Cellular death of two non-saccharomyces wine-related yeasts during mixed fermentations with saccharomyces cerevisiae

The early death of two non-Saccharomyces wine strains (H. guilliermondii and H. uvarum) during mixed fermentations with S. cerevisiae was studied under enological growth conditions. Several microvinifications were performed in synthetic grape juice, either with single non- Saccharomyces or with mixed S. cerevisiae/non-Saccharomyces inocula. In all mixed cultures, non-Saccharomyces yeasts grew together with S. cerevisiae during the first 1–3 days (depending on the initial inoculum concentration) and then, suddenly, non-Saccharomyces cells began to die off, regardless of the ethanol concentrations present. Conversely, in both non-Saccharomyces single cultures the number of viable cells remained high (ranging 107–108 CFU ml− 1) even when cultures reached significant ethanol concentrations (up to 60–70 g l− 1). Thus, at least for these yeast strains, it seems that ethanol is not the main death-inducing factor. Furthermore, mixed cultures performed with different S. cerevisiae/ H. guilliermondii inoculum ratios (3 :1; 1:2; 1: 10; 1 : 100) revealed that H. guilliermondii death increases for higher inoculum ratios. In order to investigate if the nature of the yeast–yeast interaction was related or not with a cell–cell contact-mediated mechanism, cell-free supernatants obtained from 3 and 6 day-old mixed cultures were inoculated with H. guilliermondii pure cultures. Under these conditions, cells still died and much higher death rates were found for the 6 days than for the 3 day-old supernatants. This strongly indicates that one or more toxic compounds produced by S. cerevisiae triggers the early death of the H. guilliermondii cells in mixed cultures with S. cerevisiae. Finally, although it has not been yet possible to identify the nature of the toxic compounds involved in this phenomenon we must emphasise that the S. cerevisiae strain used in the present work is killer sensitive with respect to the classical killer toxins, K1, K2 and K28, whereas the H. guilliermondii and H. uvarum strains are killer neutral

A genome-wide association study using a DNA pooling strategy identifies BBS9 and GLIS3 as novel loci influencing patient’s outcome after stroke

Stroke is a major cause of morbidity in developed countries and therefore finding adequate treatments to promote patient’s recovery is a priority task, requiring the elucidation of the molecular pathways influencing brain recovery. Few studies, however, have assessed the role of genes in stroke outcome. This study describes a pilot genome-wide association study (GWAS) to identify genetic factors contributing to patient’s outcome, using a DNA pooling design. Methods: Patient’s outcome was assessed using the modified Rankin Scale (mRS) three months after stroke. Using the 250K Affymetrix GeneChip Mapping Assay® – Nsp I, we compared SNP allele frequencies in a pool of non-disabled stroke patients (N=87, mRS=0), with a pool of severely disabled or deceased patients (N=100, mRS>=3). The 100 most interesting SNPs were selected for validation by individual genotyping. Results: 36 SNPs were validated, showing significant differences between patients with extremely good and extremely poor outcome at three months (1.7x10-4 ).This work was supported by the grant PTDC/SAU-GMG/64426/2006, Fundação para a Ciência e Tecnologia (FCT). Helena Manso and Tiago Krug were supported by FCT fellowships

Green zero-valent iron nanoparticles for the degradation of amoxicillin

In the last years, it has been proven that zerovalent iron nanoparticles, including those produced using green methods, are efficient remediation agents for a wide range of target contaminants. However, apart from the known advantages of these green nanomaterials, the knowledge of how they act on distinct contaminants is not yet fully understood and requires further investigation. The objectives of this work were to study the degradation of a common antibiotic, amoxicillin, in water and in a sandy soil using green zero-valent iron nanoparticles (gnZVIs) as reductants and as catalysts for the Fenton reaction. It represents the first study of the use of gnZVI, as alternative for the zero-valent iron nanoparticles produced with sodium borohydride, for the degradation of amoxicillin. The results of the performed tests indicate that gnZVIs have the potential to be used in remediation processes. In both chemical tests, the gnZVI was able to degrade up to 100% of amoxicillin in aqueous solutions, using an amoxicillin/ gnZVI molar ratio of 1:15 when applied as a reductant, and an amoxicillin/H2O2/gnZVI molar ratio of 1:13:1 when applied as a catalyst for the Fenton reaction. The soil tests showed that the required molar ratios for near complete degradation were higher in the reduction test (1:150) than in the gnZVI-catalyzed Fenton reaction (1:1290:73). This is possibly due to parallel reactions with the soil matrix and/or limitations of the reagents to reach the entire soil sample. The degradation efficiencies obtained in these tests were 55 and 97% for the reduction and catalyzed Fenton processes, respectively.info:eu-repo/semantics/publishedVersio

SEGURANÇA PÚBLICA, MUNICÍPIOS E SOCIEDADE: UMA REFLEXÃO SOBRE RESPONSABILIBADES

Resumo: Segurança pública, municípios e sociedade trata-se de um ensaio sobre a reponsabilidade das Administrações municipais e dos cidadãos sobre as questões de segurança pública. A abordagem é iniciada a partir da discussão sobre a relação entre o estabelecimento das redes globais do capitalismo informacional e o processo de desigualdade e exclusão social. Trata-se das atribuições legais dos municípios em relação à segurança pública e sobre os conceitos de “poder de polícia” e “poder da polícia”. Abordam-se, ainda, as questões relacionadas à participação dos cidadãos nas políticas de segurança pública e da chamada segurança cidadã. Exploram-se as possibilidades de uso do poder de polícia do município para prevenção e enfrentamento dos problemas de segurança pública, que vão além da atuação das guardas municipais. Conclui-se que, como ente público mais próximo do cidadão, o município tem um papel importante na promoção de políticas públicas de prevenção e enfrentamento dos problemas de segurança pública. Essas políticas devem ter amplo aspecto, pois muitos problemas de segurança pública estão relacionados a deficiências de infraestrutura, ordenamento do espaço urbano e promoção de direitos básicos dos cidadãos. Conclui-se também que a eficácia e eficiência das políticas de segurança pública estão relacionadas com o nível de envolvimento e participação dos cidadãos com essas políticas, devendo essa participação ser estimulada pela Administração Municipal. Palavras-chave: Segurança Pública. Segurança Cidadã. Municípios. Poder de Polícia. Gestão de Cidades. Abstract: Public Security, city and society is an article about the responsibility of municipal administrations and citizens on matters regarding public security.  The approach starts from the discussion about the relationship between establishing global networks of informational capitalism and the process of inequity and social exclusion. It is about the legal allocation of municipalities concerning public security about the concepts of “The police power” and “The power of the police”. It also addresses matters related to the participation of citizens in public security policies and the so-called citizen security. It explores the possibilities of using the police power of the city on preventing and confronting public security issues, which goes beyond the function of city guards. It has been concluded as the citizen’s closest party, the municipality has an important role in promoting public policies on preventing and confronting public security issues. These policies must have a wide aspect since many problems of public security are related to infrastructure shortcoming, urban space management, and promoting citizens’ basic rights. It has been also concluded that effectiveness and efficiency of public policies are related to the level of commitment and participation of the citizen with these policies, and this participation has to be promoted by the city administration. Keywords: Public Security. City. Power of Police. Municipal administration

Mitochondrial haplogroup H1 is protective for ischemic stroke in Portuguese patient

BACKGROUND: The genetic contribution to stroke is well established but it has proven difficult to identify the genes and the disease-associated alleles mediating this effect, possibly because only nuclear genes have been intensely investigated so far. Mitochondrial DNA (mtDNA) has been implicated in several disorders having stroke as one of its clinical manifestations. The aim of this case-control study was to assess the contribution of mtDNA polymorphisms and haplogroups to ischemic stroke risk. METHODS: We genotyped 19 mtDNA single nucleotide polymorphisms (SNPs) defining the major European haplogroups in 534 ischemic stroke patients and 499 controls collected in Portugal, and tested their allelic and haplogroup association with ischemic stroke risk. RESULTS: Haplogroup H1 was found to be significantly less frequent in stroke patients than in controls (OR = 0.61, 95% CI = 0.45-0.83, p = 0.001), when comparing each clade against all other haplogroups pooled together. Conversely, the pre-HV/HV and U mtDNA lineages emerge as potential genetic factors conferring risk for stroke (OR = 3.14, 95% CI = 1.41-7.01, p = 0.003, and OR = 2.87, 95% CI = 1.13-7.28, p = 0.021, respectively). SNPs m.3010G>A, m.7028C>T and m.11719G>A strongly influence ischemic stroke risk, their allelic state in haplogroup H1 corroborating its protective effect. CONCLUSION: Our data suggests that mitochondrial haplogroup H1 has an impact on ischemic stroke risk in a Portuguese sample

Kalirin: a novel genetic risk factor for ischemic stroke

Cerebrovascular and cardiovascular diseases are the leading causes of death and disability worldwide. They are complex disorders resulting from the interplay of genetic and environmental factors, and may share several susceptibility genes. Several recent studies have implicated variants of the Kalirin (KALRN) gene with susceptibility to cardiovascular and metabolic phenotypes, but no studies have yet been performed in stroke patients. KALRN is involved, among others, in the inhibition of inducible nitric oxide synthase, in the regulation of ischemic signal transduction, and in neuronal morphogenesis, plasticity, and stability. The goal of the present study was to determine whether SNPs in the KALRN region on 3q13, which includes the Ropporin gene (ROPN1), predispose to ischemic stroke (IS) in a cohort of Portuguese patients and controls. We genotyped 34 tagging SNPs in the KALRN and ROPN1 chromosomal region on 565 IS patients and 517 unrelated controls, and performed genotype imputation for 405 markers on chromosome 3. We tested the single-marker association of these SNPs with IS. One SNP (rs4499545) in the ROPN1-KALRN intergenic region and two SNPs in KALRN (rs17286604 and rs11712619) showed significant (P < 0.05) allelic and genotypic (unadjusted and adjusted for hypertension, diabetes, and ever smoking) association with IS risk. Thirty-two imputed SNPs also showed an association at P < 0.05, and actual genotyping of three of these polymorphisms (rs7620580, rs6438833, and rs11712039) validated their association. Furthermore, rs11712039 was associated with IS (0.001 < P < 0.01) in a recent well-powered genomewide association study (Ikram et al. 2009). These studies suggest that variants in the KALRN gene region constitute risk factors for stroke and that KALRN may represent a common risk factor for vascular diseases

FOXA1 repression is associated with loss of BRCA1 and increased promoter methylation and chromatin silencing in breast cancer

FOXA1 expression correlates with the breast cancer luminal subtype and patient survival. RNA and protein analysis of a panel of breast cancer cell lines revealed that BRCA1 deficiency is associated with the downregulation of FOXA1 expression. Knockdown of BRCA1 resulted in the downregulation of FOXA1 expression and enhancement of FOXA1 promoter methylation in MCF-7 breast cancer cells, whereas the reconstitution of BRCA1 in Brca1-deficent mouse mammary epithelial cells (MMECs) promoted Foxa1 expression and methylation. These data suggest that BRCA1 suppresses FOXA1 hypermethylation and silencing. Consistently, the treatment of MMECs with the DNA methylation inhibitor 5-aza-2'-deoxycitydine induced Foxa1 mRNA expression. Furthermore, treatment with GSK126, an inhibitor of EZH2 methyltransferase activity, induced FOXA1 expression in BRCA1-deficient but not in BRCA1-reconstituted MMECs. Likewise, the depletion of EZH2 by small interfering RNA enhanced FOXA1 mRNA expression. Chromatin immunoprecipitation (ChIP) analysis demonstrated that BRCA1, EZH2, DNA methyltransferases (DNMT)1/3a/3b and H3K27me3 are recruited to the endogenous FOXA1 promoter, further supporting the hypothesis that these proteins interact to modulate FOXA1 methylation and repression. Further co-immunoprecipitation and ChIP analysis showed that both BRCA1 and DNMT3b form complexes with EZH2 but not with each other, consistent with the notion that BRCA1 binds to EZH2 and negatively regulates its methyltransferase activity. We also found that EZH2 promotes and BRCA1 impairs the deposit of the gene silencing histone mark H3K27me3 on the FOXA1 promoter. These associations were validated in a familial breast cancer patient cohort. Integrated analysis of the global gene methylation and expression profiles of a set of 33 familial breast tumours revealed that FOXA1 promoter methylation is inversely correlated with the transcriptional expression of FOXA1 and that BRCA1 mutation breast cancer is significantly associated with FOXA1 methylation and downregulation of FOXA1 expression, providing physiological evidence to our findings that FOXA1 expression is regulated by methylation and chromatin silencing and that BRCA1 maintains FOXA1 expression through suppressing FOXA1 gene methylation in breast cancer.Oncogene advance online publication, 22 December 2014; doi:10.1038/onc.2014.421.published_or_final_versio

Lymphatic vessel density and VEGF-C expression are significantly different among benign and malignant thyroid lesions

Thyroid cancer is the most frequent endocrine neoplasia worldwide. The route for metastasis and loco-regional invasion preferentially occurs by lymphatic vessels. For this reason, the assessment of lymphatic vessel density (LVD) is supposed to represent both a prognostic parameter and also a potential therapeutic target. In order to evaluate the value of LVD in benign and malignant thyroid lesions, we analyzed 110 thyroidectomy specimens using D2-40, a specific marker for lymphatic vessels and vascular endothelial growth factor C (VEGF-C), the most potent molecule of lymphatic proliferation. LVD was significantly different between papillary and follicular carcinomas in total (p = 0.045) and peritumoral area (p = 0.042). Follicular adenoma and follicular carcinoma showed an important difference of intra- (p = 0.019) and peritumoral (p = 0.033) LVD. VEGF-C was more markedly expressed in malignancies than in benign lesions (p = 0.0001). Almost all cancers with high positive VEGF-C expression also exhibited increased peritumoral LVD (p = 0.049) when compared with the benign lesions. Indeed, the high peritumoral LVD of malignant thyroid lesions is an important finding for surgery planning and supports the practice of total thyroidectomy in malignant thyroid neoplasm's since the lymphatic peritumoral vessels definitely are an escape path for tumor cells

Expression of FOXA1 and GATA-3 in breast cancer: the prognostic significance in hormone receptor-negative tumours

The expression of additional genes, other than oestrogen receptor (ER), may be important to the hormone-responsive phenotype of breast cancer. Microarray analyses have revealed that forkhead box A1 (FOXA1) and GATA binding protein 3 (GATA-3) are expressed in close association with ERalpha, both encoding for transcription factors with a potential involvement in the ERalpha-mediated action in breast cancer. The purpose of this study was to explore if the expression of FOXA1 and GATA-3 may provide an opportunity to stratify subsets of patients that could have better outcome, among the ERalpha-negative/poor prognosis breast cancer group.The present study was supported by a research grant (SFRH/BD/15316/ 2005 to AA) financed by the Portuguese Science and Technology Foundation (FCT). The authors thank Prof. Raquel Seruca ( coordinator from the Cancer Genetics group at IPATIMUP) for scientific assistance, Dr Jose Luis Costa (postdoctorate at IPATIMUP) for critically reading the manuscript before submission, and Dr Nuno Marcos ( PhD student at IPATIMUP) for artwork assistance