[Eating Habits of People with Type 2 Diabetes Mellitus in Portugal: A Cross-Sectional Study]. / Hábitos Alimentares das Pessoas com Diabetes Mellitus Tipo 2 em Portugal: Um Estudo Transversal.

INTRODUCTION: Nutrition is a cornerstone of diabetes mellitus prevention and management; therefore, it is essential to enable patients to adopt healthy eating habits. Previous studies have not yet documented the main errors in the eating habits of Portuguese people with type 2 diabetes mellitus. This study aims to identify the main errors in the eating habits of people living with type 2 diabetes mellitus in Portugal and to evaluate its associations with sociodemographic variables. METHODS: Cross-sectional multicentric study in a convenience sample of people with type 2 diabetes mellitus in Primary Health Care Units. The UK Diabetes and Diet Questionnaire (UKDDQ) - translated and adapted, was applied from July to October 2022. Descriptive and inferential statistical analyses were conducted. RESULTS: Of the 550 participants, 52.2% were female, 68.3% were 65 years or over, 55.8 % had an education level up to the fourth grade, 24.7% had economic deprivation, and the mean time since diagnosis was 10.60 ± 8.13 years. Only 36.2% of the sample had a healthy UKDDQ score. Less than 50% of the sample had healthy scores for the items "high-fiber rice or pasta", "high-fiber bread", "butter, margarine and vegetable oils" and "vegetables and pulses". Only 8.9% of the sample had a healthy consumption of fiber. About 70.4% reported healthy scores for the consumption of "high-added-sugar foods" and 54.7% for "high-saturated fat". A statistically significant weak positive correlation was found between the UKDDQ score and age (ρ = 0.201, p < 0.001) with a more frequent choice of healthy foods with increasing age. Female respondents reported healthier habits, particularly in the consumption of "high-saturated fat" and "high-fiber foods". CONCLUSION: The majority of our sample did not take advantage of the potential benefits of healthy eating habits. The main food groups whose consumption should be emphasized or discouraged were individualized, particularly the need to encourage the consumption of high-fiber foods. Targeted educational actions must focus especially on younger and/or male patients.

Introdução: A nutrição é uma área de intervenção na prevenção e gestão da diabetes mellitus; por isso, é fulcral promover a capacitação da população para a adoção de hábitos alimentares saudáveis. Ainda que existam alguns estudos nesta área, não se conhecem os principais erros nos hábitos alimentares das pessoas com diabetes em Portugal. Os objetivos deste estudo foram identificar os principais erros nos hábitos alimentares das pessoas com diabetes mellitus tipo 2 em Portugal e avaliar a sua relação com variáveis sociodemográficas. Métodos: Estudo transversal multicêntrico, em amostra de conveniência de pessoas com diabetes mellitus tipo 2 seguidas em Unidades de Cuidados de Saúde Primários. Aplicação do UK Diabetes and Diet Questionnaire (UKDDQ) ­ traduzido e adaptado, de julho a outubro de 2022. Análise estatística descritiva e inferencial. Resultados: Amostra de 550 participantes, 52,2% do sexo feminino, 68,3% com 65 anos ou mais, 55,8% com nível de escolaridade igual ou inferior ao 1.º ciclo do ensino básico, 24,7% com insuficiência económica e tempo desde o diagnóstico médio de 10,60 ± 8,13 anos. Apenas 36,2% da amostra obteve um score UKDDQ considerado saudável. Menos de 50% obteve scores saudáveis para os itens "arroz ou massa ricos em fibras", "pão integral", "manteiga, margarina e óleos vegetais" e "vegetais e leguminosas". Somente 8,9% da amostra obteve score saudável para o consumo de fibras. Cerca de 70,4% obteve score saudável para o consumo de açúcares livres e 54,7% para o consumo de ácidos gordos saturados. Verificou-se a existência de uma correlação com significado estatístico positiva fraca entre o score UKDDQ e a idade (ρ = 0,201, p < 0,001), com escolha mais frequente de alimentos saudáveis com o aumentar da idade. As pessoas do sexo feminino reportaram hábitos alimentares mais saudáveis, particularmente no consumo de fibras e ácidos gordos saturados. Conclusão: A maior parte da nossa amostra não usufruiu do potencial efeito positivo de uma alimentação saudável. Individualizam-se grupos de alimentos cujos consumos devem ser enfatizados ou desencorajados, particularmente, a necessidade de incentivar o consumo de alimentos ricos em fibra. Ações educacionais dirigidas devem ter especial foco em pessoas mais jovens e/ou do sexo masculino.

Meniscal Bone Angle Is a Strong Predictor of Anterior Cruciate Ligament Injury.

Purpose: To evaluate the influence of lateral posterior tibial slope (LPTS) and meniscal bone angle (MBA) on primary anterior cruciate ligament (ACL) tear risk in an adult population through the LPTS-MBA ratio. Methods: A retrospective case-control study was performed with patients from a tertiary hospital who underwent primary ACL surgery and had preoperative magnetic resonance imaging (MRI). These subjects were matched by age and sex in a 1:1 ratio to patients who had an MRI without ACL tear. LPTS and MBA were measured on MRI scan. Quantitative data are presented in the median ± interquartile range (IQR). Identification of independent risk factors for primary ACL tear was performed using multivariable logistic regression. Receiver operating characteristics curves detected any variable with strong discriminative capacity. Results: In total, 95 patients with primary ACL tear confirmed on MRI were matched with 95 controls (N = 190). Nearly 80% were male subjects, with a median age of 26 years. In the ACL tear group, the median value of LPTS-MBA ratio was 0.20 (IQR 0.11-0.37) versus 0.12 (IQR 0.08-0.19) in the control group (P = .001). LPTS had a median value of 4.20° in the ACL tear group (IQR 2.05-7.35°) and 2.90° in the control group (IQR, 2.05-5.00°) (P = .026), whereas MBA was 19° (IQR, 16-24°) versus 26° (IQR, 24-30°) (P = .001), respectively. Logistic regression showed that LPTS (odds ratio 1.20, 95% confidence interval 1.03-1.42, P = .021) and MBA (odds ratio 0.78, 95% confidence interval 0.71-0.85, P = .001) were independent predictors. The area under the curve (AUC) of LPTS-MBA ratio was 0.69, greater than that of LPTS alone (AUC = 0.61) but lower than that for MBA (AUC = 0.82). Conclusions: In this study, a reduced MBA was the strongest predictive variable associated with a primary ACL tear. A threshold of 22.35° of MBA was associated with an increased risk of ACL tear, with a sensitivity of 70% and specificity of 84%. A cut-off of 0.22 of LPTS-MBA was associated with an increased risk of ACL tear, with a sensitivity of 55% and specificity of 87%. Level of Evidence: Level III, case-control study.

Relation between Selenium and Female Fertility: A Systematic Review / Relação entre o selênio e a fertilidade feminina: Uma revisão sistemática

Abstract Objective To analyze the influence of selenium in female fertility. Data sourceA search was performed in the following databases: MEDLINE, Web of Science, Scopus, SciELO, LILACS, MDPI, ScienceDirect, and Europe PMC. The descriptors selected were "selenium" AND "female" AND "fertility". The search interval was from 1996 to 2021. Study selectionThe evaluation was performed independently by two reviewers, and a third reviewer confirmed the inclusion of papers in case of divergence between the first two reviewers. Papers were selected after the title and abstract were read, and those that met the eligibility criteria had the full text read. Data collectionThe following data was extracted: author, year of publication, country, type of study, objective, method, sample size, follow-up period, patients' mean age, inclusion and exclusion criteria, and concentration of serum and capillary selenium. The data was organized in chronological order of paper publication. Data synthesisThe number of papers identified totaled 3,800, out of which 7 were included in the systematic review. The studies indicated a positive correlation between serum selenium and antioxidant concentration in the follicular fluid, reduction in antithyroid antibodies, oocyte production and follicle number. Conclusion Selenium supplementation is promising in women with this micronutrient deficiency to promote improvement of the reproductive efficiency and prevent damage to the pregnancy. Further studies on this theme are still required.

Resumo Objetivo Analisar a influência do selênio na fertilidade feminina. Fonte dos dadosUma busca foi realizada nas seguintes bases de dados: MEDLINE, Web of Science, Scopus, SciELO, LILACS, MDPI, ScienceDirect e Europe PMC. Os descritores selecionados foram "selenium" AND "female" AND "fertility". O intervalo de busca foi de 1996 a 2021. Seleção dos estudosA avaliação ocorreu de maneira independente por dois revisores, sendo que um terceiro corroborou a eleição dos artigos em casos de divergência. Os estudos foram selecionados através da leitura do título e resumo, e aqueles que contemplaram os critérios de elegibilidade foram lidos na íntegra. Coleta dos dadosOs seguintes dados foram extraídos: autor, ano de publicação, país, tipo de estudo, objetivo, método, tamanho da amostra, tempo de acompanhamento, média de idade das pacientes, critérios de inclusão e exclusão, concentração de selênio sérico e capilar. Os dados foram organizados em ordem cronológica de publicação do estudo. Síntese dos dadosForam identificados 3.800 artigos e incluídos 7 estudos na revisão sistemática. Os resultados indicaram correlação positiva entre o nível de selênio sérico e a concentração de antioxidantes no fluido folicular; diminuição dos níveis de anticorpos antitireoidianos; produção de oócitos, e número de folículos. Conclusão A suplementação de selênio é promissora em mulheres com deficiência do micronutriente, a fim de promover melhora na eficiência reprodutiva e prevenir danos na gravidez. Salientou-se a necessidade de realização de mais estudos sobre o tema.

Relation between Selenium and Female Fertility: A Systematic Review.

OBJECTIVE: To analyze the influence of selenium in female fertility. DATA SOURCE: A search was performed in the following databases: MEDLINE, Web of Science, Scopus, SciELO, LILACS, MDPI, ScienceDirect, and Europe PMC. The descriptors selected were "selenium" AND "female" AND "fertility". The search interval was from 1996 to 2021. STUDY SELECTION: The evaluation was performed independently by two reviewers, and a third reviewer confirmed the inclusion of papers in case of divergence between the first two reviewers. Papers were selected after the title and abstract were read, and those that met the eligibility criteria had the full text read. DATA COLLECTION: The following data was extracted: author, year of publication, country, type of study, objective, method, sample size, follow-up period, patients' mean age, inclusion and exclusion criteria, and concentration of serum and capillary selenium. The data was organized in chronological order of paper publication. DATA SYNTHESIS: The number of papers identified totaled 3,800, out of which 7 were included in the systematic review. The studies indicated a positive correlation between serum selenium and antioxidant concentration in the follicular fluid, reduction in antithyroid antibodies, oocyte production and follicle number. CONCLUSION: Selenium supplementation is promising in women with this micronutrient deficiency to promote improvement of the reproductive efficiency and prevent damage to the pregnancy. Further studies on this theme are still required.

OBJETIVO: Analisar a influência do selênio na fertilidade feminina. FONTE DOS DADOS: Uma busca foi realizada nas seguintes bases de dados: MEDLINE, Web of Science, Scopus, SciELO, LILACS, MDPI, ScienceDirect e Europe PMC. Os descritores selecionados foram "selenium" AND "female" AND "fertility". O intervalo de busca foi de 1996 a 2021. SELEçãO DOS ESTUDOS: A avaliação ocorreu de maneira independente por dois revisores, sendo que um terceiro corroborou a eleição dos artigos em casos de divergência. Os estudos foram selecionados através da leitura do título e resumo, e aqueles que contemplaram os critérios de elegibilidade foram lidos na íntegra. COLETA DOS DADOS: Os seguintes dados foram extraídos: autor, ano de publicação, país, tipo de estudo, objetivo, método, tamanho da amostra, tempo de acompanhamento, média de idade das pacientes, critérios de inclusão e exclusão, concentração de selênio sérico e capilar. Os dados foram organizados em ordem cronológica de publicação do estudo. SíNTESE DOS DADOS: Foram identificados 3.800 artigos e incluídos 7 estudos na revisão sistemática. Os resultados indicaram correlação positiva entre o nível de selênio sérico e a concentração de antioxidantes no fluido folicular; diminuição dos níveis de anticorpos antitireoidianos; produção de oócitos, e número de folículos. CONCLUSãO: A suplementação de selênio é promissora em mulheres com deficiência do micronutriente, a fim de promover melhora na eficiência reprodutiva e prevenir danos na gravidez. Salientou-se a necessidade de realização de mais estudos sobre o tema.

Structural properties and evaluation of the antiproliferative activity of limonene-1,2-diol obtained by the fungal biotransformation of R-(+)- and S-(-)-limonene.

Limonene-1,2-diol is a limonene oxygenated metabolite that possesses eight different stereoisomers, which could result in different biological properties. Nonetheless, the relation between its spatial configuration and biological function is still little explored. The present study aimed to perform the stereoisomers identification using nuclear magnetic resonance (NMR) investigation of the limonene-1,2-diol produced via R-(+)- and S-(-)-limonene biotransformation by Colletotrichum nymphaeae and S-(-)-limonene biotransformation by Fusarium oxysporum 152B. Besides, in vitro antiproliferative activity was evaluated against human tumor and nontumor cell lines. The NMR analysis showed that R-(+)-limonene biotransformation afforded exclusively (+)-(1S,2S,4R-limonene-1,2-diol), whereas S-(-)-limonene biotransformation afforded exclusively (-)-(1R,2R,4S-limonene-1,2-diol) independent on the fungi used. Despite no significant cytostatic effects, a possible influence of stereogenic center on the antiproliferative activity of these limonene biotransformation products was evidenced. Moreover, the lack of in vitro antiproliferative effect of limonene-1,2-diol against nontumor cells suggested a safe dose range for further in vivo evaluations, including food applications.

Lignocellulosic substrates as starting materials for the production of bioactive biopigments.

The search for sustainable processes is constantly increasing in the last years, so reusing, recycling and adding value to residues and by-products from agroindustry is a consolidated area of research. Particularly in the field of fermentation technology, the lignocellulosic substrates have been used to produce a diversity of chemicals, fuels and food additives. These residues or by-products are rich sources of carbon, which may be used to yield fermentescible sugars upon hydrolysis, but are usually inaccessible to enzyme and microbial attack. Therefore, pre-treatments (e.g. hydrolysis, steam explosion, biological pretreatment or others) are required prior to microbial action. Biopigments are added-value compounds that can be produced biotechnologically, including fermentation processes employing lignocellulosic substrates. These molecules are important not only for their coloring properties, but also for their biological activities. Therefore, this paper discusses the most recent and relevant processes for biopigment production using lignocellulosic substrates (solid-state fermentation) or their hydrolysates.

Data on the evaluation of FGF2 gene expression in Colorectal Cancer.

The data presented in this article is related with the research paper entitled "Evaluation of MGP gene expression in colorectal cancer", available on Gene journal [1]. From all the transcription factors known to regulate MGP, FGF2 is the most described in colon adenocarcinoma and colon tumor cell lines, where it was shown to: i) contribute for the invasiveness potential; and ii) promote proliferation and survival of colorectal cancer cells. These in vitro studies pose the hypothesis that FGF2 associated signaling pathways could be promoting the regulation of others genes, such as MGP, that may lead to tumor progression which ultimately could result in poor prognosis in colon adenocarcinoma.

Evaluation of MGP gene expression in colorectal cancer.

PURPOSE: Matrix Gla protein (MGP) is a vitamin K-dependent, γ-carboxylated protein that was initially found to be a physiological inhibitor of ectopic calcifications affecting mainly cartilage and the vascular system. Mutations in the MGP gene were found to be responsible for a human pathology, the Keutel syndrome, characterized by abnormal calcifications in cartilage, lungs, brain and vascular system. MGP was recently implicated in tumorigenic processes such as angiogenesis and shown to be abnormally regulated in several tumors, including cervical, ovarian, urogenital and breast. This fact has triggered our interest in analyzing the expression of MGP and of its regulator, the transcription factor runt related transcription factor 2 (RUNX2), in colorectal cancer (CRC). METHODS: MGP and RUNX2 expression were analyzed in cancer and non-tumor biopsies samples from 33 CRC patients and 9 healthy controls by RT-qPCR. Consequently, statistical analyses were performed to evaluate the clinical-pathological significance of MGP and RUNX2 in CRC. MGP protein was also detected by immunohistochemical analysis. RESULTS: Showed an overall overexpression of MGP in the tumor mucosa of patients at mRNA level when compared to adjacent normal mucosa and healthy control tissues. In addition, analysis of the expression of RUNX2 mRNA demonstrated an overexpression in CRC tissue samples and a positive correlation with MGP expression (Pearson correlation coefficient 0.636; p ≤ 0.01) in tumor mucosa. However correlations between MGP gene expression and clinical-pathological characteristics, such as gender, age and pathology classification did not provide relevant information that may shed light towards the differences of MGP expression observed between normal and malignant tissue. CONCLUSIONS: We were able to associate the high levels of MGP mRNA expression with a worse prognosis and survival rate lower than five years. These results contributed to improve our understanding of the molecular mechanism underlying MGP deregulation in cancer.

Fatores presentes na intervenção dos enfermeiros no cuidado ao doente com alterações comportamentais após traumatismo crânio encefálico: estudo numa unidade de cuidados de neurocirurgia / Factors present in nurses' intervention in the care of patients with behavioral changes after traumatic brain injury: study in a neurosurgery care unit

O cuidar faz parte de um processo inerente ao ser humano. Desde sempre, as sociedades tiveram como imperativo preservar os conhecimentos adquiridos das práticas de cuidar e transmiti-los às gerações vindouras. A enfermagem, através da incorporação de muitas dessas práticas, assim como da definição e redefinição de vários conceitos, percorreu um caminho extenso e árduo ao longo dos tempos, de forma a dar origem ao que hoje chamamos de cuidados de enfermagem. Esses cuidados de enfermagem foram tornando-se cada vez mais complexos e exigentes, definindo-se a prática dos cuidados de enfermagem ao doente crítico, como é o caso do doente com traumatismo crânio encefálico. Os traumatismos crânio encefálicos constituem um problema de saúde pública com grade impacto económico e social, mantendo-se como a principal causa de mortalidade e morbilidade entre os adultos jovens. Este estudo tem como principal objetivo aprofundar a temática relacionada com as alterações comportamentais nos doentes após traumatismo crânio encefálico, de forma a dar resposta à questão de investigação sobre "Quais os fatores presentes na intervenção dos enfermeiros no cuidado ao doente com alterações comportamentais após traumatismo crânio encefálico?". Trata-se de um estudo exploratório descritivo, com abordagem qualitativa e que decorreu durante os anos de 2017 e 2018. Contou com a participação de 17 enfermeiros que trabalham num serviço de internamento de neurocirurgia de um hospital da zona norte. Para a recolha de dados foi desenvolvido um guião de entrevista semiestruturada, recorrendo-se posteriormente à análise de conteúdo com o intuito de tratar os dados obtidos. Emergiram sete blocos temáticos centrais que permitiram dar resposta aos objetivos delineados: 1) Principais Cuidados de Enfermagem ao Doente Após TCE; 2) Importância Atribuída às Alterações Comportamentais; 3) Alterações Comportamentais mais Relevantes; 4) Impacto das Alterações Comportamentais no Profissional; 5) Estratégias Utilizadas para Lidar com o Doente com Alterações Comportamentais; 6) Contingentes ao Desenvolvimento da Proficiência no Exercício Profissional; e, 7) Vivências Marcantes da Problemática. Os resultados apontam para fatores considerados como importantes pelos enfermeiros para a proficiência no seu exercício profissional, assim como outros relativos às dificuldades encontradas aquando da prestação de cuidados. Deste modo, a temática perspetiva-se como uma área de relevo para a enfermagem, sendo necessários mais estudos de forma a que sejam otimizados os resultados em saúde esperados. Inferiu-se ainda acerca dos possíveis contributos para a prática de enfermagem e sobre as limitações do presente estudo.

Caregiving is part of an inherent process to being human. Society has always had the imperative to preserve the knowledge acquired from caregiving practices and to transmit them to future generations. Nursing, through the incorporation of many of these practices, along with the definition and redefinition of various concepts, has taken a long and arduous path through time in order to give rise to what we now call nursing care. This nursing care has become increasingly complex and demanding, leading to the definition of the practice of nursing care for critical patients, as is the case of patients with traumatic brain injury. Traumatic brain injury is a public health problem with a high economic and social impact, and remains the main cause of mortality and morbidity among young adults. This study's main objective is to deepen the knowledge on behavioral changes in patients after traumatic brain injury, in order to answer the research question "Which factors are present in the intervention of nurses in the care to patients with behavioral changes after a traumatic brain injury?". This is an exploratory descriptive study with a qualitative approach that took place between 2017 and 2018. Seventeen nurses who worked in a neurosurgical inpatient service of a hospital in the north of Portugal took part in this study. For data collection, a semi-structured interview script was developed, after which content analysis was used to treat the data obtained. Seven thematic central blocks emerged that allowed to answer the outlined objectives: 1) Main nursing care to the patient after TBI; 2) Importance Attributed to Behavioral Changes; 3) Most Relevant Behavioral Changes; 4) Impact of Behavioral Changes on the Professional; 5) Strategies Used to Deal with Patients with Behavioral Changes; 6) Contingents to Developing Proficiency in Practice; and, 7) Challenging Experiences. Results point to factors nurses consider important to achieve proficiency in their professional practice, as well as others related to difficulties encountered when providing care. Therefore, the thematic perspective is an important area for nursing, and further studies are needed to optimize the expected health outcomes. Possible contributions to nursing practice and the limitations of the present study were also discussed.

Evidences for a New Role of miR-214 in Chondrogenesis.

miR-214 is known to play a role in mammalian skeletal development through inhibition of osteogenesis and stimulation of osteoclastogenesis, but data regarding other vertebrates, as well as a possible role in chondrogenesis, remain unknown. Here, we show that miR-214 expression is detected in bone and cartilage of zebrafish skeleton, and is downregulated during murine ATDC5 chondrocyte differentiation. Additionally, we observed a conservation of the transcriptional regulation of miR-214 primary transcript Dnm3os in vertebrates, being regulated by Ets1 in ATDC5 chondrogenic cells. Moreover, overexpression of miR-214 in vitro and in vivo mitigated chondrocyte differentiation probably by targeting activating transcription factor 4 (Atf4). Indeed, miR-214 overexpression in vivo hampered cranial cartilage formation of zebrafish and coincided with downregulation of atf4 and of the key chondrogenic players sox9 and col2a1. We show that miR-214 overexpression exerts a negative role in chondrogenesis by impacting on chondrocyte differentiation possibly through conserved mechanisms.

Mutirão de saúde como ação extensiva e integrativa com a atenção primária / Health task force as extensive and integrative action with primary attention

Este trabalho buscou descrever uma das ações extensiva e interdisciplinar da LASEM da Universidade Federal dos Vales do Jequitinhonha e Mucuri, os Mutirões de Saúde. A liga acadêmica, firmada no tripé ensino, pesquisa e extensão, pretende aproximar o estudante da prática da atenção à saúde, com foco na vivência, para auxiliar o entendimento de como os fatores socioeconômicos e culturais influenciam no processo saúde-doença. Além disso, tem por objetivo descrever sua proposta e dinâmica, incentivar sua realização como uma atividade de Atenção Primária à Saúde, visto a sua importância para a formação médica. Os mutirões de saúde foram mobilizações coletivas, planejadas pela liga e pelo gestor de saúde local, com intuito de promover uma aproximação com a comunidade, permitir a aplicação do conhecimento construído nas reuniões da liga e ofertar serviços de saúde à população local. A inserção precoce do aluno de medicina na prática médica permite uma aproximação entre a instituição de ensino e a comunidade, de forma que é construído um ambiente de crescimento mútuo, tanto para a educação do aluno, quanto para a saúde da população. Isso permite que o estudante se desenvolva como agente transformador do meio. A experiência com os mutirões de saúde mostrou que são atividades eficazes para o aperfeiçoamento do ensino, para o compartilhamento de saberes entre profissionais, estudantes e a comunidade, bem como para o fortalecimento da liga acadêmica de semiologia médica dentro da instituição universitária de ensino. (AU)

This work sought to describe one of the extensive and interdisciplinary actions of LASEM of the Federal University of the Jequitinhonha and Mucuri Valleys, the Health Task Force. The academic league, signed on the tripod teaching, research and extension, aims to bring the student closer to the practice of attention to health, with a focus on the experience, to help the understanding of how socioeconomic and cultural factors influence the health-disease process. In addition, it aims to describe its proposal and dynamics, to encourage its realization as a Primary Health Care activity, given its importance for medical training. The health efforts were collective mobilizations, planned by the league and the local health manager, in order to promote a rapprochement with the community, to allow the application of knowledge built in league meetings and to offer health services to the local population. The precocious insertion of the medical student in medical practice allows an approximation between the educational institution and the community, so that an environment of mutual growth is built, both for the students education and for the health of the population. This allows the student to develop as an agent that transforms the environment. The experience with health efforts showed that they are effective activities for the improvement of teaching, for the sharing of knowledge among professionals, students and the community, as well as for the strengthening of the academic league of medical semiology within the university teaching institution. (AU)

Central role of betaine-homocysteine S-methyltransferase 3 in chondral ossification and evidence for sub-functionalization in neoteleost fish.

BACKGROUND: To better understand the complex mechanisms of bone formation it is fundamental that genes central to signaling/regulatory pathways and matrix formation are identified. Cell systems were used to analyze genes differentially expressed during extracellular matrix mineralization and bhmt3, coding for a betaine-homocysteine S-methyltransferase, was shown to be down-regulated in mineralizing gilthead seabream cells. METHODS: Levels and sites of bhmt3 expression were determined by qPCR and in situ hybridization throughout seabream development and in adult tissues. Transcriptional regulation of bhmt3 was assessed from the activity of promoter constructs controlling luciferase gene expression. Molecular phylogeny of vertebrate BHMT was determined from maximum likelihood analysis of available sequences. RESULTS: bhmt3 transcript is abundant in calcified tissues and localized in cartilaginous structures undergoing endo/perichondral ossification. Promoter activity is regulated by transcription factors involved in bone and cartilage development, further demonstrating the central role of Bhmt3 in chondrogenesis and/or osteogenesis. Molecular phylogeny revealed the explosive diversity of bhmt genes in neoteleost fish, while tissue distribution of bhmt genes in seabream suggested that neoteleostean Bhmt may have undergone several steps of sub-functionalization. CONCLUSIONS: Data on bhmt3 gene expression and promoter activity evidences a novel function for betaine-homocysteine S-methyltransferase in bone and cartilage development, while phylogenetic analysis provides new insights into the evolution of vertebrate BHMTs and suggests that multiple gene duplication events occurred in neoteleost fish lineage. GENERAL SIGNIFICANCE: High and specific expression of Bhmt3 in gilthead seabream calcified tissues suggests that bone-specific betaine-homocysteine S-methyltransferases could represent a suitable marker of chondral ossification.

Matrix Gla protein repression by miR-155 promotes oncogenic signals in breast cancer MCF-7 cells.

MGP is a protein that was initially associated with the inhibition of calcification in skeleton, soft tissues, and arteries, but more recently also implicated in cancer. In breast cancer, higher levels of MGP mRNA were associated with poor prognosis, but since this deregulation was never demonstrated at the protein level, we postulated the involvement of a post-transcriptional regulatory mechanism. In this work we show that MGP is significantly repressed by miR-155 in breast cancer MCF-7 cells, and concomitantly there is a stimulation of cell proliferation and cell invasiveness. This study brings new insights into the putative involvement of MGP and oncomiR-155 in breast cancer, and may contribute to develop new therapeutic strategies.

The controversial p.Met34Thr variant in GJB2 gene: Two siblings, one genotype, two phenotypes.

INTRODUCTION: Recent advances in molecular genetics have increased the identification of genes and mutations responsible for inherited forms of hearing loss (HL), enabling early detection of these cases. Approximately, 60% of early-onset HL cases are due to genetic causes, of which 70% are non-syndromic. Of these, 75-80% are inherited in an autosomal recessive pattern (DFNB). Mutations in GJB2 gene, coding for connexin 26 (Cx26), are the major cause of autosomal recessive hereditary HL, but some GJB2 mutations are yet of unclear or controversial significance. OBJECTIVES: The aim of the present study was to identify the etiology of hearing loss, and correlate genotype-phenotype, in two Portuguese siblings with profound and moderate non-syndromic sensorineural bilateral HL. MATERIAL AND METHODS: The affected subjects and their parents underwent audiological and genetic study. Molecular analysis of GJB2 gene was performed, searching for mutations in the coding region and receptor splicing site by automated sequencing. RESULTS: The onset and the degree of HL were different in the two affected subjects. However, the same GJB2 genotype [p.Met34Thr]+[p.Arg184Pro] was identified in both siblings. The c.551G>C (p.Arg184Pro) and c.101T>C (p.Met34Thr) missense variants were inherited from the father and mother, respectively, both heterozygous carriers of these variants. CONCLUSION: The clinical and genetic data here presented suggest that the non-syndromic sensorineural HL of these two Portuguese siblings might be due to the presence of p.Met34Thr and p.Arg184Pro variants in compound heterozygosity. If so, p.Met34Thr variant could have function as a hypomorphic allele that may cause HL depending on the opposing GJB2 allele. The observed phenotypic variability may not, however, be solely explained by variable expression of this genotype. A putative modifier gene or mutations in another HL-associated gene could probably be contributing to the severe HL in one of the siblings.

Mir-20a regulates in vitro mineralization and BMP signaling pathway by targeting BMP-2 transcript in fish.

MicroRNAs (miRNAs) are important regulators of vertebrate development but their role during skeletogenesis remains unknown. In this regard, we investigated the mineralogenic activity of miR-20a, a miRNA associated with osteogenesis, in fish bone-derived cells. Expression of miR-20a was up-regulated during differentiation and its overexpression inhibited mineralization, suggesting a role in fish tissue calcification. In this regard, a conserved miR-20a binding site was identified in bone morphogenetic protein 2 (BMP-2) 3'UTR and its functionality was evidenced through luciferase assays, and further confirmed by western-blot and qPCR. Type II BMP receptor (BMPR2) is also targeted by miR-20a in mammalian systems and evidence was collected for the presence of a binding site in fish sequences. We propose that miR-20a is a regulator of BMP pathway through specific action on BMP-2 and possibly BMPR2. Overexpression of miR-20a was also shown to up-regulate matrix Gla protein (MGP) transcript, a physiological inhibitor of calcification previously found to form a complex with BMP-2. We propose that MGP may play a role in the anti-mineralogenic effect promoted by miR-20a by decreasing availability of BMP-2. This study gives new insights into miRNA-mediated regulation of BMP-2, and sheds light into the potential role of miR-20a as a regulator of skeletogenesis.

Retinoic acid differentially affects in vitro proliferation, differentiation and mineralization of two fish bone-derived cell lines: different gene expression of nuclear receptors and ECM proteins.

Retinoic acid (RA), the main active metabolite of vitamin A, regulates vertebrate morphogenesis through signaling pathways not yet fully understood. Such process involves the specific activation of retinoic acid and retinoid X receptors (RARs and RXRs), which are nuclear receptors of the steroid/thyroid hormone receptor superfamily. Teleost fish are suitable models to study vertebrate development, such as skeletogenesis. Cell systems capable of in vitro mineralization have been developed for several fish species and may provide new insights into the specific cellular and molecular events related to vitamin A activity in bone, complementary to in vivo studies. This work aims at investigating the in vitro effects of RA (0.5 and 12.5 µM) on proliferation, differentiation and extracellular matrix (ECM) mineralization of two gilthead seabream bone-derived cell lines (VSa13 and VSa16), and at identifying molecular targets of its action through gene expression analysis. RA induced phenotypic changes and cellular proliferation was inhibited in both cell lines in a cell type-dependent manner (36-59% in VSa13 and 17-46% in VSa16 cells). While RA stimulated mineral deposition in VSa13 cell cultures (50-62% stimulation), it inhibited the mineralization of extracellular matrix in VSa16 cells (11-57% inhibition). Expression of hormone receptor genes (rars and rxrs), and extracellular matrix-related genes such as matrix and bone Gla proteins (mgp and bglap), osteopontin (spp1) and type I collagen (col1a1) were differentially regulated upon exposure to RA in proliferating, differentiating and mineralizing cultures of VSa13 and VSa16 cells. Altogether, our results show: (i) RA affects proliferative and mineralogenic activities in two fish skeletal cell types and (ii) that during phenotype transitions, specific RA nuclear receptors and bone-related genes are differentially expressed in a cell type-dependent manner.

Spectrum and frequency of GJB2 mutations in a cohort of 264 Portuguese nonsyndromic sensorineural hearing loss patients.

OBJECTIVE: To assess the spectrum and prevalence of mutations in the GJB2 gene in Portuguese nonsyndromic sensorineural hearing loss (NSSHL) patients. DESIGN: Sequencing of the coding region, basal promoter, exon 1, and donor splice site of the GJB2 gene; screening for the presence of the two common GJB6 deletions. STUDY SAMPLE: A cohort of 264 Portuguese NSSHL patients. RESULTS: At least one out of 21 different GJB2 variants was identified in 80 (30.2%) of the 264 patients analysed. Two mutant alleles were found in 53 (20%) of these probands, of which 83% (44/53) harboured at least one c.35delG allele. Twenty-seven (10.2%) of the probands harboured only one mutant allele. Subsequent analysis revealed that the GJB6 deletion del(GJB6-D13S1854) was present in at least 7.4% (2/27) of the patients carrying only one mutant GJB2 allele. Overall, one in five (55/264) of the patients were diagnosed as having DFNB1-related NSSHL, of which the vast majority (53/55) harboured only GJB2 mutations. CONCLUSIONS: This study provides clear demonstration that mutations in the GJB2 gene are an important cause of NSSHL in Portugal, thus representing a valuable indicator as regards therapeutical and rehabilitation options, as well as genetic counseling of these patients and their families.

Two portuguese cochlear implanted dizygotic twins: a case report.

Individual's hearing performance after cochlear implant (CI) is variable and depends on different factors such as etiology of deafness, age at implantation, and social/family hearing environment. Here we report the case of dizygotic twins, boy and girl, presenting with neurosensorial profound deafness prior CI (age of implantation = 3.5 years old). Both parents have severe/profound deafness, since childhood, and use sign language as primary mode of communication. Clinical and genetic characterization was performed, as well as the assessment of the auditory and oral (re)habilitation after CI, applying a battery of audiological, speech, and language tests. The twin girl and the father were homozygous for the c.35delG mutation in the GJB2 gene, while the twin boy and the mother were compound heterozygotes, both monoallelic for c.35delG and for the deletion del(GJB6-D13S1830) in the GJB6 gene. The remaining hearing impaired relatives were c.35delG homozygotes. The genetic cause of deafness was thus identified in this family. Some noteworthy differences were observed regarding twins' auditory and oral performance after CI. Subsequent follow-up of these children allowed us to conclude that those differences were most likely due to the different environment in which the twins have been living than to their different GJB2/GJB6 genotypes.

Global analysis of gene expression in mineralizing fish vertebra-derived cell lines: new insights into anti-mineralogenic effect of vanadate.

BACKGROUND: Fish has been deemed suitable to study the complex mechanisms of vertebrate skeletogenesis and gilthead seabream (Sparus aurata), a marine teleost with acellular bone, has been successfully used in recent years to study the function and regulation of bone and cartilage related genes during development and in adult animals. Tools recently developed for gilthead seabream, e.g. mineralogenic cell lines and a 4 × 44K Agilent oligo-array, were used to identify molecular determinants of in vitro mineralization and genes involved in anti-mineralogenic action of vanadate. RESULTS: Global analysis of gene expression identified 4,223 and 4,147 genes differentially expressed (fold change - FC > 1.5) during in vitro mineralization of VSa13 (pre-chondrocyte) and VSa16 (pre-osteoblast) cells, respectively. Comparative analysis indicated that nearly 45% of these genes are common to both cell lines and gene ontology (GO) classification is also similar for both cell types. Up-regulated genes (FC > 10) were mainly associated with transport, matrix/membrane, metabolism and signaling, while down-regulated genes were mainly associated with metabolism, calcium binding, transport and signaling. Analysis of gene expression in proliferative and mineralizing cells exposed to vanadate revealed 1,779 and 1,136 differentially expressed genes, respectively. Of these genes, 67 exhibited reverse patterns of expression upon vanadate treatment during proliferation or mineralization. CONCLUSIONS: Comparative analysis of expression data from fish and data available in the literature for mammalian cell systems (bone-derived cells undergoing differentiation) indicate that the same type of genes, and in some cases the same orthologs, are involved in mechanisms of in vitro mineralization, suggesting their conservation throughout vertebrate evolution and across cell types. Array technology also allowed identification of genes differentially expressed upon exposure of fish cell lines to vanadate and likely involved in its anti-mineralogenic activity. Many were found to be unknown or they were never associated to bone homeostasis previously, thus providing a set of potential candidates whose study will likely bring insights into the complex mechanisms of tissue mineralization and bone formation.