The genetic landscape of mitochondrial diseases in the next-generation sequencing era: a Portuguese cohort study.

Introduction: Rare disorders that are genetically and clinically heterogeneous, such as mitochondrial diseases (MDs), have a challenging diagnosis. Nuclear genes codify most proteins involved in mitochondrial biogenesis, despite all mitochondria having their own DNA. The development of next-generation sequencing (NGS) technologies has revolutionized the understanding of many genes involved in the pathogenesis of MDs. In this new genetic era, using the NGS approach, we aimed to identify the genetic etiology for a suspected MD in a cohort of 450 Portuguese patients. Methods: We examined 450 patients using a combined NGS strategy, starting with the analysis of a targeted mitochondrial panel of 213 nuclear genes, and then proceeding to analyze the whole mitochondrial DNA. Results and Discussion: In this study, we identified disease-related variants in 134 (30%) analyzed patients, 88 with nuclear DNA (nDNA) and 46 with mitochondrial DNA (mtDNA) variants, most of them being pediatric patients (66%), of which 77% were identified in nDNA and 23% in mtDNA. The molecular analysis of this cohort revealed 72 already described pathogenic and 20 novel, probably pathogenic, variants, as well as 62 variants of unknown significance. For this cohort of patients with suspected MDs, the use of a customized gene panel provided a molecular diagnosis in a timely and cost-effective manner. Patients who cannot be diagnosed after this initial approach will be further selected for whole-exome sequencing. Conclusion: As a national laboratory for the study and research of MDs, we demonstrated the power of NGS to achieve a molecular etiology, expanding the mutational spectrum and proposing accurate genetic counseling in this group of heterogeneous diseases without therapeutic options.

HR/MS-based lipidome analysis of rat brain modulated by tolcapone.

Lipids play key roles in the body, influencing cellular regulation, function, and signalling. Tolcapone, a potent catechol-O-methyltransferase (COMT) inhibitor described to enhance cognitive performance in healthy subjects, was previously shown to impact fatty acid ß-oxidation and oxidative phosphorylation. However, its impact on the brain lipidome remains unexplored. Hence, this study aimed to assess how tolcapone affects the lipidome of the rat pre-frontal cortex (PFC), a region of the brain highly relevant to tolcapone therapeutic effect, while evaluating its influence on operant behaviour. Tolcapone at 20 mg/kg was chronically administered to Wistar rats during a behavioural task and an untargeted liquid chromatography high-resolution mass spectrometry (LC-HR/MS) approach was employed to profile lipid species. The untargeted analysis identified 7227 features, of which only 33% underwent statistical analysis following data pre-processing. The results revealed an improved cognitive performance and a lipidome remodelling promoted by tolcapone. The lipidomic analysis showed 32 differentially expressed lipid species in tolcapone-treated animals (FC ≥ 1.2, p-value ≤ 0.1), and among these several triacylglycerols, cardiolipins and N-acylethanolamine (NAE 16:2) were found upregulated whereas fatty acids, hexosylceramides, and several phospholipids including phosphatidylcholines and phosphatidylethanolamines were downregulated. These preliminary findings shed light on tolcapone impact on lipid pathways within the brain. Although tolcapone improved cognitive performance and literature suggests the significance of lipids in cognition, this study did not conclusively establish that lipids directly drove or contributed to this outcome. Nevertheless, it underscores the importance of lipid modulation and encourages further exploration of tolcapone-associated mechanisms in the central nervous system (CNS).

Oral Health-Related Quality of Life in Celiac Portuguese Children: a cross-sectional study.

PURPOSE: Celiac Disease (CD) presents a wide variety of clinical signs and symptoms, including oral manifestations. This study pretended to characterize Oral Health-Related Quality of Life (OHRQoL) and reported oral manifestations in children with CD. METHODS: Target-population were children with CD. An online questionnaire, applied to children's parents, collected information about OHRQoL (using the Early Childhood Oral Health Impact Scale-ECOHIS), oral health behaviours, and history of oral manifestations. Data analysis included descriptive statistics, Mann-Whitney, Kruskall-Wallis, and Spearman correlation tests (α = 0.05). RESULTS: The sample included 146 celiac patients, with a mean age of 10.5 years (sd = 4.1). Mean ECOHIS score was 5.2 (sd = 6.8). The most frequently reported oral manifestations were recurrent aphthous stomatitis (46.6%), dental caries (45.2%) and dental opacity (39%). About one third of the participants mentioned improvements in oral health when a gluten-free diet was introduced. Most of the reported oral manifestations had a significant association with the ECOHIS score (p < 0.05). CONCLUSIONS: The OHRQoL of children was good, however oral manifestations had a negative impact on OHRQoL. The most reported oral manifestations were recurrent aphthous stomatitis, dental caries, and dental opacities. Oral health professionals must be aware about the heterogeneity of the disease, to recognize oral manifestations associated and their importance in the early diagnosis to reduce complications and to an improvement in the OHRQoL of these patients.

Predictors of psychotherapy dropout in patients with borderline personality disorder: A systematic review.

INTRODUCTION: Borderline personality disorder (BPD) is a highly debilitating psychiatric condition. Despite the expansion of new BPD specific forms of psychotherapy in the last few decades, high dropout rates have been reported in these treatments. Treatment discontinuation is associated with poor patient outcomes, inefficient resource utilization and the demoralization of healthcare providers. METHODS: In order to identify predictors of psychotherapy dropout among patients with BPD, a systematic search of Medline, the Cochrane Library, PsycInfo and PsycArticles was conducted. Studies included were randomized-controlled trials in which patients diagnosed with BPD were exposed to a therapeutic intervention consisted of an evidence-based psychotherapy. The quality of evidence in the studies was assessed through the use of revised Cochrane risk of bias tool. RESULTS: Six articles, incorporating four types of psychotherapy programmes, were included. Overall, the studies present low risk of attrition and reporting bias and unclear risk of selection, performance and detection bias. Patients with weaker therapeutic alliance scores and higher hostility presented with higher dropout rates. In contrast, better mindfulness skills and greater performance in specific neuropsychological domains, such as memory and executive control, were identified as predictive of lower risk of dropout. Sociodemographic variables and treatment history did not influence treatment retention. CONCLUSIONS: Factors that influence discontinuation should be taken into consideration in future treatment programmes, in an effort to optimize retention. Qualitative assessments of patients' reasons for dropping out may also help guide adjustments.

Whole-plant flint corn silage inclusion in total mixed rations for pre- and postweaning dairy calves.

Assuming that acetic acid plays a minor role in the development of ruminal epithelium of preweaning dairy calves, the fiber supply for growing calves has been neglected. More research has been done on including starch and nonfibrous carbohydrates in solid feed for preweaning calves. Accordingly, the fiber requirement of these calves is not well known, as diet recommendations vary greatly. Hence, elucidating the effects of including fiber from long particle sizes in the diet may be essential for helping calves overcome the transition challenge during weaning. Forty-five Holstein calves were used in a randomized block design, considering sex, birth date, and weight at 28 d of age, when the supply of the total mixed ration (TMR) with the inclusion of corn silage started. Three TMR with increasing whole-plant flint corn silage content (0, 10, or 20% on a dry matter basis) were compared: 0CS, 10CS, or 20CS, respectively. During the first 28 d of life, the calves were managed homogeneously and were fed 6 L/d of whole milk, a commercial calf starter pelleted, and water ad libitum. Next, the solid diet was changed to the respective solid feed treatment. Calves were gradually weaned from 52 to 56 d of age but were evaluated for an additional 14 d postweaning. Feed intake was measured daily, while body weight and metabolic indicators of intermediate metabolism were evaluated weekly. Ruminal fluid was collected at 6, 8, and 10 wk of age. Behavioral analysis was conducted on wk 7 (preweaning) and 10 (postweaning). There was a quadratic effect for dry matter intake from wk 7 to 10, with higher intake for the 10CS diet than the 0CS and 20CS diets. Consequently, the 10CS diet also promoted greater average daily gain at wk 8 and 9 compared with the 0CS and 20CS diets. However, the final body weight was not affected by the different solid diets. Silage inclusion in calves' diet positively affected time spent ruminating and chewing pre- and postweaning. Including 10% of whole-plant flint corn silage in the diets of young dairy calves is a strategy to increase total solid intake and decrease acidosis risk by increasing pH and ruminating activity around weaning.

Integral valorisation of tomato by-products towards bioactive compounds recovery: Human health benefits.

The tomato processing industry is one of the world's most important markets. This industry aims to optimise production, minimise energy costs and waste streams while ensuring high-quality products. This sector produces substantial amounts of by-products frequently disposed of as waste rather than reintroducing them with a new intent into the supply chain. However, these by-products are rich in bioactive compounds (BC), including carotenoids, fibre, which exhibit antioxidant, anti-inflammatory and chemopreventive properties, and cardiovascular protection. Reusing these compounds is favourable to reducing the environmental impact and enables the development of added-value products with various possible uses such as food and feed additives, nutraceuticals, cosmeceuticals, etc. This review summarises relevant issues towards the recovery and valorisation of BC from industrial tomato by-products within a circular economy context.

Exploring the response of a key Mediterranean gorgonian to heat stress across biological and spatial scales.

Understanding the factors and processes that shape intra-specific sensitivity to heat stress is fundamental to better predicting the vulnerability of benthic species to climate change. Here, we investigate the response of a habitat-forming Mediterranean octocoral, the red gorgonian Paramuricea clavata (Risso, 1826) to thermal stress at multiple biological and geographical scales. Samples from eleven P. clavata populations inhabiting four localities separated by hundreds to more than 1500 km of coast and with contrasting thermal histories were exposed to a critical temperature threshold (25 °C) in a common garden experiment in aquaria. Ten of the 11 populations lacked thermotolerance to the experimental conditions provided (25 days at 25 °C), with 100% or almost 100% colony mortality by the end of the experiment. Furthermore, we found no significant association between local average thermal regimes nor recent thermal history (i.e., local water temperatures in the 3 months prior to the experiment) and population thermotolerance. Overall, our results suggest that local adaptation and/or acclimation to warmer conditions have a limited role in the response of P. clavata to thermal stress. The study also confirms the sensitivity of this species to warm temperatures across its distributional range and questions its adaptive capacity under ocean warming conditions. However, important inter-individual variation in thermotolerance was found within populations, particularly those exposed to the most severe prior marine heatwaves. These observations suggest that P. clavata could harbor adaptive potential to future warming acting on standing genetic variation (i.e., divergent selection) and/or environmentally-induced phenotypic variation (i.e., intra- and/or intergenerational plasticity).

Living in endemic area for infectious diseases accelerates epigenetic age.

Inflammaging is a low-grade inflammatory state generated by the aging process that can contribute to frailty and age-related diseases in the elderly. However, it can have distinct effects in the elderly living in endemic areas for infectious diseases. An increased inflammatory response may confer protection against infectious agents in these areas, although this advantage can cause accelerating epigenetic aging. In this study, we evaluated the inflammatory profile and the epigenetic age of infected and noninfected individuals from an endemic area in Brazil. The profile of cytokines, chemokines and growth factors analyzed in the sera of the two groups of individuals showed similarities, although infected individuals had a higher concentration of these mediators. A significant increase in IL-1ra, CXCL8, CCL2, CCL3 and CCL4 production was associated with leprosy infection. Notably, elderly individuals displayed distinct immune responses associated with their infection status when compared to adults suggesting an adaptive remodelling of their immune responses. Epigenetic analysis also showed that there was no difference in epigenetic age between the two groups of individuals. However, individuals from the endemic area had a significant accelerated aging when compared to individuals from São Paulo, a non-endemic area in Brazil. Moreover, the latter cohort was also epigenetically aged in relation to an Italian cohort. Our data shows that living in endemic areas for chronic infectious diseases results in remodelling of inflammaging and acceleration of epigenetic aging in individuals regardless of their infectious status. It also highlights that geographical, genetic and environmental factors influence aging and immunosenescence in their pace and profile.

PrintThe Impact of Stigmatizing Experiences and Self-Stigma on Mental Health and Suicidal Behavior: Results from the Community of Portuguese Language Countries

Stigma is an attribute that discredits an individual from society, diminishing the person. People who have self-stigma endorse negative stereotypes about themselves, which leads to negative reactions and feelings of shame. Self-stigma affects overall health, particularly mental health, and quality of life, and can lead to suicidal behavior and poor self-esteem and self-efficacy. It has also been found to be a major barrier when it comes to seeking help from mental health professionals. This study aims (1) to assess stigmatizing experiences, self-stigma, mental health, and suicidal behavior, comparing differences between countries of residence; (2) to explore the associations between self-stigma, stigmatizing experiences, mental health variables, and suicidal behavior; and (3) to determine the predictive effect of self-stigma, number of stigmatizing experiences, and sociodemographic variables on mental health and suicidal behavior. Measurement instruments included a Sociodemographic Questionnaire, the Portuguese version of the Brief Symptom Inventory-18, the Suicidal Behaviours Questionnaire-Revised (SBQ-R), and the Paradox of Self‑Stigma scale (PaSS‑24). The results showed participants with higher levels of self-stigma and stigmatizing experiences presented significantly higher mental health issues and suicidal behavior. Correlational analyses showed significant correlations among mental health and suicidal variables and self-stigma variables. Regression analyses showed that sociodemographic variables, number of stigmatizing experiences, and self-stigma explained 25.3% of the variability in mental health issues while sociodemographic variables, number of stigmatizing experiences, and self-stigma explained 13.5% of the variability in suicidal behavior. This study was an important contribution to the knowledge regarding the relationship between stigma and mental health (AU)

Lactate modulates cardiac gene expression in mice during acute physical exercise.

The aim of the present study was to verify the role of lactate as a signaling molecule in cardiac tissue under physiological conditions. C57BL6/J male mice were submitted to acute running bouts on a treadmill at different exercise intensities (30, 60, and 90% of maximal speed - Smax) under the effect of two doses (0.5 and 5 mM) of α-cyano-4-hydroxycynnamate (CINN), a blocker of lactate transporters. Cardiac lactate levels, activity of the enzymes of glycolytic [hexokinase (HK) and lactate dehydrogenase (LDH)] and oxidative metabolism [citrate synthase (CS)], and expression of genes also related to metabolism [LDH, nuclear factor erythroid 2-related factor 2 (NRF-2), cytochrome oxidase IV (COX-IV), and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α)] were evaluated. Elevated cardiac lactate levels were observed after high intensity running at 90% of Smax, which were parallel to increased activity of the HK and CS enzymes and mRNA levels of PGC-1α and COX-IV. No changes were observed in cardiac lactate levels in mice running at lower exercise intensities. Interestingly, prior intraperitoneal administration (15 min) of CINN (0.5 mM) significantly reduced cardiac lactate concentration, activities of HK and CS, and mRNA levels of PGC-1α and COX-IV in mice that ran at 90% of Smax. In addition, cardiac lactate levels were significantly correlated to both PGC-1α and COX-IV cardiac gene expression. The present study provides evidence that cardiac lactate levels are associated to gene transcription during an acute bout of high intensity running exercise.