Recurrent copy number variations as risk factors for neurodevelopmental disorders: critical overview and analysis of clinical implications

Neurodevelopmental disorders (NDs) encompass a spectrum of neuropsychiatric manifestations. Chromosomal regions 1q21.1, 3q29, 15q11.2, 15q13.3, 16p11.2, 16p13.1 and 22q11 harbour rare but recurrent CNVs that have been uncovered as being important risk factors for several of these disorders. These rearrangements may underlie a broad phenotypical spectrum, ranging from normal development, to learning problems, intellectual disability (ID), epilepsy and psychiatric diseases, such as autism spectrum disorders (ASDs) and schizophrenia (SZ). The highly increased risk of developing neurodevelopmental phenotypes associated with some of these CNVs makes them an unavoidable element in the clinical context in paediatrics, neurology and psychiatry. However, and although finding these risk loci has been the goal of neuropsychiatric genetics for many years, the translation of this recent knowledge into clinical practice has not been trivial. In this article, we will: (1) review the state of the art on recurrent CNVs associated with NDs, namely ASD, ID, epilepsy and SZ; (2) discuss the models used to dissect the underlying neurobiology of disease, (3) discuss how this knowledge can be used in clinical practice.FEDER through the Programa Operacional Factores de Competitividade — COMPETE and by Portuguese national funds through Fundação para a Ciência e Tecnologi

Learning the biochemical basis of axonal guidance: using Caenorhabditis elegans as a model

Aim: Experimental models are a powerful aid in visualizing molecular phenomena. This work reports how the worm Caenorhabditis elegans (C. elegans) can be effectively explored for students to learn how molecular cues dramatically condition axonal guidance and define nervous system structure and behavior at the organism level. Summary of work: A loosely oriented observational activity preceded detailed discussions on molecules implied in axonal migration. C. elegans mutants were used to introduce second-year medical students to the deleterious effects of gene malfunctioning in neuron response to extracellular biochemical cues and to establish links between molecular function, nervous system structure, and animal behavior. Students observed C. elegans cultures and associated animal behavior alterations with the lack of function of specific axon guidance molecules (the soluble cue netrin/UNC-6 or two receptors, DCC/UNC-40 and UNC-5H). Microscopical observations of these strains, in combination with pan-neuronal GFP expression, allowed optimal visualization of severely affected neurons. Once the list of mutated genes in each strain was displayed, students could also relate abnormal patterns in axon migration/ventral and dorsal nerve cord neuron formation in C. elegans with mutated molecular components homologous to those in humans. Summary of results: Students rated the importance and effectiveness of the activity very highly. Ninety-three percent found it helpful to grasp human axonal migration, and all students were surprised with the power of the model in helping to visualize the phenomenon.This work has been funded by National funds, through the Foundation for Science and Technology (FCT)—project UIDB/50026/2020 and UIDP/50026/2020 and by the projects, NORTE01-0145-FEDER-000039 and NORTE-01-0145-FEDER-085468, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). This work has been also funded by ICVS Scientific Microscopy Platform, member of the national infrastructure PPBI—Portuguese Platform of Bioimaging (PPBI-POCI-01-0145-FEDER-022122). Additionally, C.V. and D.V.C. were supported by the FCT individual fellowships 2022.11176.BD and SFRH/BD/147826/2019, respectively

Análise retrospectiva do resultado do tratamento com iodo radioativo em 120 pacientes tirotóxicos por doença de Basedow-graves

We studied retrospectively the outcome of radioactive iodine therapy (RAI) in 120 thyrotoxic patients with Graves' disease (96 F/ 24 M) who were primarily treated with anti-thyroid drugs (ATD). The decision to use RAI was due either to relapse after ATD-induced remission, lack of compliance on ATD, goiter larger than 60g, old age or association with severe diseases. ATD were discontinued one week before RAI administration, calculated by the formula: glandular volume (g) vs. effective radiation (80 mCi/g of tissue) divided by 24h thyroid uptake (%). We reexamined the patients for 48 months after RAI. Medians of age, goiter size and 24h thyroid uptake were 34y (17-69), 54.5g (20-210) and 73% (21-99), respectively. After RAI, patients became gradually hypothyroid on a rate of 15%/ 6mo until the first 18mo, followed by a rate of 2-5%/ 6mo until the end of 48mo. The number of patients in thyrotoxicosis decreased continuously, being 52.2% 6mo after RAI, 25% after 18mo and 6.7% after 48mo; at the end of the study 8 patients were still thyrotoxic. The total of patients without thyrotoxicosis at the end of observation, that is, the sum of those in euthyroidism and hypothyroidism on T4 therapy (cure of thyrotoxicosis) reached 46.8% in 6mo, 63.3% in 12mo, 81.7% in 24mo, 87.5% in 36mo and 93.3% in 48mo. Transient hypothyroidism, that is, elevation of TSH higher than 4.5 mU/L until 12mo after RAI, followed by normalization of TSH was found in 7.5% of patients. The following correlations were found in this study: a) initial T3 values and absence of cure of thyroxicosis (p=0.2); b) total RAI doses in relation to time for cure of thyrotoxicosis (p=0.01); c) dose of 131l/thyroid volume in relation to time for cure of thyrotoxicosis (p=0.02). Adverse effects included exarcebation of thyrotoxicosis in 3 patients and pain in the anterior cervical area in one, all reversible and of short duration. Two patients became pregnant after RAI but both gestations and their outcomes were normal.Estudamos retrospectivamente 120 pacientes (96 F/ 24 M) portadores de tirotoxicose por doença de Basedow-Graves submetidos à terapia com iodo radioativo (131I), que receberam inicialmente, para compensação da moléstia, drogas anti-tiroidianas (DAT). A indicação terapêutica posterior de radioiodo deveu-se a uma das seguintes condições clínicas: recidiva após compensação com DAT, falta de compensação devida à baixa aderência às DAT, bócios maiores que 60g, idade avançada ou doença grave associada. As DAT foram suspensas uma semana antes da administração da dose de 131I, calculada pela fórmula: volume glandular (em g) X radiação efetiva (80 mCi/g de tecido) ¸ captação tiroideana de 24hs. Reavaliamos os pacientes por até 48 meses após a radioiodoterapia. As medianas iniciais de idade, bócio e captação tiroideana de 24hs dos 120 pacientes foram, respectivamente, 34 anos (17-69), 54,5g (20-210) e 73% (21-99). A evolução para hipotiroidismo foi gradual, com 15% dos pacientes a cada 6m até o 18º mês, seguidos de 2 a 5% dos pacientes a cada 6m até o 48º mês. O número de pacientes em tirotoxicose decresceu continuamente, representando 52,2% do total inicial após 6m, 25% após 18m e 6,7% após 48m. Ao final, 8 pacientes continuaram em tirotoxicose. O número de pacientes sem tirotoxicose após o tratamento com 131I, ou seja a soma daqueles em eutiroidismo com aqueles em hipotiroidismo com reposição com T4 (denominada por nós de cura da tirotoxicose), alcançou 46,8% dos pacientes em óm, 63,3% em 12m, 81,7% em 24m, 87,5% em 36m e 93,3% em 48m. O número de pacientes que apresentaram hipotiroidismo transitório, ou seja, elevação do TSH acima de 4,5 mU/L até 12m após a dose de 131I, com normalização posterior, foi 7,5%. Correlacionaram-se positivamente: a) valores iniciais de T3 e ausência de evolução para a cura da tirotoxicose (p=0,02); b) doses totais de 131I administradas, únicas ou múltiplas, em relação ao tempo de cura da tirotoxicose (p=0,01); c) dose de 131 l/volume glandular em relação ao tempo de cura da tirotoxicose (p=0,02). Os efeitos adversos incluíram exacerbação da tirotoxicose em 3 pacientes e dor na região cervical anterior em um, com reversão dos quadros em todos. Outras duas pacientes engravidaram após o dose terapêutica de 131I para a tirotoxicose e tiveram gestações sem intercorrência e recém-nascidos normais.Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamento de MedicinaUNIFESP, EPM, Depto. de MedicinaSciEL

Health-care guidelines and policies during the COVID-19 pandemic in Mexico: a case of health-inequalities

Background Heterogeneous government responses have been reported in reaction to COVID-19. The aim of this study is to generate an exploratory review of healthcare policies published during COVID-19 by health-care institutions in Mexico. Analyzing policies within different health sub-systems becomes imperative in the Mexican case due to the longstanding fragmentation of the health-care system and health inequalities. Data and Methods Policies purposely included in the analysis were published by four public health institutions (IMSS, ISSSTE, SSA and PEMEX) during the COVID-19 epidemic in Mexico (from February 29th to June 15th, 2020) on official institutional websites. Researchers reviewed each document and classified them into seven policy categories set by the Rapid Research Evaluation and Appraisal Lab (RREAL): public health response, health-care delivery, human resources, health-system infrastructure and supplies, clinical response, health-care management, and epidemiological surveillance. Results Policy types varied by health institution. The largest number of policies were aimed at public health responses followed by health-care delivery and human resources. Policies were mainly published during the community transmission phase. Conclusions The pandemic exposed underlying health-care system inequalities and a reactive rather than prepared response to the outbreak. Additionally, this study outlines potential policy gaps and delays in the response that could be avoided in the future

Profiling microglia in a mouse model of Machado-Joseph disease

Microglia have been increasingly implicated in neurodegenerative diseases (NDs), and specific disease associated microglia (DAM) profiles have been defined for several of these NDs. Yet, the microglial profile in Machado–Joseph disease (MJD) remains unexplored. Here, we characterized the profile of microglia in the CMVMJD135 mouse model of MJD. This characterization was performed using primary microglial cultures and microglial cells obtained from disease-relevant brain regions of neonatal and adult CMVMJD135 mice, respectively. Machine learning models were implemented to identify potential clusters of microglia based on their morphological features, and an RNA-sequencing analysis was performed to identify molecular perturbations and potential therapeutic targets. Our findings reveal morphological alterations that point to an increased activation state of microglia in CMVMJD135 mice and a disease-specific transcriptional profile of MJD microglia, encompassing a total of 101 differentially expressed genes, with enrichment in molecular pathways related to oxidative stress, immune response, cell proliferation, cell death, and lipid metabolism. Overall, these results allowed us to define the cellular and molecular profile of MJD-associated microglia and to identify genes and pathways that might represent potential therapeutic targets for this disorder.This work was supported by Fundação para a Ciência e a Tecnologia (FCT) (PTDC/NEUNMC/3648/2014) and COMPETE-FEDER (POCI-01-0145-FEDER-016818). It was also supported by Portuguese funds through FCT in the framework of the Project POCI-01-0145-FEDER-031987 (PTDC/MED-OUT/31987/2017). A.B.C. was supported by a doctoral fellowship from FCT (PD/BD/ 127828/2016). S.P.N. was also supported by FCT (PD/BD/114120/2015). Work in the JBR laboratory was financed by FEDER—Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020—Operational Programme for Competitiveness and Internationalization (POCI), Portugal 2020, and by Portuguese funds through FCT in the framework of the Project POCI-01-0145- FEDER030647 (PTDC/MED-NEU/31318/2017). This work was funded by ICVS Scientific Microscopy Platform, member of the national infrastructure PPBI (Portuguese Platform of Bioimaging) (PPBIPOCI-01-0145-FEDER-022122), and by National funds, through FCT—project UIDB/50026/2020 and UIDP/50026/2020

Homozygous Inactivating Mutation In Nanos3 In Two Sisters With Primary Ovarian Insufficiency.

Despite the increasing understanding of female reproduction, the molecular diagnosis of primary ovarian insufficiency (POI) is seldom obtained. The RNA-binding protein NANOS3 poses as an interesting candidate gene for POI since members of the Nanos family have an evolutionarily conserved function in germ cell development and maintenance by repressing apoptosis. We performed mutational analysis of NANOS3 in a cohort of 85 Brazilian women with familial or isolated POI, presenting with primary or secondary amenorrhea, and in ethnically-matched control women. A homozygous p.Glu120Lys mutation in NANOS3 was identified in two sisters with primary amenorrhea. The substituted amino acid is located within the second C2HC motif in the conserved zinc finger domain of NANOS3 and in silico molecular modelling suggests destabilization of protein-RNA interaction. In vitro analyses of apoptosis through flow cytometry and confocal microscopy show that NANOS3 capacity to prevent apoptosis was impaired by this mutation. The identification of an inactivating missense mutation in NANOS3 suggests a mechanism for POI involving increased primordial germ cells (PGCs) apoptosis during embryonic cell migration and highlights the importance of NANOS proteins in human ovarian biology.201478746

The effects of a muscle resistance program on the functional capacity, knee extensor muscle strength and plasma levels of IL-6 and TNF-α in pre-frail elderly women: a randomized crossover clinical trial - a study protocol

<p>Abstract</p> <p>Background</p> <p>With the increase in the elderly population, a growing number of chronic degenerative diseases and a greater dependency on caregivers have been observed. Elderly persons in states of frailty remain more susceptible to significant health complications. There is evidence of an inverse relationship between plasma levels of inflammatory mediators and levels of functionality and muscle strength, suggesting that muscle-strengthening measures can aid in inflammatory conditions. The purpose of this study will be verified the effect of a muscle-strengthening program with load during a ten-week period in pre-frail elderly women with attention to the following outcomes: (1) plasma levels of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α), (2) functional capacity and (3) knee extensor muscle strength.</p> <p>Methods/Design</p> <p>The study design is a randomized crossover clinical trial evaluating 26 elderly women (regardless of their race and/or social condition) who are community residents, older than 65, and classified as pre-frail according to the criteria previously described by Fried et al. (2004). All subjects will be assessed using the Timed up and go and 10-Meter Walk Test functional tests. The plasma levels of IL-6 and TNF-α will be assessed by ELISA (<it>enzyme-linked immunosorbent assay</it>) with high sensitivity kits (Quantikine^®HS, R&D Systems Minneapolis, MN, U.S.). Knee extensor muscle strength will be assessed using the <it>Byodex System 3 Pro</it>^{<it>® </it>}isokinetic dynamometer at angular speeds of 60 and 180°/s. The intervention will consist of strengthening exercises of the lower extremities at 50 to 70% of 1RM (maximal resistance) three times per week for ten weeks. The volunteers will be randomized into two groups: group E, the intervention group, and group C, the control group that did not initiate any new activities during the initial study period (ten weeks). After the initial period, group C will begin the intervention and group E will maintain everyday activities without exercising. At the end of the total study period, all volunteers will be reassessed.</p> <p>Discussion</p> <p>To demonstrate and discuss possible influences of load-bearing exercises on the modification of plasma levels of IL-6 and TNF-α and in the functional performance of pre-frail elderly women.</p> <p>Trial Registration</p> <p>ISRCTN62824599</p

Severe forms of partial androgen insensitivity syndrome due to p.L830F novel mutation in androgen receptor gene in a Brazilian family

<p>Abstract</p> <p>Background</p> <p>The androgen insensitivity syndrome may cause developmental failure of normal male external genitalia in individuals with 46,XY karyotype. It results from the diminished or absent biological action of androgens, which is mediated by the androgen receptor in both embryo and secondary sex development. Mutations in the androgen receptor gene, located on the X chromosome, are responsible for the disease. Almost 70% of 46,XY affected individuals inherited mutations from their carrier mothers.</p> <p>Findings</p> <p>Molecular abnormalities in the androgen receptor gene in individuals of a Brazilian family with clinical features of severe forms of partial androgen insensitivity syndrome were evaluated. Seven members (five 46,XY females and two healthy mothers) of the family were included in the investigation. The coding exons and exon-intron junctions of androgen receptor gene were sequenced. Five 46,XY members of the family have been found to be hemizygous for the c.3015C>T nucleotide change in exon 7 of the androgen receptor gene, whereas the two 46,XX mothers were heterozygote carriers. This nucleotide substitution leads to the p.L830F mutation in the androgen receptor.</p> <p>Conclusions</p> <p>The novel p.L830F mutation is responsible for grades 5 and 6 of partial androgen insensitivity syndrome in two generations of a Brazilian family.</p

Determinant Factors of Dividend Payments in Brazil

This study identifies factors that shaped cash disbursement distribution policies employed by Brazilian public companies listed on the Brazilian Securities, Commodities and Futures Exchange (BM&FBOVESPA) from 1995 to 2011. Relationships between Dividends/Total Assets and potential determinants discussed in the literature, including firm size, corporate governance, profitability, leverage, market to book, liquidity, investment, risk, profit growth, information asymmetry and agency conflict, are examined. The following econometric methods are employed: (1) Tobit, given the nature of the dividend data, and (2) the Generalized Method of Moments (GMM) to control for endogenous regressors. Significant positive variables found include size, return on assets (ROA), market to book, liquidity and profit growth. It can thus be inferred that larger firm size, profitability, market value, liquidity and profit growth correlate with greater firm pro pensity to distribute money to shareholders, thus supporting the theory of corporate finance. Significant negative variables found include leverage, liquidity squared, capex, beta and tag along 100%. It is thus inferred that more significantly leveraged companies that invest more heavily in fixed assets and that exhibit high liquidity, higher risk and less conflict between controlling and minority shareholders will be less likely to pay dividends to shareholders.</p