Mutagen Killing and Photoreactivation in the Anaerobic Fungus Neocallimastix frontalis EB188

Research was performed to determine if the anaerobic fungus, Neocallimastix frontalis EB 188, a common cellulolytic fungus of ruminants, is capable of photoreactivation and is susceptible to chemical and irradiational mutagenesis. Germination of zoospores and production of colony cellulase was measured after ultraviolet light (UVL), nitrosoguanidine or ethyl methyl sulfonate treatments. This fungus was susceptible to mutagen treatment and capable of photoreactivation after UVL exposure. Such procedures may be useful in the isolation of enhanced cellulase-producing strains

Enzyme-based DNA extraction from zoospores of ruminal fungi

We report here a rapid, efficient and simple method for the extraction of high molecular weight DNA from zoospores of Neocallimastix frontalis EB188. This anaerobic fungus, isolated from bovine digesta, effectively degrades plant fiber in vitro (Barichievich et al. 1990. Appl. Env. Micro. 56:43-48). Our interest in ruminal fungi stems from their ability to degrade wood materials (Joblin et al. 1989. FEMS Micro. Lett. 56:119-122) and their potential use in biomass saccharification. Zoospore DNA synthesis is of particular interest to our laboratory. It is these motile zoospores which colonize and degrade plant materials (Mountfort 1987. FEMS Micro. Rev. 46:501-508). To detail fully this metabolic event, it will be necessary to extract nucleic acids from zoospores. Such procedures have not been reported in the literature. Using acetone drying and enzymatic removal of cell walls, we have isolated high molecular weight DNA from very small amounts of culture. The procedure takes less than one hour and DNA yields are high. The DNA is readily cut with restriction endonucleases and religated efficiently but is otherwise stable. Electrophoretic analysis of the DNA confirmed the presence of repetitive sequences. This procedure will aid the study of DNA replication, DNA repair and DNA RFLP analysis of various strains using small (\u3c1 ml) cultures

Comparative analysis of mesenchymal stromal cells biological properties

The stromal progenitors of mesodermal cells, mesenchymal stromal cells (MSCs), are a heterogeneous population of plastic adherent fibroblast-like cells with extensive proliferative capacity and differentiation potential. Human MSCs have now been isolated from various tissues including bone marrow, muscle, skin, and adipose tissue, the latter being one of the most suitable cell sources for cell therapy, because of its easy accessibility, minimal morbidity, and abundance of cells. Bone marrow and subcutaneous or visceral adipose tissue samples were collected, digested with collagenase if needed, and seeded in Iscove's medium containing 5% human platelet lysate. Nonadherent cells were removed after 2-3 days and the medium was replaced twice a week. Confluent adherent cells were detached, expanded, and analyzed for several biological properties such as morphology, immunophenotype, growth rate, senescence, clonogenicity, differentiation capacity, immunosuppression, and secretion of angiogenic factors. The results show significant differences between lines derived from subcutaneous fat compared to those derived from visceral fat, such as the higher proliferation rate of the first and the strong induction of angiogenesis of the latter. We are convinced that the identification of the peculiarities of MSCs isolated from different tissues will lead to their more accurate use in cell therapy

Long-term effect of neonatal inhibition of APP gamma-secretase on hippocampal development in the Ts65Dn mouse model of Down syndrome

Neurogenesis impairment is considered a major determinant of the intellectual disability that characterizes Down syndrome (DS), a genetic condition caused by triplication of chromosome 21. Previous evidence obtained in the Ts65Dn mouse model of DS showed that the triplicated gene APP (amyloid precursor protein) is critically involved in neurogenesis alterations. In particular, excessive levels of AICD (amyloid precursor protein intracellular domain) resulting from APP cleavage by gamma-secretase increase the transcription of Ptch1, a Sonic Hedgehog (Shh) receptor that keeps the mitogenic Shh pathway repressed. Previous evidence showed that neonatal treatment with ELND006, an inhibitor of gamma-secretase, reinstates the Shh pathway and fully restores neurogenesis in Ts65Dn pups. In the framework of potential therapies for DS, it is extremely important to establish whether the positive effects of early intervention are retained after treatment cessation. Therefore, the goal of the current study was to establish whether early treatment with ELND006 leaves an enduring trace in the brain of Ts65Dn mice. Ts65Dn and euploid pups were treated with ELND006 in the postnatal period P3-P15 and the outcome of treatment was examined at ~ one month after treatment cessation. We found that in treated Ts65Dn mice the pool of proliferating cells in the hippocampal dentate gyrus (DG) and total number of granule neurons were still restored as was the number of pre- and postsynaptic terminals in the stratum lucidum of CA3, the site of termination of the mossy fibers from the DG. Accordingly, patch-clamp recording from field CA3 showed functional normalization of the input to CA3. Unlike in field CA3, the number of pre- and postsynaptic terminals in the DG of treated Ts65Dn mice was no longer fully restored. The finding that many of the positive effects of neonatal treatment were retained after treatment cessation provides proof of principle demonstration of the efficacy of early inhibition of gamma-secretase for the improvement of brain development in DS

Legionnaires' disease associated with macular rash: two cases.

Legionnaires' disease is an acute bacterial infection, generally sustained by Legionella pneumophila, which involves primarily the lower respiratory tract, although it is often associated with multi-systemic extrapulmonary manifestations. Afflicted patients may sometimes have gastrointestinal symptoms, liver function abnormalities, renal failure or central nervous system complications, while cutaneous manifestations are very uncommon and may include erythematous, maculopapular or petechial skin lesions. Pathogenesis of skin involvement in the setting of Legionnaires' disease is still uncertain, but may involve toxic or immunological mechanisms. Two exceptional cases of Legionella pneumonia complicated by diffuse, macular rash in two adult women are described, in association with severe peripheral polyneuropathy and flaccid quadriplegia in one case

Rare genetic variant burden in DPYD predicts severe fluoropyrimidine-related toxicity risk

Preemptive targeted pharmacogenetic testing of candidate variations in DPYD is currently being used to limit toxicity associated with fluoropyrimidines. The use of innovative next generation sequencing (NGS) approaches could unveil additional rare (minor allele frequency <1%) genetic risk variants. However, their predictive value and management in clinical practice are still controversial, at least partly due to the challenges associated with functional analyses of rare variants. The aim of this study was to define the predictive power of rare DPYD variants burden on the risk of severe fluoropyrimidine-related toxicity. The DPYD coding sequence and untranslated regions were analyzed by NGS in 120 patients developing grade 3–5 (NCI-CTC vs3.0) fluoropyrimidine-related toxicity and 104 matched controls (no-toxicity). The functional impact of rare variants was assessed using two different in silico predictive tools (i.e., Predict2SNP and ADME Prediction Framework) and structural modeling. Plasma concentrations of uracil (U) and dihydrouracil (UH2) were quantified in carriers of the novel variants. Here, we demonstrate that the burden of rare variants was significantly higher in patients with toxicity compared to controls (p = 0.007, Mann-Whitney test). Carriers of at least one rare missense DPYD variant had a 16-fold increased risk in the first cycle and an 11-fold increased risk during the entire course of chemotherapy of developing a severe adverse event compared to controls (p = 0.013 and p = 0.0250, respectively by multinomial regression model). Quantification of plasmatic U/UH2 metabolites and in silico visualization of the encoded protein were consistent with the predicted functional effect for the novel variations. Analysis and consideration of rare variants by DPYD-sequencing could improve prevention of severe toxicity of fluoropyrimidines and improve patients’ quality of life

Monte Belo: características da identidade regional para uma indicação geográfica de vinhos.

bitstream/CNPUV/9754/1/cir076.pdfDisponível também no formato online

Articles by Latin American Authors in Prestigious Journals Have Fewer Citations

Background: the journal Impact factor (IF) is generally accepted to be a good measurement of the relevance/quality of articles that a journal publishes. in spite of an, apparently, homogenous peer-review process for a given journal, we hypothesize that the country affiliation of authors from developing Latin American (LA) countries affects the IF of a journal detrimentally.Methodology/Principal Findings: Seven prestigious international journals, one multidisciplinary journal and six serving specific branches of science, were examined in terms of their IF in the Web of Science. Two subsets of each journal were then selected to evaluate the influence of author's affiliation on the IF. They comprised contributions (i) with authorship from four Latin American (LA) countries (Argentina, Brazil, Chile and Mexico) and (ii) with authorship from five developed countries (England, France, Germany, Japan and USA). Both subsets were further subdivided into two groups: articles with authorship from one country only and collaborative articles with authorship from other countries. Articles from the five developed countries had IF close to the overall IF of the journals and the influence of collaboration on this value was minor. in the case of LA articles the effect of collaboration (virtually all with developed countries) was significant. the IFs for non-collaborative articles averaged 66% of the overall IF of the journals whereas the articles in collaboration raised the IFs to values close to the overall IF.Conclusion/Significance: the study shows a significantly lower IF in the group of the subsets of non-collaborative LA articles and thus that country affiliation of authors from non-developed LA countries does affect the IF of a journal detrimentally. There are no data to indicate whether the lower IFs of LA articles were due to their inherent inferior quality/relevance or psycho-social trend towards under-citation of articles from these countries. However, further study is required since there are foreseeable consequences of this trend as it may stimulate strategies by editors to turn down articles that tend to be under-cited.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Latin Amer & Caribbean Ctr Hlth Sci Informat, BIREME PAHO WHO, São Paulo, BrazilUniversidade Federal de São Paulo, DIS Dept Informat Med, São Paulo, BrazilUniversidade Federal de São Paulo, DIS Dept Informat Med, São Paulo, BrazilFAPESP: 05/57665-8CNPq: 2006-0919Web of Scienc

Nuclear receptors in vascular biology

Nuclear receptors sense a wide range of steroids and hormones (estrogens, progesterone, androgens, glucocorticoid, and mineralocorticoid), vitamins (A and D), lipid metabolites, carbohydrates, and xenobiotics. In response to these diverse but critically important mediators, nuclear receptors regulate the homeostatic control of lipids, carbohydrate, cholesterol, and xenobiotic drug metabolism, inflammation, cell differentiation and development, including vascular development. The nuclear receptor family is one of the most important groups of signaling molecules in the body and as such represent some of the most important established and emerging clinical and therapeutic targets. This review will highlight some of the recent trends in nuclear receptor biology related to vascular biology