: Data from 1842 trauma orthopedic procedures were retrospectively analyzed. A total of 1421 procedures were included in the analysis. Radiation dose and time were collected to identify reference values for each surgery and compared for when the lead surgeon was a young resident, a senior resident, or a specialist. Results: The most performed surgeries requiring fluoroscopy were proximal femur short intramedullary nailing (n = 401), ankle open reduction and internal fixation (ORIF) (n = 141), distal radius ORIF (n = 125), and proximal femur dynamic hip screw (DHS) (n = 114). Surgeries using higher radiation dose were proximal femur long intramedullary nailing (mean dose area [DAP]): 1361.35 mGycm2), proximal femur DHS (1094.81 mGycm2), and proximal femur short intramedullary nailing (891.41 mGycm2). Surgeries requiring longer radiation time were proximal humerus and/or humeral shaft intramedullary nailing (02 mm:20 ss), proximal femur long intramedullary nailing (02 mm:04 ss), and tibial shaft/distal tibia intramedullary nailing (01 mm:49 ss). Senior residents required shorter radiation time when performing short intramedullary nailing of the proximal femur than young residents. Specialists required more radiation dose than residents when performing tibial nailing and tibial plateau ORIF and required longer radiation time than young residents when performing tibial nailing. Conclusions: This study presents mean values of radiation dose and time for common orthopedic trauma surgeries. Orthopedic surgeon experience influences radiation dose and time values. Contrary to expected, less experience is associated with lower values in some of the cases analyzed.

Risk factors for refracture after proximal femur fragility fracture.

Introduction: Proximal femur fragility fractures (PFFFs) are a growing worldwide concern. Recognizing the risk factors for subsequent fracture is essential for secondary prevention. This study aimed to analyze the risk factors for refracture and mortality rates in patients who suffered a PFFF. Methods: Patients aged 65 years or older with PFFF who underwent surgical treatment during the year of 2017 in the same institution were retrospectively analyzed and at least four years after the index fracture were evaluated. Results: From a total of 389 patients, 299 patients were included, with a median age of 83 years, and 81% female. Thirty-two (10.7%) suffered a refracture, with a mean time to refracture of 19.8 ± 14.80 months, being the female sex a risk factor for refracture (OR-4.69; CI [1.05-20.95]). The 1-year mortality rate was 15.4%. Seventy-three (24.4%) patients had previous fragility fractures. After the index fracture, 79% remained untreated for osteoporosis. No statistical association was found between antiosteoporotic treatment and refracture. Patients with refracture had higher prefracture functional level compared with patients without refracture (OR-1.33; CI [1.08-1.63]) and were discharged more often to rehabilitation units (31% versus 16%, P =.028). After 4 years of follow-up, patients with refracture had lower functional level compared with patients without. Chronic kidney disease was a risk factor (P = .029) for early refracture (<24 months). Conclusion: Female sex and higher prefracture functional level may increase the risk of refracture. Chronic kidney disease was associated with a shorter refracture time. Despite having a PFFF or other fragility fractures, the majority of patients remained untreated for osteoporosis.

Functional characterization of a novel PRRT2 variant found in a Portuguese patient with hemiplegic migraine.

Familial hemiplegic migraine (FHM) is a rare autosomal-dominant form of migraine with aura. Three disease-causing genes have been identified for FHM: CACNA1A, ATP1A2 and SCN1A. However, not all families are linked to one of these three genes.PRRT2 variants were also commonly associated with HM symptoms; therefore, PRRT2 is hypothesized as the fourth gene causing FHM. PRRT2 plays an important role in neuronal migration, spinogenesis, and synapse mechanisms during development and calcium-dependent neurotransmitter release. We performed exome sequencing to unravel the genetic cause of migraine in one family, and a novel PRRT2 variant (c.938C > T;p.Ala313Val) was identified with further functional studies to confirm its pathogenicity. PRRT2-A313V reduced protein stability, led to protein premature degradation by the proteasome and altered the subcellular localization of PRRT2 from the plasma membrane (PM) to the cytoplasm. We identified and characterized for the first time in a Portuguese patient, a novel heterozygous missense variant in PRRT2 associated with HM symptoms. We suggest that PRRT2 should be included in the diagnosis of HM.

Gender-Affirming Hormone Therapy: Physical and Sociopsychological Effects, Impact and Satisfaction.

Background Gender dysphoria treatment includes gender-affirming hormone therapy (GAHT). Studies are still lacking on how to characterize its effects and impact on transgender people's lives more effectively. Aim To study the physical and psychological effects of GAHT on transgender individuals, assess its impact on their lives, and rate their overall satisfaction. Methods Participants (n = 114; ages 18-62 years; median age 24.0 (21.0 - 33.0) years) included transgender adults residing in Portugal who were undergoing or had undergone hormonal therapy for at least one uninterrupted year. Participants completed an original questionnaire. For most items, an ordinal Likert-style scale ranging from 0 (worst result) to 6 (best result) was used. Descriptive statistics and non-parametric tests, including Pearson's chi-squared test, Wilcoxon signed-rank test, and Mann-Whitney U test were used to analyze categorical and continuous variables, with a significance level set at 0.05. Outcomes The outcomes included desired physical changes rating (perception and satisfaction with changes); side effects of GAHT; the sociopsychological impact of GAHT (on self-esteem, body image, psychological wellbeing, social and family relations); overall satisfaction (with treatment results and medical follow-up). Results The changes classified as the most perceptible in those undergoing masculinizing treatment (Group M) were amenorrhea (6 (5.0-6.0) points) and clitoris enlargement (6 (5.0-6.0) points). These were also the ones rated as the most satisfactory (6 (6.0-6.0) points for amenorrhea and 6 (4.0-6.0) points for clitoris enlargement). On those undergoing feminizing therapy (Group F), the alteration voted as the most perceptible was sperm production decrease (6 (2.0-6.0) points), and the ones classified as the most satisfactory were sperm production decrease (6 (4.0-6.0) points) and spontaneous erections decrease (6 (5.0-6.0) points). Side effects were reported by 89.7% of Group M (mood swings were the most common) and 96.3% of Group F (decreased libido was the most frequent). The sociopsychological impact of hormonal treatment was significantly positive in all analyzed variables (p<0.001). Overall satisfaction with treatment results and medical follow-up were rated with 5 points and 4.5 points, respectively. Clinical implications This study provides clinicians with more evidence that GAHT may improve the physical, psychological and social health of transgender people seeking medical transition. Strengths and limitations The strengths of the current study include a high participant count relative to the target population, the acquisition of data on previously unexplored variables, and the significance of being one of the few investigations of its kind conducted in Portugal. However, the study has limitations, including differences in participant characteristics, a small sample size for some variables, potential bias due to the retrospective nature of the study, individualized treatment regimens, and the inclusion of participants from different countries, which limit the generalization of the results. Conclusions This study provides further evidence that GAHT is effective, and that its physical effects are satisfactory while resulting in mostly non-severe nor life-threatening side effects. GAHT is an important therapy in gender dysphoria and has consistent results in improving numerous sociopsychological variables.

Different relationships between epilepsy syndromes and autoimmune diseases.

OBJECTIVE: Our objective was to study the relationship between epilepsy and autoimmune diseases in two different types of epilepsy: idiopathic generalized epilepsies (IGEs) and mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE-HS). The contribution of the human leukocyte antigen (HLA) system to this relationship was analyzed. METHODS: Adult patients with IGEs and MTLE-HS at a tertiary epilepsy center were consecutively enrolled between January 2016 and December 2020. RESULTS: A total of 664 patients, 422 with IGEs and 242 with MTLE-HS, were included. Patients with IGEs were 15 years younger, on average, than patients with MTLE-HS (p < .001). The frequency of autoimmune diseases was 5.5% (n = 23) and 4.5% (n = 11) in patients with IGEs and MTLE-HS, respectively (p = .716). The mean age of autoimmune disease onset was 20 ± 15.6 years in patients with IGEs and 36.7 ± 16.5 years in patients with MTLE-HS (p < .05). Clinical manifestations of autoimmune diseases preceded epilepsy onset in 30.4% of patients with IGEs (i.e., in early childhood); in the other patients, epilepsy appeared before autoimmune disease onset. In all but one patient with MTLE-HS and autoimmune diseases, the autoimmune diseases appeared after epilepsy onset from adolescence onward. SIGNIFICANCE: Our study indicates two relationship patterns: a bidirectional association between IGEs and autoimmune diseases and a unidirectional relationship between MTLE-HS and autoimmune diseases. The involvement of genetic susceptibility factors (such as the HLA system), autoinflammatory mechanisms, female sex, and antiseizure medications in these relationships are discussed.

Iron Limitation Restores Autophagy and Increases Lifespan in the Yeast Model of Niemann-Pick Type C1.

Niemann-Pick type C1 (NPC1) is an endolysosomal transmembrane protein involved in the export of cholesterol and sphingolipids to other cellular compartments such as the endoplasmic reticulum and plasma membrane. NPC1 loss of function is the major cause of NPC disease, a rare lysosomal storage disorder characterized by an abnormal accumulation of lipids in the late endosomal/lysosomal network, mitochondrial dysfunction, and impaired autophagy. NPC phenotypes are conserved in yeast lacking Ncr1, an orthologue of human NPC1, leading to premature aging. Herein, we performed a phosphoproteomic analysis to investigate the effect of Ncr1 loss on cellular functions mediated by the yeast lysosome-like vacuoles. Our results revealed changes in vacuolar membrane proteins that are associated mostly with vesicle biology (fusion, transport, organization), autophagy, and ion homeostasis, including iron, manganese, and calcium. Consistently, the cytoplasm to vacuole targeting (Cvt) pathway was increased in ncr1∆ cells and autophagy was compromised despite TORC1 inhibition. Moreover, ncr1∆ cells exhibited iron overload mediated by the low-iron sensing transcription factor Aft1. Iron deprivation restored the autophagic flux of ncr1∆ cells and increased its chronological lifespan and oxidative stress resistance. These results implicate iron overload on autophagy impairment, oxidative stress sensitivity, and cell death in the yeast model of NPC1.

Posterior Atlantoaxial Screw Placement in a Portuguese Population: A Morphometric Analysis Based on Computed Tomography Scan Measurements.

Objective The present study aims to evaluate the screw length and trajectory angles for posterior atlantoaxial fixation in a Portuguese population, through the study of cervical computed tomography (CT) scans. Methods Cervical CT scans of 50 adults were measured according to predefined screw trajectories of C1-C2 transarticular (C1C2TA), C1 lateral mass (C1LM), C2 pedicle (C2P), C2 pars and C2 laminar (C2L) screws. For each of these trajectories, screw length and angles were measured and compared between males and females. Results For the C1C2TA screw trajectory, the mean length, medial, and cranial angles were 34.12 ± 3.19 mm, 6.24° ± 3.06, and 59.25° ± 5.68, respectively, and for the C1LM screw trajectory, they were 27.12 ± 2.15 mm, 15.82° ± 5.07, and 13.53° ± 4.80, respectively. The mean length, medial, and cranial angles for the C2P screw trajectory were 23.44 ± 2.49 mm, 27.40° ± 4.88, and 30.41° ± 7.27, respectively; and for the C2 pars screw trajectory, they were 16.84 ± 2.08 mm, 20.09° ± 6.83, and 47.53° ± 6,97. The mean length, lateral, and cranial angles for the C2L screw trajectory were 29.10 ± 2.48 mm, 49.80° ± 4.71, and 21.56° ± 7.76, respectively. There were no gender differences except for the lengths of the C1C2TA ( p = 0,020) and C2L ( p = 0,001) screws, which were greater in males than in females. Conclusion The present study provides anatomical references for the posterior atlantoaxial fixation in a Portuguese population. These detailed data are essential to aid spine surgeons to achieve safe and effective screw placement.

An European overview of genetic counselling supervision provision.

Genetic testing is becoming more commonplace in general and specialist health care, and should always be accompanied by genetic counselling, according to legislation in many European countries and recommendations by professional bodies. Personal and professional competence is necessary to provide safe and effective genetic counselling. Clinical and counselling supervision of genetics healthcare practitioners plays a key role in quality assurance, providing a safe environment not only for patients but for professionals too. However, in many European countries, genetic counsellors are still an emerging professional group and counselling supervision is not routinely offered and there are no enough evidences on the impact of these insufficiencies. This study aimed to explore the current status of genetic counselling supervision provision across Europe and to ascertain factors that might be relevant for the successful implementation of counselling supervision. A total of 100 practitioners responded to an online survey; respondents were from 18 countries, with the majority working in France (27%) and Spain (17%). Only 34 participants reported having access to genetic counselling supervision. Country of origin, the existence of a regulation system and years of experience were factors identified as relevant, influencing access and characteristics of counselling supervision. Although there is a growing number of genetic counsellors trained at European level, just a few countries have implemented and required as mandatory the access to genetic counselling supervision. Nevertheless, this is essential to ensure a safe and effective genetic counselling and should be regulated at the European genetic healthcare services.

Posterior Atlantoaxial Screw Placement in a Portuguese Population: A Morphometric Analysis Based on Computed Tomography Scan Measurements / Colocação de parafusos atlantoaxiais posteriores em uma população portuguesa: Uma análise morfométrica baseada em medidas de tomografia computadorizada

Abstract Objective The present study aims to evaluate the screw length and trajectory angles for posterior atlantoaxial fixation in a Portuguese population, through the study of cervical computed tomography (CT) scans. Methods Cervical CT scans of 50 adults were measured according to predefined screw trajectories of C1-C2 transarticular (C1C2TA), C1 lateral mass (C1LM), C2 pedicle (C2P), C2 pars and C2 laminar (C2L) screws. For each of these trajectories, screw length and angles were measured and compared between males and females. Results For the C1C2TA screw trajectory, the mean length, medial, and cranial angles were 34.12 ± 3.19 mm, 6.24° ± 3.06, and 59.25° ± 5.68, respectively, and for the C1LM screw trajectory, they were 27.12 ± 2.15 mm, 15.82° ± 5.07, and 13.53° ± 4.80, respectively. The mean length, medial, and cranial angles for the C2P screw trajectory were 23.44 ± 2.49 mm, 27.40° ± 4.88, and 30.41° ± 7.27, respectively; and for the C2 pars screw trajectory, they were 16.84 ± 2.08 mm, 20.09° ± 6.83, and 47.53° ± 6,97. The mean length, lateral, and cranial angles for the C2L screw trajectory were 29.10 ± 2.48 mm, 49.80° ± 4.71, and 21.56° ± 7.76, respectively. There were no gender differences except for the lengths of the C1C2TA (p= 0,020) and C2L (p= 0,001) screws, which were greater in males than in females. Conclusion The present study provides anatomical references for the posterior atlantoaxial fixation in a Portuguese population. These detailed data are essential to aid spine surgeons to achieve safe and effective screw placement.

Resumo Objetivo O presente estudo tem como objetivo avaliar o comprimento e os ângulos de trajetória do parafuso para fixação atlantoaxial posterior em uma população portuguesa por meio do estudo de tomografia computadorizada (TC) cervical. Métodos Tomografias computadorizadas cervicais de 50 adultos foram analisadas quanto às trajetórias pré-definidas dos parafusos transarticulares C1-C2 (C1C2TA), na massa lateral de C1 (C1LM), no pedículo de C2 (C2P) e na pars de C2 e C2 laminar (C2L). O comprimento e os ângulos dos parafusos em cada uma destas trajetórias foram medidos e comparados entre homens e mulheres. Resultados O comprimento médio e ângulos medial e cranial da trajetória do parafuso C1C2TA foram de 34,12 ± 3,19 mm, 6,24° ± 3,06 e 59,25° ± 5,68, respectivamente; as medidas da trajetória do parafuso C1LM foram 27,12 ± 2,15 mm, 15,82° ± 5,07 e 13,53° ± 4,80. O comprimento médio e os ângulos medial e cranial da trajetória do parafuso C2P foram de 23,44 ± 2,49 mm, 27,40° ± 4,88 e 30,41° ± 7,27, respectivamente; as medidas da trajetória do parafuso da pars de C2 foram 16,84 ± 2,08 mm, 20,09° ± 6,83 e 47,53° ± 6,97. O comprimento médio e ângulos lateral e cranial da trajetória do parafuso C2L foram de 29,10 ± 2,48 mm, 49,80° ± 4,71 e 21,56° ± 7,76, respectivamente. Não houve diferenças entre os gêneros, à exceção do comprimento dos parafusos C1C2TA (p= 0,020) e C2L (p= 0,001), que foi maior no sexo masculino do que no feminino. Conclusão O presente estudo fornece referências anatômicas para a fixação atlantoaxial posterior em uma população portuguesa. Estes dados detalhados são essenciais para ajudar os cirurgiões de coluna a colocar os parafusos de maneira segura e eficaz.