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.

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.

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.

[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.

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.

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.

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.

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.

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.

Proliferative and mineralogenic effects of insulin, IGF-1, and vanadate in fish osteoblast-like cells.

Fish have recently been recognized as a suitable model and a promising alternative to mammalian systems to study skeletogenesis. In this regard, several fish bone-derived cell lines have been developed and are being used to investigate mechanisms associated with insulin-like action of vanadium on extracellular matrix (ECM) mineralization. Although proliferative and mineralogenic effects of vanadate, insulin-like growth factor 1 (IGF-1), and insulin have recently been evaluated in a fish prechondrocyte cell line, no data are available in fish bone-forming cells, the osteoblasts. Using fish preosteoblast cells, we showed that IGF-1, but not insulin or vanadate, stimulated cell proliferation through the mitogen-activated protein kinase (MAPK) pathway, while both IGF-1 and vanadate inhibited cell differentiation/ECM mineralization through the same mechanism. Our data also indicated that the phosphatidyl inositol-3 kinase (PI-3K) pathway stimulates differentiation/ECM mineralization in osteoblasts and could represent a way to balance MAPK pathway action. The comparison of these new data obtained in fish with those available in mammals clearly evidenced a conservation of regulatory mechanisms among vertebrate bone-derived systems, although different players are involved.

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.

Vanadate proliferative and anti-mineralogenic effects are mediated by MAPK and PI-3K/Ras/Erk pathways in a fish chondrocyte cell line.

We recently reported proliferative and anti-mineralogenic effects of vanadate on fish chondrocytes and here we investigate the signalling pathways associated with these effects. Our data show that vanadate stimulates chondrocyte proliferation through the MAPK pathway, using signalling mechanisms similar to those used by IGF-1, while it inhibits chondrocyte differentiation/mineralization through a putative PI-3K/Ras/Erk signalling, a pathway shared with insulin. Our data also suggest that vanadate impairs ECM mineralization not only by interfering with regulatory pathways but also by inhibiting enzymatic activity of ALP. Finally, this work provides additional evidence for the conservation, throughout evolution, of mechanisms regulating chondrocyte proliferation and differentiation.

Alternatively spliced transcripts of Sparus aurata insulin-like growth factor 1 are differentially expressed in adult tissues and during early development.

Spliced variants of insulin-like growth factor 1 (IGF-1), a small peptide with a critical role in metabolism and growth, have been identified in various vertebrate species. However, despite recent functional data in mammalian systems suggesting specific roles (e.g. in muscle formation) for their pro-peptides and/or E domains, their function remains unclear. In this study, three alternatively spliced variants of Sparus aurata proIGF-1 (1a, 1b, and 1c) were identified and their expression analyzed. In adult fish, IGF-1 gene expression was observed in various soft tissues (highest levels in liver) and calcified tissues, with IGF-1c being always the most expressed isoform. In developing larvae, each isoform presented a specific pattern of expression, characterized by different onset and extent and consistent with a possible role of IGF-1a and 1b during early post-hatching events (e.g. bone or muscle formation), while IGF-1c would be rather involved in early larvae formation but probably acts in concerted action with other isoforms at later stages. We also propose that, in adults, IGF-1a and 1b isoforms may have a local action, while isoform 1c would assume a systemic action, as its mammalian counterpart. This hypothesis was further supported by in silico analysis of isoform distribution, revealing that only IGF-1c/Ea isoform has been conserved throughout evolution and that other fish isoforms (i.e. 1a and 1b) may be associated with mechanisms of osmoregulation. We finally propose that IGF-1 variants may exhibit different modes of action (systemic or local) and may be involved in different developmental and adaptive mechanisms.

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.

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.

Enhanced DNA transfer into fish bone cells using polyethylenimine.

The use of in vitro cell culture systems to assess gene function largely depends on the successful transfer of DNA into target cells. Well developed in mammals, transfection methods are still to be optimized for non-mammalian cell culture systems, like fish. Here we describe a rapid, cost-efficient, and successful method to transfer DNA into a fish bone-derived cell line using polyethylenimine (PEI) as the DNA carrier. Using this method, DNA transfer was remarkably enhanced in comparison with commercially available reagents, as demonstrated by the increased activity of both luciferase and green fluorescent protein observed in the transfected cells. Its efficiency in transferring DNA into a wide range of cell types, including non-mammalian and hard-to-transfect cells, in addition to a low cost, show that PEI is a reagent of choice for nonviral vector transfection.

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.