MicroRNA-targeting in spermatogenesis: Over-expressions of microRNA-23a/b-3p and its affected targeting of the genes ODF2 and UBQLN3 in spermatozoa of patients with oligoasthenozoospermia

Background Male infertility is a multifactorial syndrome with diverse phenotypic representations. MicroRNAs (miRNAs) are small, non-coding RNAs that are involved in the post-transcriptional regulation of gene expression. Altered abundance levels of ODF2 and UBQLN3 have been reported in patients with different spermatogenic impairments. However, the transcriptional regulation of these two genes by miR-23a/b-3p is still unclear. Objectives To investigate experimentally whether miR-23a/b-3p targets the genes ODF2 and UBQLN3 and whether this targeting impacts abundance levels of ODF2 and UBQLN3 in patients with oligoasthenozoospermia. Materials and methods A total of 92 men attending a fertility clinic were included in the study, including 46 oligoasthenozoospermic men and 46 age-matched normozoospermic volunteers who served as controls. Reverse transcription-quantitative PCR (RT-qPCR), Western blot, and dual-luciferase (Firefly-Renilla) assays were used to validate the miRNAs and their target genes. Results RT-qPCR revealed that miR-23a/b-3p was more abundant and ODF2 and UBQLN3 targets were less abundant in men with impaired spermatogenesis. Besides, Western blot shows that ODF2 and UBQLN3 protein levels were reduced in men with impaired spermatogenesis. In silico prediction and dual-luciferase assays revealed that potential links exist between the higher abundance level of miR-23a/b-3p and the lower abundance level of ODF2 and UBQLN3 targets. Mutations in the miR-23a/b-3p-binding site within the 3ˊUTRs (3ˊuntranslated regions) of ODF2 and UBQLN3 genes resulted in abrogated responsiveness to miR-23a/b-3p. Correlation analysis showed that sperm count, motility, and morphology were negatively correlated with miR-23a/b-3p and positively correlated with the lower abundance level of UBQLN3, while ODF lower abundance level was positively correlated with sperm motility. Conclusion Findings indicate that the higher abundance level of miR-23a/b-3p and the lower abundance level of ODF2 and UBQLN3 targets are associated with oligoasthenozoospermia and male subfertility

Health literacy in patients with pulmonary embolism: development and validation of the HeLP (Health Literacy in Pulmonary Embolism)-Questionnaire

Background: Pulmonary embolism (PE) is a common cardiovascular disease and health literacy is necessary to deal with its consequences after the acute event. The aim of this study was to develop and validate a new questionnaire to measure PE-specific health literacy. Methods: A mixed-methods design with qualitative and quantitative elements was used in the development process. A literature review about health literacy concepts and instruments and interviews with patients with PE and clinicians were conducted. Quantitative analyses included factor analyses, item response theory with a graded partial credit model, and reliability analyses in different test and validation samples. Furthermore, convergent and known-groups validity and responsiveness were assessed. Results: The qualitative results supported a concept of PE-related health literacy with four main topics: dealing with PE-related health information, disease management, health-related selfcare, and social support. An initial item pool of 91 items was developed. Further interviews and an online survey with patients with PE (n = 1,013) were used to reduce the number of items and to confirm structural validity. Confirmatory factor analyses in the final evaluation study with patients with PE (n = 238) indicated a good model fit of the four-factor structure. The Health Literacy in Pulmonary Embolism (HeLP)-Questionnaire showed good reliability (Cronbach’s alpha: 0.82 to 0.90). All four subscales were responsive toward receiving a brochure with PE-related health information. Conclusion: The newly developed German HeLP Questionnaire comprises 23 items in four domains and showed good psychometric properties. Further evaluation of the questionnaire in different samples of patients with PE is needed

Integrated microRNA and mRNA Expression Profiling Identifies Novel Targets and Networks Associated with Ebstein’s Anomaly

Little is known about abundance level changes of circulating microRNAs (miRNAs) and messenger RNAs (mRNA) in patients with Ebstein’s anomaly (EA). Here, we performed an integrated analysis to identify the differentially abundant miRNAs and mRNA targets and to identify the potential therapeutic targets that might be involved in the mechanisms underlying EA. A large panel of human miRNA and mRNA microarrays were conducted to determine the genome-wide expression profiles in the blood of 16 EA patients and 16 age and gender-matched healthy control volunteers (HVs). Differential abundance level of single miRNA and mRNA was validated by RealTime quantitative PCR (RT-qPCR). Enrichment analyses of altered miRNA and mRNA abundance levels were identified using bioinformatics tools. Altered miRNA and mRNA abundance levels were observed between EA patients and HVs. Among the deregulated miRNAs and mRNAs, 76 miRNAs (49 lower abundance and 27 higher abundance, fold-change of ≥2) and 29 mRNAs (25 higher abundance and 4 lower abundance, fold-change of ≥1.5) were identified in EA patients compared to HVs. Bioinformatics analysis identified 37 pairs of putative miRNA-mRNA interactions. The majority of the correlations were detected between the lower abundance level of miRNA and higher abundance level of mRNA, except for let-7b-5p, which showed a higher abundance level and their target gene, SCRN3, showed a lower abundance level. Pathway enrichment analysis of the deregulated mRNAs identified 35 significant pathways that are mostly involved in signal transduction and cellular interaction pathways. Our findings provide new insights into a potential molecular biomarker(s) for the EA that may guide the development of novel targeting therapies

Characterization of micro-RNA in women with different ovarian reserve

Women undergoing infertility treatment are routinely subjected to one or more tests of ovarian reserve. Therefore, an adequate assessment of the ovarian reserve is necessary for the treatment. In this study, we aimed to characterize the potential role of microRNAs (miRNAs) as biomarkers for women with different ovarian reserves. A total of 159 women were recruited in the study and classified according to their anti-Müllerian hormone (AMH) level into three groups: (1) low ovarian reserve (LAMH, n = 39), (2) normal ovarian reserve (NAMH, n = 80), and (3) high ovarian reserve (HAMH, n = 40). SurePrint Human miRNA array screening and reverse transcription-quantitative PCR (RT-qPCR) were respectively employed to screen and validate the miRNA abundance level in the three tested groups. Compared with NAMH, the abundance level of 34 and 98 miRNAs was found to be significantly altered in LAMH and HAMH, respectively. The abundance level of miRNAs was further validated by RT-qPCR in both, the screening samples as well as in an independent set of validation samples. The abundance levels of the validated miRNAs were significantly correlated with the AMH level. The best AUC value for the prediction of the increase and decrease in the AMH level was obtained for the miR-100-5p and miR-21-5p, respectively. The level of miRNAs abundance correlates with the level of AMH, which may serve as a tool for identifying women with a different ovarian reserve and may help to lay the ground for the development of novel diagnostic approaches

Ising Spin Glasses in a Magnetic Field

Ground states of the three dimensional Edwards-Anderson spin glass are computed in the presence of an external magnetic field. Our algorithm is sufficiently powerful for us to treat systems with up to 600 spins. We perform a statistical analysis of how the ground state changes as the field is increased, and reach the conclusion that the spin glass phase at zero temperature does not survive in the presence of any finite field. This is in agreement with the droplet model or scaling predictions, but in sharp disagreement with the mean field picture. For comparison, we also investigate a dilute mean field spin glass model where an Almeida-Thouless line is present.Comment: 4 pages, 4 figures, Revte

Estudo do Pré-Tratamento Ácido e Hidrólise Enzimática da Borra de Café Visando a Produção de Bioetanol

O presente trabalho tem como objetivo avaliar a possibilidade de produzir um biocombustível, o bioetanol, valorizando resíduos de baixo valor, nomeadamente as borras de café. Temos cada vez mais assistido a uma evolução tecnológica em larga escala e a um aumento populacional, que se traduz numa vertente positiva no primeiro caso, mas devido ao crescente consumo de energia pela sociedade atual e alteração de padrões de consumo, origina uma grande dependência de utilização de combustíveis fósseis, o que levará a um previsível esgotamento das suas reservas naturais. No que respeita ao aumento populacional, este provoca um aumento da produção de resíduos. Assim, cada vez mais se torna necessário procurar recursos energéticos alternativos, de natureza renovável, designadamente através da produção de biocombustíveis a partir de resíduos de baixo valor comercial. Esta utilização de resíduos como matéria-prima para a produção de biocombustíveis traduz-se em duas vantagens: i) por um lado diminui-se a acumulação dos resíduos gerados, e ii) por outro, podem estes ser aproveitados de uma forma eficiente. O trabalho foi realizado em duas etapas fundamentais: a caraterização físico-química da borra de café e a quantificação dos açúcares formados, após aplicação do pré-tratamento ácido e hidrólise enzimática, por três métodos: DNS (ácido 3,5-dinitrosalicílico), refratometria e cromatografia líquida de alta eficiência (HPLC). Para a caraterização físico-química da borra de café analisaram-se vários parâmetros, dos quais se salientam, o poder calorífico (5082,82 kcal/kg), o teor de cinzas (1,83%), o teor de proteínas (13,74 g / 100gcafé), o teor de celulose (16,17%) e o teor de lenhina (29,04%). No pré-tratamento ácido utilizou-se concentrações (v/v) de 1% a 5% dos ácidos H2SO4, HCI, HNO3, CH3COOH e de 7% para o CH3COOH, a uma temperatura de 85 ºC, com tempo de reação de 270 min e agitação de 50 rpm. A hidrólise enzimática foi concretizada num banho a 50 ºC, durante 120 minutos, mantendo a mesma velocidade de agitação do pré-tratamento e recorrendo à enzima Viscozyme L, na proporção de 100 L enzima/g amostra. A eficiência do pré-tratamento e hidrólise enzimática foi avaliada através da quantificação dos açúcares formados. Na quantificação pelo método DNS, o ácido sulfúrico e nítrico a 3% foram os que apresentaram rendimentos superiores de açúcares redutores totais, com 43,1% e 43,2%, respetivamente, seguido do ácido nítrico a 5%, cujo rendimento igualou os 41,4%. Na determinação dos açúcares totais, realizada por refratometria, confirmou-se que o ácido sulfúrico a 5% foi aquele que apresentou um rendimento mais alto - 78,9% - seguido da concentração de 3% com 65,3%. Finalmente, no método de HPLC, o ácido sulfúrico a 5% proporcionou rendimentos mais elevados de xilose, arabinose e glucose, que tomaram os valores de 6,1%, 9,3% e 11,0%, respetivamente.The present work aims to evaluate the possibility of producing a biofuel, bioethanol, by valuing low value residues, namely coffee grounds. We have been increasingly witnessing a large-scale technological evolution and a population increase, which translates into a positive aspect in the first case, but, due to the increasing consumption of energy by the current society and changes in consumption patterns, causes a great dependence on the use of fossil fuels, which will lead to a predictable depletion of their natural reserves. As regards population growth, this causes an increase in waste production. Thus, it is increasingly necessary to look for alternative energy resources of a renewable nature, namely through the production of biofuels from low commercial value waste. This use of waste as a raw material for the production of biofuels has two advantages: (i) on the one hand, the accumulation of generated waste is reduced, and (ii) on the other, it can be used efficiently. This work was carried out in two fundamental stages: the physical-chemical characterization of coffee grounds and the quantification of the sugars formed, after application of acid pretreatment and enzymatic hydrolysis, by three methods: DNS (3,5-Dinitrosalicylic acid), refractometry and high performance liquid chromatography (HPLC). In the chemical characterization of coffee grounds, several parameters were used, such as calorific value (5082,82 cal/g), ash content (1,83%), protein (13,74 g/100gcoffee), cellulose (16,17%) and lignin (29,04%). In acid pre-treatment, 1% to 5% concentrations of H2SO4, HCl, HNO3, CH3COOH acids and 7% of CH3COOH were used (v/v) at a temperature of 85°C with a reaction time of 270 min and stirring at 50 rpm. The enzymatic hydrolysis was carried out in a bath at 50°C for 120 minutes, maintaining the same stirring speed of the pretreatment and using the enzyme Viscozyme L in the proportion of 100 μl enzyme/g sample. The efficiency of pretreatment and enzymatic hydrolysis was evaluated by quantifying the sugars formed. In the DNS method, 3% sulfuric and nitric acid were those that presented higher yields of total reducing sugars, with 43,1% and 43,2%, respectively, followed by 5% nitric acid, whose yield was equal to 41,4%. In the determination of total sugars, performed by refractometry, 5% sulfuric acid was the one that presented a higher yield – 78,9% - followed by the concentration of 3% with 65,3%. Finally, in the HPLC method, 5% sulfuric acid provided higher yields of xylose, arabinose and glucose, which took the values of 6,1%, 9,3% and 11%, respectively

Time decay of the remanent magnetization in the ±J\pm J spin glass model at T=0

Using the zero-temperature Metropolis dynamics, the time decay of the remanent magnetization in the $\pm J$ Edward-Anderson spin glass model with a uniform random distribution of ferromagnetic and antiferromagnetic interactions has been investigated. Starting from the saturation, the magnetization per spin $m$ reveals a slow decrease with time, which can be approximated by a power law:$m(t)=m_{\infty}+ ({t\over a_{0}})^{a_{1}}$, $a_{1} < 0$. Moreover, its relaxation does not lead it into one of the ground states, and therefore the system is trapped in metastable isoenergetic microstates remaining magnetized. Such behaviour is discussed in terms of a random walk the system performs on its available configuration space.Comment: 9 pages, 3 figure

Climate scenarios for Switzerland CH2018 - approach and implications

To make sound decisions in the face of climate change, government agencies, policymakers and private stakeholders require suitable climate information on local to regional scales. In Switzerland, the development of climate change scenarios is strongly linked to the climate adaptation strategy of the Confederation. The current climate scenarios for Switzerland CH2018 - released in form of six user-oriented products - were the result of an intensive collaboration between academia and administration under the umbrella of the National Centre for Climate Services (NCCS), accounting for user needs and stakeholder dialogues from the beginning. A rigorous scientific concept ensured consistency throughout the various analysis steps of the EURO-CORDEX projections and a common procedure on how to extract robust results and deal with associated uncertainties. The main results show that Switzerland?s climate will face dry summers, heavy precipitation, more hot days and snow-scarce winters. Approximately half of these changes could be alleviated by mid-century through strong global mitigation efforts. A comprehensive communication concept ensured that the results were rolled out and distilled in specific user-oriented communication measures to increase their uptake and to make them actionable. A narrative approach with four fictitious persons was used to communicate the key messages to the general public. Three years after the release, the climate scenarios have proven to be an indispensable information basis for users in climate adaptation and for downstream applications. Potential for extensions and updates has been identified since then and will shape the concept and planning of the next scenario generation in Switzerland

Upper bounds on the density of states of single Landau levels broadened by Gaussian random potentials

We study a non-relativistic charged particle on the Euclidean plane R^2 subject to a perpendicular constant magnetic field and an R^2-homogeneous random potential in the approximation that the corresponding random Landau Hamiltonian on the Hilbert space L^2(R^2) is restricted to the eigenspace of a single but arbitrary Landau level. For a wide class of Gaussian random potentials we rigorously prove that the associated restricted integrated density of states is absolutely continuous with respect to the Lebesgue measure. We construct explicit upper bounds on the resulting derivative, the restricted density of states. As a consequence, any given energy is seen to be almost surely not an eigenvalue of the restricted random Landau Hamiltonian.Comment: 16 pages, to appear in "Journal of Mathematical Physics

Perfil glucometabólico inicial en pacientes con síndrome coronario agudo y síndrome metabólico

BACKGROUND: Patients with metabolic syndrome (MetS) are at high coronary risk and beta-cell dysfunction or insulin resistance might predict an additional risk for early cardiovascular events. OBJECTIVE: This study aimed to evaluate early glucometabolic alterations in patients with MetS, but without previously known type 2 diabetes, after acute coronary syndrome. METHODS: A total of 114 patients were submitted to an oral glucose tolerance test (OGTT) 1-3 days after hospital discharge due to myocardial infarction or unstable angina. Based on the OGTT, we defined three groups of patients: normal glucose tolerance (NGT; n=26), impaired glucose tolerance (IGT; n=39), or diabetes (DM; n=49). The homeostasis model assessment (HOMA-IR) was used to measure insulin resistance; beta-cell responsiveness was assessed by the insulinogenic index at 30 min (ΔI30/ΔG30). RESULTS: Based on the HOMA-IR, patients with DM were more insulin-resistant than those with NGT or IGT (p<0.001). According to the insulinogenic index, the beta-cell responsiveness was also impaired in subjects with DM (p<0.001 vs NGT or IGT). CONCLUSION: High rates of glucometabolic alterations were found after acute coronary syndrome in patients with MetS. As these abnormalities markedly increase the risk for adverse outcomes, early OGTT among MetS patients might be used to identify those at the highest coronary risk.FUNDAMENTO: Pacientes con síndrome metabólico (SM) tienen alto riesgo coronario y la disfunción de la célula beta o la resistencia a la insulina puede prever un riesgo adicional de eventos cardiovasculares precoces. OBJETIVO: Evaluar las alteraciones glucometabólicas precoces en pacientes con SM, pero sin diagnóstico de diabetes tipo 2, tras el síndrome coronario agudo. MÉTODOS: Un total de 114 pacientes fue sometido a la prueba oral de tolerancia a la glucosa (POTG), de un a tres días tras el alta hospitalaria, y luego de infarto agudo de miocardio o angina inestable. Basado en el POTG, definimos tres grupos de pacientes: tolerancia normal a la glucosa (TNG; n=26), tolerancia alterada a la glucosa (TAG; n=39) o diabetes mellitus (DM; n=49). Se utilizó el Modelo de Evaluación de la Homeostasis (HOMA-IR) para estimarse la resistencia a la insulina; se evaluó la responsividad de la célula beta a través del índice insulinogénico de 30 minutos (ΔI30/ΔG30). RESULTADOS: Basado en el HOMA-IR, los pacientes con DM se mostraban más insulinoresistentes que los individuos con TNG o TAG (p<0,001). De acuerdo con el índice insulinogénico, la responsividad de la célula beta también estaba alterada en individuos con DM (p<0,001 vs. TNG o TAG). CONCLUSIONES: Se encontraron altas tasas de alteraciones glucometabólicas tras el síndrome coronario agudo en pacientes con SM. Como esas anormalidades incrementan acentuadamente el riesgo de desenlaces adversos, el POTG precoz se puede utilizar en pacientes con SM para identificar a los que presentan mayor riesgo coronario.FUNDAMENTO: Pacientes com síndrome metabólica (SM) têm alto risco coronariano e a disfunção da célula beta ou resistência à insulina pode prever um risco adicional de eventos cardiovasculares precoces. OBJETIVO: Avaliar as alterações glicometabólicas precoces em pacientes com SM, mas sem diagnóstico de diabete tipo 2, após síndrome coronariana aguda. MÉTODOS: Um total de 114 pacientes foi submetido ao teste oral de tolerância à glicose (TOTG), 1-3 dias da alta hospitalar, após infarto agudo do miocárdio ou angina instável. Baseado no TOTG, definimos três grupos de pacientes: tolerância normal à glicose (TNG; n=26), tolerância alterada à glicose (TAG; n=39) ou diabetes mellitus (DM; n=49). O Modelo de Avaliação da Homeostase (HOMA-IR) foi usado para estimar a resistência à insulina; a responsividade da célula beta foi avaliada através do índice insulinogênico de 30 minutos (ΔI30/ΔG30). RESULTADOS: Baseado no HOMA-IR, os pacientes com DM eram mais insulino-resistentes do que aqueles com TNG ou TAG (p<0,001). De acordo com o índice insulinogênico, a responsividade da célula beta também estava alterada em indivíduos com DM (p<0,001 vs TNG ou TAG). CONCLUSÃO: Altas taxas de alterações glicometabólicas foram encontradas após síndrome coronariana aguda em pacientes com SM. Como essas anormalidades acentuadamente aumentam o risco de desfechos adversos, o TOTG precoce pode ser utilizado em pacientes com SM para identificar aqueles que apresentam maior risco coronariano.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Federal de São Paulo (UNIFESP)UNIFESPSciEL